SERBIAN GENETICS SOCIETY
SECOND CONGRESS OF SERBIAN GENETICISTS
Sokobanja, Serbia, Serbia, November 10-13, 1999.
Chairmanship
Katarina BOROJEVIĆ
Janko DUMANOVIĆ
Ivan MIHALJEV
Bogoljub SOLDATOVIĆ
Aleksandar TUCOVIĆ
Slobodan ŽIVKOVIĆ
Program Committee
Kosana KONSTANTINOV, president
Marko ANĐELKOVIĆ
Nada BARJAKTAROVIĆ
Janoš BERENJI
Vojislav GUZINA
Mile IVANOVIĆ
Vasilije ISAJEV
Marija KRALJEVIĆ-BALALIĆ
Dragoslav MARINKOVIĆ
Draga SIMIĆ
Ljubiša TOPISIROVIĆ
Mladen VUJOŠEVIĆ
Organizing Committee
Vasilije ISAJEV, president
Jelena KNEŽEVIĆ-VUKČEVIĆ, vice-president
Marina STAMENKOVIĆ-RADAK, vice-president
Janoš BERENJI
Živorad GAJIĆ
Miodrag DIMITRIJEVIĆ
Selma KANAZIR
Desimir KNEŽEVIĆ
Miroslava MICIĆ
Snežana MLADENOVIĆ DRINIĆ
Sasa ORLOVIĆ
Slaven PRODANOVIĆ
Branka VASILJEVIĆ
Secretariat
Mirjana ŠIJAČIĆ-NIKOLIĆ, president
Dražen JELOVAC, vice-president
Jelena BEAGOJEVIĆ
Veselinka ĐORĐEVIĆ
Marina ĐURIŠIĆ
Tatjana TERZIĆ
SERBIAN GENETICS SOCIETY
SECOND CONGRESS OF SERBIAN GENETICISTS
Sokobanja, Serbia, Serbia, November 10-13, 1999.
Kosana Konstantinov
Genetics at third milenium beginning
Plenary lectures
Lj. Topisirović. M. Kojić, Dj. Fira, N. Miladinov, I. Strahinić, O. Gajić, S. Arsenijević i I. Spasojević
Molecular genetics of lactic acid bacteria is a basis for development of modern dairy biotechnology
Drinić Mladenović Snežana, Jelovac Dražen i Konstantinov Kosana
Molecular biology and information technology in plant breeding
Selma Kanazir, Sabera Ruždijić, Ljubisav Rakić
Modulation of gene expression byantisense oligonucleotides
Mladen Vujošević i Jelena Blagojević
Populational and evolutionary aspects of B-hromosomes - A case of yellow-necked mice
Marko Anđelković i Marina Stamenković-Radak
Population genetic aspect of stress
Draga Simić, Jelena Knežević-Vukčević
Plant antimutagens and cancer prevention
Roy Forster
The use of transgenic animals in carcinogenicity testing
Drinić Goran i Babić Milosav
Genetics and breeding of maize - achievements and challenges
Dragan Škorić
Achievements of sunflower breeding at the end of the 20th century
M. Ivanović, Violeta Anđelković
V.Isajev, A.Tucović
Gene pool conservation and tree breeding
Živoslav Marković, Jasmina Zdravković, Mirjana Mijatović
New approach in tomato breeding (Lycopersicon esculentum Mill.)
Desimir Knežević, Novo Pržulj
Contribution of genetics in breeding of small grains
Romac S.
The expansion of trinucleotide repeats as a genetical cause of neurological and psychiatric disease
Milena Stevanović i Marija Guc-Šćekić
Application of molecular techniques in diagnostics of chromosomal disorders
Sekcija 1. Molecular genetics
Goran Ljubijankić
Nataša Miladinov, Irena Spasojević i Ljubiša Topisirović
Zvezdana Popović, Aleksandra Đurković, Jelena Zarić i Marina Lušić
Induction of albumin synthesis in Belgrade laboratory (b/b) rat reticulocytes
Stamenković Gorana, Laketa Danijela, Gudurić Jasenka, Veljković Emilija, Krtolica-Zikić
Koviljka i Dimitrijević Bogomir
HCV genotype detection by SSCP in Yugoslav population
Vesna Lazić-Jančić, D. Kovačević i S.A. Quarrie
BSA with candidate drought-induced genes
Milena Stevanović, Nikola Arsić, Tamara Rajić, Slavica Stanojčić, Tomislav Terzin, Vesna
Verbić, Maja Vujić, Nebojša Mirković i Danica Grujić
SOX genes: Brain development and sex determination
Nikola Tanić, Nasta Dedović, Mladen Vujošević i Bogomir Dimitrijević
Branka Vasiljević, Milorad Kojić, Milojević, Sandra Vajić, Marijana Miljković,
Tatjana Ilić
Control of resistance genes in antibiotic-producing actinomycetes
Jelena Brkljačić, Jelena Avramović, Paja Šijačić, Svetlana Radović, Vesna Maksimović
The isolation of buckwheat genomic clone containing the gene for metalothionein-like protein
Sandra Vajić, Milorad Kojić, Branka Vasiljević
Comparative study of Micromonospora strains producing aminoglycoside antibiotics
Marijana Vučinić, Z. Gajić i P. Stojić
The application of genetic markers in prevention of dairy cattle mastitis
Nataša Grujičić, Sonja Pavlović, Sanja Pavlović, Zvezdana Popović
Functional analysis of the rat βbminy - globin gene promoter
Tatjana Ilić, Milorad Kojić, Goran Ljubijankić, Branka Vasiljević
Synthesis and secretion of the sgm methylase in Saccharomyces cerevisiae
Radmila Janjušević, Mirjana Lilić, Milija Jovanović, Goran Jovanović i Dragutin Savić
The regulation of cls gene expression in bacteria Escherichia coli K12
Milija Jovanović, Mirjana Lilić, Radmila Janjušević, Goran Jovanović i Dragutin Savić
Milan Kojić, Slavica Arsenijević, Olivera Gajić, Vanesa Munćan i Ljubiša Topisirović
Construction of shuttle vector pA13
Marija Mijatović, Jelena Stanković, Marijana Petakov, Gordana Jovčić, Jelena Zarić, Zvezdana Popović
Large scale of isolation of rat erythroid progenitors for gene therapy
Marijana Miljković, Nataša Milojević, Milorad Kojić, Branka Vasiljević
Two-plasmid system for isolation of regulatory and gentamicin sensitive mutants of the sgm gene
Tatjana Mitrović i Zvezdana Popović
Promoter sequences required for regulated expression of the rat βbminy - globin gene in rel cells
Snežana Mladenović Drinić i Kosana Konstantinov
Polymorphism of PCR based markers and heterosis in maize
Pavković, N., Todorović, V., Glišin, V., Ljubijankić, G.
Yeast shuttle expression vector with dominant selective marker
Tamara Rajić, Nikola Arsić, Nebojsa Mirković. Milka Sokolović and Milena Stevanović
Localisation of control elements responsible for SOX3 gene activation by retinoic acid
Spasić M., Glišin V., Ljubijankić G.
Molecular-genetic analysis of temperature-dependent posttranslational maturation of E. coli pac
Irena Spasojević, Nataša Miladinov i Ljubiša Topisirović
Slavica Stanojčić, Maja Vujić, Nikola Arsić, Tamara Rajić i Milena Stevanović
Preliminary characterisation of the three novel human SOX genes
Slavica Stanojčić, Jelena Đurović, Nikola Arsić, Nebojsa Mirković, Vesna Verbić, Milena Stevanović
Genotyping the bovine traits of economic interest
Ivana Strahinić, Milan Kojić i Ljubiša Topisirović
Analiza konjugacionih svojstava plazmida Lactococcus lactis subsp. lactis biov. diacetylactis S50
Tomislav Terzin, Milka Sokolović, Ana Basić i Milena Stevanović
Cloning of mole rats Sox2 gene using PCR method
Đorđe Fira i Ljubiša Topisirović
Biochemical and genetic characterization of staphylococcal extracellular proteinases
Ševo M., Glišin V. i Ljubijankić G.
Mechanism of posttranslational processing of the heterodimeric PAC
Paja Šijačić, Jelena Brkljačić, Svetlana Radović, Vesna Maksimović
The isolation of genomic clone containing the gene for a subunit of buckwheat seed storage protein
Sekcija 2. Population and Evolution Genetics
Svetlana Fišter
Boro P. Pavlović
Goran Živanović i Marko Anđelković
Genetic loads and coadaptivity of chromosomal inversion polymorphism in a Drosophila subobscura
Jelena Blagojević i Mladen Vujošević
S. Bojović, Ph. Heizmann, M. Barbero
Fraxinus ornus L., Sexual polymorphisme and RAPD markers
Ljiljana Vapa, Dragana Obreht, Milan Vapa*, Deana Demjen
Intra-and inter-population variability in hare (Lepus europaeus PALLAS)
Dimitrijević Miodrag, Knežević Desimir, Petrović Sofija
Gliadin allelic variation and diversity of agronomic properties in wheat
Đorđević Radiša, Zdravković Milan, Zečević Bogoljub, Sretenović-Rajačić Tatjana
"Afila" gene effects on absolute seed mass in peas (Pisum sativum L.)
Jelena Živanov-Čurlis, Stevo Najman, Estera Mrčarica
Influence of antioxidants on fertility and fecundity of D. melanogaster
Gordan Zec, Radmila Todorović, Petar Mišić, Slavica Čolić
Mirjana Ivančević, Tatjana Sretenović-Rajičić, Nenad Pavlović, Jasmina Zdravković
Janković Dragan, Janković Slađana
Atypical phenomena in walnut organogenesis
Slađana Janković
Growth habit forms in P. communis L. population in the Ibar - Kolasin region
Krajinčanić, B., Krajinčanić-Suzović V., Cukić R., Ninković D.
Population genetics investigations of PTC tasting sensibility from some localities in Serbia
Krajinčanić-Suzović V., Krajinčanić B., Čukić R., Gašić-Marušić R.
Frequency of hand clasping from some localities in Serbia
Milankov V., Vujić A., Simić S., Ludoški J.
Genetic differentiation in populations of Merodon avidus (ROSSI, 179) (Diptera: Syrphidae)
Milankov V., Vujić A., Simić S.
Olivera Milošević, Dragoslav Marinković, Slobodan Arsenijević
Stevo Najman, Jelena Živanov-Čurlis, Estera Mrčarica
Influence of antioxidants on preadult developmental duration of D. melanogaster
Dragana Obreht, Saša Orlović, Joška Erdesi, Ljiljana Vapa
Allozyme variability in Quercus robur L. and identification possibility within the population
Boro P. Pavlović i Nevenka Pavlović
Petrović Sofija, Dimitrijević Miodrag, Mladenov Novica i Kraljević-Balalić Marija
Ecological stability of yield components in wheat
Vuk Savković, Marko Anđelković
Monitoring of temporal variability of chromosomal arangements in Drosophila subobscura
Tatjana Sretenović Rajičić, Mirjana Ivančević, Jasmina Zdravković, R. Đorđević, M. Damjanović
Reactivity of cabbage (Brassica oleracea var. capitata) on paraquat in vitro
Z. Stanimirović, Jevrosima Stevanović, D. Pejović, D. Popesković
Biodiversity of the honeybee Apis mellifera, Linne (1758) - cytogenetic aspects
Z. Stanimirović, B. Soldatović, Jevrosima Stevanović
Milica Strnać, Zvonko Magić, Cedomir Radojičić i Dragoslav Marinković
IvanaTomišić, Marina Stamenković-Radak, Tatjana Terzić i Marko Anđelković
Sekcija 3. Mutagenesis and genotoxicology
Gordana Joksić, Miroslava Stanković
Micronucleus assay in to human biomonitoring
Miroslava Veličković, Jelena Blagojević i Mladen Vujošević
The effects of disturbed environment on natural populations of rodents
Branka Vuković-Gačić, Tatjana Stević, Jelena Knežević-Vukčević, Dragana Mitić, Dejan Brkić, Jelena Blagojević, Marko Anđelković, Draga Simić
Comparative study of antigenotoxic effect of sage in prokaryotic and eukaryotic test-systems
Olivera Milošević, Darko Grujičić, Dragoslav Marinković, Slobodan Arsenijević, Smilja Banković, Aleksandar Živanović, Aleksandra Dimitrijević
Ninoslav Đelić, Bogosav Soldatović, Marko Anđelković
Evaluation of genotoxic and mitogenic effects in cultures of human lymphocytes treated with insulin
Ninoslav Đelić, Bogosav Soldatović, Dijana Đelić
Influence of cloprostenol on cell cycle kinetics in cultured human lymphocytes
Biljana Burić, Z. Stanimirović, Biljana Marković
Investigation of genotoxic effects of CARISOLV™ gel
Gordana Joksić, Miroslava Stanković
Chromosome aberrations monitored in radiation workers
Dubravka Jovičić, Snežana Milačić, Radomir Kovačević, Mitar Novaković
Analysis of chromosomal status of subjects occupationally exposed to radionuclides
Jelena Knežević-Vukčević, Tatjana Stević, Dragana Mitić, Branka Vuković-Gačić, Draga Simić
Molecular mechanisms of mutagenesis inhibition by ethereal oil of sage (Salvia officinalis L.)
Biljana Marković, Z. Stanimirović, Jevrosima Stevanović, N. Jovanović
Comparative analysis of genotoxic effects of Gastrogala 10, Tiamulina S i Carbadoxa in vitro
Dragana Mitić, Tanja Berić, Branka Vuković-Gačić, Jelena Knežević-Vukčević, Ratko M. Jankov, Draga Simić
Antimutagenic potential of rosmanol-9-ethyl ether, osaine and pomiferine
Danica Mićanović, Penny Hirsch, Philip Curnow, Vera Raičević i Desimir Knežević
Colonisation of wheat by Pseudomonas and marker genes
D. Pejović, Z. Stanimirović, B. Soldatović
Analysis of genotoxic and mutagenic effect of Apitol®
Z. Stanimirović, Jevrosima Stevanović, M. Kulić, D. Pejović
Investigations of cytotoxic and genotoxic effects of Levamisole® in vivo
Jevrosima Stevanović, Z. Stanimirović, D. Pejović
Cytogenetic effect of Fumagillin-et® on human lymphocytes in vitro
Sekcija 4. Breeding of organisms
Gordana Radović, Dražen Jelovac, Jasmina Muminović
Breeding potential of early maturity local maize populations
Z. Tomić, Lugić Z., Sokolović D.
Genetic potential of seed germination in forage crops cultivars in the fifth year of their life
Branislav Dozet, Dragan Škorić, Radovan Marinković
Aleksandar Tucović, Vasilije Isajev i Milan Mataruga
Sasa Orlović, Joška Erdeši, Srbislav Radivojević, Zoran Obućina, Gojko Janjatović
Strategy and previous results of pedunculate oak (Quercus robur L.) breeding in Yugoslavia
M. Mitrović, M. Nikolić, D. Ogašanović, Z. Tešović
Selection of domestic walnut (Juglans regia L.) from natural population
Nenad Brkić, Živorad Gajić, Milovan Pušić
V. Bogdanović, Brkić. N.
Influence of systematic environmental factors on genetic indexes and rank of potencial sires
Avramov, L., Milutinović, M., Jović, S., Žunić, D., Maletić Radojka, Vujović, D.
Uvological and technological investigation of clone population in cultivar Kaberne Fran
Jovanka Atlagić
Jan Boćanski i Zoran Petrović
Jelena Bošković, Bošković M., Pešić V.
Gene-for -gene modeling for wheat hybrids resistant to Puccinia recondita tritici
Mirjana Vasić, Jelica Gvozdanović Varga. Janko Červenski
Dragana Vasić, Dragan Škorić, Gilbert Alibert
RAPD Analysis of products of fusion between cultivated sunflower Helianthus maximiliani (Schrader)
Sanja Vasiljević, Gordana Šurlan- Momirović, S. Katić, V. Mihailović, D. Lukić, T. Živanović
Mirjana Vulić
Effect of systematic factors of the environment on the variability of milk production
Jelica Gvozdanović-Varga, Mirjana Vasić
Divergence of genotipes for dry matter yield in garlic
Branislava Grbović, Vasilije Isajev
Guzina V., Orlović S., Kovačević B.
Poplar selection for superior growth
Jelena Damjanović, Maja Vračarević, Bogoljub Zečević i Slaven Prodanović
Identification of donor lines for improving fruit yield of K35 x K12 eggplant hybrid
Gordana Demić, Snežana Mladenović Drinić i Kosana Konstantinov
Protein marker polymorphism in set of maize inbred lines
Dragan J. Bukić, Dane Lukić
Alfalfa: from populations to varieties
Slađana Žilić, Irina Božović, V. Bekrić i Snežana Mladenović Drinić
Changes on proteins in different soybean varieties under effects of increased temperature
Žunić P., Bešlić Z.
Zdravković M., Čorokalo D., Đorđević R. i J. Zdravković
Jasmina Zdravković, Marković Z., Zdravković M., Mirjana Mijatović
Inheritance mode of fruit shape in tomato
Bogoljub Zečević, Dušan Stevanović, Slaven Prodanović i Radiša Đorđević
Veselinka Zečević, D. Knežević, Danica Mićanović
Variability and components of variance for plant height in different wheat cultfvars
Vasilije Isajev, Predrag Jović
Vasilije Isajev, Vera Lavadinović
I. Jevtić, Z. Stanimirović, B. Soldatović
Zoran Jerković, Radivoje Jevtić
Genetic analysis of parents in wheat breeding to obligate parasites resistance
Jestrović Zorica, Kondić Ankica, Šešek S., Denčić, S., Pavlović, M.
Callus induction from mature embryos in species of the genus Triticum L.
Joksimović J., Atlagić J., Škorić. D.
Effect of genes and combining ability for kernel yield in some sunflower inbred lines
B. Kerečki, M. Ivanović i Violeta Anđelković
Genetic control of maize grain dry-down during maturation
Kraljević-Balalić Marija
Gene effects for physiological parameters in wheat
Bojana Klašnja, Špiro Kopitović, Saša Orlović, Zoran Galić
Desimir Knežević
Level of genetic determination of small grains and future prospect
Desimir Knežević, Veselinka Zečević, Milanko Pavlović, Danica Mićanović, Miroslav Kuburović, Slaven Prodanović
Genetic determination of technological quality of wheat
Kondić Ankica i S. Šesek
Differences between homozygous and heterozygous wheat genotypes regarding their androgenic response
Borislav Kobiljski, Srbislav Denčić, Marija Kraljević-Balalić i Rončević Petar
Inheritance of kernel number per spike in crossing wheat genotypes differing in Rht genes
Branislav Kovačević
Evaluation of discrimination ability for morphometric leaf parameters of eastern cottonwood
Miroslav Kuburović, Milanko Pavlović, Milivoje Milovanović, Veselinka Zečević
Characteristics of perspective lines of winter wheat of center for small grains in Kragujevac
Lukić D., Marija Kraljević-Balalić
Breeding alfalfa genotypes for increased seed yield
Milomirka Madić, Miroslav Kuburović i Aleksandar Paunović
Genetic analysis of the grain mass per plant in barley hybrids
Dragoljub Maksimović, Dušan Urošević, Aleksandar Paunović
Study of some characteristics of barley varieties en F1 generation
Ana Marjanović-Jeromela, Radovan Marinković
Heritability of rapeseed (Brassica napus L.) yield components
Marković Z., Zdravković Jasmina, Damjanović M.
Local populations and semi-wild tomato forms as a source of genetic divergence
Milan Mataruga, Vasilije Isajev, Mirjana Šijačić-Nikolić
Mezei S., Čačić N., Nagl N., Kovačev L. i Sklenar P.
Possible use of micropropagation of dormant buds from root head of sugar beet in recurent selection
Rade Miletić1, Radoljub Petrović1, Milisav Mitrović
Rade Miletić
Fenological regularities in walnut trees population in region of Timočke krajine
Gordana Milojević, Snežana Mladenović Drinić, Goran Drinić i Kosana Konstantinov
The cluster analysis of maize inbred lines on the basis of polymorphism of proteins marker
Milivoje S., Milovanović, VladanPešić
Inheritance of grain number per spike in hexaploid tritikalea
Momčilo Milutinović, Dragan Nikolić, Milica Fotrić i Vera Rakonjac
Ratio of pollen functional ability and fruit set degree in grapevine (Vitis sp.)
Milutinović M.M., Milutinović D.M.
Coefficent of fruitfulness of apple varieties and hybrids
Milutinović. M., Žunić. P., Simić V.
Inheritance of berry skin color in seedlings of F1 generation of interspecies hybrids in grapevine
M. Mitrović, S. Milenković
Novica Mladenov, Novo Pržulj, Nikola Hristov, Veselinka Đurić
Phenotypic and genetic relationships between wheat quality
Jasmina Muminović, Dražen Jelovac, Gordana Radović
Živka Bukić, Siniša Milutinović, Danica Mladenović
The possibility of using pepper populations in the aim of further breeding
Aleksandra Nastasić, M. Stojaković, G. Bekavac, Z. Petrović, N. Vasić
Influence from S1 recurrent selection in maize synthetic NSB
Biljana Nikolić,Vasilije Isajev
Group and individual pollen variability of some 5-needle pines in function of improvement
Dragan Nikolić
Saša Orlović, Borivoj Krstić, Bojana Klašnja
Genetic and phenotypic correlation of physiological and growth parameters of poplar clones
Saša Orlović, Zagorka Petrov, Zoran Galić, Leopold Poljaković Pajnik
Mirjana Ocokoljić, Vasilije Isajev, Nada Kovačević
Boro P. Pavlović
The framework of variants of haploid chromosome complement producing a finite number of pure lines
Pavlović Nenad, Sušić Zoran, Zdravković Jasmina, Marković Živoslav, Stevanović Dušan
Variability and heritability of onion bulbs diameter
Sofija Pekić, Vesna Lazić-Jančić, Violeta Anđelković, M Ivanović, Zora Dajić, Evonne Waterman, H Rahman, A Steed i SA Quarrie
Sofija Pekić, Lora Ljubojević, Violeta Anđelković, M Ivanović, Vesna Lazić-Jančić, A. Steed i SA Quarrie
The role of ABA in drought resistance: physiological and genetical analysis in maize
Petrović, Z., J.Bocanski, Nastasić. Aleksandra, G. Bekavac, N. Vasić
Combining abilities and mode of inheritance of yield and 100-grain mass in maize (Zea mays L.)
Novo Pržulj, Novica Mladenov, Miroslav Bogdanović
Estimates of genetic effects of productive tillering in spring wheat
S. Prodanović, G. Šurlan-Momirović. D. Perović, I. Stančić, Z. Nikolić i Z. Veselinović
S. Prodanović, G. Šurlan-Momirović, B. Jovanović i M. Menkovska
Vera Rakonjac
Genetic and phenotypic correlation and path analysis in peach
Petar Rončević, Nikola Hristov i Marija Kraljević-Balalić
Effect of PEMF on grain number and grain mass per spike in spring wheat
Pavle Sklenar, Lazar Kovačev, Nikola Cačić, Snežana Mezei i Nevena Nagl
Correlations and path-coefficients analysis in sugar beets root traits
Mirjana Srebrić
Stanković G., Trifunović B.V., Trifunović V.
Accumulation of favourable alleles in two synthetic populations of maize (Zea mays L.)
Sikora V.
Heterosis for agronomic traits of broomcorn
Sušić Zoran, Zdravković Jasmina, Pavlović Nenad, Prodanović Slaven
Žarko V. Tešović, Slađana A. Nidžović, Milan M. Lukić
Žarko V. Tešović, Sladana A. Nidžović, Milan M. Lukić
Biological properties of promising apple selections
Žarko V. Tešović, Sladana A. Nidžović, Milan M. Lukić
Breeding dwarfing apple genotypes
Zorica Tomić, Sokolović, D. i Gordana Šurlan-Momirović
Inbreeding depression of productive traits in three species of the genus Agrostis L.
Zorica Tomić i Sokolović D.
Regeneration of perennial grasses seeds from the collection of the plant gene bank
Mihailo Tošić
Trifunović B.V., Trifunović V., Stanković G.
Genetic potential of yield of new ZP maize (Zea mays L.) hybrid combinations
Trivunović Snežana, Teodorović M., Petrović Milica
Genetic parameters of the fattening and reproductive traits of swedish landrace
Aleksandar Tucović, Vasilije Isajev i Mirjana Šijačić-Nikolić
Genetic -ecological bases of Ailanthus (Ailanthus altissima Swingle) adaptation in Serbia
Aleksandar Tucović, Vasilije Isajev i Mirjana Šijačić-Nikolić
Flowering of Acer saccharinum L. occurrence, frequency of polygamy and sex mosaics
Aleksandar Tucović, Vasilije Isajev i Mirjana Šijačić-Nikolić
Neoteny types in trees and shrubs and their inportace
Gordana Ćurčić, Vasilije Isajev, Mihailo Tošić
Balkan Maple generative seed orchard i near Ivanjica - pilot plot for Balkan Maple breeding
Biljana Urošević, Jasmina Radović, Zorica Tomić
Filipović M., Drinić Mladenović S., Milojević G. i Konstantinov K.
Application of protein markers in "Genetic Purity" testing of maize genome
Nikola Hristov, Novica Mladenov, Marija Kraljević-Balalić
Genetic diversity of stem traits in winter wheat
Cvikić Deian, Jasmina Zdravković, Gordana Šurlan Momirović, Sušić Zoran, Đorđević Radiša
Inheritance mode and phenotypic variability for fruit firmness in nor and rin tomato genotypes
Nikola Čačić, Lazar Kovačev, Snežana Mezei, Pave Sklenar i Nevena Nagl
Šesek S. i Ankica Kondić
Testing wheat genotypes for herbicide tolerance by in vitro culture
Lević Jelena i Tijana Petrović
Occurrence of the race R1 Exserohilum turcicum (pass.) Leonard & Sugss pathogenic to maize
Saratlić G., Rošulj M. i K. Konstantinov
Nenad Delić
Sekcija 5. Human Genetics
Koviljka Krtolica, Milica Radojković, D. Marinković, L. Ristić i Biljana Todorić
O. Miljanović, M. Kaluđerović
Chromosome abnormalities in children with mental retardation associated with dyismorphia
Gordana Anđelić, Zvonko Magić, Ljiljana Martac, Ilija Tomić, Vladislav Stepić i Vladimir Mrđa
Vesna Verbić, Danica Grujić, Milena Stevanović
Fluorescent in situ hybridization (FISH) in clinical and fundamental investigations
Slobodanka Vukosavić, Serge Przedborski
Role of synuclein in neurophatology of Parkinson’s disease
M. Guć-Šćekić, D.Radivojević, M. Đurišić, N. Arsić, M. Stevanović, D. Zdravković, K. Sedlečki, M.Banićević
Cytogenetic and molecular studies in sexual differentiation disorders
Slobodanka Grković, Vesna Ivanović Deretić
Cat Eye syndrome - case report
N. Dedović, J. Milašin, O. Josipović, M. Vukadinović i B. Dimitrijević
Molecular genetic analysis of premalignant and malignant lesions of oral cavity
Jasmina Đurković
Acanthosis nigricans - Review of a case
Ninoslav Djelić
Mitotic and proliferation indices of human lymphocytes treated with oxytocin in vitro
V. Đorđević, M. Pantić, M. Đurović, A. Novak
Turner's syndrome with de novo balanced translocation t(l;9) and ring (X) chromosome
V. Đorđević. M. Pantić, A. Novak, N. Suvajdžić, M. Čolović
M. Djurišić, M.Guć-Šćekić, T. Lalić, D. Radivojević, D. Zamurović, M. Đurić, S. Todorović
Genotype-phenotype correlation in two DMD/BMD patients with large deletions in dystrophin gene
Zarovni Nataša, Georgijević Dubravka, Radojković Dragica
Allele distribution of two VNTR and two STR loci in population of Serbia
Zamurović N., Čuljković B., Stojković, O., Major T., Keckarević D., Savić D. i Romac S.
PCR diagnostics of Charcot Marrie tooth
Ilić Vesna, Cikota Bojana, Vojvodić Danilo, Malešević Milomir, Stamatović Dragana, Magić Zvonko
Mutations in the K-ras AND p53 genes in twins before and after bone marrow transplantation
J. Jovanović, M Guć-Šćekić, M. Kuzmanović, G. Bunjevački
Cytogenetic studies of MDS in childhood
A. Krstić, F. Popić - Paljić, J. Jovanović - Privrodski, M. Obrenović, R. Aleksić, M. Kolarski, I. Kavečan. T. Redžek - Mudrinić, J. Popadić, M. Nikolić, R. Madžar, Lj. Gaćina, V. Čihi, C. Laketa, P. Kruščić, J. Rudež, T. Tarašenko, M. Grubješić
Prenatal diagnosis and health promotion
Aleksandar Krstić, Đorđe Mačvanin, Feodora Popić, Jadranka Jovanović, Ružica Aleksić, Milan Obrenović, Ivana Kavečan, Tatjana Redžek-Mudrinić, Jelena Popadić, Marko Nikolić, Milka Mitrović, Spomenka Mijić
Langdon down syndrome in Vojvodina - Epidemiology, Dermatoglyphics and New Treatments
Kečkarević P., Čuljković B., Stojković O., Vukosavić S., Savić D., Zamurović N., Major, T., Romac S.
Spinal muscular atrophy- Possibilities for prenatal diagnostics
T. Kuveljić, J. Nikoliš, Novaković, V. Bunjevački, Lj. Luković, J. Milašin, M. Krajinović, S. Radmanović, S. Branković
The role of chromosomal polymorphism in sex chromosomes meiotic nondisjunction
D. Luković, I. Novaković, J. Milašin, Lj. Luković, T. Kuveljić, V. Bunjevački, M. Krajinović i N. Ostojić
Loss of heterozygosity on chromosomes 3p and 6p in cervical carcinomas
Zvonko Magić, Bojana Cikota, Vesna Ilić, Gordana Anđelić
P53 mutations in breast cancer patients
Major T., Culiković B., Stojković O., Zamurović N., Kečkarević D., Savić D. i Romac S.
Fragile X syndrome: Molecular diagnostics
Danko Obradović, Aleksandar Krstić, Jasmina Rudež, Milan Obrenović, Ivana Kavecan, Jelena Popadić
Sex determination of human fetus by amplification of DNA molecules from amniocytes
M. Pantić, A. Novak, V. Đorđević, D. Marisavljević, N. Suvajdžić, G. Janković, *M. Stevanović, M. Čolović
Cytogenetics and fluorescence in situ hibridization in diagnosis of myelodisplastic syndromes
Sonja Pavlović, Milan Joksimović, Milica Cvorkov-Dražić, Gordana Bunjevački, Dragana Janić i Zvezdana Popović
Molecular diagnosis of inherited hemoglobinopathy – Hb lepore
Branka Popović, Jelena Milašin, Miroslava Mičić, Vitomir Konstatinović
LOH in the regions 3p21,9p21,6q21-23,17p13 AND 13q14 in squamous cell carcinomas of the lip
D. Radivojević, M. Guć-Šćekić, M. Đurišić, T. Lalić, J. Savić, P. Minić, A. Cvetković, E. Kanavakis, M.Tzetis, T. Antoniadi
Genotype/phenotype correlation (as to the pancreatic status) of Yugoslavian cf patients
Rakićević Ljiljana, Georgijević Dubravka, Janković Gordana, Miković Danijela, Radojković Dragica
Prenatal diagnostics of hemophilia a by intragenic RFLP analysis
Savić P., Čuljković B., Stojković O., Major T., Zamurović N., Kečkarević D. i Romac S.
Molecular genetics and diagnosis of myotonic dystrophy
Biljana Spremo-Potparević, Vesna Verbić, Milena Stevanović i Milan Joksimović
Biljana Todorić, Z. Magić, L. Ristić. M. Malešević i K. Krtolica
Detection of Philadelphia chromosome negative chronic myeloid leukemia by RT-PCR method
Geza Cekuš
Dyschromatopsia rate among the population of northern Backa (Voivodina)
Geza Cekuš
The ability of perceiving the smell of acetone
Bojana Cikota, Rajna Stamenov, Slavko Berger, Danilo Vojvodić, Milomir Malešević, Zvonko Magić
K-ras AND H-ras point mutations in patients with large cell lymphomas and chronic lymphadenitis
Čuljković B., Stojković, P., Vukosavić. S. i Romac S.
Human identification and patternity testing
Jan Kišgeci, Janoš Berenji
Transgenic plants - pro et contra
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENETICS AT THIRD MILENIUM BEGINNING
Kosana Konstantinov
Maize Research Institute "Zemun Polje", 11081 Zemun-Belgrade, Yugoslavia
Second Congress of Serbian Geneticists is going on at the end of 20th Century, culminating the tremendous development of our science in this century. Historically, First International Conference on this subject was held during January 1899 in London, England and Second during September 1902 in New York City. Both meetings have been named as "International Conference on Plant hybridisation". During Third Conference, held in London during 1906. Title was slightly modified to "International Conference on Hybridisation and Plant Breeding". During this meeting the term "Genetics" has been suggested for this new field of scientific inquiry. After development of all fields of science, including genetics, a serious question has been raised whether human species can sustain itself on earth. The answer is affirmative. Mankind not only can survive but also aspire to live at high quality. In order to achieve this goal, co-operation among scientific workers in all fields have to be established and geneticists can play a significant role in three basic fields:
1. Agriculture based on gene structure and function understanding;
2. Medicine based on gene structure and function understanding and
3. Environmental protection based on gene structure and function understanding.
If estimation for human population for the first half of 21.century' are about 8.9 billion the first consequence will be increase in the demand for food. By shortage in water supply desert encroachment and diminution of arable acreage could be expected. Several fundamental tasks geneticists must be involved: establishment of germplasm banks. Release of captured genetic potential and Development of new breeding technologies. In the field of human genetics, through co-operation among geneticists, physicians and pharmacologists the main contribution the understanding and treatment of single- and multiple-gene based diseases: understanding of gene role in cancer development, ageing and life expectance and introduction of routine application of gene therapy in disease treatment, particularly in the nervous system development and function.
As we have only one earth to live on, existing research should be intensified in the field of genetics covering problem of the soil, atmosphere and water contamination. With the rapid development of industry' and public transportation, damages to the environment have caused direct and serious threat to human health. Among many of important tasks in the field of the environmental protection, selection of already existing fungal strains, as well as through mutation induction of new strains, which can digest and utilise pollutants for their own use is promising. Also the selection of microorganisms for biological degradation of pollutants in water and soil. Particular task is development of highly sensitive genetic methods for testing the presence and estimating the amount of mutagens/carcinogenes in the environment.
In the coming century it could be predicted that only through the study of the genome the life phenomena themselves can be understood.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MOLECULAR GENETICS OF LACTIC ACID BACTERIA IS A BASIS FOR DEVELOPMENT OF MODERN DAIRY BIOTECHNOLOGY
Lj. Topisirović. M. Kojić, Dj. Fira, N. Miladinov, I. Strahinić, O. Gajić, S. Arsenijević i I. Spasojević
Institute of Molecular Genetics and Genetic Engineering, V. Stepe 444a, P.O.Box 794,
11001 Belgrade, Yugoslavia
Lactic acid bacteria (LAB) have been essential in food and feed fermentation for centuries. They have very broad application in food industry and agriculture. They have also a great influence on nutritional attributes of the products such as flavour, texture and others. In addition, some of them are commensal colonisers of the mammalian gastro-intestinal tract including humans, where Lactobacillus acidophilus and Lactobacillus renteri are commonly found. Some strains of LAB are probiotics, since it is believed that they have effect on human and animal health. Health benefits attributed to LAB are following: (a) control and treatment of infections in the intestine caused by enteric pathogens or infections in the urogenital tract; (b) reduction of incidence of tumour/cancer in colon or reduction of the level of serum cholesterol as well as cardiac heart disease and (c) stimulation of immune system and bowel movement. Bearing in mind these probiotic effects of LAB, an increasing interest exists for dairy products containing specific bacterial species with potential health-improving properties. These products are named "functional food".
Studying the genetic potentials of LAB isolated from the traditionally homemade fermented products produced in different geographical regions is extremely interesting. A collection of natural isolates (about 600 isolates) was made. This collection contains LAB isolated from different traditionally prepared fermented products throughout Yugoslavia. Numerous isolates were tested for the production of proteinases, exopolysaccharides, and bacteriocins. Most of those showing interesting characteristics from scientific and commercial point of view were analysed at molecular level. Among tested LAB, isolates were characterised that produces new type of proteinases or bacteriocins. In addition, such LAB could be a source of exopolysaccharides or aromogenic compounds having high commercial value. Dairy products acquire their unique characteristics from differences in the composition of starter cultures. However, genes encoding production of these components are very often located on the genetically less stabile elements, plasmids. Therefore, studying the molecular genetics of LAB especially by using genetic engineering will facilitate the construction of more stabile starter cultures. Starter cultures could be constructed by combination of genetically characterised natural isolates. On the other hand, selection of natural isolates with probiotic capacities opens a possibility to construct the starter cultures for functional food production. Taking altogether, construction of improved, reliable starter culture is a main task for molecular biotechnologists in the future.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MOLECULAR BIOLOGY AND INFORMATION TECHNOLOGY IN PLANT BREEDING
Drinić Mladenović Snežana, Jelovac Dražen i Konstantinov Kosana
Maize Research Institute "Zemun Polje", 11081 Zemun-Belgrade, Yugoslavia
In the past two decade, development of molecular genetics including molecular technology for gene transfer from heterelogous organisms into plants brought a new dimension of plant breeding. Molecular markers as universally methods for all biological systems virtually effect on success of directly examination of structure and function as well as determination of genetic polymorphism of plant genomes. The potential applications of molecular markers in plant breeding are: analysis of molecular basis of evolution; germ plasm identification, classification and management; assessing genetic diversity; identification of genes underlying agronomy important traits as yield, resistance to stress and disease and heterosis as well. High density genetic linkage maps for a number of plant species as a basis for marker assisted selection of agronomically useful traits and isolation of these genes have been established. New class of molecular markers known as "gene chips", uses microarrays of molecules immobilized on solid surfaces have recently been developed. With "gene chips" DNA belonging to thousand of genes can be arranged in small matrices (chips) and hybridized with labeled cDNA. The first biological application of DNA chip technology are analysis of gene expression and it should help to better understanding of fundamental mechanisms of plant growth and development, polymorphism detection and rapid analysis of transgenic plants.
Great move in the field of plant biotechnology are possibility to introduce genes from heterelogous systems into plant genomes resulting in plants with altered traits or in another word transgenic plants. The experiment of maize genome transformation by introducing gene coding neomycin phosphotransferaze (NPT II), chloramphenikol 0-acetiltranspheraze (CAT) and part of A. tumefeciens bacterial genome have been started during 1986 at Maize Research Institute. Both gene expression and integration of introduced genes in maize genome was determined. Integration of forcing genes induced different phenotypic changes. Changed genotypes were chosen as model system for further investigations and genes identification controlling concerned traits. Transgenic plants with genes for resistance to insects and pests, tolerance to herbicides or improvement quality are already available on market. First generation of biotechnology experiments enable transfer of single gene but by now days experiments transfer of few genes together and using plants as bioreactors are possible.
Important factor in the success of the genetic improvement of crops was development of faster and more reliable methods, which allowed easier analysis of date as well as rapid information exchange. The application of information technology and development of statistical techniques to analyze genomic information is know as "bioinformatics". By integrating genetics with informatics investigations of whole genomes aims to elucidate the structure, function and evolution of plant genomes are faster. Together these technologies as integral part of classical breeding programs contributing significantly to shorting of plant breeding process.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MODULATION OF GENE EXPRESSION BYANTISENSE OLIGONUCLEOTIDES
Selma Kanazir, Sabera Ruždijić, Ljubisav Rakić
Institut za biloška istraživanja "S.Stanković", 29. Novembar 142. Beograd
Antisense-oligonucleotides (ASON) are synthetic, single stranded DNA or RNA segments, 15-25 nucleotides in length. Their sequence is designed to be identical to antisense DNA strand and to block the expression of specific gene by binding to its DNA or RNA in a sequence-specific manner. They exist in several forms. Antigene, oligonucleotides • mat bind to DNA and inhibit transcription through formation of triplex DNA. Ribozymes, catalytic antisens RNA molecules. Antisense oligonucleotides that bind to iRNA and block translation. Aptamers. oligonucleotides that specifically interact with proteins. Selected ASON is unique and specific in its capability to inhibit the expression of single targeted gene at any level of its regulation from DNA via RNA to protein. Their ability to modulate gene expression opened a new genetic strategy for the study of physiological and pathophysiological role of a gene by loss of its function. Moreover, the completely new field in developing gene specific therapeutics, antisense therapeutics, has emerged. Since, virtually all diseases are associated with abnormal or inappropriate gene activity, antisense oligonucleotides represent a powerful therapeutic tool especially in cancer and viral infections.
The application of antisense strategy for tlie inhibition of CD44 gene expression will be discussed.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
POPULATIONAL AND EVOLUTIONARY ASPECTS OF B-HROMOSOMES -
A case of yellow-necked mice
Mladen Vujošević i Jelena Blagojević
Department of Genetics, Institute for Biological research "S. Stankovic"
29 novembra 142, 11060 Belgrade, Yugoslavia, e-mail: mladenvii@ibiss.bg.ac.yu
B-chromosomes are supernumerary to the normal chromosomal complement. They differ from the main complement by their unequal distribution in species and irregular behavior during meiosis. According to definition B-chromosomes are not essential for the survival of their carriers. They persist in spite of that and are present in more than two thousand plant and animal species. In spite of numerous investigations a general agreement on the origin, role and mode of persistence of Bs in populations' has not been reached. A number of theories exist of which the parasitic model dominates over others. This model assumes the existence of mechanism that lead to the accumulation of B-chromosomes in gametes and the establishment of a balance between the degree of transmission of B-chromosomes and the elimination of their carriers due to lowered fitness. Opposed to this model is the heterotic model which assumes the existence of adaptive advantages of B-chromosome carriers. In a large number of investigated cases the parasitic model was used to explain the mechanism mat maintain Bs in populations, whereas the heterotic model was supported by a few cases only. In populations of the yellow-necked mouse, Apodemus flavicollis, B-chromosomes are present in different frequencies that correlate with the altitude and climatological variables. Some populations remain in equilibrium for years with regard to the frequency of specimens with B-chromosomes. Nevertheless, they display seasonal variations. The mechanism of the maintenance of Bs in populations does not fit any of existing models. The effects of Bs are not noticeable at the phenotypic level. Morphometric and analysis of developmental homeostasis has uncovered the consequences of their presence in the genome. In support of this is the discovery of DNA sequences that are only found in sample of species with B-chromosomes.
Recent investigations at the molecular level suggest that B-chromosomes are very useful for studying genome evolution. Therefore ii is of great importance that these investigations are included in complex populational-evolutinary investigations.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
POPULATION GENETIC ASPECT OF STRESS
Marko Anđelković i Marina Stamenković-Radak
Faculty of Biology, University of Belgrade
The interaction between individuals of a population and their environment is of great importance for species survival and conservation. If organisms are unable to either adapt to slow environmental changes, or react to rapid ones, such as climate change, pollution, ecological catastrophies of various degree, populations, and even the species, may become extinct. The evolutive strategies of species and genetical changes at the population level differ according to the environmental effect and genetical structure of populations.
Under stress conditions, the genotypes that allow rapid establishing of individual homeostasis of the cell or organism are favored. Although selection represents the slowest mode of the response to stress, it ensures a long term adaptation of the population to stress. Stress can be environmental, caused by external factors, and genomic, caused by another genome. Many population-genetic parameters are affected by stress.
Different approaches to studying the effect of environmental changes on the development homeostasis and genetic variability of the populations, as a response to stress, are illustrated by the examples on Drosophila species. One approach is based on the variability of the trait with a known genetic basis (Amy genotypes, inversion karyotypes). It attempts to correlate a given variability with fitness in modified laboratory environments and various natural habitats.
Another approach is based on the analysis of phenotypic variance, appearing in the response to stress, which determines the type of variability. By applying selection genotypes with extreme phenolypic values can be isolated. The organisms respond to environmental stress at the level of phenotypes by behavioral, physiological, or morphological adaptations, which can appear in the form of stress avoidance or build up of resistance. In addition to the variability associated with response to stress, genetic variability for the plasticity of response can be distinguished too. It is disputable whether the extent of stress is related to the changes in the population at the level of gene pool, or via changes in plasticity'.
Prediction of the ability of a species to evolve in the response to fast environmental changes is a major task of evolutionary biology. The populations of genus Drosophila can be suitable indicators of environmental changes. Factors limiting species distributions and genetic characteristics of populations (marginal and central) represent high priority for future research.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
PLANT ANTIMUTAGENS AND CANCER PREVENTION
Draga Simić, Jelena Knežević-Vukčević
Laboratory for Microbiology, Faculty of Biology, University of Belgrade
The possibility to modulate the response of a cell to environmental and endogenous mutagens has opened a new frontier for cancer control. The concept of preventing cancer by delay, i.e. by decreasing the rate of mutation accumulation, promoted the study of plant antimutagens as possible dietary anticancerogens.
According to their mechanisms of action antimutagens are placed in two major categories: desmutagens, which prevent the DNA damage and bioantimutagens. which modulate DNA repair and replication. In recent years a number of plant desmutagens (tannins and other phenolics, mono and diterpenoids, flavonoids, vitamins, etc.) were identified and their mechanisms of action include: inhibition of endogenous formation of mutagen, inhibition of activation of promutagens, antioxidant activity and scavenging of free radicals, blocking of mutagen or competition with, induction of detoxifying enzymes or acceleration of mutagen decomposition. Moreover, many plant desmutagens were found to inhibit tumor promotion and progression. On the oilier hand, only a few plant bioantimutagens are known (cinnamaldehyde, coumann, vanillin, umbelliferone, tannic acid) and they prevent mutations by stimulating the error-free DNA repair. In our laboratory antimutagenic potential of terpenoid fractions of sage (Salvia officinalis L), as well as pure monoterpenoids, has been investigated Prokaryotic and eukaryotic tests applied measure different genetic end-points: reverse mutations, replication fidelity, recombination, induction of mutagenic repair, chromosome aberrations. Detection of bioantimutagens was performed with the new E. coli K.12 assay (Simić et al, 1997). Antimutagenic potential of antioxidants from sage was "creened with E. coli and S. typhimurium tests (Mitić et al.. 1998). The results indicate that multiple mechanisms are involved in mutagenesis inhibition by terpenoids from sage. The protective effect of sage extracts against UV-induced mutations was confirmed in S. cerevisiae D7. Preliminary experiments indicated that dietary exposure to terpenoids from sage suppressed in vivo mytomicine C induced chromosome aberration in mice. Further study will investigate the possibilities of using sage terpenoids as anlicarcinogens.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE USE OF TRANSGENIC ANIMALS IN CARCINOGENICITY TESTING
Roy Forster
Centre International de Toxicology, 27005 Evreux, France
There is much current interest in the use of transgenic approaches in the evaluation of chemical carcinogenicity. This is already reflected in international guidelines for the safety testing of new drugs (from ICH), according to which the results of transgenic assays may be presented in the evaluation of carcinogenicity. An international ring-trial coordinated by 1LS1 (International Life Sciences Institute) is underway for the evaluation of four promising transgenic mouse strains. These are (i) the heterozygous p53+/- knockout mouse which lacks one copy of the p53 gene and shows enhanced sensitivity to genotoxic carcinogens, (ii) the TG.AC strain which bears a transgene containing the rets oncogene and has been found to show an enhanced sensitivity to the induction of dermal papillomas, (iii) the rasH2 strain which carries a transgene containing the human c-ras gene, complete with regulatory sequences of DNA and (iv) the xpa mouse in which the xpa gene is interrupted and excision repair of DNA is not functional. If the evaluative ring-trial produces encouraging results for the reliability and sensitivity of these transgenic bioassays, then they offer attractive advantages. The end-point is tumour production in a whole animal, underlining the relevance of these assays. Furthermore, evaluation of the tumour response takes place when the animals are still relatively young, avoiding the confounding effects of ageing. At present, the testing strategies to be deployed with these strains are not well defined. This presentation will examine the genetic basis of the assay systems, which is relevant to strategies of utilisation.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENETICS AND BREEDING OF MAIZE
- ACHIEVEMENTS AND CHALLENGES -
Drinić Goran i Babić Milosav
Maize Research Institute, Zemun Polje, Zemun -Belgrade
Genetic improvement of plant and animal species endured through millennia, firstly as domestication and then as continuous selection for phenotypes desired by breeders and farmers. The base for scientific breeding programmes was established by discoveries referring to a sex and inheritance in plants and by a re-discovery of Mendelian inheritance principles. Breeding programmes were aimed at the achievement of economic goals. The improvement and advancement in conventional breeding are related to the progress within the following segments:
- Improvement of breeding methods
- Maintenance and broadening of genetic variability
- Development of models of quantitative genetics
Plant breeding is a slow and painstaking process, requiring not only time (backcrosses and numerous tests), but also the aim achievement is sometimes uncertain (incorporation of genes from wild relatives).
Numerous breeding strategies are characterised by common elements such as necessity to obtain the flow of new and diverse germplasm, develop new gene combinations and identification of the best ones to be used in deriving new, improved varieties and hybrids with the same principal goal - the increase of yielding potential, tolerance to economically important diseases and pests, then improvement of quality and tolerance to stress environmental conditions.
Since a genetic base for the majority of economically significant traits is not completely clarified, superior individuals, selected for the development of the succeeding generation (cycle), will be identified by testing the progeny or will be based on individual performances. The empirical results clearly confirmed such an approach. The genetic potential of maize grain yield has been annually increasing by about 100 kg ha-1 during the last 80 years. The similar improvement was achieved in some other economically important plant species (rice, wheat, soybean, cotton, etc.). Although extremely successful in maintenance and improvement of agricultural species, such approach to plant breeding did not provide the complete comprehension of genetic control and manipulation (regulation) of complex genetic phenomena.
Biotechnological studies have been very intensive during the last two decades. However, molecular biologists are chiefly able to study (analyse) simultaneously only several genes, while economically the most important traits (so called quantitative traits) of plant species are controlled by combined action of several genes. Hence, the contribution of molecular genetics to our comprehension of genetic control and phenotypic expression of these traits is far more insignificant than it was initially expected.
In the last decade of the 20th century, the determination of a structure and expression of the genome brought about a mini-revolution within research on the genome. It seems that the combination and imbuement of molecular and quantitative genetics enlighten mechanisms of genetic control, regulating inheritance of quantitative traits, and therefore it is possible to accelerate and improve applied biological research. Furthermore, it became clear that the genomic revolution could not replace activities of a breeder, but more profound comprehension of complex genetic phenomena (Heterosis, epistasis, genotype x environment interaction...) would enable more efficient identification of parents within breeding programmes and development of more efficient breeding strategies. Broadening of our perspective on the field of quantitative and molecular genetic theory is one of the essential prerequisites, and therefore, scientific experts should be qualified within a field of intersection of molecular and quantitative genetics.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
ACHIEVEMENTS OF SUNFLOWER BREEDING AT THE END OF THE 20th CENTURY
Dragan Škorić
Institute of Field and Vegetable Crops, Novi Sad
Until 1970, sunflower breeding was characterized by the development of Russian high-oil varietal populations. The discoveries of CMS sources by Leclercq (1969) and Rf genes by several authors marked the beginning of a new era in sunflower breeding - an era characterized by the utilization of heterosis and development of highly productive sunflower hybrids. A large number of productive hybrids have since been developed in our country and a number of breeding centers around the world. This has contributed to a sharp rise in total sunflower acreage worldwide and has also helped increase and stabilize yields at a higher level than before. Using wild species through interspesific hybridization, researchers have found sources of genes for resistance to the following pathogens: Plasmopara helianthi, Puccinia helianthi, Verticillium albo-atrum, Verticilllum dahliae and Orobanche cumana. Yugoslav breeders have made a significant contribution to tliis with their discovery of sources of field resistance to Phomopsis. Also, an enviable level of tolerance has been achieved in the case of several other pathogens (Sclerotinia sclerotionmi, Macrophomina phaseoli, Alternaria helianthi). Sources of high tolerance to stress (drought) have been found as well. As of 1990, new races have been appearing in a number of pathogens. In an attempt to find sources of resistance to these new races, researchers have begun using wild species. The development of hybrids for special purposes has also yielded significant results in the last ten years, during which a large number of high-oleic and confectionery hybrids have been developed. In the past seven to eight years, the major trend in sunflower breeding worldwide has been the introduction of new biotechnology methods (anther cultures, protoplast fusion, molecular markers). Sunflower scientists have also begun working on the incorporation of genes from other species into the cultivated sunflower by transgenic methods.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
EFFICIENCY OF S1, HS AND FS METHODS OF RECURRENT SELECTION IN SYNTHETIC POPULATIONS OF NARROW AND BROAD GENETIC BASE OF MAIZE (Zea mays L.)
M. Ivanović, Violeta Anđelković
Maize Research Institute, Zemun Polje, S. Bajica 1, 11080 Belgrade-Zemun
Two maize synthetic populations of narrow and broad genetic base, (B73xMo17)F2 and ZPSin 4, respectively, were used in the present study. F2 generation of the population of narrow genetic base was used as an initial cycle (progeny of two homozygous inbred lines), while the population of broad genetic base was developed by the cross of 120 inbred lines of different origin after four recombinations under random mating conditions.
Three methods of recurrent selection were simultaneously applied to both populations: S, (progenies of the first selfing), FS (full sib progenies) and HS (progenies with an unrelated inbred line F7R - used as a tester for both populations). Each population and each method of selection considered 100 randomly selected progenies planted according to the Nested design with the selection intensity of 10% cycle"1. Selection for grain yield was conducted and in such a way four cycles of selection for each synthetic, i.e. each method of selection was completed.
In order to compare efficiency of presented methods of selection in synthetics of narrow and broad genetic base, results on grain yield of all cycles of selection of synthetics per se, as well as, of progenies of certain cycles, i.e. methods of selection with two unrelated testers F7R (inbred used as a tester in the process of selection) and ZPL39. were used. Field plot experiments were set up according to the RGB design in 1995 (three locations) and 1996 (two locations), separately for the synthetics per se and HS progenies.
Results were processed by the method of the analysis of variance of the RGB design, while selection progress was expressed by the estimates of linear regression coefficients for the appropriate cycles of selection. Obtained results point out that selection progress depended on the synthetic population, as well as on the applied method of selection. The highest estimate of linear regression coefficient for the improvement of the population per se (b=0.609 t ha-1 cycle-1) was obtained for the synthetic of broad genetic base with the application of the HS method of selection. The same method of selection in progenies of the same population with the tester F7R resulted in the highest regression coefficient (b=0.9591 ha-1 cycle-1).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENE POOL CONSERVATION AND TREE BREEDING
V.Isajev, A.Tucović
Faculty of Forestry, Belgrade
Previous study of population characteristics and the effects of natural and artificial selection on the genetic structure of tree populations contribute to the conservation and enhancement of autochthonous species gene pool. The balance model of genetic structure of tree populations reveals their new properties and requires further research, especially of the relations of subpopulations, half-sib families and organisms to variable environmental factors. The interaction of these two systems is of fundamental significance for the exchange of genetic material within natural and cultural communities of trees and shrubs.
The conservation of genetic resources of populations is a part of the complex strategy of tree breeding and it is realized through different forms of conservation: in situ -natural reserves, national parks, seed stands, tree groups and individual trees, and ex situ -arboreta, live archives, seed orchards, provenance tests and progeny tests. Each strategy has its own advantages and disadvantages, both for breeding and for the conservation of biodiversity and genetic resources that makes it possible. The advantages and disadvantages are the result of the classical methods of tree and shrub improvement. They are restricted because: (1) useful properties can be improved only between the related plants, (2) the selection of the best possible combinations of characteristics lasts for a very long time and (3) in crossing, the entire genomes are crossed and not individual genes of the selected trees.
The relationship between gene pool conservation, tree improvement and rational utilization of forests is accomplished through: the balance between in situ conservation of forest genetic sources and sustainable forest management on the one hand and management of protected lands on the other hand. The conservation and management of genetic resources in situ and ex situ and the breeding activities are complementary strategies.
The focus of activities in tree gene pool conservation should be shifted from the physical protection of stands or trees to the conservation of their reproductive function and thus the potential genetic variability should be directed into free genetic variability, available to natural and directed selection.
The increased utilization of the genetic potential in tree breeding should be realized through the following activities: improved production of selected seeds of tree species, which implies the knowledge of genetic structure and variability of seed stands; establishment and management of seed orchards as specialized plantations for multiannual production of genetically good-quality seed of significant tree species, as well as through the application of the concept of new selections, by the models adapted to the requirements of forestry, which include the genetic characteristics of the species, the components of tree yield and the effect of the environment.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
NEW APPROACH IN TOMATO BREEDING (Lycopersicon esculentum Mill.)
Živoslav Marković, Jasmina Zdravković, Mirjana Mijatović
Agricultural Research Institute SERBIA, Centre for Vegetable Crops, Karadordeva 71, 11420 Smed. Palanka
The first tomato variety was breeded before 130 years and now has more man 2000. Foremost was breeded varieties, and after 1930. year F1 hybrids. Now varieties use only in late and processing production. As the genetic variability of tomato is narrow (from the reason of genetic erosion - electropheretic studies of alloenzymes show great similarities between the »European« and primitive cultivars) for amelioration of quality and diseases resistance of modern cultivars breeders use the »exotic« germlasm of wild and sami-wild relatives. Breeder teams make thats in state and private institutions of USA, Canada and France. Others benefit »elite« germplasm of new breeded and improved cultivars, to speed breeding process (many of properies, especially for disease resistance, are controlled by dominant genes). Also, breeders work on amelioration of photoperiodic activity, resistance to drought and high temperature, good emergence on low temperature, metabolism modification by change of concentration of CO2 and oxygen. Great possibilities provide application in breeding of biotechnology (somoclonal variations, fusion of protoplast, gene isolation, cloning and anther culture, use of genetic markers) and genetic engineering. Both are the challenge of new generation of tomato breeders. The first results are gained (tolerance to AMV and TMV incorporated of viral gene from oat. resistance of tomato to total herbicide Roundap by introduction of mutant for EPSP synthesis, resistance to Lepidothere and etc.).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
CONTRIBUTION OF GENETICS IN BREEDING OF SMALL GRAINS
Desimir Knežević1, Novo Pržulj2
1ARI SERBIA, Center for Small Grains, Kragujevac
2Research Institute of Field and Vegetable Crops, Novi Sad
The plant breeding is older than sedentary agriculture is based on the quite well production of non-shattering plants which have been available to these early farmers. The plant breeding began by early farmers to select crop genotypes for sedentary agriculture 10-12 thousand years ago was still in use with few refinements until 100 years when made major discoveries about sex and heredity plants (Wessman De Vries etc). However, in 1901 Menders Low of Heredity have got most importance in plant breeding, as well in the same time for breeding of grains.
The creation of new desirable genotypes began 100 years ago. Since than, plant breeding has been honed and refined in next area activity l)development of techniques and procedures; 2) collection of natural genetic variability and its use-in the present , future evaluation and plant breeding programs: 3) some techniques, such as tissue culture, colchicines treatment, induced mutation which have expanded the breadth of germplasm that the plant breeder could use. The genetics, biochemists and cell biologists in last four decades have been using sophisticated instruments and model organism to learn about the genetic material DNA, and how it functions to expression of phenotypes.
Plant breeding directs towards economic objectives. Genetics is basic to rational plant breeding but the biotechnology involves other sciences such as chemistry, plant pathology and biometry.
Traditional plant breeding involves making numerous crosses from parents, which have been selected for desirable attributes and on visual criteria and measurement of many traits. In some of derived progenies were combined the best traits from the parents. Through numerous cycles of selection, a new cultivar can be developed which carry combination of the best desirable traits from parents. The conventional plant breeding programs are long-term work and effort of breeders who are looking for optimal selection procedures. The complexity of the breeders work lies in the fact that up to 10 or more years may be necessary between the initial cross and the approval of new cultivar. The breeders permanently looking for answer on the few questions example: What resources are required for genetic progress of cereals? Which methods are most efficient in cereals breeding? How to broaden genetic variability? There is competition between breeding companies to reduce time of necessity time of cultivar creation. There are also various limitations to plant improvement through conventional breeding. Besides long term of expectation of new cultivar as a limitation factor in conventional plant breeding limitation factors are also expenses for necessity analyses of large number of plants and many requirements for plant cultivation. But in some cases also, the breeder has limited genetic variation available for its incorporation into new cultivars. The various techniques developed in molecular biology and biotechnology can be used in plant breeding. The technical advanced in recombinant DNA and molecular biology methods have been developed and should have now a major impact on the efficiency and success of conventional breeding. Those methods are contribute to increasing of our knowledge of the inheritance and are providing manipulation of responsible genes which controlled certain biological traits.
In relation to the social demand, the aims of breeding programs are stilt to increase grain yield potential, to increase resistance to diseases and pests and to improve quality and tolerance to environmental stresses.
Crop improvement is slow and tedious process involving operations which not only consume great amount of time, such as backcrossing and testing, but some of which are just very difficult to achieve, for example incorporation of wild genes or inbreeding naturally crops. The numerous of molecular and molecular genetics technologies have a great impact for direction of breeding strategy, at various specific steps assisting the breeder in making achievement to a more manageable level.
There are different breeding strategies which characterized common elements: 1) means of achieving flow of new and diverse germplasm into the program; 2) new gene combinations; 3) identification of best combinations; 4) exploitation for cultivar and hybrids development. Some biotechnical methodologies can be applied at several of those steps: a) somatic hybridization to generate new hybrids; b) direct nuclear substitutions into alien cytoplasm; c) mutation/selection and cell genetics; d) generation of somaclonal variation; e) genotype fingerprinting; 0 germplasm preservation; g) gamete breeding schemes; h) gene isolation, transfer and expression: i)molecular site directed mutagenesis in situ.
Highly specific genetic selection methods would require diagnostic biochemistry techniques involving monoclonal antibodies, molecular probes and sophisticated methods of isozyme, proteins and sequence DNA analysis. The breeders in the future need to become familiar with these methods. The development of biotechnological tools can assist the improvement of plant breeding as well as seed production and genotype protection. Extensive investigation are conducted on: a) a description of molecular markers tagging the genetic control of agronomic traits and their applications for marker assisted selection: b) the identification of the physical structure and DNA composition of chromosomes and their similarity among species and; c) gene transformation methods.
The genetic maps of crop species play important role for cultivar improvements because plant breeders are able to directly manipulate specific genes by exploiting "marker assisting selection". A molecular marker represents a way to hasten the transfer of desirable traits by earning out a selection not directly on the trait, but on the molecular markers tightly linked to it.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE EXPANSION OF TRINUCLEOTIDE REPEATS AS A GENETICAL
CAUSE OF NEUROLOGICAL AND PSYCHIATRIC DISEASE
Romac S.
Faculty of Biology
The expansion of trinucleotide repeats is the cause of 12, so far, detected hereditary neurological diseases. This type of mutations was discovered few years ago which is the first and only type of mutations found exclusively in humans. These mutations are not inherited by Mendel low and the diseases, which they cause, show the phenomenon of anticipation. Anticipation is the phenomenon describing the fact that the disease, in next generation, has an earlier age of onset and more severe clinical manifestation. Genetical basis for anticipation is the expansion of trinucleotide repeats from generation to generation. If there are more triplets in mutated gene the clinical manifestation is more severe and the disease has an earlier onset. So far all the diseases of trinucleotide repeats were/are connected to hereditary neurological diseases (Huntington chorea, Spinocerebellar ataxia type 1-7, Dentatorubral Palidoluysian atrophy, Myotonic dystrophy etc.) Many of psychiatric diseases like schizophrenia, autism, bipollar disorders, also show the phenomenon of anticipation and non-Mendelian type of inheritance, are candidates for the type of trinucleotide repeats diseases.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
APPLICATION OF MOLECULAR TECHNIQUES IN DIAGNOSTICS OF CHROMOSOMAL DISORDERS
Milena Stevanović1 i Marija Guc-Šćekić2
1 Institute of
Molecular Genetics and Genetic Engineering, Belgrade
2 Mother and Child Health Care Institute, New Belgrade
Chromosomal disorders are consequence of structural and numerical chromosome aberrations. Approximately 20% of all conceptions (embryos) have chromosomal aberration, but most of them are spontaneously aborted and frequency of chromosomes aberrations at birth is 0.6%. Not all chromosomal rearrangements are associated with phenotypic expression. It has been generally accepted that autosomal aberrations result in delay in psyhornotoral development, while sex chromosome aberrations are associated with irregular sexual differentiation and sex determination. Therefore, precise and reliable prenatal and postnatal diagnostics of chromosomal aberrations is of great importance for planning and obtaining health individuals.
In seventies, cytogenetic was revolutionized by introducing chromosome banding techniques, such as G, Q and R banding. Banding techniques, for the first time, allowed accurate differentiation of chromosomes. Chromosome banding permits more accurate diagnostics of chromosome aberrations, but this technique also have some limitations including inability to detect microchromosome rearrangements and rare mosaic karyotypes and to identify marker chromosomes. In the last few years, the fast progress has been made in more accurate diagnostics of chromosome aberration by application of new molecular techniques. FISH (Fluorescent in situ hybridization) is new highly sensitive technique, that enables visual identification of chromosomes or part of chromosomes fixed on slides. FISH has been used, as an addition to routine cytogenetic analysis, to achieve a higher sensitivity and specificity. Application of new molecular techniques such as FISH, PRTNS, PCR, RT-PCR, RFLP, LOH (lost of heterozygosity) became unavoidable in making accurate diagnosis. Therefore, fast, precise, accurate and reliable diagnosis of chromosome aberrations is possible only by combination of cytogenetics and molecular techniques.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
HETERODIMERIC PENICILLIN G AMIDASES - MOLECULAR MECHANIZAM OF POSTTRANSLATIONAL MATURATION OF INACTIVE PRECURSOR AND ITS BIOTECHNOLOGICAL IMPICATION
Goran Ljubijankić
Institute of Molecular Genetics and Genetic Engineering Vojvode Stepe 444a, PO Box 794, 11000 Beograd, Yugoslavia
The enzyme penicillin G amidase (penicillin G acylase, EC 3.5.1.11, PGA) catalyzes the hydrolysis of the acyl group of penicillin, yielding 6-aminopenicillanic acid, a key intermediate in the production of semisynthetic beta-lactam antibiotics. The main industrial source of the enzyme is Escherichia coli and that was a reason for intensive studies of this protein as well as the gene encoding it (pac). It appeared that E. coli pac gene has a specific structure which causes a synthesis of PGA as a single, biochemically inactive, polypeptide precursor. Its maturation occurs by removal of single peptide at the inner membrane, processing of the spacer peptide in the periplasm followed by formation of a biochemically active PGA composed of two different subunits (α approx. 23 kDa, and β approx. 65 kDa). The free (3 subunit carries N-tenninal serin which is a single amino-acid catalytic site. It was shown that a complex posttranslational maturation of PGA precursor was the limiting factor for a recombinant PGA overproduction in E. coli. Therefore, a investigation of the maturation process is not interesting only from a fundamental point of view, but also for industrial production of the enzyme. The following topics of our research will be discussed: the limited E. coli pac overexpression in yeast (a): the maturation as temperature-dependent process (b); characterization of a domain of precursor involved in this regulation (c); the proteolitical processing as an autocatalitic intermolecular mechanism (d).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
STRUCTURAL ANALYSIS OF THE prt GENES REGULATORY REGION IN NATURAL ISOLATE Lactococcus lactis subsp. lactis BGIS29
Nataša Miladinov, Irena Spasojević i Ljubiša Topisirović
Institute of Molecular Genetics and Genetic Engineering, V. Stepe 444a, P.O.Box 794, 11001 Belgrade, Yugoslavia
Soj Lactococcus lactis subsp. lactis BGIS29 is a natural isolate from home-made cheese. The isolate BGIS29 produces the PI-type proteinase. Regulatory domain of the prt genes was sequenced. The prtP and prtM genes expression is controlled by divergent promoters PprtP and PprtM, respectively. Transcriptional start points of both prt promoters were determined. Sequence analysis also showed that putative secondary structure could be formed. Previous results have shown that the expression of the prt genes in BGIS29 is regulated by the presence of casitone in chemically defined medium (CDM). To evaluate the role which the regulatory region has on the prt gene expression, the influence of the nucleotide sequence context on DNA bending was investigated. Ligase-mediated cyclization and gel mobility experiments together with the computer-assisted prediction of a putative DNA bending are performed. Computer-assisted prediction of the DNA bending showed that a putative center of the bending is located in the vicinity upstream of the PprtP promoter. Gel mobility experiments revealed that the PCR fragment of 350 bp containing complete regulatory region is curved. The retardation value (R) was 1.2 in the absence of and 1.4 in the presence of the MgCl2. Moreover, truncated PCR fragment (270 bp) containing both, PprtP and PprtM promoters and center of banding retained bendability. Truncated fragment is lacking 80 bp located between the prtM gene and the center of banding. This fragment has R values 1.1 and 1.5 in the absence and presence of MgCl2, respectively.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
INDUCTION OF ALBUMIN SYNTHESIS IN BELGRADE LABORATORY (b/b) RAT RETICULOCYTES
Zvezdana Popović, Aleksandra Đurković, Jelena Zarić i Marina Lušić
Institute of molecular genetics and genetic engineering, Vojvode Stepe 444a, P.O. Box 794, 11000 Belgrade, Yugoslavia
The Belgrade laboratory (b/b) rat is an autosomal mutant characterized by hemolytic anemia. An additional protein band of 68kD molecular mass, which corresponds to the rat serum albumin, characterizes b/b erythrocyte membrane. Albumin has been found in the cytoplasm of both, control (+/+, bled +/+ and phenylhydrazine treated +/+) and b/b erythrocytes. In b/b rat erythrocytes cytoplasm, amount of albumin is 5-7 times increased compared to the +/+ rat erythrocytes
Albumin synthesis is confirmed by immunoprecipitation of in vitro newly synthesized proteins with the specific polyclonal anti-human albumin antibodies. Albumin represents 35-40% of total proteins synthesized in b/b reticulocytes, 22-26% in reticulocytes of rat treated with phenylhidrazine and drops to 6-8% of total newly synthesized proteins in reticulocytes of bled rats.
Albumin synthesis in rat reticulocytes is further confirmed by the identification of albumin mRNA by RT-PCR method.
The present results confirm that albumin is synthesized, not only in liver but in rat reticulocytes, too. Albumin synthesis is inducible and increased in damaged (b/b and phenilhydrazine treated normal rat). Increased amount of albumin in damaged reticulocytes points to the specific function of albumin in stressed cells.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
HCV GENOTYPE DETECTION BY SSCP IN YUGOSLAV POPULATION
Stamenković Gorana, Laketa Danijela, Gudurić Jasenka, Veljković Emilija, Krtolica-Zikić
Koviljka i Dimitrijević Bogomir
Institut
za nuklearne nauke "Vinča", Laboratorija za radiobiologiju i
molekularnu genetiku,
Beograd, Jugoslavija
Hepatitis C virus (HCV) appears to be the major cause of chronic hepatitis (HCV). It has a positive single stranded RNA genome, which displays considerable variability precluding permanent immunity and the production of effective vaccine. On the basis of genomic sequence, HCV isolates are classified into six major genotypes and at least 15 subtypes. HCV genotypes play an important role in disease stratification and treatment modalities. These viral genotypes differ in their geographical distribution. Investigation of HCV genotype distribution in our region showed that genotypes Ib, 3a and la are the most frequent genotypes (90% of all analyzed samples). Genotyping was performed by RT-PCR method with type-specific primers which are complementary with specific sequences in core-region of HCV genome. However, genotyping based on the core segment yields frequently ambiguous results and/or are completely uninformative. On the other hand, the analysis the sequence of 5'-untranslated (NTR) region of the HCV genome also provides genotype specific information. SSCP mutational screening of NTR was optimized to yield a clear distinction between the genotypes in Yugoslavia. This is potentially valuable contribution to clinical diagnostics being suitable for routine screening and economically superior to available commercial tools.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
BSA WITH CANDIDATE DROUGHT-INDUCED GENES
Vesna
Lazić-Jančić, D. Kovačević i S.A. Quarrie
1 Maize Research Institute "Zemun Polje", Yugoslavia
2John Innes Centre, Norwich, UK
In response to drought stress, many genes are induced in plants and the question many research groups are trying to answer is to determine which of these genes have important roles to play in stressed plants. The approach normally adopted to determine the function of a gene is transformation and gene over-expression or silencing. However, interpreting the results of such studies is not always easy. An alternative approach is bulk segregant analysis (BSA). BSA using cDNAs for the gene as hybridization probes may give an answer to the question through examination of allele frequencies in two bulks of plants selected to differ specifically for the trait of interest.
We have recently used the amplified fragment length polymorphism (AFLP) technique for mRNA fingerprinting of two inbred maize lines differing in drought resistance and grown under drought stressed and control conditions. cDNA fragments prepared from leaves were identified that were specific either to a particular treatment or to a particular genotype or, in some cases, specific to a genotype only under one treatment. Using a range of primer combinations, over 100 fragments were identified and 14 of these have been cloned so far. Hybridizations to dot blots of mRNA confirmed that the majority of the cloned fragments were differentially expressed by either genotypes or treatments.
We are using BSA to screen the cDNAs that were differentially-expressed between treatments for differences in allele frequency between maize composites before and after several cycles of selection for high yield under drought conditions. Two pairs of composite populations are being tested: one pair produced entirely at CIMMYT, Mexico (Tuxpeno Sequia), and the other originating from CIMMYT and developed by the MRI in Zambia (DTP).
One AFLP-derived cDNA (M47/3C) showed allelic variation between the selected and unselected composites and this was mapped to the long arm of maize chromosome 8. We are now examining a mapping population of maize for coincidence of quantitative trait loci (QTLs) for drought responses and the M47/3C gene, and the results so far will be presented.
We are grateful to the Royal Society and UNESCO for providing financial support for this work.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
SOX GENES: BRAIN DEVELOPMENT AND SEX DETERMINATION
Milena
Stevanović, Nikola Arsić, Tamara Rajić, Slavica Stanojčić, Tomislav Terzin,
Vesna
Verbić, Maja Vujić, Nebojša Mirković i Danica Grujić
Institute of Molecular Genetics and Genetic Engineering, Belgrade, Yugoslavia
The mammalian sex determining gene SRY encodes a protein with an HMG-box, a motif found in several DNA-binding proteins, including some transcription factors. The mammalian genome contains a family of genes related to SRY and homology is restricted to the region of SRY that encodes an HMG-box. These genes have been named SOX (SRY related HMG-box). The SOX family is large comprising at least 30 genes and by similarity within the HMG-box SOX genes are grouped in several subfamilies.
Although only few SOX genes have been characterized in details, increasing evidence indicates that SOX genes have important roles in the regulation of development and mutations in SOX genes are associated with human diseases.
SRY is the only member of the SOX gene family to b directly linked to the sex-determining pathway. Expression studies suggest that many SOX genes are embryonically expressed and expression patterns indicate mat some SOX genes might have roles both in controlling development of central nervous system and sex determination.
By analogy to SRY, SOX genes are expected to have important regulatory function and further analysis of this new family could lead to better understanding of processes of development of nervous system and sex determination.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
DETECTION OF B CHROMOSOME SPECIFIC DNA MARKERS IN
POPULATIONS OF YELLOW-NECKED MOUSE Apodemus flavicollis (RODENTIA, MAMMALIA)
Nikola Tanić1, Nasta Dedović2, Mladen Vujošević1 i Bogomir Dimitrijević2
1 Institute for Biological Research "Sinisa Stankovic", Belgrade
2 Institute for Nuclear Sciences''Vinca, Belgrade
The molecular structure of the B chromosomes of Apodemus flavicollis has been analyzed by DNA comparison of animals with and without B chromosomes using AP-PCR- based DNA profiling. Mice harboring one, two, three or none B-chromosomes were examined. Chromosomal stmcture was scanned for variant bands by a series of arbitrary primers and informative ones were selected. The selection criteria were their ability to differentiate between individuals of the species, to detect markers common for both A and B chromosomes and, importantly, to differentiate between A and B chromosome set. Along with primer, profiling conditions were found to be critical for meeting the criteria. Primers and conditions of the analysis that demonstrate structural characteristics unique to the B chromosome set are described. These characteristics include variant bands as qualitative parameters and altered electrophoretic band intensities as quantitative distinctions estimated by integration of densitometric profiles of electrophoretograms. Features of DNA profiles illustrate the reliable method of analysis of B chromosomes in the context of full set of A chromosomes. It provides an easy access to B chromosome specific molecular markers. Models for the origin of yellow-necked mouse B chromosomes are discussed in the context of presented data.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
CONTROL OF RESISTANCE GENES IN ANTIBIOTIC-PRODUCING ACTINOMYCETES
Branka Vasiljević, Milorad Kojić, Nataša Milojević, Sandra Vajić, Marijana Miljković,
Tatjana Ilić
Institute of Molecular Genetics and Genetic Engineering, Belgrade
Actinomycetes have been the focus of much study because they produce a numerous medically important antibiotics. It is important to determine the mode of expression and regulation of genes governing resistance to antibiotics since those genes are usually clustered with biosynethetic genes. Concerning regulation of resistance genes in antibiotic producing actinomycetes, four different models have been described. The first model describes a complex regulatory cascade for biosynthetic genes in streptomycin-producing S. griseus. The model includes the positive A-factor signal which is transferred to strR, a regulatory gene. The resistance gene aphD is transcribed mainly by read-through from the A-factor-dependent strR promoter. The second regulatory mechanism is based on translational attenuation of MLS (macrolide, lincosamide and streptogramin) resistance genes or transcriptional attenuation (the resistance tlrA gene from the tylosin producer). The third mechanism of regulation of antibiotic resistance genes is demonstrated in a producer of novobiocin. The resistance gene from tins organism is driven by a promoter which responds to the superhelical density of DNA. The forth regulatory model is described in our laboratory and it is based on translational autoregulation. The aminoglycoside-resistance determinants from M. zionensis (sgm gene), and S. tenebrarius (kgmB gene) have been cloned and it has been shown that mechanism of resistance involves methylation of the 30S ribosomal subunit. The Sgm and KgmB methylases recognise specific sequences on 16S rRNA and also regulatory sequences on their own mRNA. In that way, if all ribosomes are modified, methylases bind to their own mRNA preventing further translation.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE ISOLATION OF BUCKWHEAT GENOMIC CLONE CONTAINING THE GENE FOR METALOTHIONEIN-LIKE PROTEIN
Jelena Brkljačić1, Jelena Avramović1, Paja Šijačić1, Svetlana Radović2, Vesna Maksimović1
1 Institute for Molecular Genetics and Genetic Engineering, Vojvode Stepe 444a, Beograd,
2 Faculty of Biology, Studentski trg 3, Beograd
Metalothioneins are small molecular weight cysteine-rich proteins, with a capacity to bind metal ions. They are found in animals, fungi and bacteria, where they take part in different physiological processes. Animal genes for metalothioneins can be induced by a variety of stimuli, such as metal ions, hormones and secondary messengers, as well as by different kinds of stresses. Metalothionein genes are found also in plants, but only a few of them were characterized at the protein level. The data about elements involved in their expression are also scarce.
A cDNA clone pBM290, 600 bp long, has been isolated from cDNA library from buckwheat seed. The complete coding region for metalothionein-like protein has been identified in this clone. Based on Southern blot of buckwheat seed genomic DNA digested with restriction enzyme EcoRI, a fragment of 5 Kb (H2) was chosen as a template for asymmetric PCR. The primer, PREX290, was derived from pBM 290 clone sequence. To single-stranded DNA, obtained by asymmetric PCR, oligo-dG tails were added to 3'-end. Further amplification was performed with two primers - PREX290 and a NotC, which contains restriction site for NotI enzyme and oligo-dC tail. PCR products, 700 bp long, were ligated into the Bluescript vector and cloned in XL 1 Blue E.coli strain. Three clones, with sequences of a gene for metalotliionein-like protein were chosen on the basis of plasmid electrophoretic mobility, PCR amplification with PREX290 and NotC primers and hybridization with pBM290 probe. The final identity of these genomic clones will be confirmed by sequencing.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
COMPARATIVE STUDY OF Micromonospora STRAINS PRODUCING AMINOGLYCOSIDE ANTIBIOTICS
Sandra Vajić, Milorad Kojić, Branka Vasiljević
Institute of Molecular Genetics and Genetic Engineering, Belgrade
Micromonospora strains which produce aminoglycoside antibiotics have high level of resistance to 4.6-disubstituted deoxystreptamine antibiotics that are structurally similar to their own products. The genes responsible for resistance in Micromonospora producers were cloned previously (the grm gene from M. purpurea, gentamicin producer, and the sgm gene from M. zionensis, producer of G-52). Both genes encode specific 16S rRNA methylases that modify target for aminoglycoside antibiotics and thus protect it. Southern hybridization experiments revealed that highly homologous genes are present also in other Micromonospora strains which produce aminoglycoside antibiotics. Genetic analyses of antibiotic producing organisms have revealed that in most cases the genes for resistance to antibiotics are clustered with their corresponding biosynthetic genes and, in some cases, with regulatory genes. This may indicate that expression of genes conferring resistance and biosynthetic genes are correlated ensuring that resistance is always operational when the organism is exposed to its own toxic product. Regulation of cloned sgm and grm resistance genes was studied. Although these genes are more then 80% identical in coding regions, their regulation is completely different. Therefore, it was interesting to analyse surrounding sequences, possible biosynthetic genes, for their homology. By hybridization with a DNA probe derived from pMK2 plasmid carrying the sgm gene on 7.2 kb fragment, chromosomal DNA of different aminoglycoside producers was analysed. Sequence analysis demonstrated that there is an open reading frame upstream of the sgm gene which show significant homology to the genes that encode aminotransferases, enzymes involved in biosynthetic pathways of different antibiotics.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE APPLICATION OF GENETIC MARKERS IN PREVENTION OF DAIRY CATTLE MASTITIS
Marijana Vučinić1, Z. Gajić1 i P. Stojić2
1 Faculty of Agriculture, Department for Livestock Production, Nemanjina 6, Zemun
2 PKB "INI Agroekonornik", Livestock Institute, Padinska Skela, Beograd
As many authors stressed, the most costly disease in dairy cattle of modern milk industry is mastitis. It is an inflammation of the mammary gland, usually caused by microbial infection in its one or more quarters. Protection of the mammary gland from microbial infection is governed by innate (nonspecific) and acquired (specific) immunity. Effectors of innate immunity are anatomical and physilogical inflammatory defense barriers, many antimicrobial substances, such as keratin, lactoferin. lysozyme. components of the complement, lactoperoxidase/thiocyanate/hydrogen peroxide system and phagocytic cells (macrophages. neutrophils and leukocytes). Effectors of acquired immunity are T- and B-lymphocytes and specific antibodies. Following microbial infection of the udder, large numbers of phagocytic cells enter the mammary gland. The number of these cells in milk is termed as Somatic Cell Count (SCC) or Somatic Cell Score (SCS) and practically serves as an indicator of alteration in the mammary gland secretion or its infection. The ability of phagocytes to ingest and kill microorganisms is a genetically regulated trait controlled by major histocompatibility complex genes (MHC genes). These genes encode histoglobulins and molecules on surface of leukocyte membranes. These molecules in cattle are known as Bovine Leukocyte Antigens (BoLA). Genes that encoded their synthesis (BoLA genes or MHC genes) influence immune response and disease resistance in cattle. Because, BoLA and Somatic Cell Score are potential genetic markers in selection of cattle to mastitis resistance.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
FUNCTIONAL ANALYSIS OF THE RAT βbminy - GLOBIN GENE PROMOTER
Nataša
Grujičić, Sonja Pavlović, Sanja Pavlović, Zvezdana Popović
Institute for Molecular Genetics and Genetic Engineering, Vojvode Stepe 444A
P.O. Box 794, 11000 Belgrade, Yugoslavia
The rat β- globin gene locus is unique for its organization. It consists of either five or three adult β- globin genes depending on haplotype and we presume that transcriptional regulation of these genes within the locus has its own specific properties. The object of our investigation is the rat βbminy - globin gene promoter..
We have analyzed the regulation of transcription of the rat βbminy globin promoter using CAT functional assay. We have cloned 900 bp long promoter region from βbminy - -globin gene upstream of the CAT reporter gene. Promoter activity was investigated in rat erythroleukemia cell line (REL), a permanent cell line arrested at the proerythroblast stage of development. Upon treatment with DMSO, REL cells c m be induced to undergo erythroid differentiation in a way that closely mimics normal red-cell maturation.
Since we decided to analyze promoter activity in stable transfection assays, we also cloned HS3 (hypersensitive site 3) from human p-LCR in the reporter construct. The reason for this is that HS3 confers gene-copy-number-dependent expression of the linked gene in REL cells, independent of the site of integration in the host genome. We analyzed the activity of the rat βbminy -globin promoter in undifferentiated and differentiated stably transfected REL cells by measuring CAT activity. In mis experiment we showed induction of transcription from βbminy -globin promoter upon erythroid differentiation of REL cells.
In order to determine which regions of this promoter are functionally important for its tissue-specific transcriptional activation and inducibility we deleted certain parts of the promoter and began the analysis of the deletion mutants using CAT assay.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
SYNTHESIS AND SECRETION OF THE SGM METHYLASE IN Saccharomyces cerevisiae
Tatjana Ilić, Milorad Kojić, Goran Ljubijankić, Branka Vasiljević
Institute of Molecular Genetics and Genetic Engineering, Belgrade
The baker's yeast Saccharomyces cerevisiae was successfully used in many cases for die expression of heterologous genes. The aim of the present work was to clone the sgm gene to yeast expression vector pGoBO under yeast promoter and secretion signal. This approach offers the advantage for purification of the Sgm methylase from media.
The sgm gene was cloned from actinomycete Micromonospora zionensis, producer of an aniinoglycoside antibiotic, as resistant determinant. The gene encodes a specific methylase that modifies 16S rRNA. It was previously shown that the expression of this gene in different Streptomyces strains, in E. coli and in Micromonospora melanosporea gives resistance to gentamicin. It was interesting to test whether the sgm gene could confer resistance to gentamicin also in eukaryotic organism such as Saccharomyces cerevisiae. Computer analysis of 16S rRNA and 15S and 18S rRNA of yeast revealed that there is high homology in regions where gentamicin can bind. Therefore, we also tested the intracellular expression of the sgm gene. Such expression of the sgm gene in S. cerevisiae could lead to successful usage of this gene as dominant marker in yeast vectors with increased host range.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE REGULATION OF cls GENE EXPRESSION IN BACTERIA Escherichia coli K12
Radmila Janjušević, Mirjana Lilić, Milija Jovanović, Goran Jovanović i Dragutin Savić
Institute for Molecular Genetics and Genetic Engineering, V. Stepe 444a, P. Fah 794, 11001 Belgrade
Escherichia coli cls gene codes for cardiolipin synthase, the enzyme that catalyses reversible phosphatidyl group transfer from one phosphatidylglycerol molecule to another to form cardiolipin (CL) and glycerol. CL is one of three major glycerophospholipides constituting biological membranes, but different from others for its tetraacyl structure and supposedly plays unique functional role. The membrane content of CL increases as E. coli cells enter the stationary phase. The specific activity of CL synthase is also about 10-fold higher in cells that are in the stationary phase than in those in exponential phase of growth. To examine cls gene expression strains containing operon (cls-lacZ+) and protein (els- 'lacZ) fusions were constructed. The same shape of curves of β-galaclosidase activities in different time points for operon and protein fusions indicate that the regulation of els gene expression is on transcriptional level. These results confirmed that the transcription of els gene increased when the cells enter the stationary phase. In addition, there is probably more than one regulatory factor involved in regulation of the cls gene transcription. The role of several E. coli global regulatory factors was examined by measuring β-galactosidase activity of an operon fusion in different regulatory mutants and by gel shift experiments. These results have revealed that some of global factors are also involved in the regulation of cls gene transcription.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
CysB ACTIVATOR ACTS AS A REPRESSOR OF THE HslJ EXPRESSION THROUGH N-ACETYL SERIN MODULATED DNA BINDING IN THE hslJ LOCUS
Milija Jovanović, Mirjana Lilić, Radmila Janjušević, Goran Jovanović i Dragutin Savić
Institute for Molecular Genetics and Genetic Engineering, V. Stepe 444a, P. Fah 794, 11001 Belgrade
CysB
protein belongs to the LysR family of regulatory proteins. It activates the
transcription of genes involved in the cysteine biosynthetic pathway as well as
some other
genes such as tauABCD, cbl, adi, lysU,. In turn, CysB negatively regulates expression of its
own gene. N-acetyl serin (NAS) is an inducer of the CysB activated
transcriptions but anti-
inducer of the autogenously controlled cysB expression. We found that CysB
protein
negatively regulates the expression of a new hslJ gene. In order to
elucidate the regulatory
role of CysB implicated in expression of the HslJ. CysB protein with His
tag on its C-
terminal region was purified near homogeneity. The functional activity of the
CysB-His
fusion protein was confirmed in vivo. Analyses of gel retardation
experiments employing
the purified CysB protein and the DNA fragment that comprises the regulatory
region and
the 5' part of the hsU gene showed that in vitro, CysB specifically
binds this fragment and
forms two complexes. Comparison of the results obtained using either the crude
cell extracts
isolated from die wild type cells or the purified CysB-His revealed that CysB
protein alone
binds this fragment, hence that no auxiliary proteins are involved in this
binding.
Furthermore, we showed that NAS decreases the efficiency of the CysB-DNA
binding in
the hsU locus acting as an anti-inducer. Therefore it might be that CysB plays
a role of the
classical repressor of the HslJ expression while the NAS binding to CysB
modulates this
negative regulation.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
CONSTRUCTION OF SHUTTLE VECTOR pA13
Milan Kojić, Slavica Arsenijević, Olivera Gajić, Vanesa Munćan i Ljubiša Topisirović
Institute for Molecular Genetics and Genetic Engineering, V. Stepe 444a, P. Fah 794, 11001 Belgrade
The smallest cryptic plasmid pAl (2820bp) of Lactobacillus plantarum A112 containing the RCR origin able to replicate in lactic acid bacteria (LAB) and E. coli was used for construction of shuttle cloning vector pA13. Vector pA13 was constructed by insertion of DNA fragment (1 kb) harbouring the erm gene from pIL253 and DNA fragment harbouring the lacZ(α) gene from M13mpl8 into pAl. Cloning DNA fragments in multiple restriction sites present in the lacZ(α) gene was facilitated by using blue/white selection of E. coli transformants. Further genetic manipulation was allowed by selection of Emr transformants in both LAB and E. coli. Results showed that vector pA13 posses high efficiency of transformation of a LAB and had a broad host range. In addition, it had pretty good stability, i.e. segregation is relatively low. The pA13 vector was used for cloning bacteriocin genes from Lactococcus lactis BGMN1-5 by making the library of bacteriocinogenic plasmid in E. coli. Recombinant clones containing the bacteriocin genes was isolated by transformation of bacteriocin-sensitive strain L. lactis IL1403 and selection for bacteriocin production and immunity. The pA13 vector and recombinant clones with bacteriocin genes were stable maintained in LAB and E.coli. Vector was also used for cloning of heterologous sgm gene (GC reach) from Micromonospora zionensis. Results showed that recombinant clones were stable in LAB as well as in E. coli. However, the expression of the sgm gene was obtained in E. coli but not in LAB.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
LARGE SCALE OF ISOLATION OF RAT ERYTHROID PROGENITORS FOR GENE THERAPY
Marija Mijatović1, Jelena Stanković2, Marijana Petakov2, Gordana Jovčić2, Jelena Zarić1, Zvezdana Popović1
1Institute for Molecular Genetics and Genetic Engineering
2 Institute for Medical Research
Hemopoiesis is studied in vitro mainly in semisolid cultures, where hemopoietic progenitors develop into discrete colonies. The fact that the cells are immobilized in the semisolid medium makes difficult or impossible to carry out analysis of growth kinetics of the developing cells.
We describe a liquid culture system that supports the growth and maturation of rat erythroid progenitors. The procedure was divided into two phases. In the first phase mononuclear cells from the peripheral blood and bone marrow were cultured in a density of 5x106 cells/ml in alpha minimal essential medium supplemented with 10% PCS, 10% PKW-SCM and 1 μg/ml cyclosporin A. The cultures were incubated at 37°C in an atmosphere of 5% CO2 in air with extra humidity.
Following a 3 days of incubation in these primary cultures, the cells were harvested, washed and recultured in alpha medium containing 30% PCS, 1% BSA, 10-5 β-mercaptoethanol and 1U/ml Epo. These secondary cultures were incubated at 37°C in an extra-humid sealed incubator flushed with a mixture of 5% O2, 7% CO2 and 88% N2.
In the first phase (Epo independent) cultures showed a sharp decrease in absolute number of erythroid colony-forming progenitors. During the second phase, the presence of Epo, a high concentration of PCS and incubation under low oxygen pressure resulted in the development of erythroid cells. This type of liquid culture can give higher cell yield with quite synchronous cell differentiation stages and easily be applied, as particularly suitable target cells, in gem therapy protocols.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
TWO-PLASMID SYSTEM FOR ISOLATION OF REGULATORY AND GENTAMICIN SENSITIVE MUTANTS OF THE SGM GENE
Marijana
Miljković, Nataša Milojević, Milorad Kojić, Branka Vasiljević
Institute of Molecular Genetics and Genetic Engineering, Belgrade
The sgm (sisomicin-gentamicin resistance methylase) gene is resistant determinant cloned from aminoglycoside producing strain Micromonospora zionensis. It encodes specific 16S rRNA methylase which protects ribosomes from inhibitory effect of aminoglycoside antibiotics. It was shown previously that the sgm gene is regulated at the translational level by negative autoregulation. Namely, Sgm methylase- binds to specific regulatory sequence in front of ribosomal binding site on its own mRNA and thus prevents its own further translation when all ribosomes are methylated (protected). Regulation of sgm gene was studied by use of sgm-lacZ fusion system. In order to study regions responsible for autoregulation or/and methylation of ribosomal RNA, a system with two plasmids was developed. The sgm-lacZ fusion was cloned on a low-copy plasmid pMS42l and cells carrying such construct were transformed with in vitro mutagenized plasmid carrying the sgm gene. Screening procedures were designed in a way that either regulatory or gentamicin sensitive mutants could be detect. Results of analysis of obtained mutants will be presented.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
PROMOTER SEQUENCES REQUIRED FOR REGULATED EXPRESSION OF THE RAT βbminy - GLOBIN GENE IN REL CELLS
Tatjana Mitrović i Zvezdana Popović
Institute of Molecular Genetics and Genetic Engineering, Belgrade, Yugoslavia
The rat β-globin locus is unique for its organization. It consists of three to five adult genes depending on haplotype. We have shown that one of these genes - βbminy is transcriptionally active in rat reticulocytes. Further, we try to elucidate the transcriptional regulation of βbminy -globin gene during erythroid differentiation. Our study is based on D5A1 REL (rat erythroleukemic) cells, homologous model system, which can successfully mimics in vitro the events of normal erythropoiesis. In order to evaluate the contribution of promoter sequences to transcriptional regulation of βbminy - globin gene, we have made hybrid promoter-CAT reporter gene constructs. The expression of these reporter plasmids was confirmed by primer-extension analysis. On the basis of stable-transfection experiments, we could define the minimum elements which are required for enhancement and induction of transcription in differentiating REL cells. We have shown that the presence of -96 bp promoter region (TATA-βDRE-CCAAT-CACCC) is sufficient for erythroid specific expression of βbminy - globin gene.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
POLYMORPHISM OF PCR BASED MARKERS AND HETEROSIS IN MAIZE
Snežana
Mladenović Drinić i Kosana Konstantinov
Maize Research Institute, 11080 Zemun Polje-Belgrade
As parental genetic divergence has been found to increase the potential for heterosis in crosses, it is suggested using of indirect measures of genetic diversity as possible predictors for the heterotic response of hybrids (polymorphism of molecular markers). The five maize inbreds (F2, ZP120, ZP 2039, W 401, EP1) and 10 hybrids were evaluated for grain yield. Heterotic effect was calculated regarding the mid-parent value and estimated heterosis for yield according Eberhart and Gardner, 1966. The parental lines were surveyed for polymorphism with randomly chosen set of 58 SSR loci and 15 AFLP primers pair resulting in a total of 1306 reproducible amplification products, 595 of them being polymorphic. Roger's genetic distance were computed for all ten combination of the five parents based on 595 marker variants (501 AFLP variants, 94 SSR allele variants) and ranged from 0.618 in cross F2 x ZP2039 to 0.841 in cross ZP120 X W401 with an average of 0.682 across all ten pairs. The coefficient of correlation between genetic distance based on AFLP/ SSR marker date and mid-parent heterosis for yield was positive and significant 0.604/0.687 and with estimated heterosis 0.578 / 0.636, respectively. The coefficient of correlation between AFLP and SSR marker date is 0.752. The results indicated that genetic diversity among parental lines is certainly related to heterosis.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
YEAST SHUTTLE EXPRESSION VECTOR WITH DOMINANT SELECTIVE MARKER
Pavković,
N., Todorović, V., Glišin, V., Ljubijankić, G.
Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 444a, Belgrade
Yeast Saccharomyces cerevisiae is one of the standard expression system in genetical engineering which shows many good features of prokaryotic expression systems and also possess mechanisms of postranslational modifications inherent to higher eukaryotic cells.
Shuttle vectors pBLITZ has been obtained by fusing selective markers and replication elements of both S. cerevisiae and E. coli. pBLITZ contains sequences responsible for replication (ORI) and maintenance of high copy number (STB, FRT, FLP) from naturally occurring yeast plasmid 2>im, E. coli plasmid pBR322 origin of replication, and selectable markers: E. coli β-lactamase gene (conferring ampicilin resistance) and auxotrophic marker LEU2 from S. cerevisiae genome. pBLITZ also contains BLITZ expression cassette that allows expression and secretion of wide range of heterologous proteins in yeast.
An obstacle for wider usage of this plasmid is lack of dominant selectable marker-gene which would increase the range of host strains and allow selection in rich media. Therefore a gene for aminoglycoside antibiotics resistance (kanamycin, geneticin...) Tn903 kanr functional in both E. coli and S. cerevisiae was cloned.
New shuttle vector with dominant selective marker has been utilized for 'construction of new plasmids for expression and secretion of heterologous proteins from prokaryotes (penicillin amidase; plasmid pGoB-2KS), and eukaryotes (human interferone-β; plasmid pG1FN).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
LOCALISATION OF CONTROL ELEMENTS RESPONSIBLE FOR SOX3 GENE ACTIVATION BY RETINOIC ACID
Tamara Rajić, Nikola Arsić, Nebojsa Mirković. Milka Sokolović and Milena Stevanović
Institute of Molecular Genetics and Genetic Engineering. Vojvode Stepe 444/A P.O. Box 794, 11000 Belgrade, Yugoslavia
Retinoids have been shown to have effects in a variety of biological processes including cell differentiation, embryogenesis and neoplasia. Retinoic acid (RA), the natural acidic derivative of vitamin A, seems to be of major importance in many aspects of CNS development. Effects of both vitamin A excess and deficiency on the developing embryos of mammals are associated with CNS defects.
SOX3 gene is a member of new family of genes, which are embryonicly expressed, and the expression is restricted to neuronal and related tissues. Expression patterns of SOX3 gene and mentally retarded patients, deleted for this gene, suggest certain role in the regulation of development of CNS.
SOX3 gene is activated by RA and the sequence responsible for RA activation resides within 400 bp fragment. Five overlapping subfragments 200-334 bp in length responsible for RA activation are made by PCR, cloned in pBLCATS vector and linked to the CAT (Chloramphenicol Acetyl Transferase) reporter gene. These constructs after transient transfection into NT2/D1 cells were assayed for the CAT activity in the presence and absence of RA.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MOLECULAR-GENETIC ANALYSIS OF TEMPERATURE-DEPENDENT POSTTRANSLATIONAL MATURATION OF E. coli PAC
Spasić M., Glišin V., Ljubijankić G.
Institute of Molecular Genetics and Genetic Engineering, V. Stepe 444a, Beograd, Yugoslavia
Penicillin G acylase (PAC) catalyses hydrolysis of penicillin G to 6-aminopenicillinic acid and phenilacetic acid (PAA), which marks this enzyme as very important for the semi synthetic antibiotics production.
Regulation of Escherichia coli pac expression is complex and encompasses different mechanisms both on transcriptional and posttranslational level. In order to acquire functional, heterodimeric enzyme, nascent PAC precursor has to undergo proteolytical processing, which represents limiting step in industrial synthesis of the enzyme both in bacteria and yeasts. In E. coli this processing takes place only below 30°C. However, Providencia rettgeri pac gene (62% homology with E. coli pac on aminoacid level) is expressed in temperature-independent manner.
In order to determine a region responsible for this difference, we constructed a recombinant plasmid carrying both pac genes. This vector, introduced into E. coli, was subjected to intraplasmid homologous recombination between two pac genes which allowed us to isolate a number of chimeric pac genes.
Further analyses encompassed clones that posses significantly changed characteristics of posttranslational processing of precursor: (1) a chimeric gene which possess temperature-independent precursor processing and (2) a group of recombinants with lowered efficiency of posttranslational maturation.
We have determined exact nucleotide positions of recombinational events by restriction analysis, PCR mapping and sequencing. That allowed us to determine region(s) that influence thermal dependencies of posttranslational processing in E. coli precursor. Our further work was aimed at precise delimiting of these regions, using technologies of genetic engineering.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MOLECULAR ANALYSIS OF THE prt GENE REGULATORY REGION OF NATURAL ISOLATE Luctobacillus paracasei subsp. paracasei BGHN14
Irena Spasojević, Nataša Miladinov i Ljubiša Topisirović
Institute for Molecular Genetics and Genetic Engineering, V. Stepe 444a, P.O.Box 794, 11001 Belgrade
Lactobadllus paracasei subsp. paracasei BGHN14 is natural isolate from homemade cheese. Preliminary analysis revealed that this strain produces the proteinase similar to the lactococcal PI-type proteinase. Hybridisation experiments as well as restriction analyses indicated that proteinase gene region have similarity with that of lactococci. In contrast to lactococci, prt genes of lactobacilli are chromosomal located. Proteolytic activity of the BGHN14 is depended on concentration of casitone in basal minimal medium (BMM). Higher casitone concentrations (1.0% and 2.0%) exhibited inhibitory effect upon the BGHN14 proteinase activity. Regulatory domain of BGHN14 was subcloned into vector pNZ273 resulting in construct pHN174, and sequenced. Results revealed that this region contains both PprtM and PprtP promoters but is shorter for 35 bp in comparison to that of lactococci. The nucleotide sequence missing in BGHN14 is forming potential steam-loop structure suspected to be involved in casitone-dependent regulation of the prt genes expression in Lactococcus lactis SKI 10, Wg2 and BGIS29. However, testing the p-glucuronidase activity in L. lactis NZ9000/pHN174 tran-formant in which the gusA gene is under control of the Pprtm promoter of BGHN14 revealed that casitone mediated regulation was at the transcriptional level.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
PRELIMINARY CHARACTERISATION OF THE THREE NOVEL HUMAN SOX GENES
Slavica Stanojčić, Maja Vujić, Nikola Arsić, Tamara Rajić i Milena Stevanović
Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 444/A, P.O. Box 794, 11000 Belgrade, Yugoslavia
The mammalian genome contains a family of genes that are related to SRY, the mammalian sex determining gene, and these genes are called SOX genes (SRY-related HMG-box genes). By analogy to SRY, these genes might be expected to play a role in the regulation of development and sex determination.
Preliminary results suggest that many SOX genes are embrionically expressed and expression patterns indicate that some of them might play some role in development of central nervous system.
A new human SOX gene, named SOX 14, has been isolated from the human genomic library. This gene has been mapped to human chromosome 3q23 region. SOX 14 HMG-box amino acid sequence shares 65,8% similarity to HMG-box of the SRY gene, and 100% similarity to HMG-box of mouse Soxl4 gene. It is predominantly expressed in foethal spinal cord, foethal brain and foethal thymus.
The other two human SOX genes, named SOX18 and SOX20, have been isolated from human foethal cDNA library. One of them, named SOX 18, shares 52,6% of amino acid similarity to HMG-box of the SRY gene and 74% amino acid similarity to amino acid sequence of mouse Sox18 gene. SOX18 has been assigned to chromosome 20q13.3 region using radiation hybrids.
The other one named SOX20 gene, shows 65,8% amino acid similarity to HMG-box of the SRY gene, and it does not appear to be a true homologue of any of known mouse Sox genes. SOX20 has been assigned to chromosome 17pl3 region using FISH and radiation hybrids. The gene is widely expressed in foethal and adult human tissues. Further analysis of these genes will provide information about their potential roles in controlling developmental processes in humans.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENOTYPING THE BOVINE TRAITS OF ECONOMIC INTEREST
Slavica Stanojčić, Jelena Đurović, Nikola Arsić, Nebojsa Mirković, Vesna Verbić, Milena Stevanović
Institute of Molecular Genetics and Genetic Engineering. Vojvode Stepe 444a P.O. Box 794, 11000 Belgrade, Yugoslavia
Caseins are predominant (80%) proteins in bovine milk. One of them, K-casein is polymorphic and the most frequent variants are A and B. The K-casein allele B is more desirable for cheese production. Milk obtained from cows with BB genotype shows higher curd firmness and higher cheese yield.
Bovine leukocyte adhesion deficiency (BLAD) is a lethal autosomal recessive disease. It is caused by point mutation in CD 18 gene. This gene encodes p subunit of glycoprotein which is responsible for adhesive interactions between leukocytes and infectious agents. The point mutation causes an amino acid substitution that makes this glycoprotein inactive. Affected animals suffer from frequent infections which recur after conventional treatment, and they die before reaching maturity.
For fast and precise detection of gene variant for K-casein and BLAD, we've introduced a DNA test based on polymerase chain reaction (PCR) and RFLP. Test included amplification of specific region of CD 18 gene, exon 4 of K-casein, which contains polymorphic sequence and later treatment of these fragments by restriction enzymes.
This PCR-RFLP DNA test enabled us to detect normal and defective CD 18 allele, to identify affected, normal and heterozygous animals and also specific allele for K-casein they had.
After analysis of CD 18 sequence and sequence of K-casein gene, we've designed oligonucleotides which have been used as primers. We have obtained fragments of expected lengths by multiplex PCR. After precipitation, PCR products were treated with restriction enzymes PstI and Hae III. Then, after double digestion, the fragments corresponding to each genotype were easily detected in 4% agarose gel.
Genotypes based on polymorphic restriction sites for both characteristics can be easily detected from banding pattern what make the test useful for the selection of traits of economic interest.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
ANALYSIS OF CONJUGATIVE ABILITIES OF PLASMIDS IN Lactococcus lactis subsp.
lactis biov. diacetylactis S50
Ivana Strahinić, Milan Kojić i Ljubiša Topisirović
Institute of Molecular Genetics and Genetic Engineering, V. Stepe 444a, P.O.Box 794, 11001 Belgrade
Lactococcus lactis subsp. lactis biovar. diacetylactis S50 is a natural isolate from starter culture for butter production. The strain S50 contains five plasmids, in range from 6.7 kb to about 300 kb (pS50-290, pS50-80, pSSO-lOa, pSSO-lOb and pS50-7). This strain produces bacteriocin with narrow antibacterial spectrum, bacteriocin S50 and cell wall-asociated proteinase of PI-type located on the large plasmid pS50-290. Other plasmids are cryptic. The strain S50 was used as a donor in conjugation experiments for transfer of plasmids to L. lactis MG1363 and IL1403 and S50 derivatives obtained by plasmid curing. The transconjugants from these crosses contain plasmid pS50-290 but also small plasmids pS50-10a, pS50-10b and pS50-7 in various combinations. These transconjugants were used as donors in further conjugations, but transfer of small plasmids was not detected. Results also showed that only plasmid pS50-290 could be transmitted in these crosses. Taking all these results together, it appeared that plasmid pS50-80 was responsible for cotransfer of small plasmids during conjugation. Hybridisation experiments revealed that plasmids pS50-7, pS50-10a and pS50-10b contain homologous nucleotide sequences with plasmid pS50-80. Homologues TaqI DNK fragments were cloned from all four plasmids and sequenced. Comparative analysis of sequences revealed that they share high degree of homology among themselves. In addition, they contain homology with oriT region of the well-known lactococcal plasmid pCI528.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
CLONING OF MOLE RATS Sox2 GENE USING PCR METHOD
Tomislav Terzin, Milka Sokolović, Ana Basić i Milena Stevanović
Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 444/A, P.O. Box 794, 11000 Belgrade, Yugoslavia
Cloning a gene for determination male sex in mammals (Sry), led to determination a whole family of related genes. Homology between them is restricted to HMG box; therefore this genes were named Sox genes (Sry related HMG box).
Sox genes are expressed during embrional development, in neural and related tissues and undifferentiated gonads. Therefore it is predicted that Sox genes are transcriptional factors that have a role in development of central nervous system and sex determination. Using PCR based approach a large number of Sox genes have been identified in different animals. Apart from mammals, Sox genes have been identified in birds, reptiles, amphibians, fishes, insects and nematodes.
Research has been shown that Sox2 gene is involved in controlling of eye development. The aim of this research is to clone and characterize, Sax2 gene from mole rat (Spalax leucodon). Mole rats eyes are rudimentary and they can distinguish only light from darkness. Because of the role of Sox2 gene in eye development this gene will be cloned and analyzed for presents of possible mutations in mole rat.
Primers specific for 5' and 3' region of human SOX2 gene are used to amplify Sox2 gene from mole rat. PCR products are cloned. By hybridization with box and SOX2 specific probe, it is shown that cloned fragment of mole rat is indeed Sox2 gene. Hybridization analysis indicate high conservation of Sox2 gene in mole rat and human. Confirmed high conservation of Sox genes makes possible to analyse the animal Sax genes using primers corresponding to human SOX genes.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
BIOCHEMICAL AND GENETIC CHARACTERIZATION OF STAPHYLOCOCCAL EXTRACELLULAR PROTEINASES
Đorđe Fira i Ljubiša Topisirović
Institute for Molecular Genetics and Genetic Engineering. V. Stepe 444a, P.O.Box 794, 11001 Belgrade
A collection of natural isolates from bacterial genus Staphylococcus is obtained from food samples, which originate from different geographical locations. The isolates have been screened for production of extracellular proteinases. Amongst 171 isolates, 55 of them were found to be proteolytically active, since their cultures proteolytically coagulate milk. Specificity of staphylococcal extracellular proteinases towards casein fractions has been analysed by electrophoresis. In the process of milk coagulation, staphylococcal proteinases, beside p-casein, significantly hydrolyzed as]- and K-casein. Biochemical characteristics of proteinases from isolates F22, F86, M104, S2007 and S2105 have been studied in detail. It was found that these proteinases have relatively low molecular masses (from 20 to 32 kDa), an that they are released from the cell envelope in the growth medium spontaneously. Their temperature optima are between 30°C and 37°C and their pH optima ranging from 6.5 to 8.7. Copper ions inhibit, but the presence of calcium ions stimulates the activity of proteinases of isolates F22, M104 and S2007. Beside casein fractions, they also hydrolyze heterologous protein substrates, such as BSA and gelatin. Experiments with specific proteinase inhibitors revealed that proteinases of isolates F22 and M104 belong to the serine group of proteinases. The S2007 and S2105 proteinases were classified as metaloproteinases whereas the type of F86 proteinase in these experiments could not be determinated. In hybridization experiments, total DNA isolated from these isolates did not hybridize to the lactococcal proteinase genes probes.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MECHANISM OF POSTTRANSLATIONAL PROCESSING OF THE HETERODIMERIC PAC
Ševo M., Glišin V. i Ljubijankić G.
Institute of molecular genetics and genetic engineering, Vojvode Stepe 444a, Belgrade
Bacterial enzyme penicillin G acylase (PAC) catalyses the hydrolysis of the acyl group of penicillin G. In bacterial cell PAC is synthesised as a single inactive cytoplasmic precursor (pre-pro-PAC). Maturation process begins by removal of signal peptide on inner membrane, followed by release of the spacer peptide and the formation of biochemical} active PAC composed of two unequal-size subunits (α and β) in the periplasm. Synthesis of active enzyme due to expression of the pac gene not only in different Gram-negative bacteria (Sizmann et oi, 1990) but also in yeast (Ljubijankic, 1995) excluded involvement in reaction of speciffic as well as unspeciffic endopeptidases, suggested that the spacer peptide release might be an autocatalytical event. To confirm this hypothesis we constructed the E. coli pac mutant gene (Δ 20 pac) that contains a deletion of the last 20 codons from the 3' terminus and encodes a biochemically inactive precursor molecule. This mutant gene was in vivo coexpressed with the intact pac gene (wt) from Gram-negative bacterium Frovidencia rettgeri in E. coli strain DH5α. Coexpression of these genes was monitored by Serratia marscescens overlay technique and by NIPAB colorimetric assay, and we observed a significant change in the whole PAC activity of the cells. Results of the periplasmic proteins analysis suggest an intermolecular interaction of two different PAC precursors in the process of autocatalytic posttranslational processing.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE ISOLATION OF GENOMIC CLONE CONTAINING THE GENE FOR A SUBUNIT OF BUCKWHEAT SEED STORAGE PROTEIN
Paja Šijačić1, Jelena Brkljačić1, Svetlana Radović2, Vesna Maksimović1
1 Institute for Molecular Genetics and Genetic Engineering, Vojvode
Stepe 444a, Beograd
2 Faculty of Biology, Studentski trg 3, Beograd
Buckwheat seed storage proteins contain three types of proteins: 13 S legumin-like globulins, 8S vicilin-like globulins and 2S albumin-like proteins. A cDNA clone pBF349, with an insert of 600 bp, has been isolated from a cDNA library from buckwheat seeds in mid-maturation stage. This insert has been shown to be a part of the gene for basic polypeptyde subunit of legumin-like storage proteins.
Based on Southern blot of buckwheat seed genomic DNA digested with restriction enzyme EcoRl, fragments 6-9 Kb (HI) were chosen as a template for asymmetric PCR with M9 primer, derived from pBF 349 clone coding sequence. To single-stranded DNA, obtained by asymmetric PCR, oligo-dG tails were added to 3'-end. Further amplification was performed with two primers - M9 and a NotC, which contains restriction site for NotI enzyme and oligo-dC tail. PCR products, 600 bp long, were ligated with the Bluescript vector and cloned in XL 1 Blue E. coli strain.
Four clones, containing sequences of a gene for polypeptyde subunit of legumin-like buckwheat storage protein were chosen on the basis of plasmid electrophoretic mobility, PCR amplification with M9 and NotC primers and hybridization with pBF349 probe. The final identity of these genomic clones will be confirmed by sequencing.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
KARYOTYPE ANALISIS AND CHROMOSOME NUMBER VARIABILITY IN
THE FISH SPECIES Cyprinus carpio L. I Carassius auratus gibelio B. (Cyprinidae, Pisces)
Svetlana Fišter
Faculty for Veterinary Medicine, University of Belgrade, Bul. JNA 18, 11000 Belgrade
The fish species Cyprinus carpio L. and Carassius auratus gibelio B. (Cyprinidae) are of tetraploid origin. According to some data from the available literature, the chromosome number in the both species in the natural bisexual population varies from 98 to 104.
The studies of wild type carp from the Danube population indicate that the diploid chromosome number was 2n=104 in the all individuals examined. There were no the individuals with 2n=98, or 2n=100 chromosomes, as some authors were reported for the carp from some waters. Karyotype of this carp consists of: 24 metacenmcs (M), 12 submetacentrics (SM), 24 subacrocentrics (SA) and 44 acrocentrics (A). The fundamental arm number (NF) equals 164.
In the fish species Carassius auralus gibelio B., in accordance with the existing data, the diploid chromosome number varies from 98 to 104, too. The bise.xuals and the gynogenetic Iryploid lines of this fish species exist in nature. Populational-genetic studies in the fishes from Yugoslav waters indicate to the chromosome number that varies among individuals in the bisexual and among gynogenetic populations. The existence of bisexual populations was affirmed in Yugoslav waters in 1989 and 1991. The basic diploid chromosome number in the fishes from Danube was 2n=100. but some differences occurred in the number of tiniest acrocentrics. The basic karyotype of 2n=100 consists of: 24 metacentrics (M). 12 submetacentrics (SM), 24 subacrocentrics (SA) and 40 acrocentrics (A). The fundamental chromosome arm number (NF) equals 160.
Variations in the number of tiniest acrocentrics in the both species probably occurred as a result of ancestral letraploidisation.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE INCLUSION OF POPULATION OR SPECIES CHROMOSOME VARIANTS IN THE COMBINATION POTENTIAL OF AN INDIVIDUAL
Boro P. Pavlović
Institute of Forestry, Kneza Viseslava 3, Belgrade, Yugoslavia
There are quite simple relations between the number of variants of haploid (V) and diploid chromosome complements (W, K) depending on the number of chromosomes in the haploid complement (n) and on the number of variants of each member (Vj) of the chromosome set presented in the group of individuals, in the population or species. Maximal potential of a diploid individual is V=2n of gamete variants and K=3n of diploid variants within the basic set W=22n of progeny variants. These relations can be used in the explanation of a wide specter of issues: origin and conservation of biodiversity, optimization of chromosome matterial partition, variation potential intra species, population or cultivated and conserved live stocks. Special attention has been devoted to the issue of minimal number of individuals that can include the maximal number of chromosome variants. This can be applied to the solving of the problem of the scope of variation contained in Noah's Ark. spaceship voyage plans or biodiversity salvage in catastrophes, but also in the conditions of everyday activities of the modem civilization. A simple and surprising finding is mat the minimal number of individuals (which can cover the maximal number of chromosome set variants) depends only on the number of variants of the maximally variant chromosome (vmax). This minimal number of diploid individuals is equal to half a number of variants of that chromosome, vmax/2, for an even, or (vmax+l)/2, for an odd number of chromosome variants. The problem of the conservation of population and species genetic diversity should be based on the study of the aspect of combinations of chromosome variants.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENETIC LOADS AND COADAPTIVITY OF CHROMOSOMAL INVERSION POLYMORPHISM IN A DROSOPHILA SUBOBSCURA
Goran Živanović i Marko Anđelković
Department of
Genetics, Institute for Biological Research "Sinisa Stankovic",
Belgrade
The most attractive way to detect the coadaptivity of chromosomal inversion polymorphism in natural populations of Drosophila might be studies on the level of the genetic load. Considering the extremely rich inversion polymorphism of Drosophila subobscura populations of this species might serve as suitable model-systems to study the connection between these two kinds of genetic polymorphism. Taking these facts into consideration an analysis was done of the inversion polymorphism and genetic loads of the O-chromosome in three populations of Drosophila subobscura sp. Observed data showed that in all three populations differences exist in the average viabilities of O-chromosome gene arrangements. The differences observed are a consequence of the uneven distribution of different viability classes among different O-chromosome gene arrangements. The uneven distribution is neither limited to the lethal viability class, but is characteristic to viability classes from the group of quasi normal’s too. The results obtained on the relation of the extent of the genetic loads and the frequencies of gene arrangements showed that the gene arrangements with a moderate and high frequency have the highest viability, i.e. the lowest extent of the genetic load. Observed results also showed that more complex gene arrangements (O3+4, O3+4+i) had the higher viabilities values than gene arrangement without inverted regions (Ost). These results point to high degree of coadaptivity of chromosomal inversion polymorphism in Drosophila subobscura.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
DOES THE PRESENCE OF B CHROMOSOMES AFFECT TAXONOMIC DETERMINATION IN THE GENUS APODEMUS (RODENTIA, MAMMALIA)?
Jelena Blagojević i Mladen Vujošević
Institute for Biological Research "Sinisa Stankovic", Belgrade
The presence of B chromosomes in the yellow-necked mouse Apodemus flavicollis was analyzed in 320 adult animals from four localities in Yugoslavia. Our aim was to assess the effects of B chromosomes on certain morphometric characteristics. The characters that were chosen are included in the taxonomic discrimination of this particular species as well as the sibling species Apodemus sylvaticus. B chromosomes were found in all localities in frequencies raging from 21.6% to 42.9%. It was established that the relative length of the tail (the body/tail ratio) was significantly increased in the presence of B chromosomes. This index demonstrates geographic variability, however, in general the studied specimens had longer bodies than tails. It was ascertained that different frequencies of animals with B cliromosomes change the slope of the regression curve of the length of foramina incisiva (FI) relative to the distance from the 3rti upper molar to the incisor (IM3). This finding questions the validity of use of this relative value in taxonomy without taking into consideration karyological data.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
Fraxinus ornus L., SEXUAL POLYMORPHISME AND RAPD MARKERS
S. Bojović, Ph. Heizmann# M. Barbero*
Srbijašume - Institute of Forestry, Kneza Višeslava 3, 11030 Beograd
tel: 553 355, e-mail: bojos@ptt.yu
#Reconnaissance Cellulaire et Amélioration des Plantes, UMR 9938-CNRS-INRA. Ecole
Normale Supérieure-Lyon. 46 allée d'ltalie, 69364 Lyon, France
*Institut Méditerranéen d'Ecologie et de Paléoécologie. CNRS-UPRES A 6034, Av.
Escadrille Normandie Niémen, 13397 Marseille, France
Flowering (or Manna) ash (Fraxinus ornus L.) is a mainly a Mediterranean tree species of wide ecological amplitude. In terms of floral biology Fraxinus ornus L. shows an extremely rare form of sex expression - androdioecy; defined by a simultaneous presence of male and hermaphrodite flowers on separate individuals (presence of male and hermaphrodite individuals in populations). It is a rare and original system, both from theoretical and from empirical points of view. The possible sex differentiation between male and hermaphrodite individuals was examined by PCR-RAPD (Polymerase Chain Reaction-Random Amplified Polymorphic DNA) molecular genetic markers.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
INTRA-AND INTER-POPULATION VARIABILITY IN HARE (Lepus europaeus PALLAS)
Ljiljana Vapa, Dragana Obreht, Milan Vapa*, Deana Demjen
Institute of Biology, Trg D. Obradovića 2, 21000 Novi Sad.
*Faculty of Agriculture, Trg D. Obradovića 8. 21000 Novi Sad
One important aspect in the complex field of genetic resources conservation is measuring of amount and distribution of genetic variation withir populations and species of wild animals. Attention should be paid to the loss of alleles caused by selective hunting, founder effect, genetic drift, poaching and reintroduction of species. In order to evaluate genetic variability in hare in northern Vojvodina, samples from four populations were analyzed by standard polyacrilamide gel electrophoresis. Twenty-three isozyme loci were analyzed, and only 8 of them were polymorphic. It was found that average heterozygosity was H=0.107 and polymorphism was P=0.043, suggesting high rate of intra and inter population homogeneity. The gene erosion is a main problem in game breeding and presented type of analysis could serve as a method for monitoring of genetic diversity.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GLIADIN ALLELIC VARIATION AND DIVERSITY OF AGRONOMIC PROPERTIES IN WHEAT
Dimitrijević Miodrag1, Knežević Desimir2, Petrović Sofija1
1 Institute of Field and Vegetable Crops, Faculty of Agriculture, Novi Sad
2 ARI SERBIA, Small Grains Research Center, Kragujevac
The sample of 77 wheat varieties was analyzed for presence of gliadin alleles Gli Bib. Gli B11, Gli D1b. and Gli D2b. Agronomic trait differences were compared, testing examined genotypes grouped due to presence or absence of these gliadin alleles in the genetic background.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
"AFILA" GENE EFFECTS ON ABSOLUTE SEED MASS IN PEAS (Pisum sativum L.)
Đorđević Radiša, Zdravković Milan, Zečević Bogoljub, Sretenović-Rajačić Tatjana
Agricurtural Research Institute "Srbija", Center for Vegetable Crops, Karađorđeva 71,
11420 Smederevska Palanka, cfvcsp@eunet.YU
The "af" gene, at its recessive homozygous stage, causes the modification of the pea leaves so that they only form vines with stipules instead of normal structure leaves. In order to investigate the effect of the "afila" genotype on the absolute seed mass of the descendants, the pea variety Filigreen of the afila type was crossed with 12 other varieties possessing normal leaf structure.
The genetic analysis of F1 and F2 generations (the segregation ratio between the normal and afila leaf type being 3:1) proved the monogenic character of this feature. The absolute seed mass in F1 generation, which was characterised by normal leaf structure, was larger in the parents of both pea types.
In F2 generation that possessed normal leaf structure, the absolute seed mass was 2% higher on the average than the one in parents, while it was 6% lower as compared to the F1 generation. The average absolute seed mass in afila pea plants was 5% lower than it was in their parents, and when compared to the F1 generation, the absolute seed mass was 12% lower. The decrease of the absolute seed mass in F2 generation of the afila genotype was conditioned by the reduction of the assimilation surface of the plant.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
INFLUENCE OF ANTIOXIDANTS ON FERTILITY AND FECUNDITY OF D. melanogaster
Jelena Živanov-Čurlis, Stevo Najman, Estera Mrčarica
Department of biology with human genetics, Medical faculty Nis
Because of importance of effects of free radicals on many living functions we used the preparation Protectin (Zdravlje Leskovac) which consist of several antioxidants in order estimate how much they can affect fertility and fecundity of D. melanogaster. In this purpose we used two lines of flies. Experimental (E) line is grown on medium having Protectin, and line is grown on standard medium served as a control (C). Protectin consists of antioxidants as alpha-tocopheryl acetate, vitamin C, beta-carotene and selenium. Parental generation was grown on medium with antioxidants also. In E line was 1,75% sterile males, and sterile females were not found in estimated pairs. In C line was found 1.75% sterile males and 2,63% sterile females. Average fecundity was 36.56 hatched flies per female in C line, and 58,22 hatched flies per female in E line. Also, it's found many unusual tiny flies in E line offspring. According to our results antioxidans from Protectin reduces sterility and increases fecundity in D. melanogaster.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
VARIABILITY AND CORRELATION ANALYSIS OF FRUIT CHARACTERISTIC OF SELECTED VINEYARD PEACH (Prunus persica L. Batsch.)
Gordan
Zec, Radmila Todorović. Petar Mišić. Slavica Čolić
"PKB INI AGROEKONOMIK" Fruit and Grape Research Station Padinska Skela, Beograd
Native populations of vineyard peach in Serbia represent a rich source of genetic variability which can contribute to the improvement of economically the most important characteristics of nowadays growing varieties and rootstock of peach. Vineyard peach seedlings are. by Misic and coll. (1988.) the most important rootstock for peach in our country. By statements of Scorca et. al. (1984.), genetic variability of the newest peach varieties in the world is extremely limited.
During 1994. and 1995.. it was investigated 12 positive genotypes of vineyard peach selected from several populations in Serbia. In this work fruit characteristics of investigated selections, are presented. Among investigated selection, fruits of round and oval shape are dominant. It is represented several groups with different peel color, mesocarp color, as well as with different leaf color before falling. From total of 12 genotypes, 6 are without additional peal color and without red coloring of mesocarp around stone, while the other 6 has the additional color and red coloring represented in a different level.
On the basis of calculated coefficient of rank correlation it was obtained statistically very important correlation between basic fruit peel color and fruit mesocarp color, between the additional peel color and mesocarp color around stone, as well as between mesocarp color around stone and leaf color before falling.
Coefficient of fruit weight variation was 15,4%, while coefficient of stone weight was 10,7%. Average seed germination ranged from 68% to 108%. The results of coefficient of rank correlation confirm the former proved correlation between pigments presence in leaf and mesocarp of peach fruit (Ackerman and Hough, 1950.). Diversity of investigated fruit and leaf characteristics of vineyard peach point to distinct variability of selected genotypes.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
CORRELATION ANALYSIS OF SOME QUANTITATIVE TRAITS OF AUTOCHTONOUS CABBAGE POPULATIONS (Brassica oleracea var. capitata L.)
Mirjana Ivančević, Tatjana Sretenović-Rajičić, Nenad Pavlović, Jasmina Zdravković
Agricultural research Institute "Srbija", Centre for Vegetable Crops, Karadjordjeva 71, 11420 Smederevska Palanka, cfvcsp@eunet.yu
In order to determine the initial material for breeding aimed at obtaining favourable agronomic features, we investigated nine local populations and one cabbage variety. The trial was set up on an experimental plot of the Centre for Vegetable Crops in Smed. Palanka by applying the method of random block system in five replications during the years 1997 and 1998. Interdependence of the traits (rosette diameter, number of leaves per rosette, plant mass, head diameter, head height, core length , stem length , and head mass) was determined by coefficient of correlation (Hadzivukovic, 1991). Coefficients of correlation demonstrated a high positive correlation ratio between the traits head mass -plant mass (r = 0.769). The lowest positive correlation ratio was found for the traits rosette diameter - core length (r = 0.142). The trait number of leaves per rosette was in negative correlation with the head diameter (r = - 0.309) and the head height (r = - 0.280).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
ATYPICAL PHENOMENA IN WALNUT ORGANOGENESIS
Janković Dragan, Janković Slađana
Faculty of Agriculture, Pristina
The knowledge of growth and development of individual organs and fruit tree as a whole is a major prerequisite for defining and proper application of cultural practices in fruit growing. In ecophysiological terms, walnut has marked specificities which are shown through organogenesis cycle, which distinguishes it from the majority of other fruit species. The proper understanding of these aspects in walnut growth and development is aggravated due to the complexity of organogenesis, pronounced proneness to cross-fertilization, heterozygosity and prevailing generative mode of propagation.
The evaluation and clarification of atypical phenomena in walnut organogenesis, which in some cases occur on individual trees every year, and in others sporadically, often induced by ecological factors, enables a better insight into the available gene fund in a certain population and a better understanding of mechanisms of expression of genes controlling walnut growth and development.
Over several years of investigation of domestic walnut population, the following atypical phenomena were recorded: late flowering, formation of racemose inflorescences containing up to 20 female flowers, formation of hermaphrodite inflorescences with fertile female flowers at the base of the inflorescence axis and male flowers near its apex, differentiation of various types of bearing branches from summer buds, formation of male bearing branches on which all the buds are differentiated into catkins (including terminal), development of catkins with a V-shaped axis, development of leaves with coalesced and deformed leaf blades, formation of fruit with three-part, four-part and five-part shell suture and corresponding kernel segmentation.
Many of these phenomena are associated with high cropping potential, hence detection and fixation of genes controlling them offer new possibilities for increasing both theoretical and real walnut yields.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GROWTH HABIT FORMS IN P. communis L. POPULATION IN THE IBAR - KOLASIN REGION
Slađana Janković
Faculty of Agriculture, Pristina
Studies were carried out in 1995-1998 on differing growth habit forms in wild pear population (P. communis L.) in the region of the Ibar Kolašin. This spontaneous population, derived by generative propagation, consists of several thousands of bearing trees aged 20-200 years and is highly heterogenous.
The evaluation of growth habit was carried out through monitoring the following parameters:
- tree vigour:
- presence and development of the central leader;
- number and development of primary scaffold branches;
- brandling angles of primary scaffold branches;
- overgrowing of primary scaffold branches by the branches of the higher order;
- characteristics of overgrowing branches.
A marked variability in all the mentioned parameters was observed within the evaluated population. Differing forms of growth habit have been determined and classified into main types and subtypes. The paper presents a detailed schematic presentation of selected growth habit forms, as well as corresponding photodocumentation.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
POPULATION GENETICS INVESTIGATIONS OF PTC TASTING SENSIBILITY FROM SOME LOCALITIES IN SERBIA
Krajinčanić1, B., Krajinčanić-Suzović V.2, Cukić R.3, Ninković D.3
1 University of Belgrade
2 Health Centre Zemun Department of Mental Health
3 Faculty of Defectology, University of Belgrade
In our investigation of somatic and genetic characteristics of children and youth in some localities of Serbia, which are geographical}' far from one another and different according to the degree and origin of the population, we investigated PTC tasting sensibility.
The investigation included 2268 children population from East. West, North West and North part of Serbia from 7-14 years old.
Mean taste threshold for PTC in Eastern Serbia (Laznica and Zagubica) is 6.04. Western (Zlatibor - Cajetina) 5.68, West river Morava (Velika Drenova) 7.58, North Western(Novi Pazar) 6,13 and Sjenica 6,77 and Northern (Surdulica) 6.95.
Comparing the data we have found out that our results are in accordance with the other authors in our country and according the frequency of recesive phenotype "non taster" and recessive alelogen (pt) certain difference exists.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
FREQUENCY OF HAND CLASPING FROM SOME LOCALITIES IN SERBIA
Krajinčanić-Suzović V.1, Krajinčanić B.2, Čukić R.3, Gašić-Marušić R.4
1 Health Centre Zemun Department of Mental Health
2 University of Belgrade
3 Faculty of Defectology University of Belgrade
4 Military Academy Belgrade
Within doing of children and young people some localities of Serbia, the whole series of genetic markers were studied beginning with polygenic height and growth structure trough colour and hair quality, eye colour the presence of Darwin's tubercle and state of lobule in concha, hand relief-dermatoglyphics up to very strong monogenic markers such as blood group systems ABO, Rh( D ) and MN, then PTC and finger overlapping.
In this paper we present results of finger overlapping from children 7-14 years old. These characteristics inherited dominant when left finger is over right and recessive when right is over left. Comparing the data we have found out that frequency of left and right type is around 50%, and in there's no greater differences in distribution between man and woman and the investigated population.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENETIC DIFFERENTIATION IN POPULATIONS OF Merodon avidus (ROSSI, 179) (DIPTERA: SYRPHIDAE)
Milankov V., Vujić A.. Simić S., Ludoški J.
University of Novi Sad, Institute of Biology Trg Dositeja Obradovica 2. 21000 Novi Sad, Yugoslavia
Natural populations of M. avidus from Mountain Dubasnica (Serbia), Morinj (Montenegro), Mountain Durniitor (Montenegro), Mavrovo lake (Macedonia) and Pindos Mountain (Greece) were assayed. Isozyme variability of twelve loci (B.C. number and locus): aldehyde oxidase (1.2.3.1; Ao), mmarate liydratase (4.2.1.2; Fum), glucosephosphate isomerase (5.3.1.9; Gpi), hexokinase (2.7.1.1; Hk), malic enzyme 1.1.1.40.; Me), phosphoglucomutase (2.7.5.1; Pgm) and superoxide dismutase (1.15.1.1.; Sod) was studied in Tris-Boric-EDTA buffer system, while aspartate amino transferases (2.6.1.1.; Aat), α-glycerophosphate dehydrogenase (1.1.1.8; Gpd-1, Gpd-2), (β-hydroxybutirate dehydrogenase (3.1.1.31; Hbd), and isocitrate dehydrogenase (1.1.1.42; Idh-2) in Tris-Citric buffer system, using the method of polyacrylamide gel electrophoresis (PAGE) according to Munstermann (1979) with a few modifications (Milankov el aL 1998).
Genetic differentiation between populations of M. avidus was evaluated on the basis of allele frequencies.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
EVOLUTIONARY RELATIONSHIPS AMONG THE MEMBERS OF THE AENEUS GROUP SPECIES OF GENUS MERODON (DIPTERA: SYRPHIDAE)
Milankov V., Vujić A., Simić S.
University of Novi Sad, Institute of Biology, Trg Dositeja Obradovića 2, 21000 Novi Sad, Jugoslavija
The genus Merodon is the second-largest European genus of family Syrphidae. More than 50 species are known on the continent, predominantly in southern and Mediterranean part. This paper deals with one monophiletic species group that includes: Merodon aeneus complex Meigen, 1822: M. cinereus (Fabricius, 1794); M. Desuturinu,s Vujic, Simic et Radenkovic, 1995, and M. funestus (Fabricius, 1794).
Natural populations of M. aeneus (Morinj, Mountain Kopaonik, Mountain Dunnitor), M. cinereus (Mountain Kopaonik, Mountain Durmitor, Mountain Sar planina. Mountain Prokletije), M. desuturinus (Mountain Kopaonik), M. funestus (Morinj) and M. sp (Morinj, Mountain Dunnitor) were assayed. Isozyme variability of fumarate hydratase (E.G. 4.2.1.2.; Fum). glucosephosphate isomerase (E.G. 5.3.1.9.; Gpi), hexokinase (E.G. 2.7.1.1.; Hk), malic enzyme (E.G. 1.1.1.40.; Me), phosphoglucomutase (E.G. 2.7.5.1.; Pgm) and superoxide dismutasc (E.G. 1.15.1.1.; Sod) was studied in Tris-Boric-EDTA buffer system, while aspartate amino transferases (E.G. 2.6.1.1.; Aat). α-glycerophosphate dehydrogenase (E.G. 1.1.1.8.; Gpd-1, Gpd-2), β-hydroxybutirate dehydrogenase (E.G. 3.1.1.31.; Hbd). isocitrate dehydrogenase (E.G. 1.1.1.42.; Idh-1, Idh-2) and malate dehydrogenase (E.G. 1.1.1.37. Mdh) in Tris-Citric buffer system using the method of polyacrylamide gel electrophoresis (PAGE) according to Munstermann (1979), Pasteour et al., 1988, with a few modifications (Milankov et al., 1998).
Evolutionary relationships among the members of the aeneus group species was evaluated on the basis of allele frequency, as well as occurrence of species-specific allozymes at diagnostic loci.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
VARIABILITY OF FREQUENCY OF SCEs AND PRI IN PERIPHERAL BLOOD LYMPHOCYTES IN CONTROL SAMPLE OF YUGOSLAV POPULATION
Olivera Milošević1, Dragoslav Marinković2, Slobodan Arsenijević3
1 Faculty of Science, Kragujevac
2 Faculty of Biology, Belgrade
3 Clinic of Gynecology, Kragujevac
In the present study, we represented baseline frequencies of sister-chromatid exchange (SCE) and prolipheration index (PRI), depending on sex and age of individuals. It was shown distribution of SCEs and PRIs between individuals, and also, between groups of individuals, as well as correlation between these two phenotypical features. Analyzed sample included 46 phenotypically healthy persons. Total average frequencies of SCE-s and PRI were 6.97±0.10 and 1.77±0.07, respectively. In group of female individuals with average age of 35 years, average SCE frequency was 7.33+0.10 SCE per person, that is statistically significantly higher (p<0.001) in comparison with sample of male persons with average age of 29 years (6.57±0.1 SCE per person). In sample of female persons average PRI was lower (1.70±0.()8) as compared with male persons (1.87±0.07), but without statistical significant. In order to establish effect of age mid sex on SCE and PRI frequencies, analyzed sample was allocated in three groups. In sample of persons with age from 40-63 years, SCE frequencies were significantly lower (p<0.001). Significant differences in PRI noted between age groups (p<0.001). SCE and PRI showed negative correlation (r=-0.01).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
INFLUENCE OF ANTIOXIDANTS ON PREADULT DEVELOPMENTAL DURATION OF D. melanogaster
Stevo
Najman, Jelena Živanov-Čurlis, Estera Mrčarica
Department of biology with human genetics, Medical faculty Niš
Many researches indicate the role of free radicals in different phases of ontogenic development, embryonic as well as aging. Drosophila melanogaster is frequent model for these researches. Using preparation Protectin (Zdravlje, Leskovac) with antioxdative vitamins we wonted to see how much antioxidants from food can modulate preadult developmental duration of D. melanogaster. In this purpose two lines of flies are investigated. Control line (C) was grown on standard medium, and experimental one (E) was on medium having Protectin, which consists of antioxidants as alpha-tocopheryl acetate, vitamin C, beta-carotene and selenium. E line is formed from flies that parental generation was grown on medium with antioxidants. First lan as were apeared third day since laying eggs in C line, but fifth day in E line. First pupas were apeared seventh day in C line, but nineth day in E line. Average duration of preadulted development in C line was 14,54 days, and in E line was 16,67 days. Differences in duration of preadult development between these two lines are statistic significant. We may conclude mat antioxidants in medium in given concentrations prolong preadult development of D. melanogaster.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
ALLOZYME VARIABILITY IN Quercus robur L. AND IDENTIFICATION
POSSIBILITY WITHIN THE POPULATION
Dragana Obreht1, Saša Orlović2, Joška Erdesi3, Ljiljana Vapa1
1 Institute of Biology, Trg D. Obradovica 2, 2 1000 Novi Sad
2 Poplar Research Institute, Antona Cehova 13, 21000 Novi Sad
3 Forest Enterprises Srbijasume, Forest District Sremska Mitrovica, 22000 Sr. Mitrovica
During 90's a broad EU supported project concerning diversity of nuclear and cvtoplasmic gene-markers in oaks was established. The obtained results showed that populations were poorly differentiated for nuclear but highly differentiated for cytoplasmic markers. Despite the fact that polymorphism of isozymes in genus Quercus is usually low, an isozyme loci variability analysis in Common oak (Quercus robur L.) from Yugoslavia was performed because it has an economic importance in our country, and there is no such information from this European region. Seventy-six common oak trees (seed orchard, fifteen years old) were analyzed originated from locality "Banov brod", near Sremska Mitrovica. The isozymes were extracted from dormant buds, and ten isozyme loci were analyzed based on standard gel electrophoresis (starch and polyacrilamide). Four loci were polymorphic with different number of presented allelic forms, and they control the expression of enzymes with low substrate specificity (EST. PX. and ACP). The variability of chosen gene-marker system didn't allow identification of genotypes within the population.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
DISTRIBUTION OF THE KNOWN DIPLOID CHROMOSOME NUMBERS OF SOME EUTHERIAN GENERA PRESENTED IN THE FAUNAS OF CONTINENTS AND OCEANS
Boro P. Pavlović i Nevenka Pavlović
Institute of Forestry, Kneza Viseslava 3, Belgrade
Faculty of Science, Mladena Stojanovica 2, Banja Luka
The data of Eutherian diploid chromosome numbers (2n) have been connected with genera and with faunistic presentation of the genus. The analysis has accomplished by geographic entirety: North America, West India, South America, Madagascar, Africa, Europe, Arctic, Asia, Southeast Asia, Philippine, New Guinea, Australia, Antarctic, Indian Ocean and Pacific. Pattern of 2n distribution is specific for nearly each analyzed area. There is some regularity in grouping of the areas on the base of maximal frequencies, averages or variations of 2n, related to the origin of fauna and to clime conditions of the area.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
ECOLOGICAL STABILITY OF YIELD COMPONENTS IN WHEAT
Petrović Sofija1, Dimitrijević Miodrag1, Mladenov Novica2 i Kraljević-Balalić Marija1
1 Institute for Field and Vegetable Crops, Faculty of Agriculture, Novi Sad
2 Institute of Field and Vegetable Crops, Novi Sad
Ecological
stability of ten diverse wheat genotypes, in four years, was studied. Traits
under the observation were the number of grains per spike, as well as, grain
weight per spike. The highest average value for the grain number per spike
exhibited variety Martonvasarska 15 (
= 58.1) in 1991/92.. in a contrast
to variety Balkan ( = 30.5) showing the lowest
mid-value in 1994-95. Mean values of grain weight per spike ranged from = 1.1g,
for variety Balkan in 1994/95., to = 2.8g, for variety NSR-2 in
1991/92 year. Eberhart and Russell (1966) model was used for stability studies.
The most stable varieties were Libelulla (bi= 1.364 and Sdi2=
1.688) and Lovrin 21 (bi = 1.005 and Sdi2=1.656)
considering grain number per spike. The best stability performance for grain
weight per spike appeared for varieties NSR-2 (bi = 1.041 and Sdi2=
0.289), as well as, Yugoslavia (bi = 0.862 and Sdi2=
0.155).
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
MONITORING OF TEMPORAL VARIABILITY OF CHROMOSOMAL ARANGEMENTS IN Drosophila subobscura
Vuk Savković2, Marko Anđelković2,1
1 Institute for biological research "S. Stankovic", Belgrade
2 Faculty of Biology. University of Belgrade
Time distribution monitoring of individuals within a certain habitat allows an insight into correlation between certain types of genetic polymorphism and specific ecological factors, which inevitably vary along with the time of day.
Inversion polymorphism in some Drosophila species demonstrates a degree of flexibility regarding a reaction to changes of ecological factors. There is an abundance of data related to rich inversion polymorphism in Drosophila subobscura, enabling different judgments on the correlation mentioned.
Bearing that in mind, we have carried out an inversion polymorphism analysis of D. subobscura individuals captured at different times of day at beech forest habitat at the Goc mountain. The time of capturing has been adjusted according to the differences of circadial activity in this species.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
REACTIVITY OF CABBAGE (Brassica oleracea var. capitata) ON PARAQUAT IN VITRO
Tatjana Sretenović Rajičić, Mirjana Ivančević, Jasmina Zdravković, R. Đorđević, M. Damjanović
Agricultural Research Institute "Serbia", Centre for Vegetable Crops, Smederevska Palanka
The aim of our study was to investigate genetic component of cabbage (Brassica oleracea var. capitata) reactivity on total herbicide paraquat in vitro. We used shoot culture of few cabbage genotypes as a model system suitable for this experiment. Examined parameters were root number, root length, shoot length, fresh and dry mass. We established analysis of variance for these parameters, wide sense heritability (h2), coefficient of phenotype variation (CVf), correlation and regression analysis.
Heritability was very high at all parameters, especially at fresh and dry mass (h2=92%). The most variable parameters are number of roots (CVf=29.36) and root length (CVf=26.57). Root number, root and shoot length are significantly correlated. At P295 and P6p genotypes, there is almost physiological dependence between these parameters (b=0.99**). Regression coefficients of dry on fresh mass are very high at all examined genotypes (from bp295=12.60 to bP6p= 17.39).
Changes induced by paraquat are highly determined by genetic component of variance, which is indicated by coefficient of heritability. Paraquat reduced all examined parameters. Similarity in profile of reaction on paraquat and very strong correlation between growth parameters indicates that these parameters are determined by structural polygenes and that these are constitutive characteristics. Differences in intensity of changes that paraquat induces in growth of shoot culture indicate that there are some differences between examined genotypes in regulatory polygenes that determine expression of these changes.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
BIODIVERSITY OF THE HONEYBEE Apis mellifera, Linne (1758) - CYTOGENETIC ASPECTS
Z. Stanimirović, Jevrosima Stevanović, D. Pejović, D. Popesković
Faculty of veterinary medicine, Belgrade
This paper presents the results of the comparative biometric studies of the chromosomes and comparative ultrastructural chromosomal analyses (the distribution of euchromatin and heterochromatin) of the two studied indigenous honeybee ecotypes (the Banate - BET ecotype and Sjenichko - Peshtersky - SET ecotype). These ecotypes originate from ecogeographically defined Yugoslav regions that differ significantly in many microgeografic and microclimatic elements.
The chromosomes for the cytogenetic analysis were obtained from nervous tissue of the cerebral ganglia of honeybee praepupaes according to the procedure established by Imai et al. (1988). G - banding of chromosomes was done by the method of Ronne (1991), which in itself presents a modification and an improvement of those established by Seabright (1971) and Verma (1998).
The conducted biometric analyses point to the existence of differences in those values pertaining to the relative chromosome length and centromere index (arm ratio). The greatest differences in the relative lengths of chromosomes were observed between the chromosomes 12, 2, 3, 1 and 6 in favour of the SET ecotype, namely 15, 14 and 11 chromosomes in favour of the BET referential ecotype. However, the monitoring of the centromere index revealed the greatest differences between the chromosomes 16, 1, 2 and 4. On the basis of these results we advanced the hypothesis that the observed chromosomal biometric differences are the result of the amplification and rearrangement of chromosome regions of the representatives of the studied honeybee ecotypes. This is what prompted us to undertake further investigations which included ultrastructural chromosome analyses. These analyses point to the existence of a marked difference in the G - band distribution on the chromosomes 1, 2, 4, 11, 12, 13, 15 and 16 of the SET honeybee ecotype with relation to the BET ecotype, thus confirming our previously advanced hypothesis.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE INVESTIGATIONS OF CHROMOSOME POLYMORPHYSM IN THE NATURAL POPULATIONS OF THE SPECIES Mus musculus, Linne (1758), ON THE TERRITORY OF THE EX YUGOSLAVIA
Z. Stanimirović, B. Soldatović, Jevrosima Stevanović
Faculty of veterinary medicine, Belgrade
The study of chromosome polymorphysm in the natural populations of the species Mus musculus, Linne (1758), was conducted at 45 sites on the territory of the ex Yugoslavia, south of the rivers Sava and Danube. The examined corpus included 458 trapped and sacrificed animals (231 males and 227 females). At the industrially more developed localities, where the degree of the environmental contamination was much higher, the trapping of the animals had to be repeated several times.
The chromosomes intended for the karyotype analysis were obtained by the direct puncture of bone marrow from long tubular bones (the femur and humerus) according to the method of Hsu and Pat on (1969), modified by Zimonjic (1990), and by the lymphocyte cell culture from the peripheral blood in vitro according to the method of Evans and O'Riordan (1996, 1997). In order to obtain prometaphasal chromosomes the synchronization with bromdeoxyuridine was performed according to the procedure of Dutrillaux and Viegas-Pequignot (1981). G-banding was done according to the method of S e a b r i g ht (1971) and Y un i s (1978), modified by R o n n e (1991).
The identification of chromosomes and chromosome bands was done on the basis of the criteria established by the Committee on Standardized Genetic Nomenclature for mice (1972, 1979) and the Cowell's photo atlas of the house mouse chromosomes.
The obtained results proved the existence of chromosome polymorphysm of Robertsonian type in the natural populations of the species Mus musculus not only in Dalmatia, Montenegro and southeastern Macedonia, as known from the existing literature (Bulic and Soldatovic. 1980; Tichy at al. 1987; Winking et al. 1988) but also in the central (bosnia and Herzegovina) and eastern parts of the ex-Yugoslavia (Serbia). At the analyzed sites in Serbia 60. 20% of the examined animals were either homozygous for the chromosomes with Robertsonian translocations; in Bosnia and Herzegovina it appeared to 59,45% and in Macedonia to 52,05% of the examined animals.
With the identification of the Robertsonian chromosomes we ascertained that, in addition to the "mutual" Rb. 5/15 chromosome already described in literature relevant for the natural populations of the species Mus musculus from Europe and North Africa (Giagia et al. 1982; Tichy at al. 1988), the mice of the natural populations from the sites in Serbia contained in their karyotype a so far unknown, new form of the Robertsonian chromosome, formed by fusion of the 4th and 19th autosomes (Rb. 4/19). In the karyotype of the animals originating from the localities in Maclwa and in the vicinity of Sremska Mitrovica was noticed the presence of a larger acrocentric chromosome, from the first pair of autosomes where, by applying the G-technique. 2 insertions were observed {1C 5 and IE (E3andE4)}.
Having in mind the fact that at the monitored localities m Yugoslavia both animals with Robertsonian translocations and mice with standard karyotype were trapped, it can be concluded that original populations of the species Mus musculus belonged to the subspecies Mus musculus musculus, and that the with the passing time, due to an introduction and migration of animals, the Robertsonian chromosomes from other populations of the "western system" (subspecies Mus musculus domesticus, Mus musculus bervirostrus, Mus musculus praetextial) were introduced into and spread among the new populations. The centres of the spreading of Robertsonian polymorphysm were, most probably, along the Adriatic coast and in southwestern Macedonia, from where animal, the carriers of the Robertsonian chromosomes, migrated towards the central parts of the ex-Yugoslavia, otherwise inhabited by the populations of Mus musculus musculus, whose representatives had the karyotype with 2n = 40 chromosomes, i.e. were monomorphic.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
THE EFFECT OF TRAUMA ON THE HAPTOGLOBIN, α2- MACROGLOBULIN, IL-1, IL-6 I TNF CONCENTRATIONS IN AO AND DA INBRED RATS
Milica Strnać1, Zvonko Magić2, Cedomir Radojičić2 i Dragoslav Marinković3
1 Institut of Patology, MMA, Belgrade
2 Institut for Medical Research. MM A, Belgrade
3 Faculty of Biology, University of Belgrade
The response of on organism to the trauma is polygenically determined and probably some genes or a group of genes are more responsible than others for the resistance to trauma. Reaction of an organism to trauma starts with acute-phase reaction (APR) that consists of the local and systemic reaction. Increased synthesis acute-phase proteins (APP) in the liver represent the most prominent aspect of APR. This is the heterogenous group of plasma proteins which helps an injured organism to restore homeostatic balance disturbed by traumatic agents. Haptoglobin (Hp) and α2-macroglobulin (α2-M) are among the most important APP rats. In this study AO (Albino Oxford) and DA (Dark Auguslin) inbred strains of rats, with different resistance to trauma, were used to compare concentrations of Hp, α2-M, IL-1, IL-6 and TNF, before and after tournique trauma. Cytokines IL-1. IL-6 and TNF are the most important inducers of APP gene expression. Our results showed that before trauma, more resistant AO rats had higher concentrations of Hp and α2-M compared to DA rats, while their concentrations in the early period after trauma were lower. The influence of genetic elements of these differences is significant. Tourniquets trauma caused early increase in the serum IL-1, IL-6 and TNF activities, which preceded increase in the Hp and α2-M concentrations but strain differences, were not found. On the basis of these results we concluded that higher basal concentrations of Hp and α2-M in the AO rats and their effect in the early period of trauma are important elements of their better response to trauma.
Published in Book of Abstracts „SECOND CONGRESS OF SERBIAN GENETICISTS“
Sokobanja, Serbia, November 10-13, 1999.
© 1999 Serbian Genetics Society
GENETIC VARIABILITY OF MORPHOMETRICAL TRAITS IN A D. subobscura POPULATION ACROSS DIFFERENT DEVELOPMENTAL TEMPERATURES
IvanaTomišić1, Marina Stamenković-Radak 2,1, Tatjana Terzić1 i Marko Anđelković2,1
1 Institute for Biological Research, Belgrade
2 Faculty of Biology, University of Belgrade
Populations of various species may be exposed to temperature changes in the environment. Populations may respond to such variations at the phenotypic and genetic levels. The array of phenotypes that develop in response to several environments is the reaction norm which is observed through the concept of genotype x environment interactions.
Reaction norms across three developmental temperatures (16°C, 19°C and 23°C) were measured for wing length, thorax length and wing/thorax length ratio, in both sexes in isofemale lines from D. subobscura population.
The adaptive significance of the observed variability in different environments is discussed.