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Institute Cytology and Genetics
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Division of Plant General Genetics


Head Academician RAS, Prof. V.K.Shumny
shumny@bionet.nsc.ru


Changes in morphological characters in transgenic tobacco plants.

Top - change in flower structure and longostyly;
Bottom - a nontransgenic tobacco plant (control)

A classification of a control sample of dicot gene mRNAs using parameter (Fi) of their 5`UTRs (high expression, F(seq)>0, low expression F(seq) <0). Expression level was accurately predicted for 77.8% of the high expression mRNAs and for 72.5% of the low expression mRNAs


Spikes

(1) wild-type barley H. geniculatum All;
(2) wheat T. aestivum L.;
(3) barley x wheat offspring, H. geniculatum x T. aestivum L. hybrids


Регенерация растений в культуре пыльников беккроссных потомков ячменно-пшеничных гибридов

FGISH patterns of mitotic chromosomes in the alloplasmic lines of wheat prod used using barley-wheat hybrids, H. geniculatum All. (2n=28) x T. aestivum L. (2n=42). FITC (green) indicates of H. geniculatum chromosomes.

A-(2n=41T.a.+1H.g.)
B-(n=20T.a.+1tT.a.+tH.g.)
C-(2n=42T.a.+2t.H.g.)
D-(2n=42T.a.+1H.g.+2tH.g.)


Awned and awnless (under the control of a single gene) wheat forms


A genetic map of the short arm of chromosome 1B in Salmon variety


Genetic microsatellite map of homoeologous group 3 wheat chromosomes. The S1, S2, and S3 sphaerococcoid wheat genes. Genetic distances are given in centiMorgans (cM). Black bars represent the centromere.



In situ hybridisation of Spelt1 repears on metaphase chromosomes of Ae. speltoides


The direction of research at the Division of Plant General Genetics, PGG, is now toward the development of the following topics:

  •  gene engineering, transgenic plants;

  •  chromosome engineering, remote hybridization, chromosome substitution;

  •  plant reproductive systems;

  •  biodiversity and genetic collections.

Researchers at the Plant Heterosis Laboratory (Deputy Head, E.V.Deineko, Ph.D.), which is within the PGG, are studying gene function in a new genetic environment of the plant genome, using a series of transgenic plant (Nicotiana tabacum, SR1) models. It was demonstrated that the expression and inheritance of the transgenes are, as a rule, stable in offspring of the transgenic plants. However, the transferred genes occasionally became silent (inactivated).The stable also occasionally reverted to the unstable state. Lines of transgenic tobacco plants, models of the unstable expression level of the marker nptII gene, were developed. Current interests center on the molecular mechanisms of transgene silencing in the developed transgenic plants.

A series of T-DNA insertion induced mutations was obtained in the transgenic tobacco plants. The mutations affect flower structure and cause pollen grain lethality. Linked inheritance of the characters longostyly and canamycin resistance was established. Abnormalities due to impairment of microsporogenesis and gametogenesis were demonstrated by cytological examination. The DNA regions flanking the T-DNA insertions in the transgenic tobacco plants are being analyzed. Current interests center on the molecular mechanisms of transgene silencing in the developed transgenic plants.

Research activities at the Plant Gene Engineering Section (Head A.V.Kochetov) are focused on study of signals controlling gene expression at the posttranscriptional level in higher plants. The performed experimental analysis of operon expression demonstrated that the distal genes in the operons are expressed at low efficiency. Studies concerned with the features of the translational activities of the enhancers of alfalfa mosaic and tobacco etch viruses indicated that they are dependent on their location within mRNA template. Comparative analysis of the mRNA characteristics of the high and low expression genes revealed enhancer-like signals potentially determining the efficiency of translation determination. Based on these data, a computer system allowing to predict transcriptional activity of mRNA in higher plant cells was developed. The system may be used for optimizing transgene expression in plant gene engineering.

Long-term research concerning targeted modification of single- and double-stranded DNAs using antisense oligonucleotide sequences was continued by the Physicochemical Biology Group (Head S.I.Oshevski). The research was based on switch-on oligonucleotide derivatives carrying various alkylating groups at their 5`-thiophosphate ends. A chemical-enzymatic method for their synthesis was developed by S.Oshevski. A novel antisense strategy - the "binary oligonucleotide reagent" - was developed. It was shown that autoligation of two oligonucleotide derivatives (carrying different reactive groups at the opposite ends and complementary to two neighboring stretches of the target) is severalfold more efficient than modification with any one of the derivatives alone, even in the presence of oligonucleotide derivatives. A set of efficient PCR primers specific to the cDNAs of each of the eleven isoforms of human proteinkinase C was developed. The set is being used for analyzing the expression levels of the isoforms CD34+ stem cells during their differentiation. This work was done in collaboration with INSERM Unite 268, France. It is intended to continue this study on cancer cells of leukemia patients.

Research at the Remote Hybridization and Tissue Culture Section (Head L.A.Pershina) is aimed at reconstructing the plant genomes. To this end, remote hybridization and tissue culture in vitro are used. A basis was established to provide a better understanding of the interspecific incompatibility mechanisms between Hordeum L. and Secale L., Hordeum L. and Triticum L. A set of genetic and biotechnological methods for overcoming incompatibility in production of new forms using barley x rye, barley x wheat hybrids was developed. Based on experimental models, the cytogenetic and genetic mechanisms providing fertility restoration in intertribe hybrids were identified. Reorganization patterns of the nuclear genomes during the formation of new rye and wheat genotypes in the presence of the cytoplasmic barley genomes were also identified. The unique series of alloplasmic euploid and aneuploid lines in common wheat developed by the group are widely used in the research.

The researchers at the Wheat Cytogenetics Section (Head L.I.Laikova, Ph.D.) have established and are maintaining a collection of aneuploids (monosomic and cytologically marked) lines of common wheat in varieties Saratovskaya 29, Diamant 1, and Chinese Spring, lines with intervarietal and alien chromosome substitutions, as well as isogenic lines. The collection presently consists of more than 100 accessions and it allows to gain insight into the cereal genome, using modern molecular and genetic methods. With this collection, chromosome localization and mapping of more than 20 wheat genes have been performed. Hitherto unknown allelism for certain genes was detected. The identified genes are now listed in the Catalog of Gene Symbols for Wheat.

Substitution and aneuploid lines have immediate implications for studies on agriculturally valuable characters that are under a complex genetic control. Thus, the role of particular wheat chromosomes in plant response to components of mineral nutrition, quality and content of gluten, drought- and frostresistance was assessed. Chromosome engineering provides starting material for breeding work. The cytogenetic collection made it possible to carry out comparative evolutionary-genetic research in relatives of common wheat that had been little studied. Morphological and physiological characters were analyzed in accessions of Triticum petropavlovskyi Udacz. et Migusch. and Triticum sphaerococcum Perciv. It was found that 13 dominant genes of the former two species are allelic to those of Triticum aestivum and that the genes have functions in common.

Research related to special and comparative genetics of wheats and their relatives is conducted at the Wheat Genetics Section (Head N.P.Goncharov, Ph.D.). It is based on the established phenotypic and genotypic collections at the diploid and tetraploid levels. Lines isogenic for the Vrn genes controlling spring versus winter habit and development rate were derived for the first time in the world from tetraploid wheat. The available Vrn isogenic lines were used to study the gene geography of the Vrn genes in tetraploid wheat species. Currently, comparative and genetic studies on gene systems, markers in the systematics of

Triticum L. and Aegilops L. genera, are under way. Molecular-genetic study on the ancestry of wheats in Siberia demonstrated their European origin. Chromosomal localization of common and hard wheat genes was done during the last five years. The short arm of chromosome 1B was mapped.

The structural and functional organization of the genomes of wheats and their relatives is under study Cereal Molecular Genetics Section( Head E.A.Salina). Research is mainly concerned with DNA repeats and their reorganization during evolution and stress conditions. The major achievements in this research area are the novel results obtained for the organization of the subtelomeric regions of the chromosome of wheats and their wild progenitors and for the mobile elements in barley. Collections of genome- and chromosome-specific markers are expanded at the laboratory. The collections are used for mapping the cereal genes and genomes, for analyzing hybrid wheat forms, both of natural and artificial origin, and also for identifying molecular markers that are closely linked to morphological characters and of economic importance. Based on the obtained results, new approaches are being developed to: (1) study at the molecular level the organization and function of agronomically valuable characters in wheats; (2) use of molecular-genetic methods in plant breeding.