1. Laboratory of Plants Heterosis
(1) Staff:
- Vladimir K. Shumny, Prof., Dr.Sci., Head of the
Laboratory
- Elena V. Deineko, Ph.D.
- Alexander V. Aksenovich, Ph.D.
- Eleonora V. Kvasova, Ph.D.
- Galina A. Pokhmelnykh, Ph.D.
- Alexander N. Sidorov, Ph.D.
- Valentina A. Godovikova, Ph.D.
- Salmas I. Ibragimova, Ph.D.
- Nadezhda A. Omelyanchuk
- Inna S. Popova
- Olga G. Smirnova
- Yuriy N. Shavrukov, Ph.D.
- Irina L. Stepanenko, Ph.D.
(2) Subjects of Research:
New approaches to plants protection from viral infection are
developed, based on genome reconstruction and using gene engineering
methods. Tobacco (Nicotiana tabacum SR1) and alfalfa stocks with human
betta-interferon gene as well as a series of transgenic tobacco
(N.Plumbaginifolia) stocks with genes of bovine and Serratium
marcescens nucleases have been obtained. Consolidation of genetic
constructions transferred into plant genome and their heritability in
transgenic progeny were proved using genetic and molecular-genetic
methods. The immunoenzyme assay methods allowed establishment of a
protein which is cross-compatible with the antibodies to
betta-interferon and has identical electrophoretic pattern with it. A
test for resistance to viruses (tobacco and alfalfa mosaik viruses)
has demonstrated that the development of symptoms of viral disease in
transgenic tobacco and alfalfa was significantly retarded. Use of
ELIZA method allowed to establish that viruses more slowly accumulate
in plants that have delayed occurrence of infection symptoms.
The transgenic plants are used as a model for the study of
stability of expression and heritability of the genetic material
transferred into genomes of these plants. It was noted that in the F3
progeny derived from self-pollination of the initial transgenic parent
the expression of neomycinphosphotransferase (nptII) marker gene is
completely lowered. Reversible nature of this phenomenon has been
demonstrated. The structure of parental transgenic tobacco population
with respect to stability of nptII gene expression was analysed. It
has been shown that about 1% of plants of the first generation derived
from self-pollination of the parental transgenic plants already loose
their gene expression. Molecular-genetic mechanisms for loss of
expression in the transgenic plants are being studied.
Caryotype instability in the progeny of wheat regenerants is
studied using genetic methods. Heritability of caryotype instability
in generations has been shown. Besides the chromosomal rearrangements,
the unstable lines display some changes in spike morphology (speltoid
and awned spike) and duration of the vegetative period. The
instability in lines was also established by heritable chimerism for
spike morphology.
Within the framework of international program on search and
introduction of new plants as alternative source of cellulose in
vitro, studies are carried out aimed at obtaining of new forms and
cloning of vegetatively propagated perennial plants of genus
Miscanthus (Miscanthius Anderss) belonging to Gramineae family. A
unique collection of various forms, ecotypes and plants-regenerants of
miscanthus was formed. The first Russian experimental plantation of
miscanthus was set up. Genetic and ecologic investigations on growth,
its possible enhancement and improvement of biological variability
were carried out.
In vitro cultivation of annual species of alfalfa has begun. The
Medicago sativa and M.orbicularis regenerated plants have been
obtained.
Associative nitrogen fixation is studied in the rhisosphere of
cereals (wheat and barley). It was established that Triticum
timopheevi is highly capable of associative nitrogen fixation in the
rhisosphere. T.timopheevi chromosomes, being introduced into common
wheat genome, increase the population of nitrogen fixing bacteria on
wheat roots. It was established that activation of nitrogen fixation
in the introgressed lines is not associated with 1A, 1G and 5G
chromosomes of T.timopheevi whose presence has been established by
biochemical markers.
A more precise genetic mapping of Pisum sativum L. is now carried
out. Genes controlling dentate leaf character in garden pea have
already been localized. Lamprech has localised Td gene for weak leaf
dentation on chromosome 4. Some investigators have demonstrated that
Td gene was linked with the markers of chromosome 3. On the basis
of analysis of two hybrid combinations we have established that Td
gene was localized in the Np-22.8-le-16.7-Td region of chromosome 4.
It has been shown that weak leaf dentation was under control of
two independent genes with manifestation of edditive effect.
Heritability of response of barley caryopsis aleuron layer to the
treatment with exogenous hybberellic acid was studied in the hybrid
barley population with different response to this treatment. The
monohybrid mode of inheritance has been noted. The locus controlling
barley caryopsis aleuron layer response to the treatment with
hybberellic acid was mapped at 9.2+0.2% distance from trd locus and at
17.9+0.2% distance from B locus. With respect to experimental error,
the localization of the given loci coinsides with that of ea-k locus
mapped to chromosome 5 of barley. These loci are supposed to be
identical. Response to the treatment with hybberellic acid is a
primary event in the chain of a wide number of pleiotropic effects of
ea-k locus on numerous characters: roots and stem growth rates, total
mass of plant, duration of the vegetative period, as well as on
preferential storage of essential amino acids in the grain.
Investigation on the variability in the reproduction systems of
alfalfa developed in the course of continuous inbreeding and selection
are carried out. A series of highly inbred stocks (35-36 generations
obtained from self-pollination) has been obtained. The variation
typical of natural alfalfa populations is maintained, irrespective of
continuous inbreeding. On the basis of self-fertile lines of different
generations the new forms have been obtained, capable of autogamy and
autotripping. These forms are exploited as initial material for
development of cultivars with increased seed productivity.
Functional significance of variability for heterochromatic
regions of chromosomes is studied in terms of maize reproduction
systems investigation. A subpopulation of low and highly nodulating
maize with and without B chromosomes has been developed. It was shown
for this species that B chromosome is responsible for panicle and
spike flowering dates.
Obtaining of seed lots and testing of maize cultivar Sibiryatchka
in a number of regions of Russia are continued.
(3) General Results 1993-1995:
- A series of transgenic plants with human betta interferon
genes and those of bovine and Serratium marcescens nucleases has been
obtained. A delay in development of symptoms of viral infection in the
transgenic alfalfa and tobacco plants was noted.
- Two transgenic tobacco lines have been isolated which differed
in their stability in nptII marker gene expression. These lines are
exploited as models for the study of molecular-genetic mechanisms of
transferred genes functioning in the transgenic plants genomes.
- A somaclonal 5AL monotelosomic line with instability in the
long arm of 5A chromosome has been obtained. The instability manifests
itself in the high frequency of rearrangements in 5A chromosome, in
variability in the characters, for whom genes localized on the long
arm of 5A chromosome are responsible, and in chimerism for these
characters.
- Annual species of alfalfa (Medicago rigidula and M.
orbicularis) and the plants belonging to sp. Miscanthus (Miscanthus
Andress) have been introduced in vitro. A unique collection of
varieties, ecotypes and regenerated miscanthus plants and the
first experimental plantation of miscanthus for
Russia have been established.
- Heritability of karyotype instability in the progeny of
regenerated plants has been shown using genetic methods.
(4) Publications:
- Deineko E.V., Rivkin M.I., Shumny V.K. (1993). Use of gene
engineering methods in selection and genetic programs for improvement
of economic characters in alfalfa. In: Peculiarities of genome
reconstruction in the higher plant populations. Novosibirsk, pp.5-15.
- Rivkin M.I., Deineko E.V. (1993). Prediction of resistance to
viruses in transgenic tobacco and alfalfa plants with human
betta-interferon genes. Rep. of Acad. Sci. 331. (Rus.)
- Deineko E.V., Rivkin M.I., Komarova M.L., et al. (1994).
Analyses of alfalfa and tobacco transgenic plants with human
betta-interferon genes for resistance to viral diseases Bio/technology
(Bull) 4:22-26.
- Deineko E.V., Zagorskaya A.A., et al. (1995). Variation in the
level of expression and inheritance of nptII marker gene in the
progeny of transgenic tobacco. Rep. of Acad. Sci. 344(3). (Rus.)
- Omelyanchuk N.A. (1994). Rearrangements in highly repeated DNA of
barley genome in vitro. Genetika 30:879-885. (Rus.)
- Aksenovitch A.V. (1995). Study on the inheritance of variation
in response of the aleurone layer of barley to the treatment with
hybberellic acid. Genetika 32(6). (Rus.)
- Stepanenko I.L., Goncharov N.P. (1995). Associative nitrogen
fixation in the rhizosphere of different wheat species. Plant Physiol.
42:837-841. (Rus.)
- Smirnova O.G., Eggi E.E. (1994). Location in linkage group 3 of
a gene coding minor vicilin polypeptide. Pisum Genetics 26:31-33.
(5) Participation in State and International Programs:
- New Trends in Genetics
- RFBR
- International program within the framework of bilateral
co-operation between Institute of Gene Engineering (Bulgaria)
and Institute of Cytology and Genetics of the Russian Academy
of Sciences
- International program sponsored by J. Soros foundation
(6) Subjects of Scientific Collaboration:
- Development of transgenic plants with resistanse to viral
infections.
- Study of molecular-genetic mechanisms of inactivation of alien
gene expression in the transgenic plant genomes;
- Search and introduction of new plants as alternative source of
cellulose.
- In vitro introduction of annual alfalfa species; study of
associative nitrogen fixation in the rhizosphere of cereals (wheat,
barley).
- Verification of Pisum sativum L. genetic map.
- Heritability of barley caryopsis aleurone layer response to
the treatment with hybberellic acid.
- Study of reproduction systems variability in alfalfa using
continuous inbreeding and selection.
- Estimation of functional significance of variability for
heterochromatic chromosome regions in relation to reproductive systems
of maize.