ASSESSMENT OF ECOLOGICAL AND GENETIC DIVERSITY OF THE GENUS VIGNA SAVI USING BIOTECHNOLOGICAL METHODS - ISSR MARKERS

The world's genetic resources of plants are the main source of improving crops for decades to come. The gene pool of plants has a hidden resource of new genes, or their combinations, including - selection-important features. The study of the potential of the plant gene pool with the main biological and economic characteristics allows to expand the genetic base of crops for the successful implementation of breeding programs in various areas. The genus Vigna has about 200 species that are grown in warm regions of the planet. The centers of origin of the species are in Africa, but mung beans, urd, azuki and rice beans are of Asian origin. The collection includes 20 specimens belonging to 7 species of the genus Vigna: V. aconitifolia (Jacq.) Marechal (willow aconitolista, mott) – 3 specimens, V. angularis (Willd.) Ohwi et Ohashi (adzuki) – 4, V. radiata (L.) R. Wilczek (mash) – 4, V. umbellata (Thunb.) Ohwi et Ohashi (vigna rice) – 4, V. unguiculata (L.) Walp. (Chinese) – 5. These are mainly local varieties, about 10 % are breeding varieties and 1% are forms that grow in nature. The multifaceted use of cultivated species of the genus Vigna contributed to their spread throughout the tropical, subtropical and temperate zones of the globe. They are economically important crops in many developing countries. Geographical analysis showed that most of the samples came from the areas of world agriculture and crop formation. most samples of V. radiata, V. mungo, V. aconitifolia, V. trilobata, V. umbellata were obtained from India and Pakistan, V. angularis – from East Asia and China, V. unguiculata – East Africa (Ethiopia, Kenya), V unguiculata – from China. However, the exact place of domestication of this culture has not been established, and in numerous sources of literature can be found different opinions of scientists on the origin of cowpea and the centers of its diversity. Recently, using the methods of molecular genetics (RAPD, AFLP and others) it was confirmed that the northern part of Africa is the center of origin of the cultured, because the wild types of West Africa are closer to cultural forms than the wild types of East and South Africa. It should be noted that the species of cowpea have a significant intraspecific polymorphism. The samples showed strong variability of morphological and economically valuable features. Such a wide range of variable variability is due to the places of cultivation of samples, different environmental conditions (plains, mountains, climate). According to the results of the evaluation of the DNA polymorphism of cowpea using molecular genetic markers, it was found that the species of cowpea involved in the study are characterized by a high level of DNA polymorphism, which averaged 78.6 %. 145 loci were identified, including 31 unique, specific to a particular sample, and 31 monomorphic, characteristic of all samples. Monomorphic loci are conserved regions of DNA that indicate the common origin of the species of cowpea involved in the work, and can be used as genus and species-specific markers. Unique loci indicate genetic divergence of the studied material and can serve as markers of certain samples. The average level of intrapopulation polymorphism of cowpea DNA (37.2–93.8 %, depending on the genotype) was established, which indicates the existence of significant variability in the studied samples of cowpea. It shows a high level of genetic divergence of cowpea species and testifies in favor of the polyphyletic theory of their origin.

Genome-Wide DNA Polymorphism Analysis and Molecular Marker Development for the Setaria italica Variety “SSR41” and Positional Cloning of the Setaria White Leaf Sheath Gene SiWLS1

Genome-wide DNA polymorphism analysis and molecular marker development are important for forward genetics research and DNA marker-assisted breeding. As an ideal model system for Panicoideae grasses and an important minor crop in East Asia, foxtail millet (Setaria italica) has a high-quality reference genome as well as large mutant libraries based on the “Yugu1” variety. However, there is still a lack of genetic and mutation mapping tools available for forward genetics research on S. italica. Here, we screened another S. italica genotype, “SSR41”, which is morphologically similar to, and readily cross-pollinates with, “Yugu1”. High-throughput resequencing of “SSR41” identified 1,102,064 reliable single nucleotide polymorphisms (SNPs) and 196,782 insertions/deletions (InDels) between the two genotypes, indicating that these two genotypes have high genetic diversity. Of the 8,361 high-quality InDels longer than 20 bp that were developed as molecular markers, 180 were validated with 91.5% accuracy. We used “SSR41” and these developed molecular markers to map the white leaf sheath gene SiWLS1. Further analyses showed that SiWLS1 encodes a chloroplast-localized protein that is involved in the regulation of chloroplast development in bundle sheath cells in the leaf sheath in S. italica and is related to sensitivity to heavy metals. Our study provides the methodology and an important resource for forward genetics research on Setaria.

In situ new variations in Puccinia triticina causing leaf rust on wheat

During the periodic purity checks of pathotypes 107-1(45R35=JCGPL) and 20(5R27=SGQPL) of Puccinia triticina in national repository at Shimla, India, mixtures of infection types were observed on the differential Lr3 and Lr15 , respectively. Single pustule isolations and further pure cultures in both the cases yielded new pathotypes designated as 57(45R39= KGTPL) and 20-2(57R27= SHKPL). Pathotype 57 was one step gain in virulence on Lr3 in 107-1 whereas 20-2 on Lr15 in mother culture of pathotype 20. The difference of virulence on one gene to each mother culture and their non occurrence in the field samples is indicative of mutation for virulence in situ . To ascertain the novelty of new pathotypes, detailed study on differentials, avirulence/virulence structure, DNA polymorphism using SSR markers, and other related information is presented in this publication. The new cultures are being maintained as PrtI 57 and PrtI 20-2 in the repository.

Polymorphism parental survey in three Indonesia improved varieties to support development of new submergence tolerant varieties

Submergence by flooding caused damage in rice growing areas and huge economic loss, and developing tolerant varieties is considered as the best approach to overcome the problem. Markers Assisted Backcrossing (MABC) approach is widely to develop Sub-1 tolerant varieties. The availability of polymorphic markers is among the most crucial requirement to implement the MABC method. This research was subjected to assess DNA polymorphism between IR64Sub1 and tree Indonesian popular varieties. A total of 136 microsatellites/simple sequence repeat markers were used to genotype tree Indonesian popular varieties; Cisantana, Angke and Mekongga and IR64-Sub1. A total of 39 markers covering 11 chromosomes were found polymorphic between IR64 Sub-1 and the three varieties, however no polymorphic markers found in chromosome 12. The lack polymorphic markers were also found in chromosome 10 and 11 between IR64 Sub 1 and Angke. With the completion of the missing markers, these 39 polymorphic SSR markers can be utilized to support the MABC program for the development of new Sub-1 tolerant variety with multiple tolerances.

Utilization of Biotechnology, Neurotransmitter and Cytogenetic Indices in Selecting Pigeon Breeds

The DNA polymorphism LDH-A and DRD4 gene, some neurotransmitter and cytogenetic indices of three sexed pigeon breeds; non-racing pigeons (wild rock), racing long and short distances pigeons (e.g: Gan aarden) and (e.g: Gansen) have been performed. The long-distance pigeon demonstrated the highest levels of neurotransmitters in brain tissues. Long distance male pigeons have a unique LDH-A polymorphism, so A changed to G, which means that changes in gene expression resulted in differences in amino acid structure K(lys) was converted to E(glu) in the LDH-A gene only in long distance male pigeons compared to other breeds. In addition to this, the DRD4 polymorphism of long distance male pigeons in which A changed to C and A changed to T indicates the presence of a unique diversity in this breed. This means that changes in gene expression resulted in overall amino acid structure differences only in long distance male pigeons when compared to other breeds, where R(arg) and L(leu) were converted to S(ser) and F(phe). Neurotransmitters are synthesised in the body from these amino acids. It concluded that measurements would be useful indices for the differentiation, genetic characterization, provide a foundation for conservation programs of the breeding and selecting racing pigeon breeders.

Geographical Structuring of Quercus robur (L.) Chloroplast DNA Haplotypes in Lithuania: Recolonization, Adaptation, or Overexploitation Effects?

We studied the maternally inherited chloroplast DNA polymorphism at three microsatellite loci of 157 Quercus robur trees from 38 native populations in Lithuania. We found high diversity of eight haplotypes from the Balkan lineage A (frequency 0.75) and the “German” subbranch of the Balkan lineage A (freq. 0.12), western and eastern Italian lineages C (freq. 0.05 and 0.06, respectively), and Iberian lineage B (freq. 0.03). The haplotypes were geographically well structured (among population differentiation index PhiPT = 0.30, the p-value < 0.001) that is unexpected for such a small territory as Lithuania. We raised a hypothesis on historical overexploitation of oaks by eliminating certain haplotypes in Lithuania, following a drastic felling of oak forests over the last few centuries.