The use of the primary structure of the ITS1-ITS2 region for species identification in some submerged aquatic macrophytes of the genus Stuckenia.

Applicability of ITS1-ITS2 primary structure for species attribution of representatives of the genus Stuckenia was experimentally tested. Analysis of the ITS1-ITS2 region sequences of S. vaginata and S. pectinata from public databases showed that they differed by insertions/deletions and single or double nucleotide substitutions. Besides, the ITS1-ITS2 region of S. pectinata was shown to be represented by two haplotype groups designated as S. pectinata type A and S. pectinata type B with good bootstrap support in phylogenetic reconstructions. In 28 samples identified as S. pectinata, S. vaginata, S. macrocarpa and S. chakassiensis on the basis of morphology, the ITS1-ITS2 region was sequenced in this study. Three groups of samples with good bootstrap support were revealed to be corresponding to S. vaginata, S. pectinata type A and S. pectinata type B. The S. vaginata group was formed by the samples identified on the basis of morphology as S. vaginata, and the S. pectinata type A group was formed by the samples identified on the basis of morphology as S. pectinata. The S. pectinata type B group was further divided into two subgroups, S. pectinata type B subgroup and S. chakassiensis subgroup. The S. chakassiensis subgroup included mainly the samples identified as such on the basis of morphology. The S. pectinata type B subgroup included samples identified on the basis of morphology as S. pectinata, S. vaginata and S. macrocarpa. We suppose that these samples were S. pectinata type B, S. macrocarpa and their hybrids.

Identification of the gene coding for seed cotyledon albumin SCA in the pea (Pisum L.) genome.

Albumins SCA and SAA are short, highly hydrophilic proteins accumulated in large quantities in the cotyledons and seed axes, respectively, of a dry pea (Pisum sativum L.) seed. SCA was earlier shown to have two allelic variants differing in mobility in polyacrylamide gel electrophoresis in acid medium. Using them, the corresponding gene SCA was mapped on Linkage Group V. This protein was used as a useful genetic and phylogeographical marker, which still required electrophoretic analysis of the protein while the DNA sequence of the corresponding SCA gene remained unknown. Based on the length, the positive charge under acidic conditions and the number of lysine residues of SCA and SAA albumins, estimated earlier electrophoretically, the data available in public databases were searched for candidates for the SCA gene among coding sequences residing in the region of the pea genome which, taking into account the synteny of the pea and Medicago truncatula genomes, corresponds to the map position of SCA. Then we sequenced them in a number of pea accessions. Concordance of the earlier electrophoretic data and sequence variation indicated the sequence Psat0s797g0160 of the reference pea genome to be the SCA gene. The sequence Psat0s797g0240 could encode a minor related albumin SA-a2, while a candidate gene for albumin SAA is still missing (as well as electrophoretic variation of both latter albumins). DNA amplif ication using original primers SCA1_3f and SCA1_3r from genomic DNA and restriction by endonuclease HindII made it possible to distinguish the SCA alleles coding for protein products with different charges without sequencing the gene. Thus, the gene encoding the highly hydrophilic albumin SCA accumulated in pea seeds, the alleles of which are useful for classif ication of pea wild relatives, has now been identif ied in the pea genome and a convenient CAPS marker has been developed on its basis.

Wild pea (Pisum sativum L. subsp. elatius (Bieb.) Aschers. et Graebn. s.l.) at the periphery of its range: Zagros Mountains.

Characteristics of wild peas and their habitats at the periphery of the range are interesting with respect to their potential importance for pre-breeding programs aimed at selection for different environmental conditions. However, wild pea diversity in peripheral regions is insufficiently represented in the existing germplasm collections. In such regions, wild pea populations are rare, small in size and suffer from climatic change and land exploitation, hence their focused search is strongly desirable. A two-week-long expedition to Iran in May 2017 revealed two small populations of the wild pea (Pisum sativum subsp. elatius) in the Zagros Mts, in Aligudarz and Khorramabad Districts of Lorestan Province, Iran, at elevations of 1841 and 1971 m a.s.l., respectively. Their habitats are briefly described. Two pea accessions derived from them, CE9 and CE10, were characterised for some visible and molecular characters. These peas appeared to belong to the evolutionary lineage B, recognised by us earlier in P. sativum as opposed to the so-called lineage AC. They contain a unique non-conservative substitution in subtype 5 of histone H1 and turned to be most related to some wild pea accessions originating from southern and south-eastern Turkey and Golan Heights. Scarce information available on wild pea occurrence in Iran suggests their existence in the south-western principal slope of Zagros Mts and southern principal slopes of Elborz and Kopet Dagh Mts. It was found that wild peas representing the evolutionary lineage B produce poorly open and poorly coloured flowers (as reported by us earlier) only in the greenhouse conditions but normally pigmented and open flowers in the wild and mesh houses at open air in Israel. Some issues of pea taxonomy are discussed.

Identification of genome compositions in allopolyploid species of the genus Elymus (Poaceae: Triticeae) in the Asian part of Russia by CAPS analysis.

The genus Elymus L., together with wheat, rye, and barley, belongs to the tribe Triticeae. Apart from its economic value, this tribe is characterized by abundance of polyploid taxa formed in the course of remote hybridization. Single-copy nuclear genes are convenient markers for identification of source genomes incorporated into polyploids. In the present work, a CAPS-marker is developed to distinguish basic St, H, and Y genomes comprising polyploid genomes of Asiatic species of the genus Elymus. The test is based on electrophoretic analysis of restriction patterns of a PCR-amplified fragment of the gene coding for beta-amylase. There are about 50 Elymus species in Russia, and most of them are supposed to possess one of three haplome combinations, StH, StY and StHY. Boreal StH-genomic species endemic for Russia are the least studied. On the basis of nucleotide sequences from public databases, TaqI restrictase was selected, as it produced patterns of restriction fragments specific for St, H, and Y haplomes easily recognizable in agarose gel. A sample of 68 accessions belonging to 32 species was analyzed. In 15 species, the earlier known genomic constitutions were confirmed, but in E. kamoji this assay failed to reveal the presence of H genome. This unusual H genome was suggested to originate from a different Hordeum species. In 16 species, genomic constitutions were identified for the first time. Fifteen accessions from Asian Russia possessed the genomic constitution StStHH, and E. amurensis, phylogenetically close to the StY-genomic species E. ciliaris, had the genomic constitution StStYY. It is inferred that the center of species diversity of the StH-genomic group is shifted to the north as compared to the center of origin of StY-genomic species, confined to China. Key words: Elymus; taxonomy; allopolyploids; genome constitution; CAPS markers.

[Comparative sequence analysis of the LEA gene fragment in Pinus sibirica du tour and Pinus pumila (Pallas) regel].

A comparative sequence analysis of the LEA (Late Embryogenesis Abundant (LEA)-like gene) intron fragment was performed in Pinus sibirica and P. pumila differing in geographic origin. It was demonstrated that in P. sibirica this fragment was represented by two types of PCR products, 224 and 202 bp in size. Similarly, in accessions of P. pumila, two PCR products of 224 and 159 bp in size were identified. Comparison of 224 bp fragments in P. sibirica and P. pumila showed that they differed in single nucleotide substitutions. Analysis of the intron fragment in a plant, which was characterized as an interspecific hybrid based on morphological characters, showed that it had fragments of 224 and 159 bp in size. The sequence of 224 bp fragment was similar to that of the corresponding fragment in P. sibirica. The structure of the short fragment was the same as the structure of the corresponding fragment in P. pumila. The data obtained are discussed in terms of the use of the sequences examined for species taxonomic classification and of an analysis of species hybridization.

[Genetic study of a new color of the beet Beta vulgaris L].

A new genetic character of the beet Beta vulgaris L., named stem color, was described and studied genetically. This character was shown to be dominant trait and monogenically inherited. The first-year beet plants with the genotype Stc/_have red leafstalks, weakly colored central rib, and colored storage root; however, the root itself is not colored. The second-year plants have a red-colored low third of the floral shoot. The plants with the genotype stc/stc are uncolored. The Stc gene was localized to the first linkage group at a distance of 17.5 +/- 2.1 crossing over units from the gene B (Bolting), which controls the annual-perennial habit of beet.

[dw2, a new mutation of beet Beta vulgaris L].

Twelve dwarf plants were found in the second hybrid generation of beet. The average height of mutant plants was 21.8 cm, their leaf blades and flowers were significantly smaller than normal, and the plants exhibited male and female sterility. This dwarfism was shown to be caused by a mutation differing from that previously described in beet, which is named dwarf2 (dw2). The experimental evidence suggests that this mutation appeared in one of the first-generation plants. Based on plant phenotype in the first hybrid generation and the number of mutant plants in the second one, this mutation is suggested to be under recessive monogenic control of the dw2 gene. The genotypic class segregation in the second hybrid generation indicates that the dw2 gene is inherited independently of genes m, at, and ap that control choricarpousness, gene male sterility, and pollen grain aggregation into tetrads.

Expression of mitochondrial genes in fertile and sterile sugar beet cytoplasms with different nuclear fertility restorer genes.

Variations in mitochondrial genome organization and in its expression between fertile, sterile sugar beet lines and fertile nuclear-restored plants were studied. Southern blot hybridization with COXI, COXII, COB and atpA mitochondrial genes as probes showed that changes in the mitochondrial genome organization of sterile lines are associated with variations in the location of COB, atpA and COXII, but not COXI. When the COXII and atpA genes were used as hybridization probes, differences in the primary structure of mitochondrial DNAs from sterile lines of different origin were revealed. Differences in the transcriptional patterns of the three mitochondrial genes (COXI, COXII and atpA) were observed between fertile and sterile sugar beet lines; COB was the only mitochondrial gene whose transcription was identical in both fertile and sterile cytoplasms. The dominant nuclear fertility restorer genes altered the transcriptional patterns of the COB and atpA without affecting those of the COXI and COXII genes; atp A expression was identical in fertile plants and nuclear-restored plants with sterile cytoplasm.