Background: Hybridization and introgression are said to occur relatively frequently in plants, and in particular among different species of willows. However, data on the actual frequency of natural hybridization and introgression is rare. Here, we report the first fine-scale genetic analysis of a contact zone shared between the three basket willow species,Salix dasyclados,S. schweriniiandS. viminalisin the vicinity of the Lake Baikal in Southern Siberia. Individuals were sampled in fourteen populations and classified as pure species or hybrids based on a set of morphological characters. They were then genotyped at 384 nuclear SNP and four chloroplast SSR loci. The STRUCTURE and NewHybrids softwares were used to estimate the frequency and direction of hybridization using genotypic data at the nuclear SNP loci.Results: As many as 19 % of the genotyped individuals were classified as introgressed individuals and these were mainly encountered in the centre of the contact zone. All introgressed individuals were backcrosses toS. viminalisorS. schweriniiand no F1 or F2 hybrids were found. The rest of the genotyped individuals were classified as pure species and formed two clusters, one withS. schweriniiindividuals and the other withS. viminalisandS. dasycladosindividuals.The two clusters were significantly genetically differentiated, withF_ST = 0.333 (0.282–0.382,p < 0.001). In contrast, for the chloroplast haplotypes, no genetic differentiation was observed as they were completely shared between the species. Based on morphological classification only 5 % of the individuals were classified as introgressed individuals, which was much less than what was detected using genotypic data.Conclusions: We have discovered a new willow hybrid zone with relatively high frequency of introgressed individuals. The low frequency of F1 hybrids indicates that ongoing hybridization is limited, which could be because of the presence of reproductive barriers or simply because the conditions are not favorable for hybridization. We further conclude that in order to get a complete picture of the species composition of a hybrid zone it is necessary to use a combination of morphological characters and genetic data from both nuclear and chloroplast markers.

With this paper we continue a new annual series, the main purpose of which is to make significant floristic findings from Russia and neighboring countries more visible in Russia and abroad. In total, this paper presents new records for 48 vascular plant species from 6 Eurasian countries, obtained during field explorations, as well as during taxonomic revisions of herbarium materials. For the first time, a new locality of Leontopodium leiolepis is recorded for Russia, Rheum uzengukuushi for China, Rorippa prolifera for Lithuania, Lappula marginata for Kyrgyzstan and Tajikistan, Anthriscus caucalis, Chenopodium ficifolium, Euphorbia prostrata for Uzbekistan, Adonis × hybrida, Potamogeton × franconicus, Solidago × niederederi for the Asian part of Russia, Echinochloa esculenta, Poa jamalinensis, Puccinellia poecilantha for Siberia, Potentilla intermedia for the Caucasus, Rhynchospora alba for the Russian part of Altai, Poa sphondylodes, Veronica beccabunga for Eastern Siberia, Asclepias syriaca for the Republic of Altai, Chimaphila umbellata, Orobanche korshinskyi, Veronica scutellata for the Republic of Buryatia, Cirsium alatum, Thalictrum simplex for the Republic of Crimea, Thymus rariflorus, Th. terekensis for the Republic of Ingushetia, Berberis thunbergii, Crataegus maximowiczii, Prunus serotina for the Republic of Mordovia, Oenothera villosa for the Republic of Tatarstan, Astragalus sulcatus, Galium mollugo for the Republic of Tyva, Phragmites altissimus for the Chelyabinsk Region, Senecio dubitabilis for the Magadan Region, Asclepias syriaca, Galatella villosa, Potentilla recta for the Novosibirsk Region, Dodartia orientalis for the Omsk Region, Viola hultenii for the Sakhalin Region, Phragmites tzvelevii for the Samara Region and the Middle Volga, Jacobaea ferganensis for the Samara Region, Carex media, Impatiens parviflora for the Tyumen Region. There are some more findings which are not new for the region but they contribute significantly to the understanding of species distribution.

Combined inoculation of legumes with rhizobia and plant growth-promoting rhizobacteria or endophytes is a known technique for increasing the efficiency of nitrogen-fixing symbiosis and plant productivity. The aim of this work was to expand knowledge about the synergistic effects between commercial rhizobia of pasture legumes and root nodule bacteria of relict legume species. Pot experiments were performed on common vetch (Vicia sativa L.) and red clover (Trifolium pratense L.) co-inoculated with the participation of the corresponding commercial rhizobial strains (R. leguminosarum bv. viciae RCAM0626 and R. leguminosarum bv. trifolii RCAM1365) and seven strains isolated from nodules of relict legumes inhabiting the Baikal Lake region and the Altai Republic: Oxytropis popoviana, Astragalus chorinensis, O. tragacanthoides and Vicia costata. The inoculation of plants with combinations of strains (commercial strain plus the isolate from relict legume) had a different effect on symbiosis depending on the plant species: the increase in the number of nodules was mainly observed on vetch, whereas increased acetylene reduction activity was evident on clover. It was shown that the relict isolates differ significantly in the set of genes related to different genetic systems that affect plant–microbe interactions. At the same time, they had additional genes that are involved in the formation of symbiosis and determine its effectiveness, but are absent in the used commercial strains: symbiotic genes fix, nif, nod, noe and nol, as well as genes associated with the hormonal status of the plant and the processes of symbiogenesis (acdRS, genes for gibberellins and auxins biosynthesis, genes of T3SS, T4SS and T6SS secretion systems). It can be expected that the accumulation of knowledge about microbial synergy on the example of the joint use of commercial and relict rhizobia will allow in the future the development of methods for the targeted selection of co-microsymbionts to increase the efficiency of agricultural legume–rhizobia systems.

Ten rhizobial strains were isolated from root nodules of a relict legume Oxytropis popoviana Peschkova. For identification of the isolates, sequencing of rrs, the internal transcribed spacer region, and housekeeping genes recA, glnII, and rpoB was used. Nine fast-growing isolates were Mesorhizobium-related; eight strains were identified as M. japonicum and one isolate belonged to M. kowhaii. The only slow-growing isolate was identified as a Bradyrhizobium sp. Two strains, M. japonicum Opo-242 and Bradyrhizobium sp. strain Opo-243, were isolated from the same nodule. Symbiotic genes of these isolates were searched throughout the whole-genome sequences. The common nodABC genes and other symbiotic genes required for plant nodulation and nitrogen fixation were present in the isolate Opo-242. Strain Opo-243 did not contain the principal nod, nif, and fix genes; however, five genes (nodP, nodQ, nifL, nolK, and noeL) affecting the specificity of plant-rhizobia interactions but absent in isolate Opo-242 were detected. Strain Opo-243 could not induce nodules but significantly accelerated the root nodule formation after coinoculation with isolate Opo-242. Thus, we demonstrated that taxonomically different strains of the archaic symbiotic system can be co-microsymbionts infecting the same nodule and promoting the nodulation process due to complementary sets of symbiotic genes.