The review article discusses advances in genome editing of pea (Pisum sativum L.) using CRISPR/Cas9 technology. Despite more than a decade of CRISPR/Cas9 application in plant biotechnology, the first successful genome editing in pea was achieved only in 2023, when researchers induced mutations in the model gene PsPDS, whose disruption leads to plant albinism. To date, CRISPR/Cas9 has also been used to introduce mutations in the PsLOX2 gene, encoding lipoxygenase, resulting in a reduced concentration of volatile compounds responsible for the undesirable odor of seeds, and in the PsBAS1 gene, leading to blocked saponin biosynthesis and improved seeds’ palatability. Researchers emphasize the need to further optimize transformation protocols to enhance their efficiency and address the low regenerative capacity of pea. The review also briefly outlines the history of CRISPR-Cas9 discovery and its development as a key genome editing tool. In addition, it examines CRISPR/Cas9 modifications that improve editing precision and their potential applications in pea genome engineering. A key aspect of the article is the discussion of CRISPR/Cas9 as a tool for modulating the specificity and efficiency of pea symbiosis with nitrogen-fixing bacteria, which may contribute to the development of resilient and productive agroecosystems.

In pea (Pisum sativum L.), alleles of the Sym2 gene determine the specificity of the interaction with nodule bacteria (rhizobia). The Sym2^A allele present in landraces from Afghanistan provides higher selectiveness toward rhizobia than the Sym2^E allele present in European cultivars. Rhizobial strains possessing the nodX gene can interact with both Sym2^A and Sym2^E peas, while strains lacking nodX can interact only with Sym2^E peas. Here, we studied the previously obtained introgression line A33.18 bearing Sym2^A in a homozygous state in the genome of the European pea cultivar ‘Rondo’. A33.18 has proved its high selectiveness in pot experiments. Genome sequencing has shown that A33.18 possesses an 18.2 Mb region inherited from Afghanistan pea with 63 genes, including 5 receptor kinase genes, among which was the Sym2 candidate gene LykX. In a field experiment, under inoculation with the nodX⁺ strain TOM, over 95% of nodules of A33.18 contained TOM, as opposed to less than 8% of nodules containing TOM in the parental European cultivar ‘Rondo’. Thus, introgression of Sym2^A enabled peas to interact specifically with the nodX⁺ strain, favoring the formation of nodules by the strain from the inoculum and protecting peas from the indigenous soil microbiota.

A review is presented on recent advancements in the identification of blackberry hybrids, including successful examples of using molecular markers and DNA barcoding for the classification of complex hybrid forms. Special attention is given to the comparative analysis of the effectiveness of such methods depending on the research objectives and resource availability. The review highlights the future need for standardizing methods and expanding genetic marker databases to improve the accuracy in the classification of blackberry hybrids .

Accurate species identification in blackberries (Rubus spp.) is difficult because of morphological similarity and frequent hybridization. We studied 56 wild accessions from the Sirius Federal Territory (Russia), representing coastal and foothill ecosystems of the Black Sea region. Multilocus DNA barcoding with the plastid rbcL gene and nuclear ITS1 and ITS2 regions revealed signals of hybridization and hidden diversity. The rbcL marker showed low variation, grouping most accessions into two clusters with several singletons, which limited its use for distinguishing species. In contrast, ITS1 and ITS2 showed higher variation, forming six clusters and eight singletons, and allowed for clear separation of taxa such as Rubus caesius L., R. irritans Focke, and R. amabilis Focke. Accession 3 carried a raspberry (closely to R. corchorifolius L.fil) plastid haplotype, pointing to a hybrid origin. We also found groups of nearby plants with identical mutations, which likely reflect clonal spread with fixed somatic changes or the persistence of recent hybrid lineages. At the same time, accessions collected up to 140 km apart did not form separate clusters, showing weak geographic structuring along the coast. The results demonstrate that multilocus barcoding can reveal not only species boundaries but also evolutionary processes among Rubus such as hybridization, clonal propagation, and early stages of speciation.

Plant regeneration capacity in in vitro culture is of considerable importance for obtaining cultivars with required traits. Somatic embryogenesis represents a potential regeneration pathway widely used in biotechnology. In vitro induction of this process is challenging in many agronomically important crops, including pea (Pisum sativum). We evaluated the potential for somatic embryo development in 20 pea cultivars under in vitro conditions. Results indicate that the protocol used is applicable for generating somatic embryos, although the majority of the investigated cultivars did not display embryogenic potential under these conditions. The obtained results may contribute to the study of the genetic regulation of somatic embryogenesis induction in Pisum sativum.