The development of an agro-industrial complex under present-day conditions is impossible to imagine without the development of agro-biotechnology, which in turn requires specialists with profound knowledge of biology, chemistry and related sciences. In this regard, training of personnel is needed to ensure active implementation of modern technologies in agricultural sciences. Until recently, such specialists have not been trained at classical universities, to which St. Petersburg State University belongs. To deal with this challenge, a Masters Program «Molecular Biology and Agrobiotechnology of Plants» has been developed and is being implemented in SPbSU. Teaching staff from eight departments of the Biological Faculty of SPbSU is involved in the creation and implementation of the Program. The Program in question is focused on familiarizing students with the modern problems, achievements, methodology of agro-biotechnology of plants, as well as on practical application of the obtained knowledge. Special attention is paid to the formation of trainees’ perceptions of the possibility and necessity of bringing plant breeding to the level of requirements and possibilities of the «post-genome era» to achieve high productivity and sustainability of agricultural production with minimal environmental risks. The Program seamlessly integrates practical exercises and students’ research work in the SPbSU facilities, as well as that performed at St. Petersburg research institutes. Much attention is paid to the development of students’ skills in conducting scientific discussions and in presenting their scientific data in different formats, for instance in English, which is very important for monitoring current scientific trends and integrating own research into world science. The Program is popular with students and many of its graduates have been employed by the leading biological and agricultural research institutes.

Background. Breeding new crops is associated with a risk of decrease (erosion) in genetic polymorphism. It is associated mainly with the fact that only a limited range of pre-existing cultivars and forms are used for breeding new cultivars. The Russian cultivars of garden pea (Pisum sativum L.) are characterized by a high level of phenotypic variability, while dynamics of polymorphism at DNA level is poorly investigated. Such an investigation is relevant from the point of view of the prospects for further breeding of the crop, and for planning strategies of polymorphism conservation in germplasm collections. Materials and methods. The material used in the study included 18 Russian cultivars bred before 1991, 22 Russian cultivars created later, 40 foreign cultivars, as well as 7 marker lines and wild-growing accessions. These lines were phenotyped and genotyped using DNA markers (14 CAPS, 8 SSR, one mitochondrial and one chloroplast). Variability level was measured as an average Jaccard coefficient resulting from pairwise comparison within each group. Additionally, we analyzed the published characteristics of cultivars included in the State Register in 1994 - 2019. Results. At the phenotypic level, the variability of grain cultivars increased, while a decrease was recorded for vegetable cultivars. A comparison with foreign cultivars has shown that their polymorphism was similar to those of Russian cultivars, except for the vegetable cultivars which are phenotypically less polymorphic than foreign ones. The DNA polymorphism demonstrated a similar tendency, although to a lesser extent. A low level of within-cultivar polymorphism was found. Productivity characteristics of cultivars included into the State Register in the early period (1995 - 2000) and those included later (2016 - 2019) do not differ significantly. Conclusions. There is no clear indication of genetic erosion in Russian cultivars of pea. We also found no reliable increase in the basic agriculturally important characters within the two decades.

Background. Genetic modification of plants is one of the promising strategies to increase their resistance to insect pests. The development of metabolic or RNA interference systems for plant protection requires appropriate models of host-insect interactions. Nicotiana tabacum L. is a classical model plant used in molecular and metabolic engineering. We consider tobacco as a model for developing protective strategies against Colorado potato beetle (Leptinotarsa decemlineata Say, CPB). Normally, tobacco is toxic for CPB due to high content of nicotine and related alkaloids in leaves. Modification of the tobacco genome could provide tobacco genotypes with altered metabolism suitable for CPB feeding. It is known that different mutations in Berberine Bridge-Like (BBL) genes cause different alterations in tobacco leaf alkaloid levels. In the current study, the Cas9/gRNA system targeting members of the BBL gene family of tobacco was used to create a line which can serve as a diet for CPB. Results. In order to obtain tobacco with modified alkaloid content, two gRNAs matching target sequences in six BBL genes were selected. Each gRNA was cloned into a gRNA/Cas9 generic vector. The created constructs were mixed and used for biolistic transformation of tobacco leaf explants together with the pBI121 plasmid harboring the kanamycin resistance gene nptII and the reporter E.coli betaglucuronidase (GUS) gene. Regenerants were selected on 100 mg/l of kanamycin and checked for transgene presence by histochemical GUS-assay. Unexpectedly, the regenerated plants displayed a variety of adverse phenotypic effects including different degree of growth and rooting inhibition, early flowering, increased number of internodes, changes in leaf shape, fusion of flowers, longostyly, and partial sterility. Only one from seven obtained calli produced a population of regenerated plants without severe phenotypic abnormalities. The NtaBBL5-14 line of clonally propagated plants was selected from this population and used for a CPB feeding experiment. It was shown that CPB larvae consume the leaves of NtaBBL5-14 line ten times more efficiently than the leaves of control plants (97±0.5% vs. 9±3% in 24 h respectively). Conclusion. The NtaBBL5-14 tobacco line is suitable for CPB feeding and can be further used as a model for studies in plant-pest interaction. The modification of other genes regulating nicotine metabolism can be a promising strategy to obtain tobacco plants edible for CPB with less pleiotropic effects.

Genome editing methods are now widely used in research aimed at studying fundamental biological processes, in particular for regulating maturation and extending shelf life of plant agricultural products. This review briefly discusses plant genome editing methods and examples of their successful application for increasing the storage life of fruits of tomato as one of the most important crops. Genome editing is one of the new areas of genetic engineering that is truly revolutionary in biotechnology. Various genome editing systems have been developed over the past decades: zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and clustered regularly located short palindromic repeats recognized by Cas9 nuclease (CRISPR/Cas9). The most common and widely used is the CRISPR/ Cas9 system, which has many advantages over other existing genome editing systems.

An important trend in the field of floriculture is the creation of new varieties of ornamental plants, among which varieties with unusual color are most in demand. To this end, traditional breeding and selection programs have been successfully applied for many years. However, currently genetic engineering is able to offer an alternative way to obtain new forms and varieties. Anthocyanins belonging to flavonoids, betalains and carotenoids are the main types of pigments that are synthesized in the plant and are responsible for the color of flower petals. The modification of pigment biosynthesis pathways using genetic engineering techniques can produce results that cannot be obtained by traditional breeding. This review presents the main advances in the application of genetic engineering techniques in floriculture using the example of flower color modification. There are several main areas of work with the genes of pigment biosynthesis. Among them, the strategy of suppressing gene expression is used most often. Expression of certain genes is suppressed to prevent pigment synthesis, or vice versa, to eliminate factors that hinder color development. The method of additional heterologous genes insertion to plants lacking them in the pathway of pigment biosynthesis is often used. Genomic editing, in particular by using the CRISPR/Cas system, is also used for color modification, but the application of this method to ornamental plants is a relatively recent innovation. Despite the rapid development of biotechnology, there are obstacles to the distribution of genetically modified plants on the world market. By addressing a number of problems, the production of transgenic ornamental plants may become economically more cost-effective and attractive than the development of new varieties exclusively through traditional breeding methods.

Precise editing of the genes of plant organisms with complex genomes has long been a difficult task. The CRISPR/Cas technology developed in the last decade has become one of the preferred tools for site-directed mutagenesis of plant genes and has quickly replaced the ZFN and TALEN systems. However, while the CRISPR/Cas system has proven to be an effective tool for modifying the genome of diploid species, its application to organisms such as cereals with complex and, in the case of common wheat, polyploid genomes is complicated by a number of obstacles. This review summarizes the main results obtained using the CRISPR/Cas system in such economically valuable cereals as common wheat Triticum aestivum L., barley Hordeum vulgare L., and maize Zea mays L., the genome structure of which increases the probability of the emergence of non-target mutations and reduces the specificity of editing. Every year the number of methodological publications on the directed mutagenesis of these crops, aimed at optimizing and improving the performance of the CRISPR/Cas system, increases exponentially, and the editing efficiency reaches 100% for maize and barley. The experimental articles are mainly aimed at improving the economically important traits of plants, such as improved yields, nutritional value and resistance to diseases and herbicides. Plant improvement is also associated with editing genes that affect pollination control, which is used in hybrid breeding. This creates the prerequisites for the creation of new maize, barley and wheat varieties, and for the saturation of existing ones with the necessary properties.