Androgenesis—Technology for Obtaining Genetically Stable Breeding Material of Capsicum annuum L.

Androgenesis in vitro is a basic method of obtaining haploid plants and DH (doubled haploid) lines of major crops such as potato, rapeseed, tomato, pepper, wheat, maize, and barley, and also many different minor crops and species with lower agricultural impact. Diploid plants appearing among androgenic regenerants are the effect of spontaneous doubling of the chromosome number in haploid cells during an embryo’s early developmental stages and are valuable fully homozygous breeding material. The subject of the presented research is spontaneous diploidization occurring in the development of androgenic, haploid pepper regenerants. In the presented experiment, the formation of diploid seeds was observed in the progeny of an androgenic, haploid plant derived in an anther culture of a hybrid (Capsicum annuum L. ATZ × Capsicum annuum L. `Corno di toro`)F2. Agromorphological and molecular analyses concerned eight diploid plants being progeny of the anther-derived haploid regenerant. Five of the plants constituted a phenotypically balanced group with valuable agromorphological features. Their genetic homogeneity was confirmed using 10 RAPD markers and 16 ISSR markers. Based on the results, it was concluded that anther-derived haploid plants of Capsicum can be the source of diploid, apomictic seeds, and the obtained offspring may constitute genetically stable, valuable breeding material.

Creation of the bank of breeding material of winter rye in vitro

Аn important issue of the selection process is the preservation of the source and obtained material, especially cross-pollinated crops, which significantly lose their viability by inbreeding. Creating a bank of genetic resources using biotechnological methods will effectively solve this problem. The aim of the work was to determine the conditions for the formation of a genetic bank of valuable winter rye materials with changes in temperature and modeling of the nutrient medium for long-term disposal of cloned plants and the use of active collection of original and created forms in the selection process. To deposit the clones, a nutrient medium, which included macro- and microelements according to the Murashige-Skuga medium was used. The nutrient substrate modified with cytokinins and carbohydrates. The clones in culture rooms at a temperature of 6–12 °С and low light intensity (2 kLk) were stored. In the course of research the conditions of creation of an active collection of plants of winter rye with use of temperature restriction and modification of a nutrient medium are defined. A consistent technological scheme for the conversion of plant material into a state of relative anabiosis has been developed. It is proved that the optimal storage temperature for samples is 6 °С. Survival of plants at the specified temperature regime after 12 months of deposition on average by genotypes at the level of 78,2 % was recorded. Modification of the nutrient medium with agar-agar at a concentration of 12,0 g/l increases the proportion of viable clones to 81,3 %, and the introduction into the substrate of an increased concentration of growth regulators, in particular 6-BAP (2,0 mg/l) and sucrose 40,0 g/l and a gradual decrease in temperature to 10 °С prolongs the period of deposition of cloned plants without changing the substrate and the shelf life of breeding material in isolated crops. Using of biotechnological methods for the preservation and reproduction of valuable material intensifies the selection process of obtaining initial samples of winter rye.

Features of morphotypic selection in populations of Serratula coronata L. cultivated in the conditions of the Moscow region

The article presents the results of morphotypic analysis of populations of perennial medicinal and fodder crops cultivated in the Moscow region (Serratula coronata L.). In the introduction, the botanical characteristics of the crowned sickle are given, the prospects and advantages of phytoecdysteroids obtained from raw materials are noted. The aim of the study is a comparative assessment of cultivated populations of S. coronata by morphotypic composition, determination of the age period for the selection of initial forms for breeding. The object of the study was the different geographical origin S. coronata three cultivated populations (Samarskaya, Belgorodskaya, Syktyvkarskaya from Rep. Komi). We have tested the application of methods for assessing varieties by uniformity, distinctness, stability for morphotypic selection in the population when creating improved, initial and breeding material. The article presents the results of the analysis of the morphotypic composition of populations by morphological characteristics and the productivity of raw materials according to long-term data (2013-2017, Moscow region, VILAR), the age period during which morphotypic selection should be carried out is established, morphotypes that are of the greatest value during selection are identified and described. The age period during which morphotypic selection should be carried out is established: 3-5 year, since the morphotype is fully formed by the 3rd year. It is established that plants in cultivated populations of Serratula coronata are represented by 2-3 morphotypes based on the height and leafiness of the generative shoot, the length of the stem leaves and the mass of the shoot. When selecting, the highest value is represented by tall morphotypes with medium-length leaves (~60-70% of them in each population), but morphotypes with medium leafiness should be selected among them (only ~41-51% of them in each population, only in the Syktyvkarskaya population – up to 69 %). The results of the study are relevant in the aspect of creating an improved, initial and breeding material of the Serratula coronata. Keywords: MEDICINAL PLANTS, SERRATULA CORONATA L., PHENOTYPIC VARIABILITY, MORPHOTYPE, POPULATION, PRODUCTIVITY, QUANTITATIVE CHARACTERISTICS, SELECTION

Virus Yellows in sugar beet – possibilities to achieve virus resistance

Virus yellows in sugar beet is caused by different virus species. Monitoring has shown that Beet yellows virus (BYV), Beet mild yellowing virus (BMYV), Beet chlorosis virus (BChV) are common and widespread, while Beet mosaic virus (BtMV) is less prevalent. The green peach aphid (Myzus persicae) is considered the main vector of these viruses. Sugar beet varieties with resistance or tolerance traits are currently not available to practical growers, therefore it is imperative to support breeding efforts with improved strategies to achieve virus resistance. For this purpose, a field test was established in which yield differences between susceptible and tolerant varieties can be generated by a 3% inoculation with BMYV-carrying aphids. A greenhouse bioassay has also been developed to distinguish susceptible and tolerant genotypes following BYV infection. Both assays pave the way for future use of natural resources such as wild forms and other breeding material to screen for virus resistance. In addition, molecular biology approaches are used to identify plant susceptibility factors of the plant-virus interaction, which will be knocked out via modern precision breeding methods to generate recessive virus resistance. Consequently, genotypes with naturally occurring mutations in the appropriate factors can be used for crossbreeding processes into elite breeding material.

Tomato hybrids with complex resistance for ground greenhouses

На фоне усиливающейся инфекционной нагрузки на культуру томата, обусловленной появлением новых болезней и новых физиологических рас традиционных патогенов, возрастает значение создания и использования в производстве гетерозисных гибридов с комплексной устойчивостью. Возрастает вредоносность новых насекомых-вредителей культуры томата. По вредоносности выходят на первое место томатная минирующая моль (Tuta absoluta) и западный цветочный трипс (Frankliniella occidentalis), который помимо нанесения прямого ущерба культуре выступает как переносчик вирусных заболеваний, в том числе вируса бронзовости (TSVW). Начатая селекционная программа на полевую устойчивость томата к трипсу позволила в 2020–2021 годах выделить восемь источников устойчивости среди образцов различных товарных групп. В результате двадцатилетней селекции созданы коммерческие гибриды с различным набором генов устойчивости. В задачи современной селекции входят этапы оценки и отбора селекционного материала на наличие генов устойчивости в генотипах методами ПЦР-диагностики, а также контроль устойчивости путем испытания материала в условиях инфекционных фонов или искусственного заражения. Важный этап – контроль товарных партий семян на присутствие генов устойчивости. Результаты ПЦР-анализа коммерческих гибридов на четыре гена устойчивости в сочетании с испытанием полевой устойчивости к кладоспориозу на сильном многолетнем инфекционном фоне приведены в табличной форме. Результаты показывают, что практически во всех товарных группах есть гибриды с устойчивостью к кладоспориозу, фузариозному увяданию, вирусу томатной мозаики (ВТоМ) и галловой нематоде. Также была обнаружена относительная устойчивость к фитофторозу у гибрида F1Изящный (Ph2\Ph2). Сегодня проводится работа по расширению числа генов для ПЦР-анализа селекционного материала. Under conditions of the increasing infectious load on the tomato crop due to the emergence of new diseases and new physiological races of traditional pathogens, the importance of creating and growing heterotic hybrids with complex resistance is increasing. The harmfulness of new insect pests of tomato culture is also increasing. In terms of harmfulness, the tomato mining moth (Tuta absoluta) and the western flower thrips (Frankliniella occidentalis) take the first place, which, in addition to causing direct damage to the culture, is a carrier of viral diseases, including Tomato spotted wilt virus. The started breeding program for the field resistance of tomato to thrips made it possible for the period 2020–2021 to identify 8 sources of resistance among the samples of various commodity groups. As a result of 20 years of selection, commercial hybrids with different sets of resistance genes have been created. The goals of modern breeding include breeding material assessment and selection for the presence of resistance genes in genotypes by PCR diagnostics method, as well as control of resistance by testing the material under infectious backgrounds or artificial infection inoculation. Another important step is the control of commercial seed lots for the presence of resistance genes. The results of PCR analysis of commercial hybrids for 4 resistance genes in combination with the test of field resistance to Cladosporium fulvum under condition of strong long-term infectious background are given in tabular form. These results show that in almost all product groups there are hybrids with resistance to ToMV, in combination with other resistances, including the relative resistance to late blight in the F1 Iziyaschniy (Ph2\Ph2) hybrid. Currently, the progress is being made towards expanding the number of genes for PCR analysis of breeding material.

Study on fusion blanket with ceramic solid as tritium breeding material

Variation of solid ceramic breeding might be one of the excellent candidates in a fusion reactor. The LiAlO2, Li4SiO4, Li2O, and Li2ZrO3 show pretty good requirements in tritium breeding capability and thermodynamic behavior. Especially for LiAlO2 and Li2ZrO3, in which they could be possible to breed without neutron multiplying needed as blanket used generally in order to reach the self-sufficiency reactor. So that, it makes up the material could be possible as high-estimation breeder material.

Creation of a new breeding material for creeping clover (Trifolium repens L.) hay-pasture type using the hybridization method

The studies were carried out in the Moscow region in 2019-2021. In order to create a new breeding material for creeping hay-pasture clover, samples of the Lodian type (giganteum variety), characterized by high peduncles and longer leaf petioles, and a low content of hydrocyanic acid (0.3 mg%), were selected as parental forms. Reciprocal crossing was carried out. Hybrids F1 and F2 in the first year of life were assessed by the main economic and biological characteristics: the length of the peduncles, the number of inflorescences on the plant, fodder and seed productivity, early maturity, winter hardiness and other indicators. Based on the assessment results, the best hybrids and individual biotypes were identified.

The breeding of spring rapeseed and brown mustard for resistance to Fusarium blight

During the growth season, the plants of rapeseed (Brassica napus L.) and brown mustard (Brassica juncea L.) are affected by various diseases: Fusarium blight, Verticillium blight, Alternaria blight, and others. The cultivation of rapeseed and mustard varieties resistant to diseases is a cost-effective and environmentally safe way to protect plants from diseases. The aim of the work was to evaluate the new breeding material of spring rapeseed and brown mustard for resistance to Fusarium blight in the form of tracheomycotic wilting of plants to continue breeding work to develop varieties of these crops. In 2017-2020, we carried out a phytopathological evaluation of new breeding samples of rapeseed and brown mustard for resistance to Fusarium blight. As a result, we selected a valuable breeding material of spring rapeseed and brown mustard resistant to Fusarium blight infection, which also exceeds the standard by economic characters. The productivity of the best selected samples is higher than the varieties Tavrion and Nika by 0.13-0.59 and 0.18-0.28 t/ha, respectively, the oil content of seeds – by 0.2-2.0 and 1.2-2.1 %, respectively. These samples will be used as donors of Fusarium blight resistance in breeding work during development of new varieties of spring rapeseed and brown mustard in the central zone of Krasnodar region.

Breeding of perennial forage crops for disease resistance

The creation of the initial breeding material of forage crops, promising in terms of disease resistance, is a difficult task that requires the solution of many related issues. Such studies should be carried out on artificial field infectious backgrounds. In FSC “VIK named after V.R. Williams” such backgrounds were created on the basis of data of long-term phytosanitary monitoring to identify the main diseases of forage crops. Techniques for creating field backgrounds and methods for laboratory assessment of resistance have been developed, which make it possible to reduce the number of samples evaluated against backgrounds, due to the rejection of susceptible to the disease. When evaluating 20 collection samples of meadow clover and awnless rump, 10 samples with increased disease resistance were identified. As a result of further breeding study of this material by methods of selection, self-pollination, cloning, promising biotypes in terms of disease resistance were obtained. On the basis of these biotypes, polycross populations of meadow clover and awnless rump were created. An assessment of the stability of these populations against artificial infectious backgrounds was carried out. In terms of resistance to fusarium, the polycross population of meadow clover exceeded the standard variety VIC 77 by 24%. The population of awnless rump exceeded the standard Fakelny cultivar by 21% on average over 2 years. A breeding program has been developed to create the initial material for meadow clover and awnless rump with increased disease resistance.

RNA-Seq of potato plants reveals a complex of new latent bacterial plant pathogens

The throughput and single-base resolution of RNA-Sequencing (RNA-Seq) have contributed to a dramatic change in diagnostics of viruses and other plant pathogens. A transcriptome represents all RNA molecules, including the coding mRNAs as well as the noncoding rRNA, tRNA, etc. A distinct advantage of RNA-Seq is that cDNA fragments are directly sequenced and the reads can be compared to available reference genome sequences. This approach allows the simultaneous and hypothesis-free identification of all pathogens in the plant. We conducted surveys for potato (Solanum tuberosum L.) -associated phytopathogenic bacteria in 56 original and GenBank RNA-seq data sets for potato breeding material. Bacteria of genera Pseudomonas, Burkholderia, Ralstonia, Xanthomonas, Agrobacterium, and species of family Enterobacteriaceae were most frequently detected in RNA sets from the studied plants. RNA-seq reads identified as Xanthomonas spp. were within X. vesicatoria, and some other species. Xanthomonas spp. covered up to 9,1% of all reads and included the major clades of these bacteria known as pathogens of solanaceous crops, but potato. Bacteria of genus Xanthomonas infect different plant species under artificial inoculation, suggesting that they are shared among wild plants and crops. Our studies indicated that a larger number of solanaceous plants can be occupied by specific Xanthomonas pathovars as endophytes or latent pathogens. Revealing bacteria distribution in the plant breeding material using RNA-seq data improves our knowledge on the ecology of plant pathogens.