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Marker-assisted selection and DH-technology utilized to accelerate fusarium-resistant cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) line development
2024
M. G. Fomicheva | G. A. Kostenko | A. S. Domblides
Relevance. The use of modern biotechnological methods in breeding, namely the doubled haploid technology and marker-assisted selection, can significantly reduce the time for creating pure lines with valuable properties. Fusarium wilt is one of the economically important diseases of white cabbage. Therefore, the development of a MAS method for selecting doubled haploids resistant to fusarium wilt would allow fast selection of resistant pure lines. Moreover, the resistance testing can be done at any plant developmental stage.Materials and methods. The response of hybrids Gertsoginya F1, Poisk 2018 F1 (resistant samples, the originator – the Agrofirm “Poisk”) and Slava 1305 (susceptible sample, the originator – Federal Scientific Vegetable Center) towards Fusarium oxysporum f. sp. conglutinans infection was determined by evaluating their growth on the inoculated soil. To test the efficiency of the FocBNUf/r marker to the polymorphic region of the FocBo1 gene, the markers were tested on resistant and susceptible genotypes. Then the markers were applied to analyze 60 doubled haploids of 12 different genotypes.Results. It was demonstrated that the FocBNUf/r marker effectively differentiated resistant and susceptible samples. Doubled haploids were obtained from 12 different breeding samples for FocBNUf/r marker-based selection of fusarium wilt resistant plants. PCR testing of doubled haploids for fusarium resistance allowed us to select 6.7-100% of plants with the fusarium resistance gene in a homozygous state in 8 genotypes. 4 genotypes of doubled haploids that did not carry the resistance gene were eliminated from the breeding program.Conclusion. The FocBNUf/r marker effectively identified resistant and susceptible samples, as well as differentiates homozygous and heterozygous plants. FocBNUf/r marker was used to select resistant doubled haploids of white cabbage at the seedling stage. Thus, it was possible not only to accelerate the production of pure lines by obtaining doubled haploids, but also to accelerate the selection of valuable samples carrying the gene of resistance to fusarium, which makes it possible to avoid the labor-intensive selection of resistant lines on inoculated soil.
Show more [+] Less [-]Creation of an interspecific hybrid of Capsicum annuum L. and C. frutescens L. using biotechnological approaches
2021
E. A. Dzhos | D. V. Shumilina | O. N. Pyshnaya | M. I. Mamedov | A. A. Baikov | A. A. Matyukina
Relevance. Pepper is a common crop both for fresh consumption and for the preparation of spices. Recently, along with the increasing popularity of C. annuum L. pepper, there is increasing interest in other species of this genus, which have a number of breeding and important properties. The most important method of enriching the gene pool of cultivated plants is distant hybridization, through which valuable traits are transferred from wild species to cultivated ones. The development of a new variety is a lengthy process, stretching over several years. In this regard, breeders have faced the challenge of obtaining pure lines to create a pepper hybrid with desired properties by applying modern biotechnological methods that will accelerate this process. One of them is the method of microspore culture, which allows mass production of haploid plants, reducing the time for creating constant parental lines.Material and methods. The aim of the work was to create an interspecific hybrid of hot pepper (C. annuum L. x C. frutescens L.) with high ornamental properties, a complex of economically valuable traits, with good taste qualities. The research was carried out in the film greenhouse of FSBSI FSVC in the Moscow region. The research material was a variety population of hot pepper Capsicum frutescens Cz-544-14, used as a paternal line, which was heterogeneous, and a pure line of C. annuum L. (Pb-551) created by classical breeding.Results. The pepper hybrid F1 Christmas bouquet was created as a result of hybridization of species parental forms obtained by different methods (biotechnological and classical). To accelerate the production of an aligned paternal form of C. frutescens L., the technology of doubled haploids through microspore culture was used. As a result, doubled haploid plants meeting the planned model (compact low habit, purple fruit colouring in technical ripeness and red in biological ripeness) were obtained. The resulting hybrid combined all the necessary economic features: high ornamentality, compactness, bouquet arrangement of fruits, high taste and aroma. Thus, the use of remote interspecific hybridization in the breeding process in combination with biotechnological approaches can accelerate the production of new forms of hot peppers that meet the demands of the market.
Show more [+] Less [-]PROMOTION OF COMPETITIVENESS OF NATIONAL DRASSICA BREEDING PROGRAM IS THE MAINEST TASK OF AGRICULTURAL SCIENCE
2017
L. L. Bondareva | S. M. Nosova
The basic research results of Russian breeding program in Brassica crops were given in the article. Conventional breeding as well as biotechnological methods were used to develop hybrids that meet all requirements of current vegetable market. Brief communication information on international scientific and practical conference “Current State and Trends in Breeding and Seed Production of Brassica crops” was given in the article.
Show more [+] Less [-]Obtaining and evaluating breeding material for the creation of F1 hybrids of Chinese cabbage (<i>B. rapa</i> ssp. <i>pekinensis</i>) with resistance to stress factors
2023
A. D. Zastavnyuk | G. F. Monakhos | S. G. Monakhos
Relevance and purpose of the study. Chinese cabbage is a vegetable crop with a high content of vitamins and low calorie content, valued for its ability to receive several crops per year and high productivity. Recently, there has been a reduction in the supply of the vegetable to Russia. There are less than 70 varieties and hybrids of the crop in the State Register of Selection Achievements, many of them are susceptible to the most important diseases. It is necessary to meet consumer demand with new commercial hybrids with resistance to stressors. The aim of the study was to evaluate and obtain genotypes with a set of valuable economic traits, including resistance to clubroot, powdery mildew, and early bolting.Materials and methods. The Chinese cabbage lines of various degrees of inbred and DH, as well as hybrid combinations from crossing these lines, were used as plant material. The lines were obtained in 2019-2020, the breeding material for them was selected on the ground infected with clubroot and with tolerance to tip burn of heads. To achieve the goal, the following methods were applied: plants cultivation; DNA isolation (CTAB method); PCR analysis; electrophoresis and visualization of results; productivity assessment and dispersion analysis; obtaining doubled haploids in culture of isolated microspores in vitro, assessment of resistance/susceptibility to powdery mildew.Results. 2 genotypes with high productivity and alignment in turnover summer-autumn 2022 and a genotype exceeding 3 standards in head weight in turnover spring-summer 2022 were recommended; as a result of molecular genotyping, 41 lines were differentiated according to the clubroot resistance gene CRb; DH plants for 4 genotypes with resistance to early bolting were obtained, their responsiveness to embryogenesis was studied; 18 lines were evaluated for resistance to powdery mildew, 7 genotypes with resistance to the disease were identified.
Show more [+] Less [-]Production of doubled haplois in Brassica purpuraria
2019
E. V. Kozar | K. S. Korottseva | O. V. Romanova | O. A. Chichvarina | L. Yu. Kan | V. A. Ahramenko | E. A. Domblides
Relevance. In recent years vegetable crop Brassica rapa ssp. chinensis var. purpuraria (synonyms: Brassica campestris L. var.purpurea Bailey; Brassica rapa L. ssp. chinensis var. purpurea) is gaining popularity as an object of genetic and molecular researches, and as an economically valuable vegetable plant due to the high content of biologically active compounds and distinctive economically valuable traits. Effective technology for development DH-plants to accelerate the breeding process for this culture has not been developed yet, so research in this area is relevant.Materials and methods. The study included two varieties from the collection of Vavilov AllRussian Research Institute of Plant Industry (VIR): No. 1301 (China) and No. 1357(Netherlands). Both protocols standard unmodified and with addition of silver nitrate (AgNO3) in the medium for embryogenesis induction were used in experiments for production of DH plants from isolated microspore in vitro. Direct chromosome counting in meristem cells and flow cytometry were used to determine the ploidy of regenerating plants.Results. As a result of the study embryogenesis in B. purpuraria culture can develop with the use of a standard protocol as well as with the addition of silver nitrate that showed a positive effect on the induction of embryogenesis. The yield of embryoids varied depending on the genotype of the individual plant within the variety accession. The highest yield of embryoids was 40 embryoids/petri dish. The main problem at the stage of regeneration is that about half of the regenerating plants occurred to be albinos and were not viable. We show a high degree of spontaneous chromosome doubling in regenerated plants (all analyzed plants were doubled haploids). In total 38 regenerated plants were obtained from accession No. 1301. It was shown that four DH-plants had self-incompatibility after self-pollination, but seed progeny from other plants was obtained. The created material was taken for genetics study and breeding work.
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