Production of Doubled Haploids in cucumber

Домблидес, Е. А.; Белов, С. Н.; Солдатенко, А. В.; Pivovarov, V. F.

doi:10.18619/2072-9146-2019-5-3-14

Cited by 5 publications

(2 citation statements)

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“…Основные достижения по получению удвоенных гаплоидов у культур семейства Cucurbitaceae были освещены в ряде обзоров [54][55][56][57][58] и опубликованы в виде протоколов, в том числе и в вышедшей в 2021 году книге «Doubled Haploid Technology» под редакцией Jose M. Segui-Simarro [10,[59][60][61][62][63].…”

Section: история открытия гаплоидов и первые гаплоиды у культур семейства Cucurbitaceaeunclassified

“…У каждого из этих методов при использовании их для тыквенных культур имеются свои преимущества и ограничения, которые были освещены в ряде обзоров [55][56][57]. Для эффективного внедрения гаплоидных технологий в селекционный процесс необходимо добиваться увеличения выхода удвоенных гаплоидов, воспроизводимости разрабатываемых методик, что невозможно сделать без тщательной отработки всех этапов технологии.…”

Section: современное состояние исследований по получению удвоенных гаплоидов C Pepo Lunclassified

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Obtaining doubled haploids of Cucurbita pepo L.

2021

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Doubled haploids have been widely used worldwide in breeding programs and fundamental research as valuable homozygous material for about 100 years. The species Cucurbita pepo L. are represented by a huge variety of forms, include highly productive vegetable crops and have a wide distribution in the world. Despite the great economic importance, the creation of effective protocols to ensure stable production of doubled haploids in this species remains an urgent task. DH plants are of interest not only because of the acceleration of the breeding process, but also because of the realization of the huge potential of gametoclonal variability inherent in this highly polymorphic species. In this review, we analyzed the main technologies used for obtaining doubled haploids in vegetable crops of C. pepo: parthenogenesis in situ stimulated by treated/irradiated pollen, gynogenesis in vitro (unpollinated ovule culture in vitro) and androgenesis in vitro (anther/microspore culture in vitro). An analysis is presented of the research carried out from the beginning of the discovery of haploid plants to the current advances and evaluation of the prospects in the field of DH plant production. The main critical factors influencing the efficiency of each technology and its individual steps are considered. The developed technology of doubled haploids obtaining using non-pollinated ovary culture in vitro is presented. This technology allows to obtain up to 55 embryoids per one cultivated ovary (28 embryoids/ 100 cultivated ovules) To introduce haploid technologies into the breeding process it is necessary to evaluate the obtained plants for ploidy level. The use of direct counting of chromosomes in apical cells may present a certain difficulty in this species due to their large number (2n=40) and their small size. Depending on the level of laboratory equipment, ploidy determination using flow cytometry of cell nuclei and counting the number of chloroplasts in stomatal guard cells in the epidermis of the abaxial side of the leaf may be more convenient methods. The prospects for the use of molecular markers for assessment for homozygosity in DH technologies used, including C. pepo, are discussed in the review.

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Section: история открытия гаплоидов и первые гаплоиды у культур семейства Cucurbitaceaeunclassified

Section: современное состояние исследований по получению удвоенных гаплоидов C Pepo Lunclassified

Obtaining doubled haploids of Cucurbita pepo L.

2021

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DH-plant production in culture of unpollinated ovules of cucumber (Cucumis sativus L.)

Домблидес

Шмыкова²,

Белов

et al. 2019

Ovoŝi Rossii

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Relevance. The development of F1 hybrids distinguishing it from cultivars by high productivity, plant uniformity in ripening date, fruit sizes and quality is the promising trend in breeding program in cucumber (Cucumis sativus L.).The aim of the study was to optimize the gynogenesis induction condition in culture of unpollinated ovules in vitro in order to broad the generation of new breeding forms and to accelerate homozygous line production.Materials and methods. Eight promising cucumber accessions from Laboratory of Cucurbit Breeding and Seed Production (FSBSI FSVC) were taken for the study. The protocol developed in Laboratory of Biotechnology (FSBSI FSVC) for production of doubled haploid in Cucurbitaceae family was used in the experiment. The medium IMC with 30 g/L sucrose and 7g/L agar supplemented with 200 mg/L ampicillin and 0.2 mg/L thidiazuron (TDZ) was applied to induce gynogenic development.Results. The half-open bud or flower was shown to be the most suitable to be taken as an explant for cultivation. Highest number of embryo-like structures in all accessions developed from ovaries 2.1-2.6 cm long. Exposure to sterilization solution of sodium hypochlorite for 15 min made ovary wall softer and ovules can be then easily extracted without traumatizing. The traumatized ovule resulted in inhibited gynogenic development. Embryoids and calli had developed in all studied cucumber accessions, but well-formed plants were only obtained in six accessions. In total 26 plants were produced. The maximum gynogenesis induction equal to 63.1% was achieved in accession 1810. Maximum number of plant produced was twelve in accession 1763, but the greatest plant outcome 7.7% of the ovules with induced gynogenesis was observed in accession 1807.

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Results and prospects for the development of scientific research in FSBSI FSVC

et al. 2021

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The article reflects the issues of the current state of scientific research at the FSBSI FSVC, shows the directions and prospects for the development of selection and seed production of vegetable crops. The main mission of the selection and seed-growing work of the FSVC is the creation and reproduction of varieties and hybrids of vegetable and melon crops of a new generation, characterized by resistance to abiotic and biotic stressors, high productivity and product quality. One of the main ways to increase the efficiency of the breeding process is the use of modern methods of biotechnology and molecular genetics, which make it possible to obtain new genotypes and reduce the time of breeding. Success has been achieved in the creation of homozygous lines of cabbage, squash, cucumber, carrots. For the first time, it was possible to complete a full cycle of obtaining doubled radish haploids in microspore culture in vitro. The institution pays special attention to immunological research on the resistance of crops and new varieties to diseases, as well as protecting plants from pests and diseases. Phytosanitary monitoring, assessment and selection of resistant genotypes are carried every years. One of the important theoretical and practical problems solved at the institution is the development of environmentally friendly products based on preparations obtained from plants with a high content of biologically active compounds with adaptogenic properties. In the center, a new direction of research in traditional breeding has received significant development - the creation of varieties of vegetable plants with an increased content of biologically active substances and antioxidants, on the basis of which functional products are created. The persistence of modern varieties and hybrids of table carrots, depending on the biochemical composition, has been studied. Methods for enriching vegetable plants with micronutrients are being developed. A large amount of scientific research is being carried out on the development of technologies for the cultivation of new varieties and hybrids, as well as technologies for their seed production. At the branches located in different soil and climatic conditions, hospitals were established to study the interaction of factors controlling soil fertility and the productivity of vegetable crops in agrocenoses (crop rotation, soil treatment systems, fertilizers and plant protection). In general, at the FSBSI FSVC, all research is aimed at developing and improving methods in breeding and seed production of vegetable crops, creating new breeding achievements and developing varietal technologies for their cultivation.

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Production of Doubled Haploids in cucumber

Cited by 5 publications

References 55 publications

Obtaining doubled haploids of Cucurbita pepo L.

Obtaining doubled haploids of Cucurbita pepo L.

DH-plant production in culture of unpollinated ovules of cucumber (Cucumis sativus L.)

Results and prospects for the development of scientific research in FSBSI FSVC

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