Obtaining doubled haploids of Cucurbita pepo L.

Домблидес, Е. А.; Ermolaev, A. S.; Белов, С. Н.

doi:10.18619/2072-9146-2021-4-11-26

Cited by 3 publications

(2 citation statements)

References 103 publications

(122 reference statements)

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“…In Cucurbitaceae species, for instance, in a summer squash (Cucurbita pepo L.) ovary/ovule culture, most regenerants doubled their genome. Although, occasional haploid, mixoploid, or polyploid regenerants were observed as well [47]. In styrian oil pumpkin (Cucurbita pepo ssp.…”

Section: Spontaneous Haploid Genome Doubling In Vegetable Cropsmentioning

confidence: 99%

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Spontaneous and Chemically Induced Genome Doubling and Polyploidization in Vegetable Crops

Fomicheva,

Kulakov,

Alyokhina

et al. 2024

Horticulturae

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Plant ploidy manipulation is often required for breeding purposes. However, there is no comprehensive review covering genome doubling in vegetable crops despite the abundance of data for a large number of vegetable species. Similar to other species, genome doubling is required in vegetable crops to obtain doubled haploids (DHs). It is also utilized for the production of polyploids to overcome interspecific hybrid sterility and improve agricultural traits. Spontaneous haploid genome duplication (SHGD) occurs in many Apiaceae, Brassicaceae, Cucurbitaceae, and Solanaceae crops, allowing for the laborious treatment with antimitotic agents to be bypassed. SHGD mechanisms are not fully understood, but existing data suggest that SHGD can occur via nuclear fusion, endoreduplication, or other mechanisms during microspore or ovule early embryogenic development. Other studies show that SHGD can occur at later developmental stages during extended plant growth in vitro or ex vitro, possibly due to the presence of phytohormones in the medium and/or diploid cell competitive advantage. For unresponsive accessions and species with rare SHGD, such as onion (Allium cepa L.) and beet cultivars (Beta vulgaris subsp. vulgaris L.), antimitotic agent treatment has to be applied. Antimitotic agent application efficiency depends on the treatment conditions, especially the agent concentration and exposure time. Also, plant developmental stage is critical for agent accessibility and plant survival. The existing methods can be used to further improve genome doubling methodology for major vegetable crops and other species.

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Section: Spontaneous Haploid Genome Doubling In Vegetable Cropsmentioning

confidence: 99%

“…Besides haploid and doubled haploid plants, triploid and polyploid regenerants can often be observed in many crops, including Brassica species, C. annuum L., D. carota L. [4], and C. pepo L. [47,48]. Possibly, triploid plants can form due to the fusion of diploid and haploid nuclei.…”

Section: Shgd Mechanismsmentioning

confidence: 99%

Spontaneous and Chemically Induced Genome Doubling and Polyploidization in Vegetable Crops

Fomicheva,

Kulakov,

Alyokhina

et al. 2024

Horticulturae

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Modern trends in the development of selection of vegetable and melon crops

et al. 2022

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The article reflects the issues of the current state of scientific research in the FSBSI FSVC, directions and prospects for the development of selection and seed production of vegetable crops are shown. Varieties have been created that realize the bioclimatic potential of the cultivation zone, meet market requirements, and are capable of producing high yields under the influence of biotic and abiotic stressors. Selection is carried out in a combination of classical methods with innovative technologies.Biotechnologists have improved the basic protocol for in vitro microspore culture for the Brassicaceae, stages of technology for obtaining doubled haploids in microspore culture in vitro for the Apiaceae are being developed. For the first time, an effective protocol for obtaining double haploids of European radish has been developed. Doubled haploids of vegetable crops of the Cucurbitaceae, Amarantháceae, Amaryllidáceae were obtained in vitro in the culture of unpollinated ovules. The Center conducts research on the increased content of biologically active substances and antioxidants. An important place is occupied by immunological studies on the resistance of vegetable crops to harmful diseases. The institution develops systems of protective measures against pests, diseases and weeds for vegetable crops based on the use of new environmentally friendly preparations. On the basis of fundamental and priority applied research of previous years, the creation of 21 varieties and hybrids of cabbage, table beet, tomato, sweet pepper, onion, chives, leek, cucumber, melon, lettuce, chervil, potato, aster, lychnis has been completed. Along with the creation of new breeding achievements, technologies for their cultivation are being improved through the use of new agricultural practices, microfertilizers, biological products and humates on vegetable crops in various soil and climatic zones of Russia. New agricultural methods are being developed in the cultivation of mushrooms. In the field of seed production, methods are being developed to increase the germination of seeds, seed production, elements of the technology for the production of mother liquors and seeds of various vegetable and melon crops; zonal seed production technologies that increase their yield. In 2021, a team of scientists received an award from the Government of the Russian Federation for scientific work "Development and implementation of innovative technologies for growing vegetables and potatoes to provide the population with environmentally friendly food".

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Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro

et al. 2022

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An important stage in doubled haploid (DH) production is to evaluate and to differentiate the ploidy level of regenerant plants at least two–three times during the technology. Therefore, rapid and reliable methods are necessary for particular species taken into the technology. In this study, Cucurbita pepo regenerants obtained through unpollinated ovule culture in vitro were evaluated including three different methods: direct chromosome counting in apical meristems, flow cytometry of the cell nucleus, and estimation of morphological parameters of the abaxial epidermis. Methods were optimized for each of three evaluations, and main criteria were determined for ploidy level differentiation. As a result, four ploidy levels, namely, 2n, 3n, 4n, and 8n, were defined among regenerant plants adapted to ex vitro conditions, while true haploids were only found among plants that remained in the in vitro culture. In total, 32.35%, 26.47%, 33.82%, 4.41%, and 2.94% of regenerant plants of courgette and patisson were diploid, triploid, tetraploid, octaploid, and aneuploid, respectively. According to results of flow cytometry of the cell nucleus, two cytotypes in diploid samples with DNA content of 2C = 1.07 ± 0.03 pg for courgette belonging to subsp. pepo and 2C = 0.95 ± 0.03 pg for patisson samples belonging to subsp. ovifera were revealed. The images of metaphase chromosomes of haploid, triploid, and tetraploid C. pepo specimens obtained using the propion–lacmoid chromosome staining method were presented for the first time. Parameters of abaxial epidermis in diploid samples of courgette and patisson grown in open-field and greenhouse conditions were described and compared. It was shown that the most robust parameter not depending on external factors was the number of chloroplasts in stomatal guard cells, which contained 9.41 to 11.31, 14.84 to 16.3, and up to 17.58 chloroplasts in diploid, triploid, and tetraploid samples, respectively. The application of several methods for estimation enables avoiding the misidentification of ploidy levels in adapted regenerant plants produced with the use of DH technology.

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Obtaining doubled haploids of Cucurbita pepo L.

Cited by 3 publications

References 103 publications

Spontaneous and Chemically Induced Genome Doubling and Polyploidization in Vegetable Crops

Spontaneous and Chemically Induced Genome Doubling and Polyploidization in Vegetable Crops

Modern trends in the development of selection of vegetable and melon crops

Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro

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