An improved protocol for carrot haploid and doubled haploid plant production using induced parthenogenesis and ovule excision in vitro

Kiełkowska, Agnieszka; Adamus, Adela; Barański, Rafał

doi:10.1007/s11627-014-9597-1

Cited by 19 publications

(12 citation statements)

References 36 publications

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“…Distant pollination of species which belong to different species or genera: for example, haploid embryos of Cichorium intybus L. were obtained a few days after pollination with Cicerbita alpina Walbr. (Dore et al 1996) and also in carrot after crossing with parsley with the simultaneous application of 2,4-dichlorophenoxyacetic acid (2,4-D) (Kiełkowska et al 2014). 3.…”

Section: Introductionmentioning

confidence: 99%

Development of haploid embryos and plants of Lactuca sativa induced by distant pollination with Helianthus annuus and H. tuberosus

Piosik¹,

Zenkteler²,

Zenkteler³

2015

Euphytica

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Haploidisation is a biotechnological method used to obtain plants with improved traits that are of use to humans. Lettuce (Lactuca sativa L.), a well-known and popular leafy vegetable, is consumed worldwide. Its haploid form would provide a good basis for producing a pure line of plants (doubled haploids) allowing new varieties to be regenerated. The main aim of this work was to develop an effective haploidisation method for this economically important species. In order to stimulate the development of haploid embryos of lettuce based on our previous experience, we conducted in vivo distant pollination with fresh pollen grains of Helianthus annuus L. or H. tuberosus L. Because the haploid proembryos obtained after pollination did not develop further (despite the presence of cellular endosperm), we implemented the technique of in vitro culture of an isolated embryo sacs (surrounded by endothelium) with parthenogenetic embryos on various, modified Murashige and Skoog media. During the in vitro culture, we observed the formation of callus tissue and, after subsequent cultures of calluses, 23 haploid L. sativa plants were regenerated. The haploid status of the regenerated plantlets was confirmed by estimation of the genome size by flow cytometry, chromosome counting in root tips, stomata cell size and by disturbances in pollen formation resulting from abnormal microsporogenesis. This paper contains the complete protocol for obtaining haploid plants of L. sativa.

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Section: Introductionmentioning

confidence: 99%

Development of haploid embryos and plants of Lactuca sativa induced by distant pollination with Helianthus annuus and H. tuberosus

Piosik¹,

Zenkteler²,

Zenkteler³

2015

Euphytica

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“…Nyman ex A.W. Hill] pollen (Kiełkowska et al., 2014). When cultivating unpollinated pumpkin ( Cucurbita mixta Pang.)…”

Section: Discussionmentioning

confidence: 99%

In vitro embryo formation and plantlet regeneration from unpollinated ovaries of Lilium L

et al. 2022

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The in vitro culture of unpollinated ovaries is one of the techniques used to induce gynogenesis. To explore the mechanism of lily gynogenesis and obtain haploids to shorten the breeding cycle, the in vitro effects of flower bud sampling time, exogenous plant growth regulators (PGRs), and different concentrations of picloram on embryo formation in unpollinated ovaries of Lilium L. 'Sorbonne' (oriental hybrids) were studied. The results indicated that flower buds ≥8 cm in length were most suitable. The PGRs picloram and 2,4-dichlorophenoxyacetic acid (2,4-D) with 6benzylaminopurine (6-BA) or kinetin were useful in inducing ovule expansion in vitro. However, most ovules did not survive on the media containing 2,4-D, whereas those grown on media containing picloram developed normally. In particular, a lower concentration of picloram (0.5 mg L −1 ) was found to be beneficial to the development of ovules into embryogenic shoots, whereas higher concentrations were more favorable to the development of shoots from the ovary wall. In total, 87 regenerated plants were obtained from unpollinated ovaries of Sorbonne. The chromosome ploidy of 20 of these was determined by counting root tip chromosomes, and eight of the regenerated plants were found to be haploids.

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“…In some plants, such as barley and rice, the induction of green plants is much higher during gynogenesis than in androgenesis. The frequency of the accidental appearance of plants with a single set of chromosomes is insignificant, up to 0.01 %, that is, it is a rare event and has limited practical value (Bohanec, 2009;Kiełkowska et al, 2014).…”

Section: Haploidymentioning

confidence: 99%

Haploid biotechnology as a tool for creating a selection material for sugar beets

Колесникова¹,

Donskikh²,

Berdnikov³

2022

Vestn. VOGiS

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Since the discovery of the phenomenon of haploidy, biotechnology has become an integral part in the successful creation of new varieties and hybrids of various plant species. In particular, these technologies are actively used in agriculture, which is concerned with increasing the volume and improving the quality of products. The integration of haploid production techniques together with other available biotechnological tools such as marker selection (MAS), induced mutagenesis and genetic engineering technologies can significantly accelerate crop breeding. This article shows the main stages in the development of biotechnology since 1921. Now they are successfully used to create doubled haploids to accelerate the selection process of various plants and, in particular, sugar beet, which is the most important sugar crop in regions with a temperate climate. There are several methods for obtaining forms with a single set of chromosomes. For sugar beets, the use of gynogenesis turned out to be expedient, since in this case the other methods turned out to be ineffective in the mass production of haploids. The article considers the stages of obtaining the H and DH lines of Beta vulgaris L., as well as the main stages of biotechnological production of homozygous breeding material of this culture. These stages include selecting parental forms – donor explants, sterilizing buds and introducing non-pollinated ovules in vitro, obtaining haploids, doubling their chromosome set, creating doubled haploids, determining ploidy at different stages, relocating the obtained plants to greenhouses and growing stecklings. A number of advantages that the technology of creating doubled haploids in vitro has in comparison with traditional methods of selection are described. It has been shown that the use of these approaches is relevant when obtaining new highly productive hybrids and varieties of agricultural plants; however, the methods for the production of homozygous forms in sugar beet still require additional research aimed at increasing the efficiency and reproducibility of each stage of the process.

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An improved protocol for carrot haploid and doubled haploid plant production using induced parthenogenesis and ovule excision in vitro

Cited by 19 publications

References 36 publications

Development of haploid embryos and plants of Lactuca sativa induced by distant pollination with Helianthus annuus and H. tuberosus

Development of haploid embryos and plants of Lactuca sativa induced by distant pollination with Helianthus annuus and H. tuberosus

In vitro embryo formation and plantlet regeneration from unpollinated ovaries of Lilium L

Haploid biotechnology as a tool for creating a selection material for sugar beets

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