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一种通过孤雌生殖胚胎发育诱导黄瓜单倍体的高效且易于使用的方案。

An efficient and easy-to-use protocol for induction of haploids in cucumber through parthenogenic embryo development.

作者信息

N Pradeepkumara, Parkash Chander, Bhatia Reeta, Munshi Anilabha Das, Rao Mahesh, Subhasmita Subhashree, Behera Tusar Kanti, Dey Shyam Sundar

机构信息

Division of Vegetable Science, ICAR-Indian Agricultural Research Institute, New Delhi, India.

ICAR- Central Institute of Temperate Horticulture, Regional Station, Dirang, India.

出版信息

Bot Stud. 2024 Oct 8;65(1):30. doi: 10.1186/s40529-024-00436-w.

Abstract

BACKGROUND

Cucumber (Cucumis sativus L.) is a model crop to study cell biology, including the development of haploids and doubled haploids in vegetable crops. In plant breeding, haploid and doubled haploids are valuable tools for developing pure homozygous inbred lines and accelerating genetic progress by reducing the time required for breeding cycles. Besides, the haploids are also valuable in genomic studies. We are reporting the induction of haploids in cucumber involving gynoecious and parthenocarpic genotypes for the first time. This study aimed to assess the efficient induction of haploids through pollination with gamma-irradiated pollen in cucumber. The effect of gamma irradiation dose on pollen viability and germination, fruit setting percentage, seed development, and haploid embryo development in cucumber hybrid genotypes were studied in detail. The goal was to utilize this information to produce haploid plants for genomics and transformation works in this model vegetable crop.

RESULTS

Pollination was done on six cucumber genotypes using varying doses of gamma rays (100, 200, 300, 400, and 500 Gy). Genotypes, doses of irradiation, and embryo developmental stage influenced the successful generation of in-vitro haploid plants. The optimal timeframe for embryo rescue was found to be 25 to 30 days after pollination. Haploid embryos were effectively induced using irradiated pollen at 400 to 500 Gy doses. Parthenogenetic plantlets were analyzed, and their ploidy level was confirmed through stomatal physiology, cytology (mitosis), and flow cytometry methods.

CONCLUSION

Through parthenogenic embryo development, it is possible to induce a large number of haploids in cucumber. This technique's power lies in its ability to streamline the breeding process, enhance genetic gain, and produce superior cultivars that contribute to sustainable agriculture and food security.

摘要

背景

黄瓜(Cucumis sativus L.)是研究细胞生物学的模式作物,包括蔬菜作物中单倍体和双单倍体的发育。在植物育种中,单倍体和双单倍体是培育纯合自交系以及通过缩短育种周期所需时间来加速遗传进展的宝贵工具。此外,单倍体在基因组研究中也很有价值。我们首次报道了在黄瓜中诱导单倍体,涉及雌性系和单性结实基因型。本研究旨在评估通过用γ射线辐照花粉授粉在黄瓜中高效诱导单倍体的情况。详细研究了γ射线辐照剂量对黄瓜杂交基因型花粉活力和萌发、坐果率、种子发育以及单倍体胚胎发育的影响。目标是利用这些信息为这种模式蔬菜作物的基因组学和转化工作培育单倍体植株。

结果

使用不同剂量的γ射线(100、200、300、400和500 Gy)对六种黄瓜基因型进行授粉。基因型、辐照剂量和胚胎发育阶段影响了体外单倍体植株的成功产生。发现胚胎拯救的最佳时间框架是授粉后25至30天。使用400至500 Gy剂量的辐照花粉有效地诱导了单倍体胚胎。对孤雌生殖的幼苗进行了分析,并通过气孔生理学、细胞学(有丝分裂)和流式细胞术方法确认了它们的倍性水平。

结论

通过孤雌生殖胚胎发育,可以在黄瓜中诱导出大量单倍体。这项技术的优势在于它能够简化育种过程、提高遗传增益,并培育出有助于可持续农业和粮食安全的优良品种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8473/11461365/f365bc737bbb/40529_2024_436_Fig1_HTML.jpg

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