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在多种物种中建立稳定、有效且通用的基因转化技术。

Establishment of a stable, effective and universal genetic transformation technique in the diverse species of .

作者信息

Sheng Xiaoguang, Yu Huifang, Wang Jiansheng, Shen Yusen, Gu Honghui

机构信息

Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.

出版信息

Front Plant Sci. 2022 Oct 12;13:1021669. doi: 10.3389/fpls.2022.1021669. eCollection 2022.

Abstract

is an economically important species, including seven cultivated variants. -mediated transformation of crops, mainly hypocotyl and cotyledon, has been achieved in the past. However, previously established transformation methods showed low efficiency, severe genotype limitation and a prolonged period for transformants acquisition, greatly restricting its application in functional genomic studies and crop improvement. In this study, we have compared the shoot regeneration and genetic transformation efficiency of hypocotyl, cotyledon petiole and curd peduncle explants from twelve genotypes of cauliflower and broccoli. Finally, an -mediated transformation method using curd peduncle as explant was established, which is rapid, efficient, and amenable to high-throughput transformation and genome editing. The average genetic transformation efficiency of this method is stable up to 11.87% and was successfully implemented in twelve different genotypes of cauliflower and broccoli and other crops with low genotype dependence. Peduncle explants were found to contain abundant cambial cells with a strong cell division and shoot regeneration ability, which might be why this method achieved stable and high genetic transformation efficiency with almost no genotype dependence.

摘要

是一种具有重要经济价值的物种,包括七个栽培变种。过去已实现了农杆菌介导的作物转化,主要是下胚轴和子叶。然而,先前建立的转化方法效率低、基因型限制严重且获得转化体的时间长,极大地限制了其在功能基因组学研究和作物改良中的应用。在本研究中,我们比较了来自12个基因型的花椰菜和西兰花的下胚轴、子叶叶柄和花球梗外植体的芽再生和遗传转化效率。最终,建立了一种以花球梗为外植体的农杆菌介导的转化方法,该方法快速、高效,适用于高通量转化和基因组编辑。该方法的平均遗传转化效率稳定高达11.87%,并成功应用于12种不同基因型的花椰菜和西兰花以及其他基因型依赖性低的作物。发现花梗外植体含有丰富的形成层细胞,具有很强的细胞分裂和芽再生能力,这可能是该方法几乎不依赖基因型就能实现稳定且高遗传转化效率的原因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87af/9597678/eeed4dfaf3cc/fpls-13-1021669-g001.jpg

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