A Stable Agrobacterium rhizogenes-Mediated Transformation of Cotton (Gossypium hirsutum L.) and Plant Regeneration From Transformed Hairy Root via Embryogenesis.
A Stable Agrobacterium rhizogenes-Mediated Transformation of Cotton (Gossypium hirsutum L.) and Plant Regeneration From Transformed Hairy Root via Embryogenesis.
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DOI：
10.3389/fpls.2020.604255
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发表时间：
2020
影响因子：
5.6
通讯作者：
Liu CL
中科院分区：
文献类型：
--
作者：
Cui ML;Liu C;Piao CL;Liu CL
遗传转化是研究作物基因功能、次生代谢途径和分子育种的有力工具。棉花(Gossypium hirsutom L.)是世界上最重要的经济作物之一。目前的棉花转化方法至少需要七个人来培养，而且是劳动密集型的，仅限于一些品种。在本研究中，我们首次实现了棉花毛状根转化植株的胚性再生。用发根农杆菌AR1193菌株接种商品品种中棉-24的子叶外植体，在T-DNA区域含有含有NPT II(新霉素磷酸转移酶)基因和GFP(绿色荧光蛋白)基因的双元载体pBI-35S：：GFP。82.6%的外植体产生了不定根，其中53%的外植体转化后表达了GFP。82%的转化毛状根产生了胚性愈伤组织，其中12%在培养7个月后再生为稳定的转化植株。通过聚合酶链式反应和Southern印迹分析证实GFP已整合到转基因棉花基因组中，并通过半定量RT-PCR检测到GFP在棉花基因组中的稳定表达。由此得到的转化植株是表型的，因此避免了Ri综合征。本文报道了一种稳定、重复性好的发根农杆菌介导的棉花遗传转化方法，为棉花基因功能分析提供了一个有用的、可靠的平台。
Genetic transformation is a powerful tool to study gene function, secondary metabolism pathways, and molecular breeding in crops. Cotton (Gossypium hirsutum L.) is one of the most important economic crops in the world. Current cotton transformation methods take at least seven to culture and are labor-intensive and limited to some cultivars. In this study, we first time achieved plantlet regeneration of cotton via embryogenesis from transformed hairy roots. We inoculated the cotyledon explants of a commercial cultivar Zhongmian-24 with Agrobacterium rhizogenes strain AR1193, harboring a binary vector pBI-35S::GFP that contained the NPT II (neomycin phosphotransferase) gene and the GFP (green fluorescent protein) gene as a fluorescent marker in the T-DNA region. 82.6% explants produced adventitious roots, of which 53% showed GFP expression after transformation. 82% of transformed hairy roots produced embryonic calli, 12% of which regenerated into stable transformed cotton plants after 7 months of culture. The integration of GFP in the transformed cotton genomes were confirmed by PCR (Polymerase chain reaction) and Southern blot analysis as well as the stable expression of GFP were also detected by semi-quantitative RT-PCR analysis. The resultant transformed plantlets were phenotypically, thus avoiding Ri syndrome. Here we report a stable and reproducible method for A. rhizogenes-mediated transformation of cotton using cotyledon as explants, which provides a useful and reliable platform for gene function analysis of cotton.
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影响因子：
5.1
作者：
Aggarwal PR;Nag P;Choudhary P;Chakraborty N;Chakraborty S
通讯作者：
Chakraborty S
DOI：
10.1073/pnas.93.23.12768
发表时间：
1996-11-12
影响因子：
11.1
作者：
John, ME;Keller, G
通讯作者：
Keller, G
影响因子：
3
作者：
Frankfater, Cheryl R.;Dowd, Michael K.;Triplett, Barbara A.
通讯作者：
Triplett, Barbara A.
影响因子：
5.2
作者：
MANO, Y;OHKAWA, H;YAMADA, Y
通讯作者：
YAMADA, Y
影响因子：
3.7
作者：
GAMBORG, OL;MILLER, RA;OJIMA, K
通讯作者：
OJIMA, K