Pistil drip following pollination: a simple in planta Agrobacterium-mediated transformation in cotton

被引：30

作者：

Chen TianZi ^{[1
]}

Wu ShenJie ^{[1
,2
]}

Zhao Jun ^{[1
]}

Guo WangZhen ^{[1
]}

Zhang TianZhen ^{[1
]}

机构：

[1] Nanjing Agr Univ, Natl Key Lab Crop Genet & Germplasm Enhancement, Nanjing 210095, Peoples R China

[2] Shanxi Acad Agr Sci, Cotton Res Inst, Yuncheng 030006, Peoples R China

来源：

BIOTECHNOLOGY LETTERS | 2010年 / 32卷 / 04期

基金：

国家高技术研究发展计划(863计划);

关键词：

Agrobacterium; Cotton; Pistil; Transformation; SOMATIC EMBRYOGENESIS; ARABIDOPSIS-THALIANA; VACUUM INFILTRATION; REGENERATION; POLLEN; TARGET; TUMEFACIENS; EMBRYO; DNA;

D O I：

10.1007/s10529-009-0179-y

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Transgenic cotton plants were developed by pistil drip inoculation in a solution containing Agrobacterium carrying a gene for resistance to the herbicide Basta (bar), 10% (w/v) sucrose, 0.05% (v/v) Silwet L-77 and 40 mg acetosyringone l(-1). Pistil drip during 17:00-19:00 on the first day of flowering resulted in 0.07-0.17% Basta-resistant plants/number of viable seeds generated, and stigma excision prior to pistil drip during this time period gave rise to a transformation efficiency of 0.46-0.93%, in contrast with 0.04-0.06% generated from pistil drip during 9:00-11:00 on the second day of flowering. PCR and Southern blot analysis confirmed the integration of the bar gene into the cotton genome, and a T1 and T2 generation herbicide resistance test consistently revealed expression and stable heritability of the bar gene in the two generations.

引用

页码：547 / 555

页数：9

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