花粉管滴液：棉花中一种简单的活体农杆菌介导转化方法。
Pistil drip following pollination: a simple in planta Agrobacterium-mediated transformation in cotton.
机构信息
National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China.
出版信息
Biotechnol Lett. 2010 Apr;32(4):547-55. doi: 10.1007/s10529-009-0179-y. Epub 2009 Dec 2.
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.
利用含有携带抗除草剂 Basta（bar）基因的农杆菌的溶液，通过柱头点滴接种的方法，培育出了转基因棉花植株，溶液中还含有 10%（w/v）蔗糖、0.05%（v/v）Silwet L-77 和 40mg 乙酰丁香酮 l(-1)。在开花第一天的 17:00-19:00 期间进行柱头点滴接种，每生成的有效种子数中就有 0.07-0.17%的植株具有抗 Basta 特性，而在此期间在柱头点滴接种之前切除柱头，则转化效率为 0.46-0.93%，相比之下，在开花第二天的 9:00-11:00 期间进行柱头点滴接种，转化效率为 0.04-0.06%。PCR 和 Southern blot 分析证实了 bar 基因已整合到棉花基因组中，T1 和 T2 代抗除草剂测试均表明，bar 基因在两代中的表达和稳定遗传。
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