【作者】 侯纯纯； 尚爱芹； 王省芬； 李志坤； 吴立强； 张桂寅； 马峙英；

【Author】 HOU Chun-chun,SHANG Ai-qin,WANG Xing-fen,LI Zhi-kun,WU Li-qiang,ZHANG Gui-yin,MA Zhi-ying(North China Key Laboratory for Crop Germplasm Resources of Education Ministry/Key Laboratory for Crop Germplasm Resources of Hebei/Agricultural University of Hebei,Baoding 071001,China)

【机构】 华北作物种质资源教育部重点实验室/河北省作物种质资源重点实验室/河北农业大学；

【摘要】 以陆地棉邯208为材料,进行茎尖分生组织再生培养,采用农杆菌介导转化外源基因,研究固化剂、植物生长调节剂和抗生素筛选对茎尖生根和成苗的作用,以及菌液浓度、侵染时间、共培养温度、共培养时间对转化率的影响。结果表明,以phytagel为固化剂,0.1mg.L-1 6-BA+0.1mg.L-1IAA最适于茎尖不定芽的诱导,1/2MSB+0.1mg.L-1 IAA+0.1mg.L-1 NAA诱导生根效果最佳。在75mg.L-1 Kan筛选下,以OD600为0.9的农杆菌菌液侵染25min,22℃共培养48h,外源基因的转化率最高。采用该转化体系将克隆的编码谷胱甘肽S-转移酶的抗黄萎病相关基因GhGST转化邯208,经PCR检测获得了转基因阳性植株。更多还原

【Abstract】 In the study,the shoot apex of Han208(Gossypium hirsutum) was selected for regeneration and transformation using Agrobacterium-mediated methods.The effects of gelling agents,plant growth regulators and antibiotics were investigated on regeneration and root-inducing rate.And the effects of Agrobacterium concentration,infection time,the time and the temperature of co-culture on the transformation rate of shoot apex were studied by orthogonal design.The results showed that adventitious buds of shoot apex were efficiently induced by 0.1 mg·L-1 6-BA +0.1 mg·L-1 IAA,and roots of shoot apex were efficiently induced by 1/2MSB +0.1 mg·L-1 IAA+0.1 mg·L-1 NAA with phytagel as gelling agents.The treatment on shoot apex by 0.9 of OD600 value of agrobacterium suspension,infection time for 25 min,and co-culturing for 48 h in 22 ℃ could obtain the highest transformation rate at 75 mg·L-1 Kan.Using the genetic transformation system,GhGST gene encoding Glutathione S-transferase Subunit was transformed into cotton variety Han208,and genetically modified plants were obtained by PCR detection.更多还原