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用农杆菌将Bt基因导入水稻愈伤组织的研究

Agrobacterium tumefaciens Bt Gene Transfer in Embryogenic Calli of Rice

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【作者】 贝丽霞王守德史芝文徐仲郭三堆

【Author】 BEI Li xia 1, WANG Shou de 2, SHI Zhi wen 2, XU Zhong 2, GUO San dui 3 (1.College of Plant Science and Technology,Heilongjiang August First Land Reclamation University,Mishan, Heilongjing 158308,China; 2. Department of Biological Engineeri

【机构】 黑龙江八一农垦大学植物科技学院!黑龙江密山158308东北农业大学生物工程系!哈尔滨150030中国农业科学院生物技术研究中心!北京100081

【摘要】 从水稻成熟胚诱导出的愈伤组织经继代培养后可以产生大量的胚性愈伤组织。通过采用农杆菌共培养法转化上述胚性愈伤组织 ,在附加羧苄青霉素Cb(5 0 0 μg/mL)和卡那霉素Km(5 0 μg/mL)的选择培养基上进行抗性愈伤组织的筛选 ,在附加羧苄青霉素Cb(5 0 0 μg/mL)和卡那霉素Km(2 5 μg/mL)的分化培养基上诱导抗性芽。GUS酶活性检测和PCR检测结果均为阳性 ,证明外源目的基因 (Bt)已在寒地水稻抗性愈伤组织中整合并表达。抗性再生植株的GUS酶活性及PCR检测正在进行中

【Abstract】 Many embryogenic calli were produced by subculture of calli induced from mature embryo of rice.The embryogenic calli were transformed after incubating the calli with Agrobacterium tumefaciens , and the resistant calli were selected on selectivemedia containing Cb (500?μg/mL)and Km ( 50?μg/mL ). The resistant shoots were induced on differentiation media containing Cb ( 500?μg/mL ) and Km (25?μg/mL).Finally,the regenerative plantlets were gained. Gus enzyme activity test and PCR test showed that the foreign gene(Bt)had been integrated and expressed in Oryza sativa L.GUS enzyme activity test and PCR test of resistant plantlets is in progress.

【基金】 黑龙江省科委资助项目
  • 【文献出处】 吉林农业大学学报 ,Journal of Jilin Agricultural University , 编辑部邮箱 ,2000年04期
  • 【分类号】Q943
  • 【被引频次】8
  • 【下载频次】185
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