用农杆菌将bt基因导入水稻愈伤组织的研究.pdfVIP

DOI :10.13327/j.j jlau.2000.04.010 吉林农业大学学报　2000, 22 (4):36 ～40 Journal of J ilin Agricultural University Bt 1 2 2 2 3 贝丽霞 　王守德 　史芝文 　徐　仲 　郭三堆 　　(1.黑龙江八一农垦大学植物科技学院　黑龙江密山 158308;2.东北农业大学生物工程系　 哈尔滨150030;3.中国农业科学院生物技 研究中心　北京100081) 　: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(500μg/mL)和卡那霉素Km(50μg/m L)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(500μg/mL)和卡那霉素Km(25μg/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。 :水稻;农杆菌;共培养;Bt 基因;遗传转化 :Q78　　　:A　　　:1000-5684 (2000)04-0036-05 Agrobacterium tumefaciens Bt GeneTransfer inEmbryogenic Calli of Rice 1 2 2 2 3 BEI Li-xia , WANG S ou-de , SHI Z i-wen , XU Z ong , GUO San-dui (1.College of Plant Science and Technology , Heilongj iang AugustFirst Land eclamation Uni- versity, Mishan, Heilongj ing 158308, China;2.Dep artment of Biological Engineering , Northest University of Agriculture, Harbin 150030, China;3 Center of Biological Tech- niques, Chinese Academy of Agricultural Sciences, Beij ing 100081, China) Abstract:Many embryogenic calli w ere produced by subculture of calli induced from mature em- bryo of rice.T e embryogenic calli were transformed after incubating t e calli w it Agrobacteri- um tumef aciens, and t e resistant calli w ere selected on selectivemedia containing Cb (500 μg/ mL)and Km (50 μg/mL).T e resistant s oots were induced on differentiation media containing Cb (500μg/mL)and Km (25 μg/mL).Finally, t e regenerative plantlets were gained.Gus en- zyme activity test and PCR test s ow ed t at t e foreign gene (Bt )ad been integrated and ex - pressed in Oryza sativa L.GUS enzyme activit