用基因枪法将Bt杀虫基因导入水稻愈伤组织的研究

Regenerative rice plants obtained by microprojectile bombardent of embrogenic calli

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。本研究采用基因枪法转化上述胚性愈伤组织 ,在附加羧苄青霉素 (5 0 0 μg· m L-1)和卡那霉素 (5 0 μg·m L-1)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 (5 0 0 μg· m L-1)和卡那霉素 (2 5 μg· m L-1)的分化培养基上诱导抗性芽。GUS酶活性检测和对基因组 DNA作 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

Many embryogenic calli were produced through subculture by calli induced from mature embryo of rice.The above-mentioned calli were transformed by using Microprojectile bombardment,and resistant calli were selected on selective media containing Cb(500 μg·mL -1 )and Km(50 μg·mL -1 ).The resistant shoots were induced on differentiation media containing Cb(500 μg·mL -1 )and Km(25 μg·mL -1 ).Finally,regenerative plantlets were obtained.the result is positive by Gus enzyme activity test and PCR test,Showing that foreign gene(Bt)had been integrated into and expressed in Oryza sativa L.resistand calli was testified. Gus enzyme activity test and PCR test of resistant plantlets are in progress.