摘要
利用包含双元载体pBI121的农杆菌LBA4404转化西瓜子叶外植体,获得转基因西瓜植株。结果表明:转化受体材料西瓜品种早花、克伦生、黑美人母本F23的分化效率没有明显差别;外植体浸染菌液10 min的处理较为合适;125 mg/L卡那霉素和300 mg/L羧苄青霉素适合于外植体筛选培养。通过PCR检测获得了3株转基因植株,转化效率为1.5%,其中只有2株整合了gus报告基因。Southern杂交和gus染色检测证明2株的基因组中整合了外源基因,报告基因在植株T0-1-3后代呈现3∶1分离。
Segments of cotyledonary explants were infected by Agrobacterium tumefaciens harboring a binary vector pBI121 and transgenic plants were obtained. The results showed that the regeneration frequency was not obviously different among three cultivars : Zaohua, Crimson sweet and female parent of Heimeiren F23. Inoculation with Agrobacterium for 10 min is optimal for transformation. The medium with 125 mg/L kanamycin and 300 mg/L carbenicillin was suitable for screening the explants after transformation. Three npt Ⅱ positive plants were obtained, but only two plants showed gus positive. The transformation frequency was 1.5 %. The results of Southern blot and GUS assay indicated that gus gene had integrated into the genome of watermelon. T0-1-3 plant was stable, inherited with a segregation of gus gene in a 3 :1 segregation ratio in plant progeny.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2007年第5期684-688,共5页
Journal of Huazhong Agricultural University
基金
武汉市科技攻关项目(2003203034)
湖北省科技攻关项目(2001AA209B05)资助
关键词
西瓜
转化
农杆菌
GUS
转基因植株
watermelon
transformation
Agrobacterium tumefaciens
gus
transgenic plant