摘要
小鼠金属硫蛋白－ＩｃＤＮＡ通过根癌农杆菌介导转入枸杞，枸杞体内的表达产物对重金属镉有一定的抗性。用带有嵌合ｍＭＴ－ＩｃＤＮＡ及ＣａＭＶ３５ｓ启动子的含非致瘤性Ｔｉ质粒载体的根癌农杆菌ＬＢＡ４４０４（ＣａＭＶ３５ｓ∶ｍＭＴ－ＩｃＤＮＡ∶ｎｏｓ）感染枸杞幼茎外植体，在含有卡那霉素１００μｇ／ｍＬ和５０μｍｏｌ／ＬＣｄ２＋的选择培养基上筛选转化的愈伤组织，并在含卡那霉素５０μｇ／ｍＬ和１００μｍｏｌ／ＬＣｄ２＋的分化培养基上得到再生小芽。继续转入选择的生根培养基进行生根筛选培养得到了完整的再生植株。对Ｃｄ２＋抗性实验、报告基因ＮＰＴ－Ⅱ、ｎｏｐａｌｉｎｅ活性检测以及Ｓｏｕｔｈｅｒｎ杂交分析结果均表明：ｍＭＴ－ＩｃＤＮＡ已导入枸杞体内并得到表达。转ｍＭＴ－ＩｃＤＮＡ枸杞对Ｃｄ２＋具有一定的抗性。
?Agrobacterium tumefaciens and transgenic Lycium barbarum L.has conferred resistance to cadmium.Young stem segments from Lycium barbarum L.can be infected by A.tumefaciens strain LBA4404 harboring a nononcogenic Ti plasmid derived vector which containing the chimaeric gene mMTI cDNA and the cauliflower mosaic virus 35s (CaMV35s) promoter as well as nopaline synthase (Non).The callus originated from selective medium (100 μmol/L Cd2++50 μg/mL Km) can differentiate into buds.The whole plant was obtained on the selective medium of growth root.The resistance test to Cd2+ and reporter NTPⅡ and nopaline activity assay as well as southern hybridization indicated that the foreign gene mMTⅠ cDNA had been integrated into the genome of Lycium barbarum L.and could be expressed in plants.The transgenic Lycium barbarum L.is resistant to Cd2+.
出处
《西北植物学报》
CAS
CSCD
1998年第2期229-236,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家"863"高新技术资助
关键词
金属硫蛋白
枸杞
根癌农杆菌
抗性
镉
metallothionein,Lycium barbarum L.,Agrobacterium tumefaciens,Cd2+ resistance
