甘蓝型油菜雄性不育基因转化体系的探索  

Exploration of the male sterile genetic transformation of Brassica napus cultivars low in erucic and glucosinolates

作　　者：万萌[1] 王敬乔[1] 寸守铣[1] 邱仕芳[1] 李根泽[1] 曾黎琼[1] 和江明[1] 

机构地区：[1]云南省农科院生物技术研究所

出　　处：《西南农业学报》1999年第1期45-50,共6页Southwest China Journal of Agricultural Sciences

基　　金：云南省基金资助项目

摘　　要：用携带ＴＡ２９—Ｂａｒｎａｓｅ雄性不育基因的农杆菌转化“双低”花培油菜，获得了大量转基因植株。用油菜的带柄子叶及下胚轴等外植体与携带载体质粒的农杆菌共培养后，将其转移到诱导愈伤组织及芽的培养基上培养，并同时进行卡那霉素的抗性筛选，２～４周后，在子叶柄基部及愈伤组织上出现小丛芽，将其切下转入Ｂ５加卡那霉素的培养基中进一步筛选并生根，再将生根的正常绿色植株移栽于花盆中。在五叶期，用５０ｍｇ／ｍ１的卡那霉素注射于真叶的叶脉中，进一步筛选，可得到完全正常的绿苗，说明卡那霉素抗性标记基因（ＮＰＴⅡ）已转入油菜植株，进一步的植株鉴定及分子生物学检测另文详述。Brassica napus cultivar H090 and H97-21,low in erucic acid and glucosinolates produced from pollen culture,were transformed by the strains of Agrobacterium tumefaciens with the male sterile gene (TA29_Barnase).A large number of transgenic plants were obtained.The petiolulate cotyledons and hypocotyl of cultivars H090 and H97-21 were transplanted on the medium with A.tumefaciens with the plasmid vector for inducing of callus and shoots,at the same time they were selected by kanamycin resistance.Shoots formed from the callus and the base of the transformed cotyledon petiole within 2 to 4 weeks,then were excised off and transferred on B 5 medium with 10mg/l kanamycin to further select the kanamycin resistant shoots and stimulate rooting.Then the rooted normal green plants were transplanted into flower pots. On the stage of five leaves,50mg/l kanamycin were injected into leaf vein to further select the kanamycin resistant plants.Normal green plants were obtained.It shows that kanamycin resistant marker gene(NPⅠ Ⅱ) were transferred to the plants.Further investigation about plants and molecular tests will be reported in another papers.

关 键 词：甘蓝型油菜 农杆菌 雄性不育 基因转化 油菜 

分 类 号：S565.403.5[农业科学—作物学] S188