甘兰型油菜双低品种抗病毒遗传转化的研究  

ANTIVIRUS GENETIC TRANSFORMATION OF BRASSICA NAPUS CULTIVARS CONTAINING LOW ERUCIC ACID AND GLUCOSINOLATES

作　　者：万萌[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》1995年第2期27-31,共5页Southwest China Journal of Agricultural Sciences

摘　　要：用携带植物抗病毒基因表达载体ｐＢＴＵ的农杆菌转化油菜花培双低品种Ｈ０９０．Ｈ１６６，获得了大量转基因植株。载体质粒ｐＢＴＵ上有卡那霉素抗性标记基因（ＮＰＴⅡ）和ＣａＭ３５Ｓ启动子控制的芜菁花叶病毒ＴｕＭＶ－ｃｐ外壳蛋白基因，用品种Ｈ０９０和Ｈ１６６的带柄子叶与携带载体质粒的农杆菌共培养后，将其转移到含有６－ＢＡ１－１０ｍｇ／ｌ的ＭＳ培养基中，７～１５天后又将其转移到含有６－ＢＡ１－１０ｍｇ／ｌ的ＭＳ加卡那霉素１０ｍｇ的培养基上培养筛选２～３周后，在子叶柄基部出现小丛芽，将其切下转入Ｂ５加卡那霉素１０ｍｇ的培养基中进一步筛选并使转化体生根，将生根的正常绿色植株移栽于花盆中，接种ＴｕＭＶ进行攻毒试验，这些植株未出现发病现象，将未发病的自交种子，播种于含有卡那霉素的Ｂ５培养基上再一次进行抗性鉴定，均未出现白化苗。结果表明：卡那霉素抗性标记基因（ＮＰＴⅡ）已转入油菜植株。H090 and H166 Brassica napus cultivars,which contained erucic acid and glucosinolates were transformed by the strains produced from pollen culture of Agrobacterium tumefaciens with the plant expression vector pBTU.A large number of transgenic plants were obtained. The plasmid vector pBTU carried a marker gene expressing kanamycin resistance and the gene of turnip yellow mosaic virus outer shell protein which was controlled by Cam35s promoter. The petiolate cotyledons of cultivars H090 and H166 were transferred on MS medium with additive of 1-10mg/1 6-BA after cocultivation with A.tumefaciens with the plasmid vector.After 7～15 days, they were transferred on MS medium with 1～10mg/1 6-BA and 10mg/1 kanamycin.Shoots formed from the base of the tranformed cotyledon petiole within 2 to 3 weeks or so.Then the shoots were excised off and transferred on B5 medium with 10mg/1 kanamycin to further select the kanamycin resistant shoots and stimulate rooting.The normally rooting green plants were transplanted into flower pots. No symptoms of TuMV were found in these plants after inoculation of the virus. Finally, the inbred seeds were sowed on B5 medium with kanamycin.The kanamycin resistance of the plants was detected.The results indicated that at last the kanamycin resistant marker gene was introduced into the plants.

关 键 词：花叶病毒 抗病性 遗传转化 甘蓝型油菜 双低品种 

分 类 号：S565.403.4[农业科学—作物学]