论文部分内容阅读
用含β－１，３－葡聚糖酶和几丁质酶串联基因双价载体质粒ＰＢＬＧＣ的根癌农杆菌侵染甘蓝型油菜（Ｂｒａｓｉｃａｎａｐｕｓ）带柄子叶和下胚轴的方法，初步建立了抗真菌病害油菜高效基因转化育种体系．研究了不同种类和不同浓度的激素对带柄子叶和下胚轴不定芽诱导的影响，试验结果表明经卡那霉素（ｋａｎ）筛选，含６ｍｇ／Ｌ６－ＢＡ的ＭＳ培养基中带柄子叶的出苗率及绿苗分化率最高，分别为１００％和２５％；下胚轴在含１ｍｇ／Ｌ６－ＢＡ和０．１ｍｇ／Ｌ２，４－Ｄ的ＭＳ培养基中出芽率及绿苗分化率最高，分别为１００％和２３％．绿苗在含５０ｍｇ／Ｌｋａｎ的诱根培养基中培养后移栽于花盆，套袋自交，自交种子所得无菌苗的带下胚轴子叶再经含５０ｍｇ／Ｌｋａｎ的ＭＳ培养基筛选培养，结果小苗全部生根成活，初步证明目的基因已转入油菜植株中，经诱导不定芽阶段和生根阶段两次ｋａｎ筛选是有效的
The method of inoculating the cotyledon and hypocotyls of Brasenia canadensis with A. tumefaciens containing β-1,3-glucanase and chitinase tandem gene vector PBLGC was preliminarily established Antifungal disease rape high efficiency gene transformation breeding system The effects of different kinds and concentrations of hormones on the induction of adventitious buds of cotyledons and hypocotyls with stems were studied. The results showed that cotyledons with 6mg / L 6-BA in kanamycin- The highest germination rate and the highest rate of green seedling were 100% and 25% respectively. The germination rate and the rate of green plantlet differentiation in hypocotyls of MS medium containing 1mg / L 6-BA and 0.1mg / L 2,4-D Highest, respectively 100% and 23%. Green shoots were cultured in rooting medium containing 50 mg / L kan, then transplanted in flower pots, bagged self-selfing, subcultured hypocotyl cotyledon cotyledons and screened by MS medium containing 50 mg / L kan Culture, the results of seedling all rooting alive, initially proved that the target gene has been transferred to rape plants, adventitious buds induced by induction and rooting stage two kan screening is effective