根癌农杆菌(Agrobacterium tumefaciens)介导的马铃薯高效遗传转化体系筛选及优化

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马铃薯是重要的块茎类作物,根癌农杆菌(Agrobacterium tumefaciens)介导的遗传转化方法是马铃薯基因工程育种的重要技术,研究其遗传转化体系和转化效率有助于提高分子育种效率。针对利用根癌农杆菌介导的2种外植体遗传转化马铃薯普通栽培品种,通过筛选获得最佳转化体系和转化率。以4种马铃薯普通栽培品种(Favorita、Shepody、Atlantic、甘农薯2号)的茎段和试管薯薄片为受体材料,对根癌农杆菌介导的不同遗传转化体系的分化率和转化率进行分析。结果表明:Favorita、甘农薯2号和Atlantic茎段经S2(MS;6-BA 2.5mg·L~(-1),2,4-D0.6mg·L~(-1),Carb 400mg·L~(-1),Kan 50mg·L~(-1))和M2(MS;6-BA 2.5mg·L~(-1),IAA 0.25 mg·L~(-1),2,4-D0.25mg·L~(~(-1)),Carb 400mg·L~(-1),Kan 50mg·L~(-1))培养后获得的转化率显著高于其他培养基。Favorita茎段最大转化率显著高于试管薯薄片转化率;甘农薯2号和Shepody试管薯薄片转化率显著高于茎段转化率。茎段转化体系的最佳诱导愈伤培养基为S2,最佳分化培养为基为M2。Favorita适合采用茎段转化体系,甘农薯2号和Shepody适合采用试管薯薄片转化体系。不同品种的愈伤组织诱导培养基和分化培养基的差别主要是由基因型差异引起的,在提高遗传转化效率中应该针对品种进行转化体系的筛选。 Potato is an important tuber crop. Agrobacterium tumefaciens-mediated genetic transformation is an important technique in potato genetic engineering. Studying its genetic transformation system and transformation efficiency can help to improve molecular breeding efficiency. Aiming at the common transformation of common potato cultivars using two kinds of explants mediated by Agrobacterium tumefaciens, the optimal transformation system and the conversion rate were obtained through screening. The differentiation rate and transformation rate of Agrobacterium tumefaciens-mediated different genetic transformation systems were studied with stem segments and tube-tuber slices of four common potato cultivars (Favorita, Shepody, Atlantic, Gannongshu No.2) Analyze. The results showed that there were significant differences in the yield of Favorita, Gannongshu No.2 and Atlantic stems by S2 (MS; 6-BA 2.5 mg · L -1, 2,4-D 0.6 mg · L -1, Carb 400 mg · L -1) (-1), Kan (50 mg · L -1) and M2 (MS; 6-BA 2.5 mg · L -1, IAA 0.25 mg · L -1) D 0.25 mg · L -1, Carb 400 mg · L -1, Kan 50 mg · L -1) were significantly higher than those of other media. The maximum conversion rate of Favorita stems was significantly higher than that of tube potato slices. The conversion of Gannongshu No.2 and Shepody tube potato slices was significantly higher than that of stems. Stem transformation system of the best callus induction medium for the S2, the best differentiation based on the M2. Favorita is suitable for stem transformation system. Gannongshu No.2 and Shepody are suitable for tube-potato slice transformation system. Differences between callus induction medium and differentiation medium of different varieties are mainly caused by genotypic differences. In order to improve the efficiency of genetic transformation, the transformation system should be screened for the varieties.
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