SUN Xin,ZHAO Jian-ru,YAN Fan,et al.Optimization on Cotyledonarynode Genetic Transformation System of Soybean Cultivar Jilin 47[J].Soybean Science,2013,32(03):291-294.[doi:10.11861/j.issn.1000-9841.2013.03.0291]
吉林47大豆子叶节遗传转化体系的优化
- Title:
- Optimization on Cotyledonarynode Genetic Transformation System of Soybean Cultivar Jilin 47
- Keywords:
- Jilin 47; Cotyledonarynode; Browning; Shoot elongation
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- ?为检测不同基因型大豆对于根癌农杆菌的敏感性,使用携带pCAMBIA1301质粒(含GUS基因)的农杆菌EHA105对10个性状优良的大豆基因型进行侵染,共培养后进行GUS染色,筛选得到对根癌农杆菌较敏感的基因型吉林47、吉林35和绥农10号;而后以吉林47子叶节为外植体进行大豆遗传转化体系的优化,在重悬及共培养期间添加不同浓度巯基化合物,发现400 mg·L-1L-Cys、248 mg·L-1硫代硫酸钠和300 mg·L-1DTT组合对于褐化的抑制效果最好;对芽伸长阶段的关键激素GA3和IAA进行比较研究,得到最佳配比为1.0 mg·L-1GA3、0.5 mg·L-1IAA。
- Abstract:
- ?After infected by EHA105 caring plasmid pCAMBIA1301,the GUS staining of 10 soybean genotypes was carried out to detect the sensitivity for Agrobacterium tumefaciens.Jilin 47,Jilin 35 and Suinong 10 were screened out as sensitive genenotypes.Jilin 47 was chosen as the genotype for optimization of soybean cotyledonary genetic transformation due to its good performance on shoot regeneration.Amending thiol compounds in liquid and solid cocultivation medium could decrease the browning rate of explants and the best combination of 400?mg·L-1 L-Cys, 248 mg·L-1 sodium thiosulfate and 300 mg·L-1DTT were obtained through orthogonal test.The highest elongation rate was got when 1.0 mg·L-1 GA3 and 0.5 mg·L-1IAA were added to shoot elongation medium.
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