不同基因型小豆胚尖再生能力筛选和培养条件优化

Screening on Regeneration Capacity of Embryonic Tip from Different Genotypes of Adzuki Bean(Vigna angularis) and Optimizing on Culture Condition

本研究以47份不同基因型小豆的胚尖为外植体,采用不同6-苄氨基嘌呤（6-benzylaminopurine,6-BA）和吲哚乙酸（indole butytic acid,IBA）激素浓度配比,筛选再生能力强的小豆资源,优化胚尖再生条件。通过最小显著差数法（least significant difference,LSD）多重比较不同基因型间和不同培养基小豆再生率和平均再生芽数,结果显示：激素浓度配比相同时,不同基因型间小豆胚尖再生能力差异显著;基因型和培养基间互作差异显著。筛选出9份再生能力较好且培养基不敏感的小豆资源DR033、DR158、DR177、京农5号、DR-088、DR074、京农21号、DR039和DR049,其平均再生芽数分别为3.79、3.33、4.06、2.93、2.73、2.46、2.70、2.32和2.35,再生率分别为92.48%、87.04%、80.69%、94.15%、96.21%、100%、88.35%、84.02%和84.67%。0.2 mg/L6-BA＋0.2 mg/L IBA胚尖芽诱导培养基具有较好的通用性,胚尖再生周期短,从接种到获得3-5 cm高度再生芽仅需要约40 d时间。DR158、京农5号、京农6号和京农21号适宜的潮霉素B筛选浓度为7.5 mg/L,DR033和DR177则分别为5.0 mg/L和10.0 mg/L。本研究建立了效率高的小豆胚尖再生体系,有助于小豆遗传转化和基因功能分析。

Efficient regenerating accessions and optimized culture condition were screen by using embryonic tips explants of 47 adzuki beans in different genotypes and different hormone concentration ratio of BAP and IBA. Regeneration rate and the average number of regenerated shoots among different genotypes of adzuki bean and media were analyzed by LSD multiple comparisons. The results showed that regenerative capacity of different genotypes in adzuki bean had obvious differences and interaction between genotypes and culture media was significantly different as well. Nine accessions with higher regenerating ability and insensitive to the medium, which included DR033, DR158, DR177, Jingnong5, DR088, DR074, Jingnong21, DR039, and DR049 were scre- ened out with their average numbers of regenerated buds being 3.79, 3.33, 4.06, 2.93, 2.73, 2.46, 2.70, 2.32, and 2.35, respectively. The regeneration rates were calculated as 92.48%, 87.04%, 80.69%, 94.15%, 96.21%, 100%, 88.35%, 84.02%, and 84.67%, respectively. The regeneration cycle of embryonic tip spanned only 40 days from and embryonic tip planted in the medium to 3-5 cm cluster shoots harvested. The hormone concentration pro-portion with 0.2 mg/L 6-BA＋0.2 mg/L IBA exhibited a higher universality for bud induction of embryonic tip explants. The optimal hygromycin B selecting concentration for DR158, Jingnong5, Jingnong6, and Jingnong21 was 7.5 mg/L, that of DR177 and DR033 were 5.0 mg/L and 10.00 mg/L, respectively. Efficient regenerating system of embryonic tip in adzuki bean was constructed in this reseach. It might facilitate practical studies for transformation and gene functional analysis of adzuki bean.