不同基因型甜高粱愈伤组织与丛生芽诱导条件优化

【目的】探讨不同培养基类型和激素配比对不同基因型甜高粱的愈伤组织和丛生芽诱导的影响,为建立高效稳定的甜高粱遗传转化体系奠定基础。【方法】以不同基因型甜高粱辽甜三号和北甜三号的成熟种子为外植体,在MS培养基中添加不同浓度2,4-D及2,4-D与NAA、6-BA或KT配比组合,对愈伤组织和丛生芽进行诱导;分别在B5、N6、MS和1/2MS培养基中添加不同2,4-D、NAA、6-BA组合,对愈伤组织进行诱导。【结果】适宜北甜三号愈伤组织和胚性愈伤组织诱导的最佳培养基为MS＋2.0mg/L2,4-D＋0.1mg/LNAA,辽甜三号为MS＋2.0mg/L2,4-D＋0.2mg/LNAA,可获得质量较好的愈伤组织,愈伤组织块大且色泽鲜艳。适于两个基因型甜高粱丛生芽诱导的最佳基本培养基为MS＋1.0mg/L6-BA＋0.2mg/LNAA＋0.5mg/L2,4-D,但不同基因型的诱导率有明显差异,以辽甜三号的诱导效果较好。【结论】不同浓度2,4-D与其他激素配比对两个基因型甜高粱的愈伤组织诱导有明显影响,以对辽甜三号的诱导效果较好;在MS、B5、N6、1/2MS基本培养基中,MS可作为愈伤组织诱导的最佳基本培养基。

Optimization of callus and cluster buds induction in different genotypes of sweet sorghum

Objective]The present experiment was conducted to investigate the effects of different culture media and hormone combinations on induction of callus and cluster buds in different genotypes of sweet sorghum to facilitate the genetic transformation studies. Method]The mature seeds of two different genotypes of sweet sorghum （Liaotian 3 and Beitian 3） were used as explants. Different concentrations of 2,4-D and its combinations with NAA, 6-BA or KT were added to MS medium to induce the callus and cluster buds. Different combinations of 2,4-D, NAA and 6-BA were also added to different media, viz., B5, N6, MS and 1/2MS for inducing the callus, respectively. Result]The best hormone combinations for induction of callus and embryo callus for varieties Beitian 3 and Liaotian 3 was MS＋2.0 mg/L 2,4-D＋0.1 mg/L NAA and MS＋2.0 mg/L 2,4-D＋0.2 mg/L NAA, respectively. The induced callus was large in size and showed better quality. The best hormone combinations for the induction of cluster buds in two different genotypes was MS＋1.0 mg/L 6-BA＋0.2 mg/L NAA＋0.5 mg/L 2,4-D, however, the induction of cluster buds in variety Liaotian 3 was better. Conclusion]Combinations of different concentrations of 2,4-D with other different hormones showed significant and positive effects on induction of callus in both genotypes of sweet sorghum, which were more pronounced in Liaotian 3. Amongst MS, B5, N6 and 1/2MS medium, MS medium may be used as the best basic medium for induction of callus.