小麦成熟胚诱导胚性愈伤组织的影响因素

Study on the Conditions of Wheat Embryonic Callus Induction from Mature Embryos

为加快小麦成熟胚在转基因研究中的利用,以陕麦159成熟胚为外植体,通过胚性愈伤组织诱导,建立了一套有效的小麦愈伤组织诱导体系。研究表明,不同消毒处理、2,4-D浓度和接种方法对胚性愈伤出愈率的影响有极显著差异,不同浸种时间和培养基类型对胚性愈伤组织出愈率的影响无显著差异。采用次氯酸钠初次消毒20min,升汞二次消毒5min,消毒彻底,对种子损害较小;浸种时间18～20h易于成熟胚的剥取,且诱导的愈伤组织质量较好;MS培养基较1/2MS培养基诱导愈伤组织效果好,MS培养基2,4-D浓度为4mg.L-1时(30g蔗糖+7g琼脂)胚性愈伤组织率最高,愈伤组织质量较好;完整胚诱导胚性愈伤组织率最高达84.17%,碎胚诱导胚性愈伤组织率最高达94.17%。

An effective wheat callus induction system via inducing embryonic callus was set up using mature embryos of wheat variety Shaanmai 159 as exophyte.The results showed that the seeds were thoroughly disinfected and got less damage after dipping into sodium hypochlorite for 20 min,then 5 min in mercury bichloride.It was easy to strip the mature embryo and get better callus when the dipping time was between 18 h to 20 h.Better callus was got on MS medium than on 1/2 MS medium,when adding 4 mg/L 2,4-D into MS medium,the callus would be better,meanwhile the inducing efficiency was the highest.Compared with complete embryos,the ratio of embryonic callus induction was increased from 84.167% up to 94.167% by using broken embryos.So,the system was that disinfecting seeds first with sodium hypochlorite for 20 minutes,treated with mercury bichloride for 5 minutes,and then dipping the seeds into water 18h to 20h for stripping embryos,finally breaking the embryos on MS medium with 2,4-D（4 mg·L-1）,30 g sucrose and 7 g agar.