摘要
针对甘蓝组织培养过程中再生体系难度大,培养效率低的问题,本研究以白花芥蓝下胚轴为外植体,对离体培养关键影响因素——无菌苗光/暗培养时间、NaClO消毒时间、激素配比对甘蓝种子、不定芽分化及再生的影响进行了比较分析.结果表明,75%无水乙醇 40 s+5%NaClO 15 min消毒后,将种子暗培养 3 d,再光培养 3 d,无菌苗生长最好;预培养时培养基最佳激素组合是 0.05 mg L-1 NAA(α-萘乙酸)+4 mg L-1 6-BA(6-苄氨基嘌呤);筛选培养基的最佳激素组合为 0.05 mg L-1 NAA+4 mg L-1 6-BA+250 mg L-1 Cb(羧苄青霉素),愈伤发生率 91.1%,不定芽分化率57.3%;生根培养基MS+30 g L-1 蔗糖+7 g L-1 琼脂+0.1 mg L-1 NAA+0.1%活性炭,不定根产生数量多,获得植株根系发达、生长势强.本研究建立的甘蓝组织培养优化体系能快速获得再生植株,有助于加速甘蓝良种的繁育与评价.
Abstract
Addressing the challenges of a difficult regeneration system and low culture efficiency in the tissue culture process of Brassica oleracea L.,this study focused on using the hypocotyl of Brassica albograbra Bailey as explants,which was identified as a key influencing factor in in vitro culture.A comparative analysis was conducted to assess the effects of light/dark culture time,NaClO disinfection time,and hormone ratio on the differentiation and regeneration of Brassica oleracea L.seeds and adventitious buds.The results showed that the best growth of sterile seedlings was achieved when the seeds were disinfected with 75%anhy-drous ethanol for 40 s+5%NaClO 15 minutes.The seeds were then cultured in the dark for 3 d and then,followed by 3 d in the light.The optimal hormone combination for the pre-culture medium was 0.05 mg L-1 NAA+4 mg L-1 6-BA.In the screening cul-ture medium,the optimal hormone combination was 0.05 mg L-1 NAA+4 mg L-1 6-BA+250 mg L-1 Cb(Carbenicillin)inci-dence of 91.1%and a differentiation rate of adventitious buds of 57.3%.For the rooting medium MS+30 g L-1 sucrose+7 g L-1 agar+0.1 mg L-1 NAA+0.1%activated carbon yielded a large number of adventitious roots with well-developed root systems and strong growth potential.The optimized tissue culture system established in this study for Brassica oleracea L.allows for the rapid production of regenerated plants.This system is beneficial for accelerating the breeding and evaluation of improved varieties of Brassica oleracea L.
基金项目
国家自然科学基金项目(32272150)
重庆市自然科学基金项目(CSTB2022NSCQ-LZX0034)
重庆市自然科学基金项目(CSTB2022TIAD-KPX0010)
国家科技期刊平台
NSTL
万方数据