珊瑚樱丛芽高效诱导体系及植株再生研究

Efficient induction system of adventitious shoots and plant regeneration in Solarium pseudo-capsicum

目的以珊瑚樱Solanum pseudo-capsicum种子为实验材料,建立珊瑚樱高效、稳定的丛芽诱导体系,优化组织培养条件,筛选出适宜的植株再生途径。方法将消毒后的种子接种于空白MS、OM和WPM培养基中,培养45 d后获得无菌苗;在此基础上,选取茎尖、带芽茎段和叶片为外植体材料,通过正交试验研究不同基本培养基和不同植物激素种类及其质量浓度对愈伤组织诱导、丛芽发生及植株再生的影响。结果适宜珊瑚樱生长的基本培养基为WPM;3种外植体中叶片是最适宜诱导愈伤组织的材料,在WPM+6-BA 0.5 mg/L+NAA 0.01 mg/L中可在7d内诱导出分化能力极强的愈伤组织,得愈率为100%,10d内即分化为大量绿色丛芽,愈伤组织丛芽分化率亦达100%;带芽茎段在上述培养基中亦能诱导出愈伤组织,得愈率为81.7%,20 d后开始分化出丛芽;而茎尖可以诱导出愈伤组织,但无再分化能力。适宜丛芽增殖培养基为WPM+6-BA 0.5 mg/L+NAA0.5 mg/L+KT 1.0 mg/L,30d增殖系数6.0以上。无菌苗生根则在1/2WPM+NAA0.01 mg/L中进行,30 d后可获得生长健壮的再生植株,生根率100%。生根苗移栽至排水良好的沙土中,保温保湿培养25 d后,成活率90%以上。结论为保持珊瑚樱优良品种特性、种苗繁殖提供了有效途径,也为其优良品种的培育和遗传转化研发奠定了良好基础。

Objective To investigate the conditions for improving the efficiency of plant regeneration system for Solanum pseudo-capsicum via the induction of adventitious shoots. Methods Sterilized seeds of S. pseudo-capsicum were cultured on MS, OM, and WPM basal media for germination and seedling growth. Effectiveness of different combination of tissue culture media and growth regulators on the callus induction, adventitious shoots production, and plant regeneration were compared, using the explants excised from the seedlings of 45 d. Results WPM is the best medium for the growth of S. pseudo-capsicum. Leaf is the best explant for callus induction with the frequency up to 100% cul^Lred on WPM medium containing 0.5 mg/L 6-BA and 0.01 mg/L NAA after 7 d and the callus divided into green adventitious shoots whose frequency reached 100% after 10 d; The callus whose frequency was 81.7% could be induced from the stem with buds cultured on the same medium as mentioned above, the adventitious shoots generated 20 d later. However, the stem tip induced callus without capacity of differentiation. The optimal medium for adventitious shoots proliferation is WPM ＋ 0.5 mg/L 6-BA ＋ 0.5 mg/L NAA ＋ 1.0 mg/L KT, the proliferation coefficient is far more 6.0 after 30 d. The optimal medium for plantlet rooting is 1/2 WPM ＋ 0.01 mg/L NAA, the strong plant regeneration occurred 30 d later with the rooting rate up to 100%. Over 90% plantlets survived after transplanting into sand with constant temperature and humidity condition for 25 d. Conclusion The current study provides an effective solution for maintaining the improved seeds trait and sprouts multiplication of S. pseudo-capsicum, which is helpful for the research of cultivation and genetic transformation.