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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 establish an efficient and stable bud-inducing system for Coral saurinum by using Solanum pseudo-capsicum seeds as experimental materials, optimize tissue culture conditions and screen suitable plant regeneration pathways. Methods The sterilized seeds were inoculated into blank MS, OM and WPM medium, and the sterile seedlings were obtained after 45 days of culture. On the basis of this, the shoot tips, bud stem segments and leaves were used as explant materials, The effects of different basic media and different plant hormones and their concentrations on the callus induction, shoot generation and plant regeneration were studied by cross test. Results The best medium for the growth of coral was WPM. The leaf explants were the most suitable explants for inducing callus. Within 7 days, WPM + 6-BA 0.5 mg / L + NAA 0.01 mg / L Induced a very strong differentiation ability of callus, the cure rate was 100%, within 10d that is divided into a large number of green buds, callus bud bud differentiation rate of 100%; with bud stem segment in the above medium also Can induce callus, the cure rate was 81.7%, after 20 days began to differentiate buds; and shoot tips can induce callus, but no redifferentiation. The suitable bud proliferation medium was WPM + 6-BA 0.5 mg / L + NAA 0.5 mg / L + KT 1.0 mg / L, and the multiplication coefficient of 6.0 was above 30d. Aseptically rooted at 1 / 2WPM + NAA0.01 mg / L, 30 d after the growth of robust regenerated plants, rooting rate of 100%. The rooting seedlings were transplanted into the well-drained sandy soil and the survival rate was over 90% after incubation for 25 days. Conclusion It is an effective way to maintain the characteristics of fine coral species and seedling propagation, and lay a good foundation for the cultivation and genetic transformation of its fine varieties.