多胺、香豆素对黑果枸杞植株再生体系褐化、玻璃化的影响

Effects of polyamines and coumarins on browning,vitrification in Lycium ruthenicum plant regeneration system

以优质‘大果’黑果枸杞(Lycium ruthenicum)幼嫩叶片经初代培养获得的愈伤组织为试材,在再生体系的不同培养阶段分别添加腐胺(Put)、精胺(Spm)及香豆素(CM)三种不同外源诱导子,研究对愈伤组织增殖、不定芽诱导增殖、壮苗生根及褐化、玻璃化("两化")的影响。结果表明:在愈伤组织继代增殖培养中,Put最佳浓度为15 mg·L^(-1),增殖倍数最高6.8,褐化指数降至0级;在不定芽诱导和增殖培养中,Spm能提高不定芽的分化率和增殖倍数,并能防止玻璃化苗发生和愈伤组织在分化过程中的褐变,不定芽诱导培养的最佳浓度为20 mg·L^(-1),分化率达68.3%,不定芽增殖培养的最佳浓度为30 mg·L^(-1),增殖7.2倍;在壮苗培养中,适宜浓度CM能有效降低组培苗玻璃化率,提高丛芽质量,最适浓度为15 mg·L^(-1),壮苗生根效果最佳,移栽成活率达94.6%。试验构建了促生长、控"两化"的植株再生技术新体系。

This study use callus obtained from the primary culture of the young leaves of the high-quality Lycium ruthenicum as material.Three exogenous materials,putrescine(Put),spermine(Spm)and coumarin(CM),were added in different culture stages of the regeneration system.The effects of these exogenous materials on callus proliferation,adventitious bud induction or proliferation,seedling,rooting,browning and vitrification were investigated.The callus proliferation coefficient was increased to 6.8 times when the optimal concentration of 15 mg·L^(-1) Put was added in the subculture medium,meanwhile,the browning index was reduced to 0 level.The differentiation rate and proliferation coefficient of adventitious buds could be increased greatly by Spm.Simultaneously,the adding of Spm could prevent the occurrence of vitrification at different levels and reduce the browning of callus during differentiation.The differentiation rate and proliferation coefficient of adventitious buds was 68.3%and 7.2 times when Spm was added at the concentration of 20 and 30 mg·L^(-1) in adventitious bud induction and proliferation culture,respectively.In seedling cultivation,adding CM at appropriate concentration of 15 mg·L^(-1) can effectively reduce the vitrification rate of tissue cultured seedlings,improve the quality of clump buds,and the survival rate of transplanting reached 94.6%.This study establishes a new regeneration system that promotes plant growth and controls the browning and vitrification.