美洲黑杨与大青杨杂种无性系离体培养和叶片再生体系的建立

Establishment of Leaf Regeneration System and in vitro Culture of Hybrid Clones of Populus deltoids Bartr. and Populus ussuriensis Kom

以美洲黑杨与大青杨杂种(Populus deltoids Bartr.×Populus ussuriensis Kom.)无性系的茎段作为外植体,研究了杂种无性系的离体培养及叶片再生体系,探讨了不同激素组合对杂种无性系不定芽的诱导、分化、增殖以及生根的影响。结果表明:杂种无性系茎段的最佳消毒时间为5min;最佳分化培养基为MS+6-BA2.0mg/L+NAA0.1mg/L+ZT0.5mg/L;MS+6-BA0.5mg/L+NAA0.05mg/L培养基可对不定芽实现增殖与复壮;最佳生根培养基为MS+NAA0.3mg/L。5个杂种无性系中,无性系177的分化能力最强;叶片近轴面向上放置培养,其不定芽再生能力显著高于叶片近轴面向下放置培养;叶片、茎段及根段的分化能力大小依次为叶片>茎段>根段。本研究优化了美洲黑杨与大青杨杂种的组织培养体系,为杂种无性系快速繁殖和遗传转化研究提供参考。

Stems of hybrid clone of Populus deltoids and P. ussuriensis were used as explants to study its in vitro culture and leaf regeneration system, and to explore effect of different prescriptions of hormone on induction, differentiation, propagation of adventitious bud, and rooting. The results showed that, the optimal sterilization time was 5 min, the optimum medium for multiplication was MS ＋ 6-BA 2 mg/L ＋NAA 0.1 mg/L ＋ ZT 0.5 mg/L, medium with MS ＋ 6-BA 0.5 mg/L ＋ NAA 0.05 mg/L might achieve the proliferation and restoration effect on adventitious bud, and the optimal medium for rooting was MS ＋ NAA 0.3 mg/L. Among the five hybrid clones, No.177 showed the best differentiation potential, its regeneration ability for leaf adventitious shoot was significantly higher when cultured with the abaxial side upward than with the adaxial side downward, its corresponding differentiation ability with leaf, stem and root segment come out to be in a descending order. This study had optimized the tissue cultural system of hybrid clone of P. deltoids and P. ussuriensis, result of which had laid groundwork foundation for its rapid propagation and genetic transformation analysis.