野葛叶片和茎段高频再生体系的建立

Establishment of High-frequency Regeneration System from Leaves and Stems of Pueraria lobata

探讨几种因子对野葛叶片和茎段高频再生体系建立的影响。采用植物组织培养、正交实验和单因子实验的方法。野葛叶片和茎段的最佳消毒方式为70%酒精处理30 s后再用0.1%HgCl2处理15 min;野葛叶片愈伤组织诱导的最佳培养基为MS+NAA 1.0 mg.L-1+2,4-D 2mg.L-1,野葛茎段愈伤组织诱导的最佳培养基为MS+NAA0.5 mg.L-1+6-BA1.0 mg.L-1+2,4-D 2 mg.L-1;暗培养更有利于野葛愈伤组织的诱导;野葛叶片和茎段愈伤组织诱导的最佳蔗糖浓度均为30 g.L-1;野葛叶片愈伤组织的最佳出芽培养基为MS+NAA 1.0 mg.L-1+6-BA3.0 mg.L-1,而野葛茎段愈伤组织的最佳出芽培养基为MS+NAA0.5 mg.L-1+KT2 mg.L-1;光照培养更有利于野葛叶片和茎段愈伤组织芽的再分化;野葛叶片愈伤组织再生芽生根的最佳培养基为MS+NAA0.5 mg.L-1+PP3330.5 mg.L-1,而野葛茎段愈伤组织再生芽生根的最佳培养基为MS+NAA0.5 mg.L-1+PP3333.0 mg.L-1;野葛叶片和茎段愈伤组织再生芽生根的最佳蔗糖浓度均为30 g.L-1;叶片再生苗移栽的最佳PP333浓度为1.0 mg.L-1,茎段再生苗移栽的最佳PP333浓度为3.0 mg.L-1;叶片和茎段再生苗的最佳移栽基质均为蛭石:珍珠岩(2:1)。

To illustrate the effects of several factors on the high-frequency regeneration plant system of Pueraria lobata leaves and stems. Plant tissue culture, orthogonal design and single factor treatment were applied. The best disinfection way of P. lobata leaves and stems was firstly using 70% ethyl alcohol to process 30 s, then using 0.1% HgC12 to process 15 min again ; The best culture medium of P. lobata leaf callus induction was MS ＋ NAA 1.0 mg ·L^-1 ＋ 2,4-D 2 mg·L^-1 ; The best culture medium of P. lobata stem callus induction was MS ＋NAA 1.0 mg ·L^-1 ＋6-BA 1.0 mg·L^-1 ＋2,4-D 2 mg·L^-1; The dark culture was more advantageous for P. lobata callus induction; The best sucrose concentration of P. lobata leaf and stem callus induction was 30 g ·L^-1 sucrose. The best culture medium of P. lobata leaf callus redifferentiation was MS ＋ NAA 1.0 mg - L- 1 ＋ 6-BA 3.0 mg·L^-1. The best culture medium of P. lobata stem callus redifferentiation was MS ＋ NAA 0.5 mg ·L^-1 ＋ KT 2 mg ·L^-1 ; The light culture was more advantageous for P. lobata leaf and stem callus redifferentiation ; The best culture medium of P. lobata leaf callus regenerated buds rooting was MS ＋ NAA 0.5 mg ·L^-1 ＋ PP333 0.5 mg ·L^-1 ＋ 30 the g·L^-1 sucrose. The best culture medium of P. lobata stem callus regenerated buds rootingwas MS ＋ NAA 0.5 mg·L^-1 ＋ PP333 3.0 mg·L^-1 ＋ 30 the g ·L^-1sucrose ; 1.0 mg ·L^-1 PP333 and transplant matrix of vermiculite： perlite （2：1） could remarkably improve the transplant survival rate ofP. lobata leaf regeneration seedling, 3.0 mg ·L^-1 PP333 and transplant matrix of vermiculite： perlite （2：1） could remarkably improve the transplant survival rate of P. lobata stem regeneration seedling.