外源激素对杜衡叶柄愈伤组织诱导与分化的影响

Effects of exo-phytohormones on induction and differentiation of callus from Asarum forbesii petiole

以杜衡（Asarum forbesii Maxim.）叶柄为外植体,采用L9（3^4）正交实验设计分别研究了培养基中外源激素的种类及质量浓度对杜衡叶柄愈伤组织诱导和分化的影响,并据此筛选出适宜的诱导及分化培养基。结果表明：在杜衡叶柄愈伤组织的诱导过程中,培养基中细胞分裂素的种类（1.00 mg·L^-1 6-BA、1.00 mg·L^-1KT和1.00 mg·L^-1ZT）和NAA质量浓度（0.00、0.10和0.30 mg·L^-1）的影响效应均不显著,而2,4-D质量浓度（0.10、0.50和1.00 mg·L^-1）则有显著影响（P〈0.05）;在愈伤组织的分化培养过程中,NAA（0.10、0.30和0.50 mg·L^-1）的影响效应大于6-BA（1.00、3.00和5.00 mg·L^-1）和IBA（0.01、0.05和0.10 mg·L^-1）。综合比较结果显示,适宜于杜衡叶柄愈伤组织诱导的培养基为添加1.00 mg·L^-1 6-BA、0.30 mg·L^-1 NAA和1.00 mg·L^-1 2,4-D的MS培养基（含6.5 g·L^-1琼脂和30 g·L^-1蔗糖,pH5.8~pH6.0）,在此培养基上愈伤组织诱导率达到83.33%,且愈伤组织生长速度快、颗粒紧密;适宜于杜衡愈伤组织分化和不定芽增殖的培养基为添加3.00 mg·L^-1 6-BA、0.10mg·L^-1 IBA和0.30 mg·L^-1 NAA的MS培养基（含6.5 g·L^-1琼脂和30 g·L^-1蔗糖,pH5.8-pH6.0）,在此培养基上愈伤组织的分化率最高（达到53.33%）,增殖系数也最高（3.13）。

Using petiole of Asarum forbesii Maxim.as an explant,effects of type and concentration of exo-phytohormones in culture media on induction and differentiation of callus were researched by L9（34） orthogonal experiment,and the suitable induction and differentiation media were also selected on the basis of the research results.The results show that in induction process of callus from A.forbesii petiole,cytokinin type（1.00 mg·L^-1 6-BA,1.00 mg·L^-1 KT and 1.00 mg·L^-1 ZT）and NAA concentration（0.00,0.10 and 0.30 mg·L^-1） in media have no obvious effects,but 2,4-D concentration（0.10,0.50 and 1.00 mg·L^-1） has a significant effect（P〈0.05）.In differentiation process of callus,effect of NAA（0.10,0.30 and 0.50 mg·L^-1） is stronger than those of 6-BA（1.00,3.00 and 5.00 mg·L^-1） and IBA（0.01,0.05 and 0.10 mg·L^-1）.The comprehensive analysis results show that the optimal medium suitable for callus induction from A.forbesii petiole is MS medium added 1.00 mg·L^-1 6-BA,0.30 mg·L^-1 NAA and 1.00 mg·L-1 2,4-D（containing 6.5 g·L^-1 agar and 30 g·L^-1 sucrose,pH 5.8-pH 6.0）.And in this medium,induction rate of callus reaches 83.33% and callus grows fast and forms compact cell masses.The optimal medium suitable for callus differentiation and propagation of adventitious bud of A.forbesii is MS medium added 3.00 mg·L^-1 6-BA,0.10 mg·L^-1 IBA and 0.30 mg·L^-1 NAA（containing 6.5 g·L^-1 agar and 30 g·L^-1 sucrose,pH 5.8-pH 6.0）.And the differentiation rate of callus is the highest with a percentage of 53.33%,and the multiplication coefficient is also the highest with a value of 3.13 in the optimal differentiation medium.