华北落叶松胚性愈伤组织诱导影响因子的研究  

Factors Affecting Induction of Embryogenic Callus of Larix principis-rupprechtii

作　　者：韩登媛 李旦 赵健[1] 李慧[1] 张金凤[1] 

机构地区：[1]林木育种国家工程实验室/林木花卉遗传育种教育部重点实验室/国家林业局树木花卉育种与生物工程重点开放实验室/北京林业大学生物科学与技术学院,北京100083

出　　处：《林业科学研究》2013年第4期454-458,共5页Forest Research

基　　金：国家自然科学基金项目"科研训练及科研能力提高项目"(J1103516);国家林业局林业公益性行业专项"重要乡土树种核心种质评价及高效育种共性技术研究"(201004009)";北京市与中央在京高校共建项目"华北落叶松遗传资源收集评价及新品种选育(GJ2011-2)"

摘　　要：以成熟合子胚为外植体,用完全随机、正交设计等试验方法加之组培技术来研究影响华北落叶松(Larix princi-pis-rupprechtii)胚性愈伤组织诱导的几种主要因子。结果表明:诱导胚性愈伤组织过程中的最佳灭菌方式为2%Na-ClO灭菌30 min,剥皮、去胚乳处理。其中污染率和NaClO的灭菌时间呈负相关:NaClO灭菌时间越长,污染率越低。诱导胚性愈伤组织的最佳琼脂浓度为4 g·L-1,且过高和过低的琼脂浓度不仅降低了其诱导率,提高了愈伤组织的褐化水平,也影响了外植体的发育形态。外源激素的正交试验结果表明:2,4-D是主要影响因子,对诱导率的影响显著,其次是KT,最后是6-BA。最佳的植物外源激素配比是2,4-D 1.8 mg·L-1+6-BA 0.3 mg·L-1+KT 0.3 mg·L-1。Taking mature zygotic embryos of Larix principis-rupprechtii as explants to induce embryogenic callus.Several factors influencing the induction were assayed by using completely random design,orthogonal design together with tissue culture technology in order to explore embryogenic callus induction in the early stage in larch and optimize its somatic embryogenesis system.The results show that the best way of disinfection during the induction of embryogenic callus is peeling,removing endosperm after disinfection of 30 minutes with 2% NaClO.The contamination rate and disinfection time are negatively correlated： the longer the sterilization time,the lower the contamination rate.The optimal agar concentration of embryogenic callus induction is 4 g.L-1,and the higher or lower of the agar concentration will not only reduce the induction rate,increase the level of callus browning,but also affect the development of explants.The results of orthogonal design for plant growth exogenous hormone show that the 2,4-D is the key factor and has a significant influence,followed is KT and 6-BA.The best combination of plant growth exogenous hormone is 2,4-D 1.8 mg.L-1 ＋ 6-BA 0.3 mg.L-1 ＋ KT 0.3 mg.L-1.

关 键 词：华北落叶松 体细胞胚胎发生 诱导 培养基 胚性愈伤组织 

分 类 号：S722.3[农业科学—林木遗传育种]