华北落叶松胚性愈伤组织诱导影响因子的研究

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 principis-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％ NaClO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4g·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－t,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.