文献类型：期刊文献

中文题名：侧柏古树组织培养体系的建立

英文题名：Establishment of Tissue Culture System of the Ancient Trees of Platycladus orientalis

作者：常二梅[1] 江泽平[1] 刘建锋[1] 贾子瑞[1] 李香[2] 张健强[3] 赵秀莲[1]

第一作者：常二梅

机构：[1]林木遗传育种全国重点实验室,国家林业和草原局林木培育重点实验室,中国林业科学研究院林业研究所,北京100091;[2]北京市绿地养护管理事务中心,北京102211;[3]邯郸市林业工程项目中心,河北邯郸057650

年份：2023

卷号：36

期号：4

起止页码：41-49

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：“十四五”国家重点研发计划课题(2022YFD2200103);国家自然科学基金面上项目(31870664);河北省林业和草原局科学技术研究项目“邯郸市古树名木资源调查和复壮技术研究(2001029)”。

语种：中文

中文关键词：侧柏;古树;组织培养;褐化;生根

外文关键词：Platycladus orientalis;ancient tree;tissue culture;browning;rooting

分类号：S722.43

摘要：[目的]攻克侧柏古树在组织培养中外植体消毒不彻底、褐化严重、分生能力弱等技术难点,建立侧柏古树的组织培养体系。[方法]以树龄约3000年侧柏古树当年生的新梢为外植体,经过消毒灭菌处理获得无菌外植体。通过筛选适宜的培养基和激素,建立侧柏古树组织培养体系。[结果]外植体用0.3%HgCl2消毒10 min的灭菌效果最好,污染率为25.81%,成活率为54.82%。侧柏古树组织培养基的糖源用3%蔗糖的效果比3%葡萄糖更佳。抗氧化剂与吸附剂试验结果表明,在外植体启动培养基中加入3.00 g·L^(?1)活性碳,外植体褐化率最低。侧柏古树组织培养的最佳初代培养基为:1/2MS+NAA 0.2 mg·L^(?1)+6-BA 0.1 mg·L^(?1),不定芽诱导率为32.05%,芽生长快,嫩绿且粗壮。最佳的增殖培养基为:1/2MS+6-BA 0.05 mg·L^(?1)+NAA 0.2 mg·L^(?1)+KT 0.1 mg·L^(?1),增殖系数为1.93,绿色芽生长快且粗壮。最佳的生根培养基为:WPM+NAA 0.5 mg·L^(?1)+IBA 0.5 mg·L^(?1),生根率达到9.12%,每株平均生根数为2.67,平均根长为1.23 cm。总共获得24株生根苗,2个月后2株有红根出现,嫩芽继续生长,成功获得侧柏古树组织培养生根苗。[结论]侧柏古树培养基的选择、生长素与细胞分裂素的比例是侧柏古树增殖培养和不定根形成的重要因素,此实验为其他柏科树木组织培养提供了参考。
[Objective]This study aims to establish a tissue culture system for ancient Platycladus orientalis for overcoming the difficulties such as non-thoroughness of disinfection and sterilization,sever browning,and weak meristematic ability.[Method]In this study,the new shoots of about 3000-year-old P.orientalis were used as explants,and aseptic explants were obtained through disinfection and sterilization.The tissue culture system for ancient P.orientalis was established by selecting the appropriate culture medium and hormone.[Result]Disinfecting the explants from ancient P.orientalis using 0.3%HgCl2 for 10 min showed the best sterilization effect,with a pollution rate of 25.81%and the survival rate of 54.82%.The effect of using 3%sucrose as the sugar source for the tissue culture medium of ancient P.orientalis was better than that of 3%glucose.The results of antioxidant and adsorbent tests demonstrated that the browning rate of explants was the lowest under the addition of 3.00 g·L^(?1) active carbon to the explant initiation medium.The optimal primary culture medium for tissue culture of ancient P.orientalis was 1/2MS+NAA 0.2 mg·L^(?1)+6-BA 0.1 mg·L^(?1),with the induction rate of adventitious buds of 32.05%,rapid bud growth,as well as tender green and robust buds.The optimal proliferation medium was 1/2MS+6-BA 0.05 mg·L^(?1)+NAA 0.2 mg·L^(?1)+KT 0.1 mg·L^(?1),with the proliferation coefficient reaching 1.93,and rapid growth of green and robust buds.The optimal rooting culture medium was WPM+NAA 0.5 mg·L^(?1)+IBA 0.5 mg·L^(?1),which achieved a rooting rate of 9.12%,with an average number of roots per plant of 2.67,and an average root length of 1.23 cm.A total of 24 rooting seedlings were obtained.Two months later,red roots appeared in 2 plants,and the tender buds continued to grow.The tissue-cultured rooting seedlings of ancient P.orientalis were successfully obtained.[Conclusion]The selection of culture medium and the ratio of auxin to cytokinin are important factors for the proliferation and adventitious root formation of ancient P.orientalis.This study provides a reference for tissue culture of other Cupressaceae trees.