文献类型：期刊文献

中文题名：极小种群野生植物波叶杜鹃组培快繁体系的构建

英文题名：Construction of Tissue Culture and Rapid Propagation System for Rhododendron hemsleyanum,a Plant Species with Extremely Small Populations

作者：柳文[1,2] 杨琳祥[1,2] 木正春[3] 马永鹏[4] 谢孟[1,2] 万有名[1] 李正红[1] 马宏[1,5]

第一作者：柳文

机构：[1]中国林业科学研究院高原林业研究所,云南昆明650233;[2]南京林业大学风景园林学院,江苏南京210037;[3]贡山县林业和草原局,云南贡山673500;[4]云南省极小种群野生植物综合保护重点实验室,中国科学院昆明植物研究所,云南昆明650201;[5]国家林业和草原局资源昆虫培育与利用重点实验室,云南昆明650233

年份：2024

卷号：39

期号：4

起止页码：99-109

中文期刊名：云南农业大学学报（自然科学版）

外文期刊名：Journal of Yunnan Agricultural University:Natural Science

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD_E2023_2024】；

基金：云南省重大科技专项(202302AE090018);云南省重点研发计划(202202AE090012);兴滇人才支持计划青年人才专项。

语种：中文

中文关键词：波叶杜鹃;极小种群野生植物;Box-Behnken;组织培养;快速繁殖

外文关键词：Rhododendron hemsleyanum;plant species with extremely small populations;Box-Behnken;tissue culture;rapid propagation

分类号：S685.210.43

摘要：【目的】建立极小种群野生植物波叶杜鹃(Rhododendron hemsleyanum)的组培快繁体系,探究其种子萌发、增殖壮苗、生根诱导、移栽基质等不同环节的最佳条件。【方法】选择颗粒饱满、大小基本一致的种子作为萌发材料,探究光暗条件以及不同灭菌预处理对种子萌发的影响。在增殖壮苗和诱导生根试验中,以种苗培养的继代苗为外植体,以WPM为基本培养基,分别采用L9(33)正交试验设计和Box-Behnken试验设计响应面法,研究增殖壮苗和生根诱导的最适条件。在炼苗及移栽试验中,以成功诱导生根的组培苗为材料,炼苗9 d后移栽至5种不同的基质中,分析波叶杜鹃在不同移栽基质中的成活率。【结果】75%乙醇处理1 min+1%NaClO溶液处理10 min的种子萌发效果最好,萌发率为66.67%。种苗生长的最佳培养基为WPM+1.0 mg/L 6-BA;增殖壮苗的最佳培养基为WPM+0.5 mg/L NAA+1.5 mg/L ZT,pH 4.5,增殖系数为3.13。在生根诱导培养基中,横置于培养基表面的茎尖生根良好,植入培养基的茎尖均无不定根形成,且培养40 d时,植入培养基的外植体基部均腐化变黑;最佳生根培养基为1/2 WPM+0.5 mg/L NAA+1.0 mg/L IBA,生根率达88.00%。移栽基质V_(腐殖土)∶V_(珍珠岩)∶V_(蛭石)=2∶1∶1最适合组培苗移栽,成活率达80.00%。【结论】本研究成功构建了波叶杜鹃的组培快繁体系,为其他杜鹃花科木本植物的组织培养提供了参考,也为今后开展回归引种、复壮野生种群和合理开发利用奠定基础。
[Purpose]To establish a tissue culture system for Rhododendron hemsleyanum,a plant species with extremely small populations,and to investigate the optimal conditions for seed germination,seedling proliferation,root induction,and transplant substrate.[Methods]The seeds with plump grains and similar size were selected as germination materials,the effects of light and dark conditions and different sterilization pretreatment on seed germination were studied.For the seedling proliferation and root induction experiments,the progeny seedlings were used as explants,and WPM was used as the basic culture medium,the optimal conditions for seedling proliferation and root induction were studied by a L9(33)orthogonal experimental design and Box-Behnken response surface methodology.For the hardening and transplant experiments,the successfully root-induced test seedlings were transplanted into five different substrates after nine days,and the survival rate of R.hemsleyanum in different transplant substrates was analyzed.[Results]The best seed germination results were achieved with one minute treatment with 75%ethanol and ten minutes treatment with 1%NaClO solution,resulting in a germination rate of 66.67%.The best growth medium for seedlings was WPM+1.0 mg/L 6-BA;the optimal culture medium for seedling proliferation was WPM+0.5 mg/L NAA+1.5 mg/L ZT,and the pH value was 4.5,with a proliferation coefficient of 3.13.In the root induction culture medium,stem tips placed horizontally on the surface of the medium showed good rooting,while those inserted into the medium did not form adventitious roots,and after 40 days of culture,the basal part of the explants implanted in the culture medium decayed and turned black.The optimal rooting medium was found to be 1/2 WPM+0.5 mg/L NAA+1.0 mg/L IBA,resulting in a rooting rate of 88.00%.The most suitable transplant substrate for tissue-cultured seedlings was V_(humus soil)∶V_(perlite)∶V_(vermiculite)=2∶1∶1,with a survival rate of 80.00%.[Conclusion]This study successfully establishes a tissue culture rapid propagation system for R.hemsleyanum,providing a reference for tissue culture of other woody plants in the Ericaceae family.It also lays the foundation for future activities such as reintroduction,revitalization of wild populations,and sustainable development and utilization.