西南桦节间茎段植株再生体系的建立  

Establishment of Plant Regeneration System from Internodes of Betula alnoides

作　　者：王欢 郭俊杰[1] 王春胜[1] 尹海锋[1] 曾杰 WANG Huan;GUO Jun-jie;WANG Chun-sheng;YIN Hai-feng;ZENG Jie(Research Institute of Tropical Forestry,Chinese Academy of Forestry,Key Laboratory of National Forestry and Grassland Administration on Tropical Forestry Research,Guangzhou 510520,Guangdong,China;Nanjing Forestry University,Nanjing 210037,Jiangsu,China)

机构地区：[1]中国林业科学研究院热带林业研究所,热带林业研究国家林草局重点实验室,广东广州510520 [2]南京林业大学,江苏南京210037

出　　处：《林业科学研究》2023年第6期115-125,共11页Forest Research

基　　金：“十三五”国家重点研发计划项目(2016YFD0600604)。

摘　　要：[目的]探究西南桦间接器官发生过程的愈伤组织诱导、不定芽分化、生根诱导等各阶段的适宜培养基配方,建立西南桦高频再生体系,为其遗传转化和良种繁育提供理论依据和技术支持。[方法]以西南桦TC2号无性系的节间茎段为外植体开展愈伤组织诱导、不定芽分化以及生根培养基筛选试验,并优化了预培养条件,揭示了愈伤组织诱导阶段基本培养基、激素浓度、暗培养时间,愈伤组织分化阶段激素组合,以及预培养条件等因素对不定芽分化的影响。[结果](1)预培养的适宜条件为:弱光培养(1000 lx)15 d后转至暗培养7 d,再正常光照(2000 lx)培养8 d,该条件下可获得适度黄化的植株,其平均株高和节间长度分别为6.6 cm和3.1 cm;(2)适合西南桦节间茎段愈伤组织诱导的培养基为WPB5+1.0 mg·L^(-1) TDZ+0.2 mg·L^(-1) NAA+20 g·L^(-1)蔗糖+5.8 g·L^(-1)琼脂(pH5.8),适宜暗培养时间为15 d;(3)适合愈伤组织分化的培养基为WPM+0.8 mg·L^(-1)6-BA+0.5 mg·L^(-1) GA3+30 g·L^(-1)蔗糖+5.8 g·L^(-1)琼脂(pH5.8);(4)采用上述最优方案,其茎段诱导的愈伤组织分化率和净增殖系数为88.9%和6.2以上,平均每个预培养植株可产生56.8个不定芽;(5)适宜生根培养基为WPM+0.1 mg·L^(-1) NAA+20 g·L^(-1)蔗糖+5.8 g·L^(-1)琼脂(pH5.8),培养30 d后生根率可达100%。[结论]本研究完整构建了高效稳定、重复性好的西南桦节间茎段高频再生体系,该体系不仅愈伤组织分化率和增殖系数较高,还可提升节间茎段取材的效率,为今后开展西南桦组培快繁以及利用基因工程进行其遗传性状改良奠定了基础。[Objective]To explore the optimal media components at stages for callus induction,adventi-tious bud differentiation,rooting induction of indirect organogenesis,a high-frequency regeneration system of Betula alnoides was established for providing theoretical evidence and technical support for genetic transformation and multiplication of elite cultivars of this species.[Methods]The internode stem segment of Betula alnoides TC2 clone was used to conduct callus induction,adventitious bud differentiation and rooting medium screening experiments,and the pre-culture conditions were optimized,revealing the fun-damentals of callus induction stage.The effects of basic media,application of hormones,and dark culture at callus induction stage,hormone combination at adventitious bud differentiation stage,and preculture conditions on adventitious bud differentiation were assessed.[Results](1)The suitable conditions for pre-culture were:15 days under low illumination(1000 lx),then 7 days in dark and 8 days under normal illu-mination(2000 lx),and properly yellowing plantlets could be obtained under these conditions with the mean height and internode length reaching 6.57 cm and 3.07 cm,respectively;(2)The medium suitable for callus induction of internodes was WPB5+1.0 mg·L^(-1) TDZ+0.2 mg·L^(-1) NAA+20 sucrose+5.8 g·L^(-1)Agar(pH5.8)for the callus induction,and the optimal dark culture time was 15 days;(3)The medium suitable for callus differentiation was WPM+0.8 mg·L^(-1)6-BA+0.5 mg·L^(-1) GA3+30 sucrose+5.8 g·L^(-1)Agar(pH5.8);(4)Using the above optimal scheme,the differentiation rate and net proliferation coefficient of the inter-node segments were 88.89%and more than 6.2,respectively,and 56.8 adventitious buds were obtained eventually per preculture plantlet on average;and(5)The reasonable rooting medium was the Wood Plant Medium supplemented with 0.2 mg·L^(-1) NAA+20 g·L^(-1) sucrose,and the rooting rate could reach 100%after 30 days.[Conclusion]A high-frequency regeneration system of B.alnoides is completely establis

关 键 词：西南桦 节间茎段 预培养条件 再生体系 

分 类 号：S722.3[农业科学—林木遗传育种]