元宝枫组织培养与快速繁殖技术

Tissue culture and rapid propagation technology of Acer truncatum

【目的】元宝枫是我国特有的木本油料植物,非常适合开发利用,但其组培离体繁殖生根非常困难,建立合理的完整组培离体再生体系,在短时期内获得大量种苗和当年生带芽茎段,为该树种优良种苗规模化生产提供新途径,为选育元宝枫良种、种质资源保存与推广等提供研究基础。【方法】以元宝枫当年生带芽茎段和种子为试材,通过正交设计筛选法和单因素筛选法对其灭菌方式及腋芽诱导、继代增殖、生根培养基进行设计筛选。在MS、1/2MS和NN69基础培养基中分别添加不同质量浓度的6-BA、NAA和TDZ激素,通过不同配比找出元宝枫各阶段的最适培养基。【结果】全年中4月份为带芽茎段最佳采集时间,经洗衣粉溶液轻刷表面并冲洗半小时后,以75%酒精30 s+0.1%HgCl_(2) 10 min浸泡处理的灭菌效果最佳。再生体系各阶段最适培养基均为MS基础培养基,在其基础上再添加不同质量浓度的激素可以诱导嫩芽分化成不同器官。启动培养阶段,6-BA对腋芽诱导影响最大,最适培养基诱导率可达73.33%;继代增殖阶段,TDZ对不定芽分化影响最大,增殖系数最高可达4.31;生根阶段,采用单因素筛选法,发现添加0.2 mg·L^(-1) IBA元宝枫生根率和生根条数均为最高,达93.33%和4.3。【结论】元宝枫最适培养基为:1)启动培养:MS+0.06 mg·L^(-1) 6-BA和0.06 mg·L^(-1) NAA,诱导率为73.33%;2)增殖培养:MS+0.30 mg·L^(-1) TDZ和0.05 mg·L^(-1) NAA;3)生根培养:MS+0.2 mg·L^(-1) IBA。

【Objective】Acer truncatum is a unique woody oil plant in China,which is very suitable for development and utilization,but it is very difficult to propagate and root ex vivo in tissue culture,establish a reasonable complete tissue culture ex vivo regeneration system,obtain a large number of seedlings and the current year’s bud stem segment in a short period of time,which will provide a new way for the large-scale production of excellent seedlings of this species,and provide a research basis for the selection and breeding of A.truncatum seeds,the preservation and promotion of germplasm resources,etc.【Method】The germ segments and seeds of A.truncatum were used as the test materials,and the sterilization method and axillary bud induction,sub-proliferation and rooting medium were designed and screened by orthogonal design screening method and univariate screening method.Different concentrations of 6-BA,NAA and TDZ hormones were added to MS,1/2MS and NN69 basal medium,respectively,and the most suitable medium for each stage of A.truncatum was found through different ratios.【Result】The best collection time for bud stem segments in April of the year,after brushing the surface and rinsing for half an hour with washing powder solution,the sterilization effect was best with 75%alcohol 30 s+0.1%HgCl_(2) soaked for 10 min.The most suitable medium at each stage of the regeneration system was MS basal medium,and the addition of different concentrations of hormones on the basis could induce the differentiation of young shoots into different organs.At the beginning of the culture stage,6-BA had the greatest effect on axillary bud induction,and the induction rate of optimal medium could reach 73.33%.In the secondary proliferation stage,TDZ had the greatest effect on adventitious bud differentiation,with a proliferation coefficient of up to 4.31,and a univariate screening method was used in the rooting stage,and it was found that 0.2 mg·L^(-1) IBA had the highest rooting rate and rooting number,reaching 93.33%and 4.3.【Conclusion】The optimal medium for A.truncatum is 1)Initiation culture:MS+0.06 mg·L^(-1)6-BA and 0.06 mg·L^(-1) NAA,induction rate was 73.33%;2)proliferation culture:MS+0.3 mg·L^(-1) TDZ and 0.05 mg·L^(-1) NAA;3)Rooting culture:MS+0.2 mg·L^(-1) IBA.