“水城小黄姜”芽茎尖组培脱毒与快繁体系的建立  

Establishing the System of Detoxification and Rapid Propagation in"Shuicheng Small Yellow Ginger"

作　　者：吕金丽 吕树鸣 张冬莲 罗凯 吴迪 李伟[1,3] LüJinli;LüShuming;Zhang Donglian;Luo Kai;Wu Di;Li Wei(College of Agriculture,Guizhou University,Guiyang,550025;Liupanshui Academy of Agricultural Sciences,Liupanshui,553000;Vegetable Research Institute,Guizhou University,Guiyang,550025)

机构地区：[1]贵州大学农学院,贵阳550025 [2]六盘水市农业科学研究院,六盘水553000 [3]贵州大学蔬菜研究院,贵阳550025

出　　处：《分子植物育种》2024年第4期1170-1178,共9页Molecular Plant Breeding

基　　金：贵州省教育厅项目(黔教合KY字[2019]018号);贵州省农业农村厅(第一批农村产业革命蔬菜产业发展项目);贵州省科技厅项目(黔科合支撑[2016]2548号);六盘水市科技局农业攻关项目(52020-2019-N-01);六盘水市平台项目(52020-2018-03-09)共同资助。

摘　　要：为建立“水城小黄姜”芽茎尖组培脱毒快繁体系,本试验以“水城小黄姜”芽茎尖为外植体,研究不同消毒时间、不同诱导分化培养基、不同增殖培养基及不同生根培养基对茎尖消毒、愈伤组织诱导、不定芽增殖、生根的影响,并对组培苗进行脱毒效果的检测。结果表明:“水城小黄姜”芽茎尖采用75%医用酒精消毒60 s+10%次氯酸钠(NaClO)消毒20 min处理效果最好,污染率7.7%,成活率达87.2%;不同培养基配比对茎尖诱导分化效果有差异,以MS+6-BA 3.5 mg/L+NAA 0.1 mg/L效果最佳,愈伤诱导率达75.0%;最适增殖培养基为MS+6-BA 3.0 mg/L+NAA 0.3 mg/L,增殖系数大,长势旺盛、茎粗芽壮、叶绿,组培苗生长好,且可与生根协同接续;适宜生根的培养基为MS+NAA 0.10 mg/L,生根数多,平均根长适中。用双抗体夹心酶联免疫法(ELISA)测定组培苗中烟草花叶病毒(TMV)、黄瓜花叶病毒(CMV),获得了“水城小黄姜”脱毒核心苗,且脱毒率达93.75%。因此,本研究认为通过“水城小黄姜”芽茎尖脱毒组培快繁体系的建立,可为“水城小黄姜”种苗的脱毒快繁及工厂化育苗提供参考。To establish a tissue culture and rapid propagation system of"Shuicheng Small Yellow Ginger",taking the shoot tip of"Shuicheng Small Yellow Ginger"bud as the explant,the effect of shoot tip disinfection,callus induction,adventitious bud proliferation,rooting and domestication were studied with different disinfection time,different differentiation medium,different proliferation medium and different rooting medium on the shoot tip disinfection,callus induction,adventitious bud proliferation,and tested the virus-free effect of tissue culture seedlings.The results showed that the shoot tip of"Shuicheng Small Yellow Ginger"sprouts was sterilized with 75%medical alcohol for 60 s+10%sodium hypochlorite(NaClO)for 20 min,and the effect was the best,with a pollution rate of 7.7%and a survival rate of 87.2%.The different medium had the different effects of the shoot tip differentiation,MS+6-BA 3.5 mg/L+NAA 0.1 mg/L had the best effect,and the callus induction rate reached 75.0%.The most suitable proliferation medium was MS+6-BA 3.0 mg/L+NAA 0.3 mg/L which had large proliferation coefficient,vigorous growth,thick stems and strong buds,green leaves,tissue cultured seedlings growed well,and can be coordinated with rooting.The suitable medium for rooting was MS+NAA 0.10 mg/L,with a large number of roots,the average root length was moderate.The double antibody sandwich enzyme-linked immunoassay(ELISA)was used to determine the tobacco mosaic virus(TMV)and cucumber mosaic virus(CMV)in the tissue cultured seedlings,the core plantlets of"Shuicheng Small Yellow Ginger"detoxified was obtained,and the detoxification rate was over 93.75%.Therefore,this study believes that the establishment of a virus-free tissue culture and multiplication system for the shoot tip of"Shuicheng Small Yellow Ginger"can provide references for the detoxification and rapid propagation of"Shuicheng Small Yellow Ginger"seedlings and industrial seedlings.

关 键 词：小黄姜 组织培养 茎尖 脱毒 病毒检测 

分 类 号：S632.5[农业科学—蔬菜学]