半夏茎尖脱毒培养及病毒检测  

Virus-free culture and virus detection of stem tip of Pinellia ternata

作　　者：王宝霞 齐永红 肖雅尹 崔丽艳 王德富[1] 牛颜冰[1] WANG Bao-Xia;QI Yong-Hong;XIAO Ya-Yin;CUI Li-Yan;WANG De-Fu;NIU Yan-Bing(College of Life Sciences,Shanxi Agricultural University,Taigu,Shanxi 030801,China;Fruitry Station of Shanxi Province,Taiyuan 030001,China)

机构地区：[1]山西农业大学生命科学学院,山西太谷030801 [2]山西省果业工作站,太原030001

出　　处：《植物生理学报》2018年第12期1813-1819,共7页Plant Physiology Journal

基　　金：国家自然科学基金(31601612)~~

摘　　要：以半夏(Pinellia ternata)茎尖为外植体,研究不同生长调节物质浓度配比对半夏茎尖脱毒苗诱导效果的影响。在单因素试验基础上,分别选取6-苄氨基腺嘌呤(6-BA)和2,4-二氯苯氧乙酸(2,4-D)三个最优诱导浓度设计L29(3)正交试验,以确定茎尖愈伤组织诱导的最适培养基,并利用分子生物学方法对脱毒苗进行病毒检测,结果表明:当6-BA浓度为1.5 mg·L^(-1), 2,4-D浓度为2.0 mg·L^(-1)时,愈伤组织诱导率最高。正交试验得出诱导愈伤的最佳培养基配方为MS+1.5 mg·L^(-1) 6-BA+2.0 mg·L^(-1) 2,4-D+7 g·L^(-1)琼脂+30 g·L^(-1)蔗糖,并可一次性成苗。该培养基所诱导的脱毒苗经斑点酶联免疫吸附法(DOT-ELISA)和反转录PCR检测后,烟草花叶病毒(Tobacco mosaic virus,TMV)、黄瓜花叶病毒(Cucumber mosaic virus, CMV)和大豆花叶病毒(Soybean mosaic virus, SMV)脱毒率均达到100%。Absrtact: The stem tips were taken as explants to study the induction effects of different hormone concentrations on virus-free stem tips of Pinellia ternata seedlings. On the basis of single factor experiment, L9(32) orthogonal test of 6-benzylamino adenine(6-BA) and 2,4-dichlorophenoxyacetic acid(2,4-D) with three optimal concentrations was conducted to screen the optimal medium of callus induction, followed by molecular detection of the virus condition of virus-free seedlings. The results showed that the callus induction rate was the highest when the concentration of 6-BA was 1.5 mg·L^-1 and 2,4-D was 1.8 mg·L^-1. The optimum medium for callus induction was MS+1.5 mg·L^-1 6-BA+1.8 mg·L^-1 2,4-D+7 g·L^-1 agar+30 g·L^-1 sugar, which could induce the seedlings at one step. After reverse transcription-polymerase chain reaction(RT-PCR) and dot-enzymelinked immunosorbent assay(DOT-ELISA) detection, the virus-free rate of Tobacco mosaic virus(TMV), Cucumber mosaic virus(CMV) and Soybean mosaic virus(SMV) reached 100%.

关 键 词：半夏 茎尖培养 脱毒苗 病毒检测 

分 类 号：S567.239[农业科学—中草药栽培]