茎尖培养法脱除菊花B病毒的研究  

Study on the elimination of Chrysanthemum virus B by using meristem-tip culture

作　　者：尤燕平[1] 彭诗怡[1] 朱文莹[1] 王靓[1] 蒋甲福[1] 陈素梅[1] 房伟民[1] 管志勇[1] 陈发棣[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095

出　　处：《南京农业大学学报》2013年第5期144-148,共5页Journal of Nanjing Agricultural University

基　　金：江苏省科技支撑计划项目(BE2011325);国家农业科技成果转化项目(2010GB2360063);农业部948重点滚动项目(2011-G17);国家公益性行业(农业)科研专项(200903020);中央高校基本科研业务费专项资金项目(KYZ201112;Y0201100252)

摘　　要：以感染菊花B病毒(CVB)的菊花品种‘寒小白’为试材,分别切取长度小于0.3 mm、0.3～0.7 mm和0.7～1.0 mm的茎尖接种于MS+2.0 mg·L-16-BA+0.5 mg·L-1NAA的培养基中,以及第1次切取0.7～1.0 mm菊花茎尖接种于MS培养基中,培养后再切取0.3～0.7 mm的菊花茎尖接种于MS+2.0 mg·L-16-BA+0.5 mg·L-1NAA培养基中(二次茎尖培养法),结合RT-PCR方法鉴定脱毒效果,研究茎尖长度及培养方式对茎尖诱导分化成活率和脱毒效果的影响。结果表明:茎尖大小与茎尖诱导成活率成正比,与脱毒效果成反比,即茎尖长度小于0.3 mm时,成活率最低,仅为6%,而脱毒率最高,可达66.67%;当茎尖长度为0.7～1.0 mm时,茎尖成活率最高,为90%,脱毒率最低,仅为6.67%。在一次茎尖培养法的基础上,运用二次茎尖培养法对CVB进行脱毒培养,既保证了较高的成活率(60%),又保证了较高的脱毒率(43.33%)。Chrysanthemum variety＇ Harbxiaobai ＇ plants infecting Chrysanthemum virus B （ CVB ） were made CVB-free plants by using meristem-tips culture. Different sizes（ 〈0.3 mm,0.3-0.7 mm,0.7-1.0 mm）of shoot tips were cultured on MS medium with 2.0 mg-L^-1 6-BA and 0.5 mg＇L-~ NAA,0.7-1.0 mm of shoot tips were first cultured on MS medium,and then 0.3-0.7 mm shoot tips cut from the plants were cultured on MS medium with 2.0 rag. L^-1 6-BA and 0.5 rag. L-l NAA （secondary culture）. In the secondary culture,tips of 0.7-1.0 mm were first cultured on MS medium,and then tips with 0.3-0.7 mm cut from the plants were cultured on MS medium with 2.0 mg-L^-1 6-BA and 0.5 mg-L^-1 NAA. The plants were indexed by RT-PCR. The result showed that in the first tissue culture, the size of shoot tip was proportional to the livability rate and in inverse proportion to the elimination of CVB,meristem- tips with length less than 0.3 mm, produced the lowest re-growth plants （6%）, but obtained the most virus-free plants （66.67%） ; meristem-tips 0.7-1.0 mm in length, produced the most re-growth plants （90%）, but obtained the least virus-free plants （6.67%）. Secondary tissue culturing was the most effective method to the elimination of CVB,since 60% plants survived and 43.33% virus-free plants can be obtained by this way.

关 键 词：菊花 菊花B病毒 检测 脱毒 

分 类 号：S682.1[农业科学—观赏园艺]