费乌瑞它马铃薯茎尖分化成苗的培养基筛选与病毒的RT―PCR检测.docVIP

费乌瑞它马铃薯茎尖分化成苗的培养基筛选与病毒的RT―PCR检测 　　摘要：以分别携带3种病毒（PVX、PVY、PLRV）和纺锤块茎类病毒（PSTVd）的费乌瑞它（Favorita）马铃薯（Solanum tuberosum）无菌苗为材料，38 ℃、4 h热处理4周，剥离带1个叶原基的茎尖，接种至不同浓度激素组合的24种MS固体培养基上，24 ℃、16 h培养15 d后统计茎尖的愈伤组织诱导率和分化成苗率;反转录PCR（RT-PCR）检测茎尖分化再生苗的脱毒率。结果表明，最适宜费乌瑞它马铃薯茎尖分化成苗的培养基为MS+0.075 mg/L 6-BA +0.05 mg/L NAA +0.10 mg/L GA3，愈伤组织诱导率为75%，分化成苗率为47.5%;再生苗3种病毒PVX、PVY和PLRV的脱毒率分别为65%、90%和100%，纺锤块茎类病毒PSTVd的脱毒率为10%。 　　关键词：费乌瑞它马铃薯（Solanum tuberosum）;茎尖;分化成苗;培养基;RT-PCR检测 　　中图分类号：S532 文献标识码：A 文章编号：0439-8114（2014）20-4988-04 　　DOI：10.14088/j.cnki.issn0439-8114.2014.20.060 　　Screening the Culture Medium of Favorita Potato Meristem Differentiation 　　Seedlings and RT-PCR Detection of Virus 　　FENG Guang-hui， DU Hu-ping， LI Xia-long， KANG Fu-ren 　　（College of Life Science，Yulin University， Yulin 719000，Shaanxi，China） 　　Abstract： Aseptic seedlings of Favorita potato carrying three kinds of potato viruses（PVX，PVY，PLRV） and potato spindle tuber viroid （PSTVd） were treated with 38 ℃，4 hours for 4 weeks， stripped with oneleaf primordia of potato meristem，inoculated with 24 kinds of MS solid media of different hormone combinations. The callus induction rate of meristem and differentiation seedling rate after cultured by 24 ℃，16 hours for 15 days were determined statistically. Reverse transcription PCR （RT-PCR） was used to detect the virus-free rate of meristem regeneration. The results showed that the best meristem differentiation medium was MS+0.075 mg/L 6-BA +0.05 mg/L NAA+0.10 mg/L GA3 with the callus inductionrate of 75% and differentiation seedling rate of 47.5%. The rate of detoxification of regenerated plantlets of three potato virus PVX，PVY and PLRV were 65%，90% and 100%. The virus-free rate of potato spindle tuber viroid was 10%. 　　Key words： Favorita potato; meristem; differentiation seedling; culture medium; RT-PCR detection 　　脱除马铃薯（Solanum tuberosum）病毒和纺锤块茎类病毒（以下简称类病毒）主要采用茎尖组织培养法，并结合热处理[1]、低温疗法[2]和病毒唑法[3]等提高脱毒效果。其中，热处理结合茎尖组织培养法是一种应用较广的马铃薯脱毒方法，在葡萄（Vitis vinifera）、草莓（Fragaria×ananas