重庆卷丹病毒检测与脱毒方法研究

Virus Detection of Lilium lancifolium Thunb.in Chongqing and Study on Its Detoxification Technology

为建立一种卷丹(Lilium lancifolium Thunb.)的脱毒快繁技术。以重庆南川种植的卷丹为材料,通过RT-PCR检测表明,其主要含百合斑驳病毒(Lily mottle virus,LMoV)和百合无症病毒(Lily symptomless virus,LSV)。采用38℃和4℃高低温交替处理卷丹珠芽,通过连续2次直剥1.0~1.5 mm的茎尖接种到10 mg/L病毒唑培养基中培养,检测脱毒效果。结果表明,该方法能有效脱除LMoV和LSV病毒,脱毒率分别为86%和88%,茎尖成苗率均为90%。利用无毒幼苗外层鳞片在培养基(MS+1.5 mg/L 6-BA+0.2 mg/L NAA+30 g/L蔗糖)中进行组培效果最佳,芽诱导系数达6.35;不定芽在培养基(MS+2.5 mg/L 6-BA+0.3 mg/L NAA+30 g/L蔗糖)中增值最显著,增殖系数为3.39;鳞片在添加50 g/L蔗糖的继代培养基中生根效果最优,生根率为98.72%。可获取大量脱毒幼苗,方法简便,可操作性强,为重庆卷丹产业发展提供技术支持。

The purpose of this study is to establish a detoxification and rapid propagation technique for Lilium lancifolium Thunb.the RT-PCR method was used to dectect Lily mottle virus(LMoV)and Lily symptomless virus(LSV)in L.lancifolium Thunb.in Nanchuan,Chongqing.The bulbils of L.lancifolium Thunb.were treated with higher(38℃)and lower(4℃)temperatures alternatively,then the 1.0-1.5 mm shoot tips were stripped and cultured on the medium supplemented with 10 mg/L ribavirin.After the shoot tips grew into seedlings,1.0-1.5 mm shoot tips were stripped again and cultured on the same medium to detect the detoxification effect.The results showed that LMoV and LSV viruses could be removed and the detoxification rate was 85%.In addition,shoot tip survival rate was significantly improved to more than 90%.The outer scales of the virus-free seedlings were cultured on the MS medium supplemented with 1.5 mg/L 6-BA and 0.2 mg/L NAA,and the bud induction coefficient could reached 6.35.Adventitious bud proliferation rate was 3.39 when cultured on the MS medium supplemented with 2.5 mg/L 6-BA and 0.3 mg/L NAA.The best rooting medium was MS medium supplemented with 50 g/L sucrose,and the rooting rate was 98.72%.The results in the paper provide a simple and effective method for propagating a large number of virus-free seedlings,which is important for the development of Lilium industry in Chongqing.