不同外植体对草莓病毒脱除效果及病毒的多重RT-PCR检测

EFFECT OF EXPLANTS ON STRAWBERRY VIRUS ELIMINATION OF STRAWBERRY AND VIRUSES DETECTION BY MULTIPLEX RT-PCR

以草莓茎尖和花药为外植体,多重RT-PCR为检测方法研究了不同培养材料对3种草莓病毒,即斑驳病毒(strawberry mottle virus,SMoV)、轻型黄边病毒(strawberry mild-yellow edge virus,SMYEV)和镶脉病毒(strawberry vein-band virus,SVBV)的脱除情况。结果表明,花药愈伤组织再生苗途径对3种病毒的脱除率为100%。而不同大小茎尖脱除病毒效果不同:0.2mm茎尖可完全脱除3种病毒,但成苗率低;0.5mm茎尖可有效脱除SMoV和SVBV,但对SMYEV脱除不彻底;0.5mm茎尖成苗率明显高于0.2mm茎尖,但较难脱除SMYEV。综合病毒脱除率和成苗率,对花药再生频率较高的草莓基因型(如红颊)可以采用花药愈伤组织再生途径进行病毒脱除,而对于花药再生困难的基因型可以考虑选用茎尖分生组织途径获取脱毒苗。

The effects of explant, culture approach of both anther and shoot tip on strawberry virus elimination was studied. Three strawberry viruses, strawberry mottle virus（SMoV）, strawberry mild-yellow edge virus （SMYEV）and strawberry vein-band virus（SVBV）were detected with multiplex RT-PCR technique. The results showed that the elimination rates of three viruses were 100% by anther culture. The size of shoot tip influenced affected the efficiency of viruses elimination no virus was found from explant of 0.2mm size tip. The SMoV and SVBV could be eliminated effectively in 0.5mm shoot tip, however SMYEV was difficult to eliminate. The shoot forming rate of 0.2mm shoot tip was lower than that of 0.5mm shoot tip. Considering the rate of virus elimination and the rate of regeneration of plantlets, it was feasible to eliminat strawberry viruses using anther as explant for genotype with high regeneration ability of an ther such as Benihope, and using shoot tip as explant suitable for genotype as Toyonoka, which has low regeneration ability of anther.