[Objective] The objective of this experiment is to explore a simple-operate method to eradicate viruses of Chongming Daffodil (Narcissus tazetta var.chinensis),and detect the effect of virus-free technology.[Method] The treatments of DHT+thermotherapy and DHT+virazole were used to eliminate viruses in bulblets in vitro of Chongming Daffodil,and the effect of treatments were tested by indicator plant and RT-PCR.[Result]Thermotherapy treatment had effect on the growth and survival rate of bulblets in vitro.Thermotherapy with DHT treatment decreased the content of viruses,and when the concentration of DHT was less than 15 mg·L-1,there was less inhibitory effect on growth.Virazole at 30 mg·L-1+DHT 30 mg·L-1 had good inhibitory effect on NMaV,and there was a very low level of virus detected by RT-PCR.NDV and NCLV had higher sensitivity to two antiviral compounds,and could be mostly eradicated.[Conclusion] There was a negative effect on thermotherapy treatment.DHT with thermotherapy treatment could decrease the content of viruses detected by indicator plant.The content of NMaV in bulblets in vitro treated with 30 mg L-1DHT +30 mg L-1vrazole was reduced to a very low level,and the treatment had no impact on bulblet's survival.NDV-free and NCLV-free plants were obtained by the treatment of combination with 10 mg L1DHT and 10 mg L1virazole,this result was proved by RT-PCR detection.%[目的]探索操作简单且高效的方法脱除或减弱崇明水仙携带的病毒,为脱毒球生产奠定基础.[方法]采用DHT+热处理、DHT+病毒唑处理等化学药剂与物理手段结合的方法,对崇明水仙试管球进行离体脱毒研究,脱毒结果采用指示植物法和RT-PCR检测法验证.[结果]热处理影响崇明水仙试管球生长和存活；热处理+DHT处理后,病毒含量降低,DHT浓度小于15 mg.L-1,对试管球生长抑制作用较小.DHT30 mg·L-1+病毒唑30 mg.L-1处理,对崇明水仙NMAV病毒,起到了很好的抑制作用,用RT-PCR技术只检测到了极其微弱的病毒残留；NDV和NCLV对两种药剂较敏感,在10 mg·L-1时,RT-PCR检测不到病毒的特异谱带.[结论]热处理对崇明水仙试管球生长和存活有不利影响；DHT+热处理能明显降低病毒含量；DHT30 mg·L-1+病毒唑30 mg·L-1处理能将NMaV含量降低到极微弱的水平,不影响试管球存活,可通过增加两种药剂的浓度或多次继代来彻底脱除;DHT10 mg·L-1+病毒唑10 mg·L-1处理可以显著降低NDV和NCLV病毒含量, RT-PCR检测,没有出现病毒的特异谱带.
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