滁菊’病毒脱除及脱毒苗品质分析

[目的] 本文旨在比较3种脱毒方法对‘滁菊’病毒脱除的效果，并对‘滁菊’脱毒苗和非脱毒苗在遗传稳定性、产量和品质等方面的表现进行分析和比较。[方法] 采用电镜观察和巢式PCR两种方法检测‘滁菊’8种（类）病毒；比较茎尖分生组织培养、热处理结合茎尖分生组织培养及病毒唑结合茎尖分生组织培养3种脱毒方法的脱毒率，筛选出确定‘滁菊’病毒脱除的最适方法。采用SRAP分子标记技术检测脱毒苗和非脱毒苗的遗传稳定性，并以总黄酮、绿原酸以及鲜花产量等为检测指标，比较脱毒苗与非脱毒苗的产量品质。[结果] 病毒检测结果表明：侵染‘滁菊’的主要（类）病毒为菊花B病毒（CVB）和菊花矮化类病毒（CSVd）。热处理结合茎尖分生组织培养和病毒唑结合茎尖分生组织培养脱毒效果较单独采用茎尖分生组织培养脱毒效果好且差异显著，当2种（类）病毒CVB和CSVd都脱除时，脱毒率能达到35.21%、36.62%。3种脱毒方式均能保持‘滁菊’的遗传稳定性。采用脱毒‘滁菊’种苗生产，其在单株产量和品质上都显著优于非脱毒苗‘滁菊’，花直径、单株花数、单株产量分别比非脱毒苗高4.5%、21.2%、24.0%。[结论] 茎尖分生组织培养结合热处理或病毒唑处理脱除‘滁菊’病毒效果明显，采用脱毒‘滁菊’种苗生产能有效提高产量和品质。
[Objectives] We compared the effects of three different virus-elimination methods and made analysis and comparison in genetic stability, yield and quality of virus-free and non-virus-free seedlings.[Methods] Eight viruses or viroid infecting ‘Chuju’ were detected by electron microscopy and nested PCR;the effects of three virus-elimination methods were compared:apical meristem culture, thermotherapy combined with meristem culture, and ribavirin combined with meristem culture. We screened out the best method to remove the virus. The genetic stability of virus-free and non-virus-free seedlings was determined by SRAP molecular marker. The virus-free and non-virus-free seedlings were compared with the total flavonoids, chlorogenic acid and the amount of flowers as the detection index.[Results] The results of virus detection showed that the main viruses or viroid were Chrysanthemum virus B (CVB)and Chrysanthemum stunt virus (CSVd). Both methods of thermotherapy combined with meristem culture and ribavirin combined with meristem culture represented a significantly higher effects on virus-elimination than the method of apical meristem culture. When CVB and CSVd were removed, the induction rate reached 35.21%, 36.62%, and all of the three different virus-elimination methods could maintain genetic stability in ‘Chuju’. After using the virus-free ‘Chuju’ seedlings for production, the yield and quality per plant were both significantly better than non-virus-free seedlings, and flower diameter, the number of flowers per plant and yield per plant were 4.5%, 21.2% and 24.0% higher than non-virus-free seedlings, respectively.[Conclusions] The thermotherapy and chemical reagents combined with meristem culture are better than other methods for virus-elimination, and the yield and quality of seedling produced by the virus-free seed could be improved effectively