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目的研究栽培藏红花的病毒病原。方法综合运用病毒粒子形态和细胞病理学电镜观察、DAS-ELISA检测、RT-PCR检测及序列测定等技术进行病原鉴定。结果透射电镜负染色观察到病株汁液含有600～900 nm的线状病毒粒子;超薄切片观察到病株细胞质内有大量线状病毒粒子、II型风轮状内含体和电子致密无定型体,符合菜豆黄花叶病毒Beamyellow mosaic virus(BYMV)的病理学特征。应用BYMV抗体进行病株DAS-ELISA检测结果为阳性,应用马铃薯Y病毒属特异性引物Sprimer和M4对病株进行RT-PCR检测结果为阳性,对阳性结果进行分子克隆及序列测定发现目标序列与BYMV有99%的同源性。结论综合检测结果判明侵染浙江藏红花的病毒病原为BYMV。
Objective To study the virus pathogen of saffron cultivation. Methods The pathogen was identified by morphological and cytopathological examination of virus particles, DAS-ELISA, RT-PCR and sequencing. Results Negative transmission electron microscopy showed that the strain juice contained linear virus particles of 600-900 nm. In the ultrathin section, a large number of linear virions were observed in the cytoplasm of the diseased plants. The type II rotundity inclusions and electron dense amorphous In line with the pathological features of Beamyellow mosaic virus (BYMV). The result of DAS-ELISA using BYMV antibody was positive. The results of RT-PCR test showed that the positive results were positive by RT-PCR using the specific primers of potato Y virus genus Sprimer and M4. The positive results were analyzed by molecular cloning and sequence analysis. BYMV has 99% homology. Conclusion The results of the comprehensive test showed that the virus pathogen in Zhejiang saffron was BYMV.