水鬼蕉感染HCRV不同发病时期的病毒粒体分布与亚细胞病变特征  

Virions distribution and sub-cellular pathological characteristics at different stages of Hymenocallis littoralis infected with Hippeastrum chlorotic ringspot orthotospovirus

作　　者：张涵 王茂森 王田田 朱若萌 童金金 李妤 ZHANG Han;WANG Maosen;WANG Tiantian;ZHU Ruomeng;TONG Jinjin;LI Yu(College of Biology,China Agricultural University,Beijing 100193,China;Biotechnology and Germplasm Resources Institute,Yunnan Academy of Agricultural Sciences/Yunnan Province Key Laboratory of Agricultural Biotechnology,Kunming 650205,China)

机构地区：[1]中国农业大学生物学院,北京100193 [2]云南省农业科学院生物技术与种质资源研究所/云南省农业生物技术重点实验室,昆明650205

出　　处：《植物病理学报》2023年第4期563-569,共7页Acta Phytopathologica Sinica

基　　金：国家自然科学基金-云南联合资助重点项目(U1802235)。

摘　　要：采集疑似朱顶红褪绿环斑病毒(HCRV)侵染的水鬼蕉(Hymenocallis littoralis)不同发病时期的叶部组织,通过HCRV-N基因特异性引物的RT-PCR扩增克隆与测序分析,确定由HCRV侵染。进一步应用负染色、超薄切片制样及透射电子显微镜观察,结果显示:叶部无症、褪绿、黄化和坏死不同发病时期的HCRV粒体在细胞内呈现散布或聚集的特征,散布的病毒粒体包被双层膜结构,发病后期聚集的病毒粒体呈现串珠状,聚集于管状囊泡中。在侵染潜伏期,水鬼蕉细胞核、叶绿体等亚细胞结构较完整;发病早期和中期,细胞核结构完整,叶绿体基粒片层消解,线粒体增加且结构较完整;发病后期,大部分细胞坏死,液泡中残存内溶物,少部分细胞的液泡中含有串珠状病毒粒体聚集于管状囊泡,是HCRV区别于同属其他病毒的明显特征,可作为诊断鉴定的依据。In this study,leaf tissues of Hymenocallis littoralis(Jacq.)Salisb.suspected to be infected with Hippeastrum chlorotic ringspot orthotospovirus(HCRV)were collected at different stages of the disease.The infection of HCRV was confirmed by RT-PCR amplification,cloning and sequencing analysis of HCRV-N gene-specific primers.Further,negative staining,ultra-thin section sample preparation and transmission electron microscopy(TEM)analyses were performed.The results showed that HCRV virions at different stages(symptomless or with symptoms of chlorosis,yellowing and necrosis)showed the characteristics of dispersion or aggregation.The dispersed virions with a double-membrane structure,and virions in the late stage of diseased cells showed a beaded shape and gathered in tubular vesicles.During the incubation period of virus infection,the nucleus,chloroplast and other subcellular structures remained intact.At the early and middle stages of the diseased cells,the nucleus structure was intact,the chloroplast grana lamella dissolved,and the number of mitochondria increased exhibiting an intact structure.At the late stage of the infection,most of the cells were necrotic,and there were residual endosomes in the vacuoles,and the vacuoles of a few cells contained beaded virions clustered in tubular vesicles,suggesting that the characteristics of HCRV were obviously different from viruses of the same genus and could be used as the basis for diagnosis and identification.

关 键 词：朱顶红褪绿环斑病毒 水鬼蕉 病毒粒体 亚细胞结构 

分 类 号：S432.41[农业科学—植物病理学]