张金珠 邹承武 黄俊源 邓崇岭 陈保善 张木清

摘  要：調查了广西南宁、崇左、柳州、桂林、玉林、河池、贺州及防城港等主要柑橘种植区的柑橘碎叶病和叶斑病的发生情况，并对其病原柑橘碎叶病毒（Citrus tatter leaf virus， CTLV）、苹果茎沟病毒（Apple stem grooving virus， ASGV）和柑橘叶斑病毒（Citrus leaf blotch virus， CLBV）进行了RT-PCR检测、病毒序列测定和遗传多样性分析。结果发现，在所有疑似柑橘病毒病样品中，CTLV检出率为7.3%，CLBV检出率为5.5%，ASGV检出率为3.0%。其中在南宁、桂林和玉林3个主要柑橘种植区采集的疑似柑橘病毒病样品的3种病毒病检出率合计分别为21.2%、28.6%和7.6%，且存在多种病毒复合侵染的现象。将克隆到的上述3个病毒分离物的病毒片段，分别与已报道的相应病毒的代表性分离物的病毒片段进行系统进化分析，结果表明，ASGV和CTLV的中国分离物在进化关系上均呈寄主相关性，ASGV的日本分离物则与地理位置相关性更强，而韩国和印度的分离物未表现出明显的寄主相关性和地理位置相关性。柑橘和苹果的ASGV和CTLV分离物均表现与地理位置相关，而梨的ASGV和CTLV分离物却与地理位置和寄主均不相关;CLBV分离物则呈明显的寄主相关性。本研究首次报道了广西柑橘碎叶病和柑橘叶斑病的发生情况及其病原的遗传多样性与系统进化关系，为柑橘病毒病的诊断和防治提供参考依据。

关键词：柑橘;病毒病调查;病毒检测;遗传多样性分析

中图分类号：S436.661.1      文献标识码：A

Abstract： The occurrence of citrus tatter leaf disease and citrus leaf spot disease were investigated in the main planting areas of citrus in Nanning， Chongzuo， Liuzhou， Guilin， Yulin， Hechi， Hezhou， and Fangchenggang of Guangxi. RT-PCR detection， viral genome sequencing and genetic diversity analysis were carried out for the pathogenic viruses， including Citrus tatter leaf virus （CTLV）， Apple stem grooving virus （ASGV）， and Citrus leaf blotch virus （CLBV）. The results showed that the detection rates of CTLV， ASGV， and CLBV in suspected viral samples collected from the ci-trus-producing areas in Nanning， Guilin， and Yulin was 21.2%， 28.6%， and 7.6%， respectively. Additionally， the mixed infection of multiple viruses was detected. Among the samples tested， the detection rate of CTLV was the highest （7.3%）， followed by CLBV （5.5%）， and ASGV （3.0%）. The genetic diversity analysis and phylogenetic analysis were carried out by comparing the sequences of the three virus isolates cloned in this study with those of several repre-sentative viruses reported. The results demonstrated that both ASGV and CTLV isolates in China presented host correlation in the evolutionary relationship， while ASGV isolates in Japan showed a stronger association with geographical location. Still， the strains in South Korea and India did not show significant host or geographical location correlation. The isolates of ASGV and CTLV on citrus and apple presented a relationship with geographic location， while their isolates on pear showed no association with geographic location or host. Moreover， all strains of CLBV showed a distinct relationship with the host. It is the first time to report the occurrence of citrus tatter leaf disease and citrus leaf spot disease in Guangxi and the genetic diversity of their pathogenic viruses， which would provide a reference for the diagnosis and control of citrus virus diseases.

Keywords： citrus; viral disease investigation; virus detection; analysis of genetic diversity

DOI： 10.3969/j.issn.1000-2561.2021.05.030

近年来广西柑橘产业发展迅猛，种植面积和产量均居全国首位。但是生产上大量使用嫁接苗，容易导致柑橘病毒病流行，严重威胁柑橘产业的健康发展[1]。……