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作 者:谢丽雪[1] 张立杰[1] 张小艳[1] 郑姗[1] 李韬[1] XIE Lixue;ZHANG Lijie;ZHANG Xiaoyan;ZHENG Shan;LI Tao(Fruit Research lnstitute,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China)
机构地区:[1]福建省农业科学院果树研究所,福州350013
出 处:《园艺学报》2018年第8期1587-1594,共8页Acta Horticulturae Sinica
基 金:国家重点研发计划资助项目(2016YFF0203200);福建省现代农业水果产业技术体系岗位项目(闽农科教[2017]129);福建省农业科学院新兴特色果类创新团队项目(STIT2017-2-4);福建省农业科学院院管A类项目三农业一融合项目(A2017-15)
摘 要:为明确东亚西番莲病毒(East Asian passiflora virus,EAPV)在福建西番莲上的发生情况,采用血清学和RT-PCR检测技术对采集的西番莲病样进行检测,并对阳性样品PCR产物进一步克隆、测序和序列分析。结果表明,11份样品的马铃薯Y病毒属(Potyvirus)病毒血清学检测结果为阳性,大豆花叶病毒(Soybean mosaic virus,SMV)血清学检测结果均为阴性;应用Potyvirus属通用引物从11份样品中扩增得到预期大小约660 bp的目的片段,序列比对发现,其中10份阳性样品与已报道的夜来香花叶病毒(Telosma mosaic virus,TeMV)核苷酸序列高度一致,1份样品与已报道的EAPV核苷酸序列一致性超过98.3%。针对EAPV疑似样品(命名为FJBXG-P1),利用特异性引物扩增得到预期大小约955 bp的目的片段,获得的外壳蛋白(Coat protein,CP)基因序列全长870 bp(Gen Bank登录号:MG650164)。序列比对发现,EAPV分离物FJBXG-P1的CP基因序列与已报道的EAPV核苷酸序列、氨基酸序列一致性分别为81.0%~97.6%和84.0%~96.9%;系统发育分析结果表明,38个EAPV分离物在系统发育关系上聚为两簇(GroupⅠ和GroupⅡ),其中GroupⅠ为AO株系,GroupⅡ为IB株系,FJBXG-P1分离物属于AO株系。福建西番莲上EPAV的检出,是该病毒在中国大陆地区发生的首次报道。In order to investigate the occurrence of East Asian passiflora virus(EAPV)on Passiflora edulis in Fujian Province,the suspected samples of Passiflora edulis were detected by serological and RT-PCR technology,and the positive samples were further cloned,sequenced and analyzed. The results showed that the serological test of 11 samples were positive for detection of Potyvirus,but all were negative for detection of Soybean mosaic virus(SMV). The expected fragments of about 660 bp in size were amplified from 11 samples using universal degenerate primers for detection of virus species from the genus Potyvirus. Results of sequence determination and analysis revealed that the sequences of 10 samples were highly consistent with those of the reported Telosma mosaic virus(TeMV)isolates,and the sequence of one sample shared more than 98.3% nucleotide identity with those of the reported EAPV isolates. For the suspected EAPV sample(named FJBXG-P1),the expected size of ≈ 955 fragment was amplified using specific primers. The total length of CP gene of this EAPV-positive sample was 870 nucleotides(Gen Bank accession number:MG650164). The sequence of CP gene of FJBXG-P1 were 81.0%–97.6% and 84.0%–96.9% identity with the reported nucleotide sequence and amino acid sequence of EAPV,respectively. Phylogenetic analysis showed that 38 EAPV isolates were clustered into two groups(GroupⅠ and GroupⅡ),in which GroupⅠ was AO strain and Group Ⅱ was IB strain. FJBXG-P1 isolate in this study belonged to AO strain. To the best of our knowledge,EPAV infected Passiflora edulis in Fujian Province is the first report of the virus in China's Mainland.
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