摘要
番木瓜环斑病毒(Papaya ringspot virus,PRSV)与番木瓜畸形花叶病毒(Papaya leaf distortion mosaic virus,PLDMV)存在混合感染的现象,近年来混合感染发病率逐年递增,是一种新的威胁番木瓜种植业的病毒病害。运用RT-PCR和RACE方法从混合感染样品中克隆获得2种病毒基因组全长c DNA,分别命名为PRSV-LM和PLDMV-LM,Gen Bank登录号分别为KT633943和KT633944,基因组序列大小分别为10 325、10 153 nt(不包括3′端的poly A)。序列分析表明:PRSV-LM与单独感染样品中的PRSV海南分离物HN(Gen Bank登录号:EF183499)的核苷酸序列和氨基酸序列相似性最高,为94%;PLDMV-LM与单独感染样品中的PLDMV海南分离物Hainan-DF(Gen Bank登录号:JX974555)核苷酸和氨基酸序列相似性最高,分别高达99%和97%。进化树分析显示,PRSV-LM和PLDMV-LM分别与单独感染样品的PRSV和PLDMV海南分离物有共同的进化起源。进一步利用实时荧光定量PCR对混合感染样品的PRSV与PLDMV病毒积累量进行分析,结果表明,混合感染样品中PLDMV病毒积累量大约是PRSV含量的100倍。研究结果为进一步探究PRSV/PLDMV混合感染发病机制奠定基础。
The incidence of the mixed infection between Papaya ringspot virus（PRSV） and Papaya leaf distortion mosaic virus（PLDMV） increases in recent years, and it is becoming a new threat to papaya farming. Two full-length c DNAs, named as PRSV-LM（total 10 325 nucleotieds, not including the poly（A） tail, Gen Bank accession number： KT633943） and PLDMV-LM（total 10 153 nucleotieds, not including the poly（A） tail, Gen Bank accession number： KT633944）, were cloned from mixed infection using RT-PCR and rapid-amplification of c DNA ends（RACE） techniques. Sequence analysis indicated that the similarity between PRSV-LM and PRSV single infectious isolate HN（Gen Bank accession number： EF183499） was 94% at the full-length nucleotide sequences and amino acid sequences, meanwhile PLDMV-LM and PLDMV single infectious isolate Hainan-DF（Gen Bank accession number：JX974555） reached up to 99% and 97%. To further understanding the hypotaxis between mixed infection and single infection, phylogenetic trees were constructed. It indicated that, PRSV-LM and PLDMV-LM isolate respectively, had a common evolutionary origin with Hainan single infection isolates. Further analysis of Real-Time PCR indicated that,in mixed infection samples, the quantitation of PLDMV-LM was approximately 100 times than that of PRSV-LM.This study found a foundation for deep understanding the pathogenesis of PRSV and PLDMV mixed infection.
出处
《热带作物学报》
CSCD
北大核心
2016年第4期742-751,共10页
Chinese Journal of Tropical Crops
基金
国家自然科学基金(No.31171822
No.31371918)
海南省重大科技专项(No.ZDZX2013023)
