Identification, characterisation and detection of a new tospovirus on sweet pepper
Identification, characterisation and detection of a new tospovirus on sweet pepper
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DOI：
10.1111/aab.12084
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发表时间：
2014-01-01
影响因子：
2.6
通讯作者：
Jan, F. -J.
中科院分区：
文献类型：
--
作者：
Cheng, Y. -H.;Zheng, Y. -X.;Jan, F. -J.
2009年在台湾观察到一种假定的病毒引起的疾病，在甜椒的叶子和果实上显示斑点和变形。用西瓜银斑驳病毒（WSMoV）血清群中的番茄斑萎病毒（Tospovirus）抗血清对有症状的甜椒叶片的粗汁液进行酶联免疫吸附测定（ELISA），结果呈阳性反应。从有症状的甜椒中分离的病毒培养物TwPep3通过电子显微镜显示直径约80 - 100 nm的等距病毒粒子。接种TwPep3的甜椒幼苗表现出与田间观察到的症状相似的症状。用番茄斑萎病毒属L RNA保守区的简并引物可以扩增出TwPep3分离物0.9kb的病毒基因组，而用黄瓜花叶病毒属、马铃薯Y病毒属和黄瓜花叶病毒属的简并引物则不能扩增出该病毒基因组。TwPep3（KF383955）与WSMoV和番茄坏死环斑病毒（TNRV）的部分LRNA序列同源性分别为75.1%和82.4%。为了确定分离物TwPep3的分类学关系，对分离物KF383956的S RNA进行了全序列测定。序列分析表明，TwPep3的N基因与5个与该病毒关系最密切的TNRV分离物的N基因核苷酸和氨基酸序列同源性分别为82 - 83%和87.9 - 89%。此外，TwPep3和TNRV的NSs蛋白和S RNA的氨基酸和核苷酸序列同源性分别为78.7%和77.6%。根据我们的研究结果，分离物TwPep3应被认为是一个新的番茄斑萎病毒，我们建议将其命名为辣椒褪绿斑点病毒（PCSV）。针对N基因设计的特异性引物成功地应用于PCSV的逆转录-聚合酶链反应诊断。
A putative virus-induced disorder showing mottling and deformation on leaves and fruits of sweet pepper was observed in Taiwan in 2009. Crude sap of symptomatic sweet pepper leaves reacted positively in enzyme-linked immunosorbent assay (ELISA) with antisera to tospoviruses in the Watermelon silver mottle virus (WSMoV) serogroup. A virus culture, TwPep3, isolated from the symptomatic sweet pepper revealed isometric virions measuring about 80-100nm in diameter via electron microscopy. Seedlings of sweet pepper inoculated with TwPep3 showed similar symptoms as those observed in the field. A 0.9-kb viral genome of isolate TwPep3 could be amplified using degenerate primers for L RNA conserved region of tospoviruses but not using those of tobamoviruses, potyviruses and Cucumber mosaic virus. The partial L RNA sequence of isolate TwPep3 (KF383955) shared 75.1% and 82.4% nucleotide identities with those of WSMoV and Tomato necrotic ringspot virus (TNRV), respectively. To determine the taxonomic relationship of isolate TwPep3, the full length of S RNA (KF383956) was sequenced. Sequence analyses showed that the nucleocapsid (N) gene of TwPep3 shared 82-83% and 87.9-89% nucleotide and amino acid sequence identities, respectively, with those of five isolates of TNRV, the most closely related tospovirus. Moreover, the NSs proteins and S RNAs of TwPep3 and TNRV shared only 78.7% and 77.6% amino acid and nucleotide sequence identities, respectively. Based on our results, isolate TwPep3 should be considered as a new tospovirus and we propose the name Pepper chlorotic spot virus (PCSV). A specific primer pair designed in the N gene was successfully used in reverse transcription-polymerase chain reaction for PCSV diagnostic work.