Phytopathology. 2006 May;96(5):437-46. doi: 10.1094/PHYTO-96-0437.
ABSTRACT Serological analysis of diseased chickpea and faba bean plantings with yellowing and stunting symptoms suggested the occurrence of an unknown or uncommon member of the family Luteoviridae in Ethiopia. Degenerate primers were used for reverse transcriptase-polymerase chain reaction amplification of the viral coat protein (CP) coding region from both chickpea and faba bean samples. Cloning and sequencing of the amplicons yielded nearly identical (96%) nucleotide sequences of a previously unrecognized species of the family Luteoviridae, with a CP amino acid sequence most closely related (identity of approximately 78%) to that of Groundnut rosette assistor virus. The complete genome (5,900 nts) of a faba bean isolate comprised six major open reading frames characteristic of polero-viruses. Of the four aphid species tested, only Aphis craccivora transmitted the virus in a persistent manner. The host range of the virus was confined to a few species of the family Fabaceae. A rabbit antiserum raised against virion preparations cross-reacted unexpectedly with Beet western yellows virus-like viruses. This necessitated the production of murine monoclonal antibodies which, in combination with the polyclonal antiserum, permitted both sensitive and specific detection of the virus in field samples by triple-antibody sandwich, enzyme-linked immunosorbent assay. Because of the characteristic field and greenhouse symptoms in chickpea, the name Chickpea chlorotic stunt virus is proposed for this new member of the genus Polerovirus (family Luteoviridae).
摘要 对埃塞俄比亚出现叶片发黄和植株矮小症状的患病鹰嘴豆和蚕豆种植园进行血清学分析,表明在这些地区有一种尚未被认识或不常见的布罗克病毒科成员。利用简并引物,通过反转录-聚合酶链式反应(RT-PCR),从鹰嘴豆和蚕豆样本中扩增病毒外壳蛋白(CP)编码区。对扩增产物进行克隆和测序,得到了一个以前未被识别的布罗克病毒科新种的近相同(96%)核苷酸序列,其 CP 氨基酸序列与花生丛矮病毒(Groundnut rosette assistor virus)最为接近(同一性约为 78%)。一株蚕豆分离物的完整基因组(5900 个核苷酸)包含了与多粒病毒(polero-viruses)特征一致的 6 个主要开放阅读框。在所测试的 4 种蚜虫中,只有桃蚜(Aphis craccivora)以持久方式传播该病毒。该病毒的宿主范围局限于少数豆科植物。针对病毒粒子制剂制备的兔抗血清出人意料地与西方黄条叶甲类病毒(Beet western yellows virus-like viruses)发生交叉反应。这就需要制备鼠源单克隆抗体,并用该单克隆抗体与多克隆抗血清结合,通过三抗体夹心酶联免疫吸附试验(ELISA),实现对田间样本中病毒的灵敏、特异检测。鉴于在鹰嘴豆上的特征性田间和温室症状,建议将这种新的多粒病毒属(Polerovirus)成员命名为鹰嘴豆花叶矮缩病毒(Chickpea chlorotic stunt virus)。