Lecoq H, Dufour O, Wipf-Scheibel C, Girard M, Cotillon A C, Desbiez C
INRA, Station de Pathologie Végétale, Domaine St Maurice, BP94, 84143, Montfavet cedex, France.
LNPV, Domaine St Maurice, BP94, 84143, Montfavet cedex, France.
Plant Dis. 2007 Jul;91(7):909. doi: 10.1094/PDIS-91-7-0909C.
During the fall of 2003, mild mosaic symptoms were observed in melon (Cucumis melo L.) plants grown in glasshouses near Eyragues (southeastern France) resembling those caused by the Bemisia tabaci transmitted Cucumber vein yellowing virus (CVYV, genus Ipomovirus, family Potyviridae). In addition, large numbers of B. tabaci were observed to be colonizing these crops. The identification of CVYV was established through differential host range reaction, immunosorbent electron microscopy (IEM), and reverse transcription (RT)-PCR experiments. Crude sap from symptomatic leaves was used to inoculate differential host plants. Mild mosaic symptoms were observed on melon, and cucumber developed vein-clearing symptoms typical of CVYV. No symptoms were observed in Chenopodium quinoa, C. amaranticolor, Nicotiana benthamiana, N. tabacum, and Vigna sinensis. Numerous, slightly flexuous, elongated virus particles were observed in infected plant extracts; these particles were decorated by a polyclonal antiserum raised against a Sudanese CVYV isolate. To confirm CVYV identification, total RNA extracts (TRI-Reagent, Sigma Chemical, St. Louis, MO) were obtained from the original symptomatic melon tissues. RT-PCR was carried out using CVYV-specific primers CVYV-CP-5': 5'-GCTTCTGGTTCTCAAGTGGA-3' and CVYV-CP- 3': 5'-GATGCATCAGTTGTCAGATG-3' designed according to the partial sequence of the coat protein gene of CVYV-Isr (GenBank Accession No. AF233429) (2). A 540-bp fragment corresponding to the central region of CVYV coat protein was amplified from total RNA extracted from symptomatic but not from asymptomatic melon tissue. Direct sequencing was done on RT-PCR products (GenBank Accession No. EF441272). The sequence was 95 and 99% identical to that reported for CVYV isolates from Israel and Spain, respectively. CVYV was first described in Israel and has recently emerged as the cause of important diseases in Spain and Portugal (1,3). Shortly after detecting CVYV during 2003, efforts were made to eradicate the virus in susceptible crops. CVYV was not detected again during intensive surveys conducted in southeastern France during 2004, 2005, and 2006, suggesting that the CVYV detected during 2003 resulted from an accidental introduction and that the virus has not become established in France. References: (1) I. M. Cuadrado et al. Plant Dis. 85:336, 2001. (2) H. Lecoq et al. J. Gen. Virol. 81:2289, 2000. (3) D. Louro et al. Plant Pathol. 53:241, 2004.
2003年秋季，在法国东南部埃拉格附近温室种植的甜瓜（Cucumis melo L.）植株上观察到轻度花叶症状，类似于由烟粉虱传播的黄瓜叶脉黄化病毒（CVYV，甘薯病毒属，马铃薯Y病毒科）引起的症状。此外，还观察到大量烟粉虱在这些作物上定殖。通过鉴别寄主范围反应、免疫吸附电子显微镜（IEM）和逆转录（RT）-PCR实验确定了CVYV的身份。用有症状叶片的粗汁液接种鉴别寄主植物。在甜瓜上观察到轻度花叶症状，黄瓜出现了CVYV典型的叶脉褪绿症状。在藜麦、苋色藜、本氏烟草、烟草和豇豆上未观察到症状。在受感染的植物提取物中观察到许多稍有弯曲的细长病毒粒子；这些粒子被针对苏丹CVYV分离物产生的多克隆抗血清标记。为了确认CVYV的身份，从最初有症状的甜瓜组织中提取总RNA提取物（TRI试剂，西格玛化学公司，密苏里州圣路易斯）。使用根据CVYV-Isr外壳蛋白基因部分序列设计的CVYV特异性引物CVYV-CP-5′：5′-GCTTCTGGTTCTCAAGTGGA-3′和CVYV-CP-3′：5′-GATGCATCAGTTGTCAGATG-3′进行RT-PCR。从有症状但无症状的甜瓜组织中提取的总RNA中扩增出一个与CVYV外壳蛋白中心区域相对应的540 bp片段。对RT-PCR产物进行直接测序（GenBank登录号EF441272）。该序列与以色列和西班牙报道的CVYV分离物序列分别有95%和99%的同一性。CVYV最早在以色列被描述，最近在西班牙和葡萄牙成为重要病害的病因（1,3）。在2003年检测到CVYV后不久，就努力在易感作物中根除该病毒。在2004年、2005年和2006年对法国东南部进行的密集调查中未再次检测到CVYV，这表明2003年检测到的CVYV是偶然引入的，并且该病毒在法国尚未定殖。参考文献：（1）I.M.Cuadrado等人，植物病害，85:336，2001年。（2）H.Lecoq等人，普通病毒学杂志，81:2289，2000年。（3）D.Louro等人，植物病理学，53:241，2004年。
