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危地马拉甜瓜中检测到的疾病的病毒病因

Viral Etiology of Diseases Detected in Melon in Guatemala.

作者信息

Jordá C, Font M I, Martínez-Culebra P, Tello J

机构信息

Departamento de Ecosistemas Agroforestales, Universidad Politécnica de Valencia, Cno. de Vera s/n, 46022 Valencia, Spain.

Departamento Producción Vegetal, Universidad de Almería, Cañada de San Urbano s/n 04120. Almería, Spain.

出版信息

Plant Dis. 2005 Mar;89(3):338. doi: 10.1094/PD-89-0338A.

Abstract

At the beginning of 1999, 30 melon (Cucumis melo L.) plots on several farms (1,500 ha) in the Zacapa Valley of Guatemala were visited, and melon plants with two different symptomologies were observed. One group of plants exhibited stem necrosis at the crown level, and less frequently, small necrotic spots on leaves. Some plants exhibited necrosis of veins and yellow areas that evolved into interveinal necrosis and often expanded into large necrotic interveinal lesions. Roots were poor and lacked secondary rootlets. In some cases, wilt and plant death were detected. Affected plants appeared as localized patches in various areas of the plots on farms that were visited. Double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) serological analyses were carried out with 34 symptomatic plants. In these plants, a mixture of the crown and root were analyzed with two repetitions and two lots of different Melon necrotic spot virus (MNSV) polyclonal antisera (Loewe No. 07097 and Sanofi No. 70217). All 34 plants were positive for this virus. These results were confirmed using reverse transcription-polymerase chain reaction (RT-PCR) with specific MNSV primers (1). Spores of Olpidium bornovanus, the vector of MNSV, were seen on all ELISA-positive plants after staining rootlets with potassium hydroxide and neutralization with hydrochloric acid. In the same fields, another group of melon plants showed yellowing, curling, and mottling of leaves. Leaves collected from five symptomatic plants gave positive results in triple-antibody sandwich-ELISA using a Tomato yellow leaf curl begomovirus antiserum (DSMZ AS-0421 and DSMZ AS-0546/2). In 2001, these results were confirmed using PCR with degenerate primers that amplify the core region of most begomovirus coat protein genes (P. Martínez-Culebras, M. I. Font, and C. Jordá, unpublished). A 560-bp DNA fragment was amplified in these symptomatic melon samples. Three of the PCR products were sequenced and each showed 99% identity with the Melon chlorotic leaf curl virus isolate from Guatemala (GenBank Accession No. AF325497). Only one mixed infection of MNSV and MCLCV was found. During the four years subsequent to 1999, the number of melon plants showing both types of symptoms has increased. This study provides information on the current status of virus diseases in melon crops in Guatemala, and to our knowledge, this is the first report of MNSV in Guatemala. Reference: (1) B. Gosalvez et al. J. Virol. Methods 113:87.

摘要

1999年初,对危地马拉萨卡帕山谷几个农场(1500公顷)的30块甜瓜(黄瓜属甜瓜种)地进行了考察,观察到有两种不同症状的甜瓜植株。一组植株在茎基部出现坏死,叶片上偶尔出现小坏死斑。一些植株叶脉坏死,出现黄色区域,进而发展为脉间坏死,常扩展成大片脉间坏死病斑。根系发育不良,缺少次生根。在某些情况下,检测到植株萎蔫和死亡。在考察的农场中,受影响的植株在地块的不同区域呈局部斑块状分布。对34株有症状的植株进行了双抗体夹心酶联免疫吸附测定(DAS-ELISA)血清学分析。在这些植株中,对茎基部和根系混合样本进行了两次重复分析,并使用了两批不同的甜瓜坏死斑病毒(MNSV)多克隆抗血清(罗威公司07097号和赛诺菲公司70217号)。所有34株植株对该病毒检测呈阳性。使用MNSV特异性引物通过逆转录-聚合酶链反应(RT-PCR)证实了这些结果(1)。在用氢氧化钾对根进行染色并用盐酸中和后,在所有ELISA阳性植株上均发现了MNSV载体——瓜类油壶菌的孢子。在同一地块,另一组甜瓜植株表现出叶片发黄、卷曲和斑驳。从5株有症状的植株上采集的叶片,使用番茄黄化曲叶双生病毒抗血清(德国微生物和细胞培养物保藏中心AS-0421和德国微生物和细胞培养物保藏中心AS-0546/2)进行三抗体夹心ELISA检测呈阳性。2001年,使用简并引物进行PCR扩增,证实了这些结果,这些引物可扩增大多数双生病毒外壳蛋白基因的核心区域(P. 马丁内斯-库莱布拉斯、M. I. 丰特和C. 霍尔达,未发表)。在这些有症状的甜瓜样本中扩增出一个560碱基对的DNA片段。对其中3个PCR产物进行了测序,每个产物与来自危地马拉的甜瓜褪绿曲叶病毒分离株(GenBank登录号AF325497)的同源性均为99%。仅发现1例MNSV和甜瓜褪绿曲叶病毒混合感染。在1999年之后的四年里,表示两种症状的甜瓜植株数量有所增加。本研究提供了危地马拉甜瓜作物病毒病现状的信息,据我们所知,这是危地马拉关于MNSV的首次报道。参考文献:(1)B. 戈萨尔韦斯等人,《病毒学方法杂志》113:87。

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