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墨西哥哈利斯科州马铃薯中重组马铃薯Y病毒株的首次报道

First Report of Recombinant Potato virus Y Strains in Potato in Jalisco, Mexico.

作者信息

Quintero-Ferrer A, Karasev A V

机构信息

Department of PSES, University of Idaho, Moscow, ID 83844-2339.

出版信息

Plant Dis. 2013 Mar;97(3):430. doi: 10.1094/PDIS-10-12-0919-PDN.

Abstract

Potato virus Y (PVY) is a serious problem for potato production worldwide. The virus reduces both tuber yield and quality, and recent spread of recombinant strains of PVY in potato production areas is largely credited with the spread of potato tuber necrotic ringspot disease (PTNRD) (1). In Mexico, recombinant strains of PVY were reported in at least two states, Chihuahua (4) and the State of Mexico (3); however, no surveys have been conducted in other potato-producing areas, and the spectrum of PVY isolates circulating in the country has remained uncharacterized. In October 2011, a small-scale survey of seed potato was conducted in the state of Jalisco, Mexico, to identify PVY isolates present in fields. Twelve seed potato fields were inspected visually. These represented various generations of seed potato, from nuclear to G2. Leaf samples were collected from plants displaying mosaic, crinkling, and yellowing symptoms, and were tested for PVY. Fifty samples were collected from cultivars Fabula, Mondial, Fianna, Gigant, Caesar, and Adora. Of the 50 leaf samples collected, seven were PVY-positive using the Immuno-strip Kit (Agdia, Elkhart, IN), and six of these were determined to have a N-serotype according to the typing by the Pocket Diagnostics lateral flow kit (Forsite Diagnostics, Ltd., York, UK). PVY-positive samples came from cultivars Fabula (2 with N serotype), Mondial (4 with N serotype), and Fianna (1 with O serotype). Extracts of the seven PVY-positive leaf samples were applied to Whatman FTA cards (Sigma, St. Louis, MO), dried, and transported to the Plant Virology Laboratory at the University of Idaho for further characterization. All samples immobilized on FTA cards were subjected to RNA extraction and standard reverse transcriptase (RT)-PCR typing using a set of PVY-specific primers (2) to determine the strain type. All PVY isolates were recombinant. The six N-serotype samples were found to contain recombinant PVY isolates and produced characteristic bands of 181 and 452 bp in RT-PCR, which indicated the presence of two recombination junctions in the HC-Pro/P3 and VPg regions typical of European PVY isolates. The one O-serotype sample was identified as a recombinant PVY isolate, and produced 181 and 689 bp bands in RT-PCR, which indicated the presence of one recombination junction in the HC-Pro/P3 region. Sequence analysis of RT-PCR products amplified from five samples with N serotype identified them as PVY isolates, and from the one with O serotype identified it as PVY isolate. Sequence comparisons confirmed that N serotype samples contained PVY isolates most closely related to typical PVY sequences (Accession No. EF026075), while the O serotype sample contained the PVY isolate most closely related to PVY from Europe (HE608963). The data obtained suggest the presence of two different types of PVY recombinants, PVY and PVY, in seed potato in Jalisco. Additional surveillance for these recombinant isolates may be needed, as well as a survey of their effects on tuber quality in production areas. This is the first report of recombinant isolates of PVY often associated with PTNRD circulating in seed potato in Jalisco, Mexico. References: (1) S. M. Gray et al. Plant Dis. 94:1384, 2010. (2) J. H. Lorenzen et al. Plant Dis. 90:935, 2006. (3) V. R. Ramirez-Rodriguez et al. Virol. J. 6:48, 2009. (4) L. Robles-Hernandez et al. Plant Dis. 94:1262, 2010.

摘要

马铃薯Y病毒(PVY)是全球马铃薯生产面临的一个严重问题。该病毒会降低块茎产量和品质,近期PVY重组毒株在马铃薯产区的传播在很大程度上导致了马铃薯块茎坏死环斑病(PTNRD)的蔓延(1)。在墨西哥,至少在奇瓦瓦州(4)和墨西哥州(3)报道了PVY重组毒株;然而,尚未在其他马铃薯产区进行调查,该国境内传播的PVY分离株谱系仍未得到鉴定。2011年10月,在墨西哥哈利斯科州对种薯进行了小规模调查,以确定田间存在的PVY分离株。对12块种薯田进行了目视检查。这些田代表了从原原种到G2代的不同世代种薯。从表现出花叶病症状、皱缩和黄化症状的植株上采集叶片样本,并对其进行PVY检测。从法布拉、蒙迪阿尔、菲安娜、吉甘特、凯撒和阿多拉等品种中采集了50个样本。在采集的50个叶片样本中,使用免疫试纸条试剂盒(Agdia公司,印第安纳州埃尔克哈特)检测出7个样本PVY呈阳性,根据口袋诊断侧向流动试剂盒(Forsite Diagnostics有限公司,英国约克)的分型,其中6个样本被确定为N血清型。PVY阳性样本来自法布拉品种(2个N血清型)、蒙迪阿尔品种(4个N血清型)和菲安娜品种(1个O血清型)。将7个PVY阳性叶片样本的提取物应用于沃特曼FTA卡(Sigma公司,密苏里州圣路易斯),干燥后运往爱达荷大学植物病毒学实验室进行进一步鉴定。固定在FTA卡上的所有样本都进行了RNA提取,并使用一组PVY特异性引物(2)进行标准逆转录酶(RT)-PCR分型,以确定毒株类型。所有PVY分离株均为重组株。6个N血清型样本被发现含有重组PVY分离株,在RT-PCR中产生了181和452 bp的特征条带,这表明在HC-Pro/P3和VPg区域存在两个典型欧洲PVY分离株的重组位点。1个O血清型样本被鉴定为重组PVY分离株,在RT-PCR中产生了181和689 bp的条带,这表明在HC-Pro/P3区域存在1个重组位点。对5个N血清型样本扩增的RT-PCR产物进行序列分析,确定它们为PVY分离株,对1个O血清型样本扩增的RT-PCR产物进行序列分析,确定它为PVY分离株。序列比较证实,N血清型样本所含的PVY分离株与典型PVY序列(登录号EF026075)关系最为密切,而O血清型样本所含的PVY分离株与来自欧洲的PVY(HE608963)关系最为密切。获得的数据表明,在哈利斯科州的种薯中存在两种不同类型的PVY重组体,即PVY和PVY。可能需要对这些重组分离株进行进一步监测,以及调查它们对产区块茎品质的影响。这是关于与PTNRD相关的PVY重组分离株在墨西哥哈利斯科州种薯中传播的首次报道。参考文献:(1)S.M.格雷等人,《植物病害》94:1384,2010年。(2)J.H.洛伦岑等人,《植物病害》90:935,2006年。(3)V.R.拉米雷斯-罗德里格斯等人,《病毒学杂志》6:4&2009年。(&)L.罗夫莱斯-埃尔南德斯等人,《植物病害》94:1262,2010年。

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