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老挝首次发现斯里兰卡木薯花叶病毒和木薯花叶病。

First report of Sri Lankan cassava mosaic virus and Cassava Mosaic Disease in Laos.

作者信息

Chittarath Khonesavane, Jimenez Jenyfer, Vongphachanh Pinkham, Leiva Ana Maria, Sengsay Somkhit, Lopez-Alvarez Diana, Bounvilayvong Touy, Lourido Derlyn, Vorlachith Viengvilay, Cuellar Wilmer Jose

机构信息

Department of Agriculture, Plant Protection Center, Vientiane Capital, Lao People's Democratic Republic;

International Center for Tropical Agriculture, 67609, Virology Laboratory, Cali, Colombia;

出版信息

Plant Dis. 2021 Jan 12. doi: 10.1094/PDIS-09-20-1868-PDN.

Abstract

Cassava ( Crantz) has been traditionally grown as a subsistence crop in Laos, but in recent years cassava cultivation in this country has expanded and is becoming a 'cash crop' for farmers (Malik et al., 2020). This also means that cassava vegetative seed (stakes) is rapidly multiplied and distributed. One of the most important diseases affecting cassava in the world is the Cassava Mosaic Disease (CMD), caused by several species of begomoviruses and disseminated by infected stakes or vectored by the whitefly (Legg et al., 2014). Sri Lankan cassava mosaic virus (SLCMV), a bipartite begomovirus, is the virus species causing CMD in Southeast Asia (SEA) and is widespread in Cambodia, Vietnam, Thailand and south China (Siriwan et al., 2020). During field surveys on July 12 to 14, 2020, the team in south Laos, surveyed 8 fields along the border with Cambodia, in the southern provinces of Attapeu and Champassack and identified CMD symptoms (Supplementary Figure 1A) in only one of the fields, located at Kong District of the Champassack province (GPS coordinates 13.94325, 105.99102). From these 8 fields, samples were collected from every third plant in an X pattern. Photographs from each sampled plant were taken and uploaded into CIAT's PestDisPlace platform (https://pestdisplace.org), for CMD symptom confirmation (Supplementary Figure 1B). Leaf samples were sent to the laboratory for PCR using primers SLCMV-F 5'-ATGTCGAAGCGACCAGCAGATATAAT-3' and SLCMV-R 5'-TTAATTGCTGACCGAATCGTAGAAG-3' targeting the AV1 gene (Dutt et al., 2005), following the protocol described in Siriwan et al. (2020) and primers SLCMV-B-F1 5'-ACCGGATGGCCGCGCCCCCCTCT-3' and SLCMV-B-606R 5'-CACCTACCCTGTTATCGCTAAG-3' targeting part of the BV1 gene. Out of 60 samples collected for the field in Kong district, eleven (18.3%) resulted PCR positive to SLCMV (to DNA-A and DNA-B) but only four plants (6.7%) showed symptoms of CMD (see Supplementary Figure 1B and 1C). None of the samples in the other seven fields had CMD symptoms nor was SLCMV detected in any of these plants. Furthermore, the presence of CMD symptoms in the old leaves of the plants in the affected field suggests that the virus was introduced with contaminated stakes. The complete bipartite genome of one isolate (Champ1), was amplified by Rolling Circle Amplification and sequenced with the nanopore MinION technology as described by Leiva et al. (2020). The sequences were submitted to GenBank under accession nos MT946533 (DNA-A) and MT946534 (DNA-B). A phylogenetic tree for SLCMV and a link to the open SLCMV Nextstrain map (Hadfield et al., 2018) is included in Supplementary Figure 2. The sequences of the DNA-A and DNA-B components of the Champ1 isolate were nearly identical to those of anisolate of SLCMV from Ratanakiri, Cambodia (99.72% for DNA-A and 99.82 for DNA-B; Wang et al., 2016). Phylogenetic analysis (Supplementary Figure 2), grouped isolate Champ1 with those that form the cluster of SEA isolates that contain the shorter version of the rep gene (Siriwan et al., 2020). This short version of rep present a deletion of 7 amino acids at the C-terminus, which is involved in host responses to SLCMV (Wang et al., 2020). The confirmation of CMD and SLCMV in the border between Laos and Cambodia should be followed by disease containment and management strategies, particularly given that the majority cassava varieties grown in Laos are from neighbor countries, most of which have already reported the presence of CMD. Acknowledgements We thank all staff from the CIAT's Cassava Program and the Plant Protection Center of Laos in Vientiane. We acknowledge financial support from the Australian Centre for International Agricultural Research (ACIAR) and the CGIAR Research Program on Roots, Tubers and Bananas (RTB) (https://www.cgiar.org/funders/).

摘要

木薯(Crantz)在老挝一直作为自给作物种植,但近年来该国的木薯种植面积有所扩大,正成为农民的“经济作物”(Malik等人,2020年)。这也意味着木薯营养繁殖体(种茎)迅速繁殖和传播。世界上影响木薯的最重要病害之一是木薯花叶病(CMD),由多种双生病毒引起,通过受感染的种茎传播或由粉虱传播(Legg等人,2014年)。斯里兰卡木薯花叶病毒(SLCMV)是一种双分体双生病毒,是在东南亚(SEA)引起CMD的病毒种类,在柬埔寨、越南、泰国和中国南方广泛分布(Siriwan等人,2020年)。在2020年7月12日至14日的实地调查中,老挝南部的团队对阿塔佩省和占巴塞省与柬埔寨接壤的8个田地进行了调查,仅在占巴塞省孔县(GPS坐标13.94325,105.99102)的一块田地中发现了CMD症状(补充图1A)。从这8个田地中,按照X形从每三株植物中采集样本。对每个采样植物拍照并上传到国际热带农业中心(CIAT)的PestDisPlace平台(https://pestdisplace.org),以确认CMD症状(补充图1B)。将叶片样本送往实验室,按照Siriwan等人(2020年)所述的方案,使用靶向AV1基因的引物SLCMV-F 5'-ATGTCGAAGCGACCAGCAGATATAAT-3'和SLCMV-R 5'-TTAATTGCTGACCGAATCGTAGAAG-3'以及靶向BV1基因部分区域的引物SLCMV-B-F1 5'-ACCGGATGGCCGCGCCCCCCTCT-3'和SLCMV-B-606R 5'-CACCTACCCTGTTATCGCTAAG-3'进行PCR检测。在孔县采集的60个样本中,有11个(18.3%)对SLCMV(DNA-A和DNA-B)的PCR检测呈阳性,但只有4株植物(6.7%)表现出CMD症状(见补充图1B和1C)。其他7个田地的样本均未出现CMD症状,且在这些植物中均未检测到SLCMV。此外,受影响田地中植物老叶上出现CMD症状表明病毒是通过受污染的种茎引入的。按照Leiva等人(2020年)所述,通过滚环扩增技术扩增了一个分离株(Champ1)的完整双分体基因组,并使用纳米孔MinION技术进行测序。序列已提交至GenBank,登录号分别为MT946533(DNA-A)和MT946534(DNA-B)。补充图2中包含了SLCMV的系统发育树以及开放的SLCMV Nextstrain图谱链接(Hadfield等人,2018年)。Champ1分离株的DNA-A和DNA-B组分序列与来自柬埔寨腊塔纳基里的一个SLCMV分离株的序列几乎相同(DNA-A为99.72%,DNA-B为99.82%;Wang等人,2016年)。系统发育分析(补充图2)将Champ1分离株与形成包含较短rep基因版本的SEA分离株簇的那些分离株归为一组(Siriwan等人,2020年)。这种较短版本的rep在C末端缺失了7个氨基酸,这与宿主对SLCMV的反应有关(Wang等人,2020年)。鉴于老挝种植的大多数木薯品种来自邻国且其中大多数已报告存在CMD,在老挝和柬埔寨边境确认CMD和SLCMV后,应采取病害控制和管理策略。致谢我们感谢国际热带农业中心木薯项目和万象老挝植物保护中心的所有工作人员。我们感谢澳大利亚国际农业研究中心(ACIAR)和国际农业研究磋商组织根茎、块茎和香蕉研究计划(RTB)(https://www.cgiar.org/funders/)的资金支持。

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