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洪都拉斯茄子感染“番茄伪菌 Liberibacter solanacearum”的首次报告。

First Report of 'Candidatus Liberibacter solanacearum' Infecting Eggplant in Honduras.

作者信息

Munyaneza J E, Sengoda V G, Aguilar E, Bextine B R, McCue K F

机构信息

USDA-ARS, Yakima Agricultural Research Laboratory, 5230 Konnowac Pass Road, Wapato, WA 98951.

Zamorano University, Km 30 carretera a Danlí, Tegucigalpa, Honduras.

出版信息

Plant Dis. 2013 Dec;97(12):1654. doi: 10.1094/PDIS-06-13-0641-PDN.

Abstract

In May of 2012, eggplant (Solanum melongena) plants in an experimental research plot located at Zamorano in the Department of Francisco Morazán, Honduras, were observed with symptoms that included leaf chlorosis and cupping, overall stunting, and production of small and malformed fruits. The research plot was planted next to a commercial tomato field heavily infested with the psyllid Bactericera cockerelli, a vector of 'Candidatus Liberibacter solanacearum' (1,2,3). This bacterium severely affects potato and other solanaceous species and is the putative causal agent of zebra chip disease (2,3). The plot was planted with the eggplant variety 'China' and about 25% of the plants were symptomatic. A total of 10 eggplant samples, including five symptomatic and five asymptomatic plants, were collected from the plot. Total DNA was extracted from the leaf tissue of each of the collected plants with the cetyltrimethylammonium bromide (CTAB) buffer extraction method (1). The DNA samples were then tested by PCR using specific primer sets OA2/OI2c and OMB 1482f/2086r to amplify a portion of 16S rDNA and the outer membrane protein (OMB) genes, respectively, of 'Ca. L. solanacearum' (1,2). OMB gene and 16S rDNA fragments of 605 and 1,168 bp, respectively, were amplified from the DNA of two of the five (40%) symptomatic plants with each primer set, indicating the presence of 'Ca. L. solanacearum.' No 'Ca. L. solanacearum' was detected in the five asymptomatic plants with either primer sets. DNA amplicons with both primer sets were cloned from the DNA of the two 'Ca. L. solanacearum'-positive plant samples and four clones of each of the four amplicons were sequenced. BLASTn analysis of the 16S rDNA resulted in two independent but related consensus sequences (deposited in GenBank as Accession Nos. KF188224 and KF188225) and were 99% similar to each other. The two sequences showed 99 to 100% identity to a number of 16S rDNA sequences of 'Ca. L. solanacearum' in Genbank, including accessions HM245242, FJ811596, and KC768319. For the OMB amplicons, a single consensus sequence was obtained following clone sequencing and was deposited in GenBank as accession KF188229. BLASTn analysis of the sequence indicated that it was 100% identical to several OMB sequences of 'Ca. L. solanacearum' in GenBank, including accessions KC768331 and CP002371. To our knowledge, this is the first report of 'Ca. L. solanacearum' associated with eggplant in Honduras. Eggplant is an economically important commodity in Central America and serious damage to this crop due to this plant pathogen could expand throughout the region, especially if its insect vector B. cockerelli is not properly managed. 'Ca. L. solanacearum' has also caused millions of dollars in losses to potato and several other solanaceous crops in the United States, Mexico, Central America, and New Zealand (2,3). In addition, this bacterium severely damages carrot crops in Europe, where it is transmitted to carrot by the psyllids Trioza apicalis and B. trigonica (3,4). It is imperative that both 'Ca. L. solanacearum' and its insect vectors be effectively monitored and managed to minimize their threat to economically important vegetable crops in many parts of the world. References: (1) J. M. Crosslin et al. Southwest. Entomol. 36:125, 2011. (2) L. W. Liefting et al. Plant Dis. 93:208, 2009. (3) J. E. Munyaneza. Am. J. Pot. Res. 89:329, 2012. (4) J. E. Munyaneza et al. J. Econ. Entomol. 103:1060, 2010.

摘要

2012年5月,在洪都拉斯弗朗西斯科·莫拉桑省萨莫拉诺的一个实验研究地块中,茄子(茄属茄科)植株出现了叶片褪绿和卷曲、整体发育不良以及果实小且畸形等症状。该研究地块紧邻一片受木虱科克雷氏木虱严重侵染的商业化番茄田,科克雷氏木虱是“番茄细菌性溃疡病菌”的传播媒介(1,2,3)。这种细菌会严重影响马铃薯及其他茄科植物,是疑似导致斑马薯片病的病原体(2,3)。该地块种植的是“中国”茄子品种,约25%的植株出现了症状。从该地块共采集了10份茄子样本,其中包括5份有症状植株样本和5份无症状植株样本。采用十六烷基三甲基溴化铵(CTAB)缓冲液提取法,从每份采集植株的叶片组织中提取总DNA(1)。然后使用特异性引物对OA2/OI2c和OMB 1482f/2086r,通过聚合酶链式反应(PCR)对DNA样本进行检测,分别扩增“番茄细菌性溃疡病菌”的16S核糖体DNA(rDNA)的一部分和外膜蛋白(OMB)基因(1,2)。使用每个引物对,从5份有症状植株中的2份(40%)的DNA中扩增出了分别为605 bp和1,168 bp的OMB基因片段和16S rDNA片段,表明存在“番茄细菌性溃疡病菌”。使用任何一个引物对,在5份无症状植株中均未检测到“番茄细菌性溃疡病菌”。从两个“番茄细菌性溃疡病菌”阳性植株样本的DNA中克隆出了使用两个引物对扩增得到的DNA扩增子,并对每个扩增子的4个克隆进行了测序。对16S rDNA进行的BLASTn分析产生了两个独立但相关的共有序列(在GenBank中登记为登录号KF188224和KF188225),它们彼此相似度为99%。这两个序列与GenBank中多个“番茄细菌性溃疡病菌”的16S rDNA序列具有99%至100%的同一性,包括登录号HM245242、FJ811596和KC768319。对于OMB扩增子,克隆测序后获得了一个单一的共有序列,并在GenBank中登记为登录号KF188229。对该序列进行的BLASTn分析表明,它与GenBank中多个“番茄细菌性溃疡病菌”的OMB序列具有100%的同一性,包括登录号KC768331和CP002371。据我们所知,这是洪都拉斯首次报道“番茄细菌性溃疡病菌”与茄子相关。茄子是中美洲一种具有重要经济价值的作物,这种植物病原体对该作物造成的严重损害可能会在整个地区蔓延,特别是如果其昆虫传播媒介科克雷氏木虱没有得到妥善控制。“番茄细菌性溃疡病菌”还在美国、墨西哥、中美洲和新西兰给马铃薯及其他几种茄科作物造成了数百万美元的损失(2,3)。此外,这种细菌在欧洲严重损害胡萝卜作物,在欧洲它通过木虱属顶梢木虱和三角木虱传播给胡萝卜(3,4)。必须对“番茄细菌性溃疡病菌”及其昆虫传播媒介进行有效监测和管理,以尽量减少它们对世界许多地区具有重要经济价值的蔬菜作物的威胁。参考文献:(1)J. M. Crosslin等人,《西南昆虫学家》36:125,2011年。(2)L. W. Liefting等人,《植物病害》93:208,2009年。(3)J. E. Munyaneza,《美国马铃薯研究杂志》89:329,2012年。(4)J. E. Munyaneza等人,《经济昆虫学杂志》103:1060,2010年。

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