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关于乌拉圭酸腐链霉菌引起马铃薯疮痂病的首次报告。

First Report Regarding Potato Scab Caused by Streptomyces acidiscabies in Uruguay.

作者信息

Lapaz M I, Verdier E, Pianzzola M J

机构信息

Cátedra de Microbiología, Departamento de Biociencias, Facultad de Química, Universidad de la República, Uruguay.

Dirección General de Servicios Agrícolas, Departamento Laboratorios Biológicos, Ministerio de Ganadería Agricultura y Pesca, Uruguay.

出版信息

Plant Dis. 2012 Jul;96(7):1064. doi: 10.1094/PDIS-02-12-0203-PDN.

Abstract

Potato scab disease is caused by gram-positive filamentous bacteria in the genus Streptomyces. A great variety of species cause this disease, but Streptomyces scabies is the most ancient of these pathogens and can be found in a worldwide distribution, whereas S. turgidiscabies and S. acidiscabies are newly emerged pathogens (2). During the autumn of 2010, potato (Solanum tuberosum) crops had large economic losses by common scab, corresponding to 29% of the total potato-cultivation area (according to our survey), which was unusual in Uruguay. Specifically, the disease was very aggressive and the tubers showed particularly deep scab lesions. We isolated the Streptomyces species present in these particular scab lesions of tubers collected in July 2010 from one of the three potato cultivation areas (San José). A total of 19 Streptomyces spp. strains were isolated and identified using classical and molecular techniques. Morphological characteristics of colonies and microscopic structure of the mycelium were observed (1). Molecular characterization by conventional PCR was carried out using primers directed to specific regions of the 16S rRNA gene for the genus Streptomyces, Aci1: (5'-TCACTCCTGCCTGCATGGGCG-3') and Aci2: (5'-CGACAGCTCCCTCCCACAAG-3'). Also, regions of two pathogenicity genes, namely txtAB and nec1, were amplified and confirmed by sequencing (2). Additionally, melanin production and pathogenicity of the isolates was determined by inoculation of potato discs (1). Six of the 19 strains succeeded in PCR amplification with primers specific to Streptomyces acidiscabies, which has white, aerial hypha and flexuous spore chains. These strains did not produce melanin on tyrosine agar media. The amplified fragments for 16S rRNA and pathogenicity genes from one representative strain 61 were sequenced. BLASTn analysis of the 16S rRNA gene sequence obtained of the strain 61 (Accession No. JN206667) showed the highest similarity (100%) with S. acidiscabies type strain 84-01-182 (GenBank Accession No. FJ007427.1). Pathogenicity of the isolate was tested on tuber slices. The isolate was grown on YME for 5 to 7 days at 28°C and agar plugs from the sporulating colonies were inverted onto excised tuber tissue. Disks were incubated at 28°C in the dark and the presence of necrosis was evaluated after 5 days (1). All tuber slice assays were repeated three times. The noninoculated control tuber slices did not show any necrosis, while those inoculated with the strain did. To our knowledge, this is the first report of S. acidiscabies causing potato scab disease in Uruguay. References: (1) D. H. Park et al. Plant Dis. 87:1290, 2003. (2) L. A. Wanner. Phytopathology 96:1361, 2006.

摘要

马铃薯疮痂病由链霉菌属的革兰氏阳性丝状细菌引起。多种链霉菌会引发这种病害,但疮痂链霉菌是这些病原菌中最古老的一种,在全球范围内均有分布,而肿胀链霉菌和酸腐链霉菌是新出现的病原菌(2)。2010年秋季,乌拉圭马铃薯(Solanum tuberosum)作物因普通疮痂病遭受了巨大经济损失,据我们调查,这相当于马铃薯种植总面积的29%,这种情况在乌拉圭并不常见。具体而言,该病极具侵袭性,块茎上出现了特别深的疮痂病变。我们从三个马铃薯种植区之一(圣何塞)采集了2010年7月带有这些特殊疮痂病变的块茎,从中分离出了链霉菌。共分离出19株链霉菌菌株,并采用经典和分子技术进行了鉴定。观察了菌落的形态特征和菌丝体的微观结构(1)。使用针对链霉菌属16S rRNA基因特定区域的引物Aci1:(5'-TCACTCCTGCCTGCATGGGCG-3')和Aci2:(5'-CGACAGCTCCCTCCCACAAG-3')进行常规PCR分子鉴定。此外,对两个致病基因txtAB和nec1的区域进行了扩增,并通过测序进行了确认(2)。另外,通过接种马铃薯圆片测定了分离株的黑色素产生情况和致病性(1)。19株菌株中有6株成功用酸腐链霉菌特异性引物进行了PCR扩增,酸腐链霉菌具有白色气生菌丝和弯曲的孢子链。这些菌株在酪氨酸琼脂培养基上不产生黑色素。对一株代表性菌株61的16S rRNA和致病基因的扩增片段进行了测序。对菌株61(登录号JN206667)获得的16S rRNA基因序列进行BLASTn分析,结果显示与酸腐链霉菌模式菌株84-01-182(GenBank登录号FJ007427.1)的相似度最高(100%)。在块茎切片上测试了该分离株的致病性。将分离株在28°C的YME培养基上培养5至7天,从产孢菌落上切下的琼脂块倒置在切除的块茎组织上。圆片在28°C黑暗条件下培养,5天后评估坏死情况(1)。所有块茎切片试验均重复进行了三次。未接种的对照块茎切片未出现任何坏死,而接种该菌株的切片出现了坏死。据我们所知,这是酸腐链霉菌在乌拉圭引起马铃薯疮痂病的首次报道。参考文献:(1)D. H. Park等人,《植物病害》87:1290,2003年。(2)L. A. Wanner,《植物病理学》96:1361, 2006年。

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