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番茄斑萎病毒在得克萨斯州导致马铃薯块茎坏死的首次报道

First Report of Tomato spotted wilt virus Causing Potato Tuber Necrosis in Texas.

作者信息

Crosslin J M, Mallik I, Gudmestad N C

机构信息

USDA-ARS Vegetable and Forage Crops Research Unit, Prosser, WA 99350.

Department of Plant Pathology, North Dakota State University, Fargo 58105.

出版信息

Plant Dis. 2009 Aug;93(8):845. doi: 10.1094/PDIS-93-8-0845A.

Abstract

In the summer of 2008, potato (Solanum tuberosum L.) tubers (cvs. FL1867, FL2053, and FL1922) from commercial fields near Dalhart, TX were observed with distinct external erumpent rings and severe internal discolorations including blotches, spots, and dry, cork-like tissue. The presence of rings suggested the possible involvement of one or more viruses. Nucleic acid from seven of eight symptomatic tubers received in Washington (cvs. FL1867 and FL1922) tested positive for Tomato spotted wilt virus (TSWV) by reverse transcription (RT)-PCR with primers TSWV 1 and 2 (3). Similarly, tubers (cvs. FL1867 and FL2053) received in North Dakota tested positive for TSWV with forward (S1983) and reverse (S2767) primers of Tsompana et al. (4). The 777-bp amplicon obtained with primers TSWV 1 and 2 and the 803-bp amplicon obtained with primers S1983 and S2767 were cloned and three clones of each were sequenced. Analysis of the consensus sequences and BLAST comparisons confirmed the Washington and North Dakota sequences were indeed TSWV in origin and were each 98 to 99% identical to the corresponding nucleocapsid region of a number of TSWV isolates and most closely related to an isolate detected in eastern black nightshade from Colorado (GenBank No. AY777475). The deduced amino acid sequences of the 777-bp nucleocapsid open reading frame differed from AY777475 at only two residues in each of the Washington and North Dakota sequences. The Washington and North Dakota derived sequences were deposited with GenBank (Nos. FJ882069 and FJ882070, respectively). None of the eight symptomatic tubers tested positive for Tobacco rattle virus (TRV), Alfalfa mosaic virus (AMV), or the necrotic strains of Potato virus Y (PVY) by RT-PCR. Mechanical transmission tests were conducted by grinding symptomatic tissue of a TSWV-positive FL1867 tuber in 10 volumes of 30 mM potassium phosphate buffer, pH 8.0, containing 10 mM of sodium diethyldithiocarbamate and 10 mM of sodium thioglycollate and rub inoculated onto Carborundum-dusted leaves of Samsun NN tobacco. Approximately 10 days after inoculation, chlorotic-necrotic rings were present on the inoculated leaves and circular necrotic lesions developed on the upper leaves. Dark stem lesions were also present on inoculated tobacco, and after 3 weeks, the upper leaves developed severe, spreading lesions. Tissue from the symptomatic tobacco tested positive for TSWV by RT-PCR (primers TSWV 1 and 2) and also with a TSWV-specific ImmunoStrip (Agdia, Inc., Elkhart, IN), but tested negative for TRV, AMV, and PVY by RT-PCR. TSWV has been reported on field-grown potatoes in North Carolina (1) and has been reported on potatoes in Australia (2) and in other parts of the world (referenced in 1). To our knowledge, this is the first report associating TSWV with tuber necrosis on potatoes in Texas. References: (1) J. A. Abad et al. Am. J. Potato Res. 82:255, 2005. (2) L. J. Latham and R. A. C. Jones. Aust. J. Agric. Res. 48:359, 1997. (3) R. Navarre et al. Am. J. Potato Res. 86:88, 2009. (4) M. Tsompana et al. Mol. Ecol. 14:53, 2005.

摘要

2008年夏天,在得克萨斯州达尔哈特附近的商业田地里,观察到马铃薯(Solanum tuberosum L.)块茎(品种FL1867、FL2053和FL1922)有明显的外部隆起环,内部严重变色,包括斑点、斑块以及干燥的、类似木栓的组织。环的出现表明可能涉及一种或多种病毒。通过使用引物TSWV 1和2进行逆转录(RT)-PCR,华盛顿收到的8个有症状块茎中的7个(品种FL1867和FL1922)的核酸检测出番茄斑萎病毒(TSWV)呈阳性(3)。同样,北达科他州收到的块茎(品种FL1867和FL2053)使用Tsompana等人(4)的正向引物(S1983)和反向引物(S2767)检测TSWV呈阳性。用引物TSWV 1和2获得的777 bp扩增子以及用引物S1983和S2767获得的803 bp扩增子被克隆,每个克隆3个并进行测序。对共有序列的分析和BLAST比较证实,华盛顿和北达科他州的序列确实源自TSWV,并且与许多TSWV分离株的相应核衣壳区域的同一性均为98%至99%,与在科罗拉多州东部龙葵中检测到的一个分离株关系最为密切(GenBank编号AY777475)。华盛顿和北达科他州推导的777 bp核衣壳开放阅读框的氨基酸序列与AY777475相比,在华盛顿和北达科他州的序列中每个仅在两个残基处不同。源自华盛顿和北达科他州的序列已存入GenBank(分别为FJ882069和FJ882070号)。通过RT-PCR,8个有症状的块茎中没有一个检测出烟草脆裂病毒(TRV)、苜蓿花叶病毒(AMV)或马铃薯Y病毒(PVY)坏死株呈阳性。通过将TSWV阳性的FL1867块茎的有症状组织在10倍体积的30 mM磷酸钾缓冲液(pH 8.0)中研磨进行机械传播试验,该缓冲液含有10 mM二乙基二硫代氨基甲酸钠和10 mM巯基乙酸钠,并摩擦接种到撒有金刚砂的三生NN烟草叶片上。接种后约10天,接种叶片上出现褪绿坏死环,上部叶片上出现圆形坏死斑。接种的烟草上也出现深色茎斑,3周后,上部叶片出现严重的、扩展的病斑。有症状烟草的组织通过RT-PCR(引物TSWV 1和2)检测TSWV呈阳性,并且使用TSWV特异性免疫试纸条(Agdia公司,印第安纳州埃尔克哈特)检测也呈阳性,但通过RT-PCR检测TRV、AMV和PVY呈阴性。TSWV已在北卡罗来纳州的田间种植马铃薯上被报道(1),并且在澳大利亚(2)和世界其他地区的马铃薯上也有报道(在1中引用)。据我们所知,这是TSWV与得克萨斯州马铃薯块茎坏死相关的首次报道。参考文献:(1)J. A. Abad等人,《美国马铃薯研究杂志》82:255,2005年。(2)L. J. Latham和R. A. C. Jones。《澳大利亚农业研究杂志》48:359,1997年。(3)R. Navarre等人,《美国马铃薯研究杂志》86:88,2009年。(4)M. Tsompana等人,《分子生态学》14:53,2005年。

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