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新西兰灯笼果(酸浆属酸浆)中马铃薯纺锤块茎类病毒的首次报道。

First Report of Potato spindle tuber viroid in Cape Gooseberry (Physalis peruviana) in New Zealand.

作者信息

Ward L I, Tang J, Veerakone S, Quinn B D, Harper S J, Delmiglio C, Clover G R G

机构信息

Plant Health and Environment Laboratory, MAF Biosecurity New Zealand, P.O. Box 2095, Auckland 1140, New Zealand.

出版信息

Plant Dis. 2010 Apr;94(4):479. doi: 10.1094/PDIS-94-4-0479A.

Abstract

In February 2009, 10 cape gooseberry plants (Physalis peruviana) grown from seed on a domestic property in Christchurch, New Zealand, showed severe leaf distortion, fasciation and etiolation of growing tips, and weak flowering. Symptoms were first observed in the emerging seedlings. No virus particles were observed in sap from infected plants with the electron microscope. Total RNA was isolated from leaves of the 10 plants with a Qiagen RNeasy Plant Mini Kit (Valencia, CA). All 10 plants tested positive for Potato spindle tuber viroid (PSTVd) by real-time reverse transcription (RT)-PCR (1) and by RT-PCR with PSTVd-specific primers (3) and generic pospiviroid primers (4). For both conventional PCRs, the expected 359-bp amplicons were sequenced directly and sequences were aligned together to create a consensus sequence (GenBank Accession No. FJ797614). BLASTn analysis showed 98% nucleotide identity to PSTVd (EU862231, DQ308556, X17268, and AY532801-AY532804). Sap from one of the infected plants was mechanically inoculated onto healthy P. peruviana, Solanum lycopersicum 'Rutgers', Chenopodium amaranticolor, C. quinoa, Cucumis sativum 'Crystal Apple', Gomphrena globosa, Nicotiana benthamiana, N. clevelandii, N. occidentalis '37B', N. tabacum 'WB', N. sylvestris, and Phaseolus vulgaris 'Prince'. After 4 weeks, the leaves of the 'Rutgers' tomato plants were showing severe distortion, purpling, and necrosis of mid-veins and P. peruviana plants were showing distortion of newly emerging apical leaves. Healthy control P. peruviana were asymptomatic. Symptoms appeared milder than that observed in the original P. peruviana plants, but this may be related to different environmental conditions or age or growth stage of the plants when inoculated. All other indicator plants were symptomless, but along with P. peruviana, tested positive for PSTVd by real-time RT-PCR (1). The presence of PSTVd was further confirmed in one original symptomatic and the mechanically inoculated P. peruviana plants and in the indicator plants by dot-blot hybridization with a digoxygenin-labeled synthetic ssRNA probe specific to the full-length PSTVd genome. PSTVd has been reported in New Zealand previously in commercial glasshouse crops of tomatoes and peppers (2), but was eradicated and so remains a regulated pest. The plants were grown from seeds imported from Germany and it is possible that the infection was seedborne. PSTVd was reported in young cape gooseberry seedlings in Germany and Turkey but the infection was asymptomatic (5). Symptoms were associated with the PSTVd-infected cape gooseberry in New Zealand. To our knowledge, this is the first report of the viroid in domestically grown plants in New Zealand, and only the second report of PSTVd in cape gooseberry worldwide. Our findings suggest that this species is an emerging host for PSTVd and that dissemination of seed may provide a pathway for international movement of the viroid. References: (1) N. Boonham et al. J. Virol. Methods 116:139, 2004. (2) B. S. M. Lebas et al. Australas. Plant Pathol. 34:129, 2005. (3) A. M. Shamoul et al. Can. J. Plant Pathol. 19:89, 1997. (4) J. T. H. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004. (5) J. T. H. Verhoeven et al. Plant Dis. 93:316, 2009.

摘要

2009年2月,在新西兰克赖斯特彻奇一处住宅内,10株由种子培育而成的灯笼果植株(酸浆属秘鲁酸浆)出现严重的叶片扭曲、枝条扁化、生长点黄化以及开花不良的情况。症状最初在新出土的幼苗上被观察到。用电镜观察感染植株的汁液,未发现病毒粒子。使用Qiagen RNeasy植物小提试剂盒(加利福尼亚州瓦伦西亚)从这10株植株的叶片中提取总RNA。通过实时逆转录(RT)-PCR(1)以及使用针对马铃薯纺锤块茎类病毒(PSTVd)的特异性引物(3)和通用马铃薯纺锤类病毒引物(4)进行RT-PCR,这10株植株均检测出PSTVd呈阳性。对于这两种常规PCR,将预期的359 bp扩增子直接测序,并将序列比对在一起以创建一个共有序列(GenBank登录号FJ797614)。BLASTn分析显示与PSTVd(EU862231、DQ308556、X17268以及AY532801 - AY532804)具有98%的核苷酸同一性。将一株感染植株的汁液机械接种到健康的秘鲁酸浆、番茄品种‘罗格斯’、苋色藜、藜麦、黄瓜品种‘水晶苹果’、千日红、本氏烟草、克利夫兰烟草、西方烟草‘37B’、烟草品种‘WB’、野生烟草以及菜豆品种‘王子’上。4周后,‘罗格斯’番茄植株的叶片出现严重扭曲、发紫以及中脉坏死的情况,而秘鲁酸浆植株新长出的顶端叶片出现扭曲。健康对照的秘鲁酸浆没有症状。症状比在最初的秘鲁酸浆植株上观察到的要轻,但这可能与接种时不同的环境条件、植株年龄或生长阶段有关。所有其他指示植物均无症状,但与秘鲁酸浆一样,通过实时RT-PCR(1)检测出PSTVd呈阳性。通过用针对全长PSTVd基因组的地高辛标记合成单链RNA探针进行斑点杂交,在一株最初有症状的和机械接种的秘鲁酸浆植株以及指示植物中进一步证实了PSTVd的存在。此前在新西兰的商业温室种植的番茄和辣椒作物中已报道过PSTVd(2),但已被根除,因此它仍然是一种受管制的有害生物。这些植株是由从德国进口的种子培育而成,感染有可能是种子传播的。在德国和土耳其的幼灯笼果幼苗中曾报道过PSTVd,但感染是无症状的(5)。在新西兰,症状与感染PSTVd的灯笼果有关。据我们所知,这是该类病毒在新西兰国内种植植物中的首次报道,也是全世界灯笼果中PSTVd的第二例报道。我们的研究结果表明,该物种是PSTVd的一个新出现的寄主,种子传播可能为该类病毒的国际传播提供了一条途径。参考文献:(1)N. Boonham等人,《病毒学方法杂志》116:139,2004年。(2)B. S. M. Lebas等人,《澳大拉西亚植物病理学》34:129,2005年。(3)A. M. Shamoul等人,《加拿大植物病理学杂志》19:89,1997年。(4)J. T. H. Verhoeven等人,《欧洲植物病理学杂志》110:823,2004年。(5)J. T. H. Verhoeven等人,《植物病害》93:316,2009年。

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