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华盛顿州大丽轮枝菌引起的茄子嫁接苗黄萎病的首次报道

First Report of Verticillium Wilt Caused by V. dahliae on Grafted Solanum aethiopicum in Washington.

作者信息

Johnson S, Miles C, Inglis D A

机构信息

Washington State University, Mount Vernon Northwestern Washington Research and Extension Center, 16650 State Route 536, Mount Vernon, 98273.

出版信息

Plant Dis. 2013 Jun;97(6):840. doi: 10.1094/PDIS-09-12-0870-PDN.

DOI:10.1094/PDIS-09-12-0870-PDN
PMID:30722609
Abstract

Solanum aethiopicum L., previously S. integrifolium Poir. (4), has been used as a rootstock for commercial, grafted eggplant production throughout Asia (3). In August 2010 and 2011, symptoms of Verticillium wilt were observed on 'Epic' eggplant (S. melongena L.) grafted onto S. aethiopicum at two sites with a history of the disease: one in the irrigated, dryland Columbia Basin of eastern Washington near Eltopia, and the other in maritime western Washington near Mount Vernon. Interveinal chlorosis, V-shaped necrotic lesions, and wilting were evident at both sites in both years. Each year, stems of 20 symptomatic plants from each field site were cut at the soil line to a 20-cm length, surface sterilized for 5 min in a 10% bleach solution, rinsed in tap water for 30 s, cut longitudinally, and incubated in moisture chambers for 4 weeks at room temperature in the dark. Microsclerotia that formed in the stems were typical of those produced by V. dahliae. One isolate, 'MVEgg301', from an infected stem at Mount Vernon, developed dark microsclerotia and verticillate conidiophores in a radiating pattern, typical of Verticillium on half-strength potato dextrose agar (1/2 PDA) medium. The internal transcribed spacer (ITS) region of rDNA amplified by PCR assay using primers ITS6 and ITS4 from mycelia sampled directly from 1/2 PDA media revealed a 100% match with ITS rDNA sequences of >50 V. dahliae accessions in GenBank (1). Pathogenicity of 'MVEgg301' was assessed in two tests. In both, 12 each of non-grafted and grafted (with cv. Epic) S. aethiopicum plants were inoculated with 'MVEgg301' by cutting approximately 5 mm off the root tips and dipping the remaining roots in a suspension of 10 conidia/ml for 5 s. Similarly, 12 each of non-grafted and grafted S. aethiopicum plants were cut and dipped similarly in sterile water as controls. Chlorosis, necrosis, and wilting were observed in 11 of the 12 inoculated, non-grafted plants and 8 of the 12 inoculated, grafted plants in Test I. The same symptoms were observed in 10 of the 12 inoculated, non-grafted plants and 10 of the 12 inoculated, grafted plants in Test II. V. dahliae was reisolated and confirmed from symptomatic, inoculated non-grafted and grafted plants using the stem assay and direct PCR assay described above. Chlorosis, necrosis, and wilting were observed in one non-grafted control plant in Test I, and two non-grafted and four grafted control plants in Test II. The symptoms were mild and likely due to nutritional deficiencies; microsclerotia were not observed in any assayed water-inoculated plant stems. Although there are several reports of V. albo-atrum infecting S. aethiopicum in the United States (2), to our knowledge, this is the first report of V. dahliae causing Verticillium wilt on this eggplant species. This finding is significant because S. aethiopicum is used as a rootstock for control of soilborne diseases like Verticillium wilt in commercial grafted eggplant production (3). References: (1) G. Calmin et al. Biotechnol. Biotechnol. Equ. 21:40, 2007. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. American Phytopathological Society, St. Paul, MN, 1989. (3) M. Oda. Food Fert. Technol. Ctr. Ext. Bul. 480:1, 1999. (4) PBI Solanum Project. 2012. Solanaceae Source. Accessed at http://www.nhm.ac.uk/solanaceaesource/ . Natural History Museum, London, 29 Aug. 2012.

摘要

埃塞俄比亚茄(Solanum aethiopicum L.),之前被称为全缘叶茄(S. integrifolium Poir.)(4),在亚洲各地一直被用作商业嫁接茄子生产的砧木(3)。2010年8月和2011年,在两个有黄萎病病史的地点,观察到嫁接到埃塞俄比亚茄上的“Epic”茄子(S. melongena L.)出现了黄萎病症状:一个地点在华盛顿州东部靠近埃尔托皮亚的灌溉旱地哥伦比亚盆地,另一个地点在华盛顿州西部靠近弗农山的沿海地区。这两年在两个地点都明显出现了叶脉间黄化、V形坏死斑和萎蔫现象。每年,从每个田间地点选取20株有症状的植株,在土壤线处将茎干剪至20厘米长,在10%的漂白剂溶液中进行5分钟的表面消毒,用自来水冲洗30秒,纵向切开,然后在黑暗的保湿箱中于室温下培养4周。茎干中形成的微菌核是大丽轮枝菌产生的典型微菌核。从弗农山一株受感染茎干上分离得到的一个菌株“MVEgg301”,在半强度马铃薯葡萄糖琼脂(1/2 PDA)培养基上形成了深色微菌核和呈辐射状的轮状分生孢子梗,这是黄萎病菌的典型特征。使用引物ITS6和ITS4通过PCR分析从直接取自1/2 PDA培养基的菌丝体中扩增出的rDNA内部转录间隔区(ITS),与GenBank中50多个大丽轮枝菌菌株的ITS rDNA序列100%匹配(1)。在两项试验中评估了“MVEgg301”的致病性。在这两项试验中,通过将根尖切下约5毫米,并将剩余根系浸入每毫升含10个分生孢子的悬浮液中5秒,对12株未嫁接和12株嫁接(品种为Epic)的埃塞俄比亚茄植株接种“MVEgg301”。同样,将12株未嫁接和12株嫁接的埃塞俄比亚茄植株的根系类似地切下并浸入无菌水中作为对照。在试验I中,12株接种的未嫁接植株中有11株以及12株接种的嫁接植株中有8株出现了黄化、坏死和萎蔫现象。在试验II中也观察到了相同的症状,12株接种的未嫁接植株中有10株以及12株接种的嫁接植株中有10株出现了这些症状。使用上述茎干分析和直接PCR分析方法,从有症状的接种未嫁接和嫁接植株中重新分离并确认了大丽轮枝菌。在试验I中,1株未嫁接对照植株出现了黄化、坏死和萎蔫现象;在试验II中,2株未嫁接和4株嫁接对照植株出现了这些症状。这些症状较轻,可能是由于营养缺乏所致;在任何经检测的接种水的植株茎干中均未观察到微菌核。尽管在美国有几篇关于黑白轮枝菌感染埃塞俄比亚茄的报道(2),但据我们所知,这是首次关于大丽轮枝菌导致该茄子品种发生黄萎病的报道。这一发现具有重要意义,因为在商业嫁接茄子生产中,埃塞俄比亚茄被用作控制诸如黄萎病等土传病害的砧木(3)。参考文献:(1) G. Calmin等人,《生物技术与生物技术设备》21:40,2007年。(2) D. F. Farr等人,《美国植物和植物产品上的真菌》。美国植物病理学会,明尼苏达州圣保罗,1989年。(3) M. Oda,《食品肥料技术中心推广通报》480:1,1999年。(4) PBI茄属项目。2012年。茄科植物资源。访问于http://www.nhm.ac.uk/solanaceaesource/ 。伦敦自然历史博物馆,2012年8月29日。