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美国太平洋西北地区由核盘菌引起的啤酒花菌核病

Sclerotinia Wilt of Hop (Humulus lupulus) Caused by Sclerotinia sclerotiorum in the Pacific Northwest United States.

作者信息

Kropf S M, Putnam M L, Serdani M, Twomey M C, Woods J L, Gent D H

机构信息

Department of Botany and Plant Pathology, Oregon State University, Corvallis, 97331.

Department of Crop and Soil Science, Oregon State University, Corvallis, 97331.

出版信息

Plant Dis. 2012 Apr;96(4):583. doi: 10.1094/PDIS-12-11-1039.

Abstract

In June 2009, wilted hop bines were observed in a yard in Marion County, OR. The wilt was associated with a stem rot that occurred ~1 m from the ground near the point where bines are tied together for horticultural purposes. Samples of affected stems were submitted to the Oregon State University Plant Clinic. White hyphae and large, black sclerotia were present on the stems, with a clear delineation between healthy and diseased tissue. The pathogen was identified as Sclerotinia sclerotiorum based on morphological characters. In June 2011, bine wilting was observed on the same farm but in a different hop yard (cv. Nugget) ~10 km from the 2009 occurrence. Affected plants had upward curled leaves with necrotic margins or wilted bines that were severed at the soil line. Wilted bines tended to have smaller diameters than bines with foliar symptoms only. Of 100 plants examined, 75% displayed some foliar symptoms and 66% had at least one bine that was wilted. Yield loss was estimated at 10 to 20% due to bine wilting before cone development. Unlike the 2009 occurrence, wilted bines did not display aerial signs of S. sclerotiorum. Rather, water-soaked lesions covered in white, cottony mycelium were apparent on affected stems 2.5 to 5 cm below the soil surface, some bearing large, irregularly shaped sclerotia. Isolations made onto potato dextrose agar yielded isolates with rapid growth rates and morphological characters consistent with S. sclerotiorum (1). DNA was extracted (2) and pathogen identity was confirmed by PCR amplification and sequencing of the internal transcribed spacer regions from isolates SS001 and SS002 as described before (4). The amplicons were sequenced bidirectionally and consensus sequences were 100% similar to S. sclerotiorum (GenBank No. AAGT01000678.1). Two nucleotide polymorphisms were present that differentiated the sequences from those of 12 S. trifoliorum accessions in GenBank that could be aligned (2). Greenhouse assays utilizing a toothpick inoculation procedure (3) were conducted to fulfill Koch's postulates. Stems of five 4-week-old hop plants of cv. Agate were pierced with a toothpick colonized with S. sclerotiorum. Five control plants were similarly inoculated with toothpicks without the fungus. Inoculated plants developed symptoms similar to those observed in the field within 11 days; four of five plants inoculated with isolate SS001 and two of five plants inoculated with isolate SS002 completely wilted. S. sclerotiorum was reisolated from all inoculated plants but not the control plants. To our knowledge, this is the first report of Sclerotinia wilt on hop in Oregon or the Pacific Northwest (1), where nearly all commercial hop production occurs in the United States. The disease appears to be localized to a limited number of yards, although given the widespread distribution and host range of S. sclerotiorum, it is plausible that the disease may occur in other yards. Recurrent outbreaks and spread of the disease among yards on the affected farm suggests that Sclerotinia wilt has the potential to become a perennial problem on hop and efforts to limit the introduction of S. sclerotiorum into other yards are warranted. References: (1) D. H. Gent. Page 32 in: Compendium of Hop Diseases and Pests. The American Phytopathological Society, St. Paul, MN, 2009. (2) E. N. Njambere et al. Plant Dis. 92:917, 2008. (3) M. L. Putnam. Plant Pathol. 53:252, 2004. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

摘要

2009年6月,在俄勒冈州马里恩县的一个院子里发现了枯萎的啤酒花藤蔓。这种枯萎与一种茎腐病有关,该病发生在距地面约1米处,靠近为园艺目的而将藤蔓绑在一起的位置。将受影响茎干的样本提交给俄勒冈州立大学植物诊所。茎干上出现了白色菌丝和大型黑色菌核,健康组织和患病组织之间界限清晰。根据形态特征,该病原菌被鉴定为核盘菌。2011年6月,在同一个农场,但在距离2009年发病地点约10公里的另一个啤酒花院子(品种为Nugget)里观察到藤蔓枯萎。受影响的植株叶片向上卷曲,边缘坏死,或者藤蔓枯萎,并在土壤线处折断。枯萎的藤蔓直径往往比仅表现出叶片症状的藤蔓小。在检查的100株植株中,75%表现出一些叶片症状,66%至少有一根藤蔓枯萎。由于在球果发育前藤蔓枯萎,估计产量损失为10%至20%。与2009年的情况不同,枯萎的藤蔓没有表现出核盘菌的气生症状。相反,在土壤表面以下2.5至5厘米处的受影响茎干上,可见覆盖着白色棉絮状菌丝体的水渍状病斑,有些病斑上带有大型、形状不规则的菌核。在马铃薯葡萄糖琼脂培养基上进行分离培养,得到的分离物生长速度快,形态特征与核盘菌一致(1)。提取DNA(2),并如前所述(4)通过对分离物SS001和SS002的内部转录间隔区进行PCR扩增和测序来确认病原菌身份。扩增产物进行双向测序,得到的一致序列与核盘菌(GenBank编号:AAGT01000678.1)100%相似。存在两个核苷酸多态性,将这些序列与GenBank中12个三叶草核盘菌分离物的序列区分开来(2)。利用牙签接种程序(3)进行温室试验以验证科赫法则。用定殖有核盘菌的牙签穿刺5株4周龄的玛瑙品种啤酒花植株的茎干。同样用没有真菌的牙签接种5株对照植株。接种的植株在11天内出现了与田间观察到的症状相似的症状;接种分离物SS001的5株植株中有4株、接种分离物SS002的5株植株中有2株完全枯萎。从所有接种的植株中重新分离出了核盘菌,但对照植株中未分离到。据我们所知,这是俄勒冈州或太平洋西北地区啤酒花上核盘菌枯萎病的首次报道(1),美国几乎所有的商业啤酒花生产都集中在该地区。尽管鉴于核盘菌分布广泛且寄主范围广,但该病似乎局限于少数院子,不过它有可能在其他院子里发生。该病在受影响农场的院子之间反复爆发和传播,表明核盘菌枯萎病有可能成为啤酒花上的一个常年性问题,因此有必要努力限制核盘菌传入其他院子。参考文献:(1)D. H. Gent。载于《啤酒花病虫害简编》第32页。美国植物病理学会,明尼苏达州圣保罗,2009年。(2)E. N. Njambere等人。《植物病害》92:917,2008年。(3)M. L. Putnam。《植物病理学》53:252,2004年。(4)T. J. White等人。《PCR协议:方法与应用指南》。学术出版社,圣地亚哥,1990年。

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