Gachango E, Kirk W, Hanson L, Rojas A, Tumbalam P
Department of Plant Pathology, Michigan State University, East Lansing 48823.
Plant Dis. 2011 Sep;95(9):1194. doi: 10.1094/PDIS-04-11-0291.
Fusarium dry rot of potato (Solanum tuberosum L.) is a postharvest disease caused by several Fusarium species and is of worldwide importance. Thirteen species of Fusarium have been implicated in fungal dry rots of potatoes worldwide. Among them, eight species have been reported in the northern United States (2). In Michigan potato production, F. sambucinum was the predominant species reported to be affecting seed potato in storage and causing seed piece decay after planting (3). Some previous identifications of F. sambucinum as dry rot may have been F. torulosum since F. torulosum was previously classified within F. sambucinum (4). To further investigate this, dry rot symptomatic tubers were collected from Michigan seed lots in the summers of 2009 and 2010. Small sections from the margins of necrotic regions were cut with a scalpel, surface sterilized in 0.5% sodium hypochlorite for 10 s, rinsed twice in sterile distilled water, and blotted with sterile filter paper. The tissue pieces were plated on half-strength potato dextrose agar (PDA) amended with 0.5 g/liter of streptomycin sulfate and incubated at 23°C for 5 to 7 days. Cultures resembling Fusarium species were transferred onto water agar, and single hyphal tips from actively growing isolates were removed and plated either on carnation leaf agar (CLA) or on half-strength PDA to generate pure cultures. Among the Fusarium isolates obtained, five isolates were identified as F. torulosum (GenBank Accessions Nos. JF803658-JF803660). Identification was based on colony and conidial morphology on PDA and CLA, respectively. These features included slow growth (2.8 ± 0.2 cm in 5 days), white mycelium that became pigmented with age, narrow concentric rings, red or white pigmentation on agar, macroconidia (32.4 ± 0.4 μm average length) with five septa, a pointed apical cell, and a foot-shaped basal cell (4). The identity was confirmed through DNA extraction followed by amplification and sequencing of the translation elongation factor (EF-1α) gene region (1). The Fusarium-ID.v (1) and the NCBI database were used to obtain the closest match (99%) to previously sequenced materials (GenBank Accession No. AJ543611). Pathogenicity testing was done on disease-free potato tubers cv. Red Norland. Tubers were surface sterilized for 10 min in 0.5% sodium hypochlorite and rinsed twice in distilled water. Three tubers per isolate were injected with 20 μl of a conidial suspension (10 conidia/ml) made from F. torulosum cultures grown on PDA for 7 to 10 days. Control tubers were injected with 20 μl of sterile distilled water. All tubers inoculated with F. torulosum developed typical potato dry rot symptoms consisting of a brown and dry decay. There was no disease incidence on the control tubers. F. torulosum was reisolated from the symptomatic tubers. To our knowledge, this is the first report of F. torulosum causing potato dry rot in the United States. References: (1) D. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) L. E. Hanson et al. Phytopathology 86:378, 1996. (3) M. L. Lacy and R. Hammerschmidt. Fusarium dry rot. Extension Bulletin. Retrieved from http://web1.msue.msu.edu/msue/iac/onlinepubs/pubs/E/E2448POT , 23 May 2010. (4) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Wiley-Blackwell, Hoboken, NJ, 2006.
马铃薯(Solanum tuberosum L.)的镰刀菌干腐病是一种由多种镰刀菌引起的采后病害,在全球范围内具有重要影响。全球有13种镰刀菌与马铃薯的真菌干腐病有关。其中,美国北部已报道了8种(2)。在密歇根州的马铃薯生产中,接骨木镰刀菌(F. sambucinum)是据报道影响贮藏种薯并在种植后导致种薯切块腐烂的主要菌种(3)。之前一些将接骨木镰刀菌鉴定为干腐病菌的结果可能有误,实际应为串珠镰刀菌(F. torulosum),因为串珠镰刀菌以前被归类在接骨木镰刀菌内(4)。为进一步研究此问题,于2009年和2010年夏季从密歇根州的种薯批次中收集了有干腐病症状的块茎。用手术刀从坏死区域边缘切取小块组织,在0.5%次氯酸钠中进行表面消毒10秒,在无菌蒸馏水中冲洗两次,并用无菌滤纸吸干。将组织块接种在添加了0.5克/升硫酸链霉素的半强度马铃薯葡萄糖琼脂(PDA)上,于23°C培养5至7天。将类似镰刀菌属的培养物转移到水琼脂上,从生长活跃的分离物中切取单个菌丝尖端,接种到香石竹叶琼脂(CLA)或半强度PDA上以获得纯培养物。在所获得的镰刀菌分离物中,有5个分离物被鉴定为串珠镰刀菌(GenBank登录号:JF80365至JF803660)。鉴定分别基于在PDA和CLA上的菌落及分生孢子形态。这些特征包括生长缓慢(5天内生长2.8±0.2厘米)、白色菌丝体随时间变色、狭窄的同心环、琼脂上的红色或白色色素沉着、具有5个隔膜的大型分生孢子(平均长度32.4±0.4微米)、尖形顶端细胞和足形基部细胞(4)。通过DNA提取,随后对翻译延伸因子(EF - 1α)基因区域进行扩增和测序来确认其身份(1)。使用Fusarium - ID.v(1)和NCBI数据库获得与先前测序材料(GenBank登录号:AJ543611)最接近的匹配(99%)。对无病马铃薯品种红诺兰(Red Norland)块茎进行致病性测试。将块茎在0.5%次氯酸钠中表面消毒10分钟,在蒸馏水中冲洗两次。每个分离物接种3个块茎,注射20微升由在PDA上培养7至10天的串珠镰刀菌培养物制成的分生孢子悬浮液(10个分生孢子/毫升)。对照块茎注射20微升无菌蒸馏水。所有接种串珠镰刀菌的块茎都出现了典型的马铃薯干腐病症状,表现为褐色干腐。对照块茎未发病。从有症状的块茎中重新分离出串珠镰刀菌。据我们所知,这是串珠镰刀菌在美国引起马铃薯干腐病的首次报道。参考文献:(1)D. Geiser等人,《欧洲植物病理学杂志》110:473,2004年。(2)L. E. Hanson等人,《植物病理学》86:378,1996年。(3)M. L. Lacy和R. Hammerschmidt,《镰刀菌干腐病》,推广公告,检索自http://web1.msue.msu.edu/msue/iac/onlinepubs/pubs/E/E2448POT,2010年5月23日。(4)J. F. Leslie和B. A. Summerell,《镰刀菌实验室手册》,Wiley - Blackwell出版社,新泽西州霍博肯,2006年。