Wharton P S, Tumbalam P, Kirk W W
Department of Plant Pathology, Michigan State University, East Lansing 48824.
Plant Dis. 2006 Nov;90(11):1460. doi: 10.1094/PD-90-1460B.
Fusarium dry rot is one of the most important diseases of potato (Solanum tuberosum L.), affecting tubers in storage and whole seed or seed pieces after planting (2). Fusarium sambucinum Fuckel (teleomorph Giberella pulicaris) is the most common pathogen causing dry rot of stored tubers in North America. (4). Cut seed potato tubers of cvs. FL1879 and Pike with severe sprout rot were collected in Michigan during May 2006. As well as having rotted sprouts, all diseased tubers had dry rot. When diseased sprouts were cut in half, brown, necrotic lesions could be seen spreading down the center of the sprout in vascular tissue and at the base of the sprout in tuber tissue. Pathogen isolations were made from both infected tuber tissue and diseased sprouts on potato dextrose agar (PDA). In both cases, only F. sambucinum was isolated from diseased sprout and tuber tissue. Identification of the pathogen was based on colony and conidial morphology. This included white, fluffy mycelium on the surface and crimson coloration of the colonies viewed from the underside of PDA plates and large distinctive macroconidia (3). Identification was confirmed by comparison of ITS (internal transcribed spacer) sequence data with reference isolates. The ITS region of rDNA was amplified by polymerase chain reaction (PCR) with primers ITS1/ITS4 and sequenced. BLASTn analysis (1) of the sequence obtained showed a 100% homology with F. sambucinum Fuckel. For inoculum production, isolates were grown on PDA at 8°C for 14 days prior to inoculation. Pathogenicity was tested in potato tubers of cv. FL1879 with a single isolate collected from diseased sprouts. Whole seed tubers with 4 mm long sprouts were cut in half longitudinally with a sterile knife to ensure that seed pieces had viable sprouts. The cut surfaces of seed pieces were spray inoculated with 200 ml of conidial suspension (1 × 10 conidia ml) over the entire cut surface to give a final dosage of approximately 1 ml per seed piece. Care was taken to limit inoculum spray to the cut surface so that sprouts were not inoculated. Seed pieces (40 per replicate × 4 replicates) were then placed in plastic boxes (30 × 15 × 10 cm) and incubated in the dark at 18°C and 95% relative humidity for 30 days in a controlled environment chamber. As a control, cut seed pieces were spayed with sterile distilled water and incubated as above. All tubers inoculated with the pathogen developed typical Fusarium dry rot symptoms consisting of a brown, dry decay of tuber tissue with mycelial lined cavities. Sprouts on inoculated tubers developed symptoms that were observed in the initially collected seed pieces, and F. sambucinum was reisolated from all infected sprouts. The noninoculated control tubers did not develop any symptoms of dry rot. The results of the pathogenicity tests indicate that F. sambucinum caused sprout rot on potato seed pieces. Since only the cut surfaces of tubers were inoculated, it is assumed that infection of sprouts is systemic through the tuber. To our knowledge, this is the first report of F. sambucinum causing a sprout rot of developing sprouts on seed tubers in the United States. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) L. E. Hanson et al. Phytopathology 86:378, 1996. (3) P. E. Nelson et al. Pages 118-119 in: Fusarium Species: An Illustrated Manual for Identification. The Pennsylvania State University, University Park and London, 1983. (4) G. A. Secor and B. Salas. Fusarium dry rot and Fusarium wilt. Pages 23-25 in: Compendium of Potato Diseases. 2nd ed. W. R. Stevenson et al., eds. The American Phytopathological Society, St. Paul, MN, 2001.
镰刀菌干腐病是马铃薯(Solanum tuberosum L.)最重要的病害之一,会影响贮藏期的块茎以及种植后的整粒种薯或种薯切块(2)。接骨木镰刀菌Fuckel(有性型为赤霉属Giberella pulicaris)是北美引起贮藏块茎干腐病最常见的病原菌(4)。2006年5月在密歇根州采集了严重芽腐病的cv. FL1879和Pike品种的切块种薯。除了芽腐烂外,所有患病块茎都有干腐病。将患病芽切成两半时,可以看到褐色坏死病斑沿芽的维管束组织中心向下蔓延,并在块茎组织中芽的基部出现。从感染的块茎组织和患病芽上在马铃薯葡萄糖琼脂(PDA)上进行病原菌分离。在这两种情况下,从患病芽和块茎组织中仅分离到接骨木镰刀菌。病原菌的鉴定基于菌落和分生孢子形态。这包括PDA平板下表面观察到的菌落表面白色蓬松菌丝体和深红色,以及大型独特的大分生孢子(3)。通过将ITS(内转录间隔区)序列数据与参考分离株进行比较来确认鉴定。用引物ITS1/ITS4通过聚合酶链反应(PCR)扩增rDNA的ITS区域并进行测序。对获得的序列进行BLASTn分析(1)显示与接骨木镰刀菌Fuckel有100%的同源性。为了生产接种物,在接种前将分离株在8°C的PDA上培养14天。用从患病芽中收集的单个分离株在cv. FL1879的马铃薯块茎上测试致病性。用无菌刀将带有4毫米长芽的整粒种薯纵向切成两半,以确保种薯切块有活力的芽。种薯切块的切割面用200毫升分生孢子悬浮液(1×10个分生孢子/毫升)喷洒在整个切割面上,使每个种薯切块的最终剂量约为1毫升。注意将接种物喷雾限制在切割面上,以免芽被接种。然后将种薯切块(每个重复40个×4个重复)放入塑料盒(30×15×10厘米)中,在可控环境箱中于18°C和95%相对湿度下黑暗中培养30天。作为对照,将切割的种薯切块用无菌蒸馏水喷雾并按上述方法培养。所有接种病原菌的块茎都出现了典型的镰刀菌干腐病症状,包括块茎组织褐色、干腐,有菌丝排列的空洞。接种块茎上的芽出现了最初收集的种薯切块中观察到的症状,并且从所有感染的芽中重新分离到接骨木镰刀菌。未接种的对照块茎没有出现任何干腐病症状。致病性测试结果表明接骨木镰刀菌导致马铃薯种薯切块芽腐病。由于仅对接种块茎的切割面进行接种,推测芽的感染是通过块茎系统感染的。据我们所知,这是美国关于接骨木镰刀菌导致种薯上发育中的芽芽腐病的首次报道。参考文献:(1)S. F. Altschul等人,《核酸研究》25:3389,1997。(2)L. E. Hanson等人,《植物病理学》86:378,1996。(3)P. E. Nelson等人,《镰刀菌属:鉴定图解手册》第118 - 119页。宾夕法尼亚州立大学,大学公园和伦敦,1983。(4)G. A. Secor和B. Salas,《镰刀菌干腐病和镰刀菌枯萎病》。《马铃薯病害简编》第2版第23 - 25页。W. R. Stevenson等人编。美国植物病理学会,明尼苏达州圣保罗,2001年。