Merlington A, Hanson L E, Bayma R, Hildebrandt K, Steere L, Kirk W W
Department of Plant, Soil and Microbial Science, Michigan State University, East Lansing, MI, 48824.
Department of Plant, Soil and Microbial Science and USDA-ARS, 494 PSSB, Michigan State University, East Lansing, MI, 48824.
Plant Dis. 2014 Jun;98(6):843. doi: 10.1094/PDIS-09-13-0919-PDN.
Fusarium dry rot of potato (Solanum tuberosum L.) is a postharvest disease caused by several Fusarium spp. Thirteen Fusarium spp. have been implicated in dry rot of potatoes worldwide. Among them, 11 species have been reported causing potato dry rot of seed tubers in the northern United States (1). Historically, Fusarium sambucinum was the predominant species in Michigan potato production (3). Dry rot symptomatic tubers (n = 972) were collected from Michigan commercial potato storage facilities in 2011 and 2012 to determine the composition of Fusarium spp. Sections were cut from the margins of necrotic tissue with a sterile scalpel and surface disinfested in 0.6% sodium hypochlorite for 10 s, rinsed twice in sterile distilled water, and dried on sterile filter paper. The tissue sections were plated on half-strength potato dextrose agar (PDA) amended with 0.5 g/liter of streptomycin sulfate. Dishes were incubated at 23°C in the dark for 7 days. Putative Fusarium isolates were transferred onto water agar and hyphal tips from the margin of actively growing cultures were removed with a sterile scalpel and plated to carnation leaf agar (CLA) and half-strength PDA to generate pure cultures. Seven hundred and thirty Fusarium isolates were collected using these techniques. Preliminary identification of the 730 isolates was based on colony and conidial morphology on PDA and CLA, respectively. While F. oxysporum and F. sambucinum were isolated as expected from prior reports (3), three isolates of F. proliferatum were also identified. On CLA, macroconidia of F. proliferatum were sparse, slender, and mostly straight, with three to five septae (4). Microconidia were abundant, usually single celled, oval or club-shaped in short chains or false heads on monophialides and polyphialides (4), and chlamydospores were absent. On PDA, abundant white mycelium was produced and turned violet with age. Koch's postulates were confirmed through pathogenicity testing on disease-free potato tubers cvs. Atlantic and Russet Norkotah. Tubers were surface disinfested for 10 min in 0.6% sodium hypochlorite and rinsed twice in distilled water. Three tubers of each cultivar per isolate were wounded at the apical end of the tuber to a depth of 4 to 10 mm with a 4 mm diameter cork-borer. Tubers were inoculated by inserting a mycelial plug from a 7-day-old culture grown on PDA into the wound and incubating the tubers at 20°C for 21 days. All Fusarium isolates were tested. Control tubers were inoculated by inserting a water agar plug. Pathogenicity and virulence testing were replicated three times and repeated. Tubers inoculated with F. proliferatum developed typical potato dry rot symptoms but no dry rot symptoms were observed on control tubers. Fusarium proliferatum was re-isolated from symptomatic tubers, confirming Koch's postulates. To our knowledge, this is the first report of F. proliferatum causing potato dry rot in Michigan. References: (1) E. Gachango et al. Plant Dis. 96:1767. (2) D. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (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.)的镰刀菌干腐病是由多种镰刀菌引起的一种采后病害。全球有13种镰刀菌与马铃薯干腐病有关。其中,有11种已报道在美国北部引起种薯的马铃薯干腐病(1)。从历史上看,接骨木镰刀菌(Fusarium sambucinum)是密歇根州马铃薯生产中的主要菌种(3)。2011年和2012年从密歇根州商业马铃薯储存设施中收集了有干腐病症状的块茎(n = 972),以确定镰刀菌的种类组成。用无菌手术刀从坏死组织边缘切取切片,在0.6%次氯酸钠中进行表面消毒10秒,在无菌蒸馏水中冲洗两次,并在无菌滤纸上干燥。将组织切片接种在添加了0.5 g/升硫酸链霉素的半强度马铃薯葡萄糖琼脂(PDA)上。培养皿在23°C黑暗中培养7天。将疑似镰刀菌分离物转移到水琼脂上,用无菌手术刀从活跃生长的培养物边缘切取菌丝尖端,接种到香石竹叶琼脂(CLA)和半强度PDA上以获得纯培养物。使用这些技术收集了730个镰刀菌分离物。对这730个分离物的初步鉴定分别基于在PDA和CLA上的菌落和分生孢子形态。虽然尖孢镰刀菌(F. oxysporum)和接骨木镰刀菌如先前报道(3)所预期的那样被分离出来,但也鉴定出了3个轮枝镰刀菌(F. proliferatum)分离物。在CLA上,轮枝镰刀菌的大型分生孢子稀少、细长且大多为直形,有三到五个隔膜(4)。小型分生孢子丰富,通常为单细胞,椭圆形或棍棒形,在单瓶梗和多瓶梗上呈短链或假头状(4),并且没有厚垣孢子。在PDA上,产生大量白色菌丝体,随着时间推移会变成紫色。通过对无病马铃薯品种大西洋(Atlantic)和褐皮诺科塔(Russet Norkotah)块茎进行致病性测试,证实了柯赫氏法则。将块茎在0.6%次氯酸钠中进行表面消毒10分钟,然后在蒸馏水中冲洗两次。每个分离物对每个品种的三个块茎在块茎顶端用直径4毫米的钻孔器钻至深度为4至10毫米进行创伤。通过将在PDA上培养7天的菌丝块插入伤口对块茎进行接种,并将块茎在20°C下培养21天。对所有镰刀菌分离物进行了测试。对照块茎通过插入水琼脂块进行接种。致病性和毒力测试重复进行三次。接种轮枝镰刀菌的块茎出现典型的马铃薯干腐病症状,但对照块茎未观察到干腐病症状。从有症状的块茎中再次分离出轮枝镰刀菌,证实了柯赫氏法则。据我们所知,这是轮枝镰刀菌在密歇根州引起马铃薯干腐病的首次报道。参考文献:(1)E. Gachango等人,《植物病害》96:1767。(2)D. Geiser等人,《欧洲植物病理学杂志》110:473,2004。(3)M. L. Lacy和R. Hammerschmidt,《镰刀菌干腐病》,推广公告。从http://web1.msue.msu.edu/msue/iac/onlinepubs/pubs/E/E2448POT获取,2010年5月23日。(4)J. F. Leslie和B. A. Summerell,《镰刀菌实验室手册》,Wiley - Blackwell,新泽西州霍博肯,2006。