Peters R D, Drake K A, Gudmestad N C, Pasche J S, Shinners-Carnelley T
Agriculture and Agri-Food Canada, Charlottetown, PE Canada C1A 4N6.
Department of Plant Pathology, North Dakota State University, Fargo.
Plant Dis. 2008 Dec;92(12):1707. doi: 10.1094/PDIS-92-12-1707B.
Early blight of potato (Solanum tuberosum L.) caused by Alternaria solani Sorauer is a frequent concern for potato growers in Canada. Management of early blight has relied on foliar fungicides that often include quinone outside inhibitor (QoI) fungicides such as azoxystrobin. In recent years, isolates of A. solani with reduced sensitivity to QoI fungicides, conferred by the presence of the F129L mutation (in the cytochrome b gene causing amino acid substitution of phenylalanine with leucine at position 129), have become widespread in potato-production areas of the United States, leading to a reduced efficacy of these products (3). Observations of reduced fungicide efficacy, following application of QoI fungicides to commercial fields in Manitoba, Canada in 2007, prompted an examination of the fungicide sensitivity of isolates of A. solani collected from fields in this province. Nine isolates of A. solani were obtained from potato foliage with typical early blight symptoms from four fields in Manitoba using standard protocols (2). Isolates were maintained on clarified V8 agar (1) and identified to species level based on conidial morphology (4). The sensitivity of each isolate to azoxystrobin was determined by assessing conidial germination on water agar plates amended with 0, 0.001, 0.01, 0.1, 1.0, or 10.0 mg/liter of azoxystrobin with protocols described previously (1). Two reference isolates of A. solani from North Dakota with known sensitivities to azoxystrobin and one isolate from Prince Edward Island (PEI), Canada, (a province yielding only isolates sensitive to azoxystrobin in previous surveys; R. D. Peters, unpublished data) were included in the assays. Calculated effective concentration (EC) values (azoxystrobin concentration inhibiting conidial germination by 50%) were determined for each isolate response from two replications of the assays. The reference isolates of A. solani from North Dakota were sensitive or had reduced sensitivity to azoxystrobin with mean EC values of 0.02 and 0.2 mg/liter, respectively. The isolate from PEI was sensitive to azoxystrobin with a mean EC value of 0.04 mg/liter. By contrast, isolates of A. solani from Manitoba had reduced sensitivity to azoxystrobin with mean EC values from 0.2 to 0.8 mg/liter. Real-time PCR analysis of each isolate was performed (2) and confirmed the presence of the F129L mutation in the Manitoba isolates and the isolate with reduced sensitivity to azoxystrobin from North Dakota. The F129L mutation was absent in the azoxystrobin-sensitive wild-type isolates from PEI and North Dakota. To our knowledge, this is the first report of isolates of A. solani with reduced sensitivity to azoxystrobin in Canada. Since cross resistance among QoI fungicides has been demonstrated in A. solani isolates with the F129L mutation (3), adoption of resistance management strategies, including alternating QoI fungicides with fungicides having different modes of action and further monitoring pathogen populations for QoI sensitivity in Canadian production areas, is recommended. References: (1) J. S. Pasche et al. Plant Dis. 88:181, 2004. (2) J. S. Pasche et al. Plant Dis. 89:269, 2005. (3) J. S. Pasche and N. C. Gudmestad. Crop Prot. 27:427, 2008. (4) J. Rotem. The Genus Alternaria: Biology, Epidemiology, and Pathogenicity. The American Phytopathological Society, St. Paul, MN, 1994.
由链格孢菌(Alternaria solani Sorauer)引起的马铃薯早疫病是加拿大马铃薯种植者经常面临的问题。早疫病的防治一直依赖于叶面杀菌剂,其中通常包括醌外抑制剂(QoI)类杀菌剂,如嘧菌酯。近年来,由于存在F129L突变(细胞色素b基因中导致第129位氨基酸由苯丙氨酸替换为亮氨酸),对QoI类杀菌剂敏感性降低的链格孢菌分离株在美国马铃薯产区广泛传播,导致这些产品的药效降低(3)。2007年,在加拿大曼尼托巴省的商业田地施用QoI类杀菌剂后观察到药效降低,促使人们对从该省田地采集的链格孢菌分离株的杀菌剂敏感性进行研究。使用标准方案(2)从曼尼托巴省四个田地的具有典型早疫病症状的马铃薯叶片中获得了九株链格孢菌分离株。将分离株保存在澄清的V8琼脂(1)上,并根据分生孢子形态鉴定到种的水平(4)。按照先前描述的方案(1),通过评估在添加了0、0.001、0.01、0.1、1.0或10.0毫克/升嘧菌酯的水琼脂平板上的分生孢子萌发情况,测定每个分离株对嘧菌酯的敏感性。测定中包括了两株来自北达科他州的对嘧菌酯敏感性已知的链格孢菌参考分离株以及一株来自加拿大爱德华王子岛(PEI)的分离株(在之前的调查中该省仅产生对嘧菌酯敏感的分离株;R. D. Peters,未发表数据)。针对测定的两次重复中每个分离株的反应,确定计算得出的有效浓度(EC)值(抑制分生孢子萌发50%的嘧菌酯浓度)。来自北达科他州的链格孢菌参考分离株对嘧菌酯敏感或敏感性降低,平均EC值分别为0.02和0.2毫克/升。来自PEI的分离株对嘧菌酯敏感,平均EC值为0.04毫克/升。相比之下,来自曼尼托巴省的链格孢菌分离株对嘧菌酯的敏感性降低,平均EC值为0.2至0.8毫克/升。对每个分离株进行了实时PCR分析(2),证实曼尼托巴省的分离株以及来自北达科他州对嘧菌酯敏感性降低的分离株中存在F129L突变。来自PEI和北达科他州的对嘧菌酯敏感的野生型分离株中不存在F129L突变。据我们所知,这是加拿大首次报道对嘧菌酯敏感性降低的链格孢菌分离株。由于在具有F129L突变的链格孢菌分离株中已证明QoI类杀菌剂之间存在交叉抗性(3),建议采用抗性管理策略,包括将QoI类杀菌剂与具有不同作用方式的杀菌剂交替使用,并进一步监测加拿大产区病原菌群体对QoI的敏感性。参考文献:(1)J. S. Pasche等人,《植物病害》88:181,2004年。(2)J. S. Pasche等人,《植物病害》89:269,2005年。(3)J. S. Pasche和N. C. Gudmestad,《作物保护》27:427,2008年。(4)J. Rotem,《链格孢属:生物学、流行病学和致病性》。美国植物病理学会,明尼苏达州圣保罗,1994年。