Gevens A J, Seidl A C
Department of Plant Pathology, University of Wisconsin, Madison, WI 53706.
Plant Dis. 2013 Jan;97(1):152. doi: 10.1094/PDIS-09-12-0825-PDN.
Potato (Solanum tuberosum) crops are grown on over 25,090 ha in Wisconsin annually. Late blight, caused by Phytophthora infestans (Mont.) deBary, is a potentially devastating disease that affects tomato and potato crops in Wisconsin every few years when inoculum is introduced and weather conditions favor disease. Incidence and severity of late blight are highly variable in these few years due to differences in pathogen clonal lineages, their timing and means of introduction, and weather conditions. Prevention of this disease through prophylactic fungicide application can cost producers millions of dollars annually in additional chemical, fuel, and labor expenses. Populations of P. infestans in the U.S. have recently undergone significant genetic change, resulting in isolates with unique clonal lineages and epidemiological characteristics (1). In 2010, late blight epidemics were of low severity in discrete portions of a few fields and were seen exclusively on potato in two counties of central Wisconsin. Symptoms included water-soaked to dark brown circular lesions with pale green haloes accompanied by white fuzzy pathogen sporulation typically on leaf undersides in high humidity conditions. Infected plants were collected by professional crop consultants and submitted to the authors at the University of Wisconsin Vegetable Pathology Laboratory in Madison, Wisconsin. Eight isolates of P. infestans were generated from individual leaf samples, representing separate fields, by removing sporangia from sporulating lesions and placing onto Rye A agar amended with rifampicin and ampicillin. Axenic, single zoospore-derived cultures of isolates were generated from parent cultures and maintained on Rye A agar for further characterization. Mycelium was coenocytic with hyphal diameter of 5 to 8 μm (n = 50). Sporangia were limoniform to ovoid, semi- to fully papillate, caducous, had short pedicels, and were 36.22 × 19.11 μm (height × width; n = 50). The average length-width ratio was 1.91. Allozyme banding patterns at the glucose-6-phosphate isomerase (Gpi) locus indicated a 100/100/111 profile, consistent with the US-24 clonal lineage (3,4). Mating type assays confirmed the isolates to be A1 and intermediate insensitivity to mefenoxam was observed in vitro (4). Genomic DNA was extracted with a phenol:chloroform:isoamyl alcohol solution and restriction fragment length polymorphism (RFLP) analysis was performed using the RG-57 probe on a representative isolate and resulted in banding patterns consistent with US-24 (2,3). Clonal lineages of P. infestans documented in Wisconsin in previous epidemics included US-8 in the mid-1990s and US-1 in the 1970s. The US-24 (A1) clonal lineage was very widespread in the U.S. in 2010 and its presence in Wisconsin in the same year as identification of US-22 (A2) posed great concern for potential sexual recombination, oospore production, and soil persistence. Fortunately, the opposite mating types were separated spatiotemporally. To the best of our knowledge, this is the first report of the P. infestans clonal lineage US-24 causing late blight on potato in Wisconsin. References: (1) K. Deahl. (Abstr.) Phytopathology 100:S161, 2010. (2) S. B. Goodwin et al. Curr. Genet. 22:107, 1992. (3) Hu et al. Plant Dis. 96:1323, 2012. (4) A. C. Seidl and A. J. Gevens. (Abstr.) Phytopathology 101:S162, 2011.
威斯康星州每年种植马铃薯(Solanum tuberosum)的面积超过25,090公顷。由致病疫霉(Phytophthora infestans (Mont.) deBary)引起的晚疫病是一种具有潜在毁灭性的病害,每隔几年,当接种体传入且天气条件有利于病害发生时,就会影响威斯康星州的番茄和马铃薯作物。在这几年中,由于病原菌克隆谱系、引入的时间和方式以及天气条件的差异,晚疫病的发病率和严重程度变化很大。通过预防性施用杀菌剂来预防这种病害,每年会使生产者在额外的化学品、燃料和劳动力费用上花费数百万美元。美国致病疫霉的种群最近发生了重大的遗传变化,产生了具有独特克隆谱系和流行病学特征的分离株(1)。2010年,晚疫病在少数田地的离散区域中严重程度较低,仅在威斯康星州中部的两个县的马铃薯上出现。症状包括水浸状至深褐色的圆形病斑,周围有浅绿色晕圈,在高湿度条件下,病斑下面通常伴有白色绒毛状病原菌孢子形成。专业作物顾问收集了受感染的植株,并将其提交给位于威斯康星州麦迪逊的威斯康星大学蔬菜病理实验室的作者。从单个叶片样本(代表不同田地)中产生了8个致病疫霉分离株,方法是从产生孢子的病斑上取下孢子囊,并将其置于添加了利福平和平氨苄青霉素的黑麦A琼脂上。从亲本培养物中产生了分离株的无菌、单游动孢子衍生培养物,并在黑麦A琼脂上保存以进行进一步鉴定。菌丝体是多核的,菌丝直径为5至8μm(n = 50)。孢子囊柠檬形至卵形,半乳头状至全乳头状,脱落,有短梗,大小为36.22×19.11μm(高×宽;n = 50)。平均长宽比为1.91。葡萄糖-6-磷酸异构酶(Gpi)位点的等位酶带型显示为100/100/111谱型,与US-24克隆谱系一致(3,4)。交配型测定证实分离株为A1,并且在体外观察到对甲霜灵有中等程度的不敏感性(4)。用苯酚:氯仿:异戊醇溶液提取基因组DNA,并使用RG-57探针在一个代表性分离株上进行限制性片段长度多态性(RFLP)分析,结果产生了与US-24一致的带型(2,3)。在威斯康星州以前的疫情中记录的致病疫霉克隆谱系包括20世纪90年代中期的US-8和20世纪70年代的US-1。2010年,US-24(A1)克隆谱系在美国非常普遍,并且在与US-22(A2)鉴定同年出现在威斯康星州,这引起了人们对潜在有性重组、卵孢子产生和土壤持久性的极大关注。幸运的是,相反的交配型在时空上相互分离。据我们所知,这是致病疫霉克隆谱系US-24在威斯康星州引起马铃薯晚疫病的首次报道。参考文献:(1)K. Deahl。(摘要)植物病理学100:S161,2010。(2)S. B. Goodwin等人。当代遗传学22:107,1992。(3)Hu等人。植物病害96:1323,2012。(4)A. C. Seidl和A. J. Gevens。(摘要)植物病理学101:S162,2011。