Wharton P S, Nolte P, Kirk W W, Dangi S, Gevens A J
Department of Plant, Soil and Entomological Sciences, University of Idaho, Aberdeen.
Department of Plant, Soil and Microbial Science, Michigan State University, East Lansing.
Plant Dis. 2015 Mar;99(3):417. doi: 10.1094/PDIS-02-14-0196-PDN.
Late blight, caused by Phytophthora infestans (Mont.) de Bary, is a destructive disease of potato (Solanum tuberosum) and tomato (S. lycopersicum) in the United States. Prior to 2007, the US-8 clonal lineage was the predominant genotype in the United States (4). Since 2007, a significant genetic change in the population of P. infestans occurred in the eastern United States with the appearance of new isolates with unique genotypes and epidemiological characteristics (3). These new genotypes US-22, US-23, and US-24 are sensitive to metalaxyl and represent mating types A2, A1, and A1, respectively (1,2). Prior to 2012, only US-8 had been documented in Idaho (5). In 2013, late blight was discovered in late August on potato crops (cv. Russet Norkotah) in Bingham and Madison counties, ID. Infected foliage (four samples from Bingham County and five from Madison) was sent to Michigan State University and the University of Wisconsin for confirmation of P. infestans and characterization of the isolates. Five sections from the leading edge of lesions were excised with a sterilized scalpel and placed on potato tuber slices ('Dark Red Norkotah'). Pathogen sporulation on the excised lesions was enhanced by incubation in plastic boxes lined with moistened paper towels for 5 days at 18°C in the dark. The sporulating lesions were transferred onto pea agar medium (160 g peas, 5 g sucrose, 15 g agar, 700 ml distilled water) amended with 50 mg/ml vancomycin. Ten pure cultures were obtained for each of 4 isolates per county by hyphal tipping. Cellulose acetate electrophoresis was conducted to determine Gpi allozyme genotype of the 4 isolates (4). The allozyme banding patterns were 100/100 at the Gpi locus, consistent with previously reported analyses of the US-23 genotype (1,2). Genomic DNA was extracted from 10 pure cultures using the DNeasy Plant Mini Kit (Qiagen, Germantown, MD), and SSR analyses were performed. Microsatellite markers Pi02, Pi4B, Pi63, PiG11, and D13 were used in SSR analyses. Pi02, Pi4B, and Pi63 had alleles of 162/164, 213/217, and 270/279 bp in size, respectively which is consistent with the reference US-23 genotype (1). However, heterozygosity was detected at locus D13 in the Idaho genotype with allele size of 134/210 bp and an additional allele of 140/155/176 bp at locus PiG11. This is different from the standard US-23 genotype (homozygous alleles 134/134 at locus D13 and two alleles 140/155 at locus PiG11). These allele changes indicate the isolates may be variants of US-23 isolates as all other phenotypic characteristics were similar to those of reference US-23 isolates. The Idaho genotypes were sensitive to metalaxyl both in vitro on rye A agar medium amended with metalaxyl at <0.1 ppm, and in vivo on Ridomil treated foliage tests at <0.1 ppm (1,2). Mating type assays confirmed the pathogen to be the A1 mating type. In the 2009 and 2010 late blight epidemics in the eastern United States, US-23 was the predominant genotype, but to our knowledge this genotype has never been reported previously in Idaho. Thus, this is the first known report of P. infestans genotype US-23 causing late blight on potato in Idaho, indicating a change in the population of P. infestans. In Idaho, the source of this genotype remains unknown, although infected tomatoes have been implicated in the widespread dissemination of this genotype of P. infestans in the eastern United States. References: (1) G. Danies et al. Plant Dis. 97:873, 2013. (2) C. Hu et al. Plant Dis. 96:1323, 2012. (3) K. Deahl. (Abstr.) Phytopathology 100:S161, 2010. (4) S. B. Goodwin et al. Plant Dis. 79:1181, 1995. (5) USAblight. Recent US Genotypes. Online: www.usablight.org/node/52 , retrieved 3 January 2014.
晚疫病由致病疫霉(Phytophthora infestans (Mont.) de Bary)引起,是美国马铃薯(Solanum tuberosum)和番茄(S. lycopersicum)的一种毁灭性病害。2007年之前,US-8克隆谱系是美国的主要基因型(4)。自2007年以来,美国东部致病疫霉种群发生了显著的遗传变化,出现了具有独特基因型和流行病学特征的新分离株(3)。这些新基因型US-22、US-23和US-24对甲霜灵敏感,分别代表交配型A2、A1和A1(1,2)。2012年之前,爱达荷州仅记录到US-8(5)。2013年8月下旬,在爱达荷州宾厄姆县和麦迪逊县的马铃薯作物(品种为褐皮诺科塔)上发现了晚疫病。将感染的叶片(来自宾厄姆县的4个样本和来自麦迪逊县的5个样本)送往密歇根州立大学和威斯康星大学,以确认致病疫霉并对分离株进行鉴定。用消毒手术刀从病斑前沿切取5个切片,置于马铃薯块茎切片(‘深红色诺科塔’)上。将切下的病斑在衬有湿纸巾的塑料盒中于18°C黑暗条件下培养5天,以增强病原体在病斑上的产孢。将产孢的病斑转移到添加了50 mg/ml万古霉素的豌豆琼脂培养基(160 g豌豆、5 g蔗糖、15 g琼脂、700 ml蒸馏水)上。通过菌丝尖端法,每个县的4个分离株各获得10个纯培养物。进行醋酸纤维素电泳以确定4个分离株的Gpi等位酶基因型(4)。在Gpi位点的等位酶条带模式为100/100,与先前报道的US-23基因型分析结果一致(1,2)。使用DNeasy植物微量提取试剂盒(Qiagen,马里兰州日耳曼敦)从10个纯培养物中提取基因组DNA,并进行SSR分析。SSR分析中使用了微卫星标记Pi02、Pi4B、Pi63、PiG11和D13。Pi02、Pi4B和Pi63的等位基因大小分别为162/164、213/217和270/279 bp,这与参考US-23基因型一致(1)。然而,在爱达荷州基因型的D13位点检测到杂合性,等位基因大小为134/210 bp,在PiG11位点还有一个额外的等位基因140/155/176 bp。这与标准的US-23基因型不同(D13位点为纯合等位基因134/134,PiG-- 11位点为两个等位基因140/155)。这些等位基因变化表明这些分离株可能是US-23分离株的变体,因为所有其他表型特征均与参考US-23分离株相似。爱达荷州基因型在添加了<0.1 ppm甲霜灵的黑麦A琼脂培养基上的体外试验以及在<0.1 ppm甲霜灵处理的叶片体内试验中均对甲霜灵敏感(1,2)。交配型测定证实该病原体为A1交配型。在美国东部2009年和2010年的晚疫病流行中,US-23是主要基因型,但据我们所知,此前在爱达荷州从未报道过该基因型。因此,这是致病疫霉基因型US-23在爱达荷州导致马铃薯晚疫病的首次已知报道,表明致病疫霉种群发生了变化。在爱达荷州,该基因型的来源仍然未知,尽管受感染的番茄被认为与这种致病疫霉基因型在美国东部的广泛传播有关。参考文献:(1)G. Danies等人,《植物病害》97:873,2013年。(2)C. Hu等人,《植物病害》96:1323,2012年。(3)K. Deahl,(摘要)《植物病理学》100:S161,2010年。(4)S. B. Goodwin等人,《植物病害》79:1181,1995年。(5)USAblight. 美国近期基因型。在线:www.usablight.org/node/52,2014年1月3日检索。