Reeser P W, Sutton W, Hansen E M
Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331.
Plant Dis. 2008 Aug;92(8):1252. doi: 10.1094/PDIS-92-8-1252B.
Tanoak (Lithocarpus densiflorus) is a principal host of Phytophthora ramorum, cause of sudden oak death (SOD), in the western United States (1). In the course of SOD surveys in southwestern Oregon, other Phytophthora species were encountered to be causing stem cankers on tanoak that were indistinguishable from those caused by P. ramorum. In Oregon, SOD is subject to quarantine and eradication. Aerial surveys are flown two or more times a year to locate symptomatic tanoaks, which are then examined from the ground to determine the cause of death. Isolations on selective media were attempted from all trees with stem cankers typical of Phytophthora. Phytophthora species were identified by morphological features and DNA sequencing of either internal transcribed spacer (ITS) or the mitochondrial COX spacer region. ITS sequences were compared with validated GenBank records, and COX spacer sequences were compared with known reference isolates in the OSU collection. From 2001 through 2006, Phytophthora spp. were isolated from 482 of 1,057 tanoak stem cankers sampled. P. ramorum was isolated from 359 cankers, P. nemorosa was isolated from 102 cankers, P. gonapodyides was isolated from six cankers, P. cambivora was isolated from four cankers (all A1 mating type), P. siskiyouensis was isolated from four cankers, P. pseudosyringae was isolated from two cankers, P. cinnamomi was isolated from one canker (mating type A2), and P. taxon "Pgchlamydo" was isolated from one canker. Three cankers yielded isolates that were not identified but were closely related to P. pseudosyringae based on ITS sequence. No Phytophthora spp. were cultured from the remaining cankers. One isolate from each species identified (except P. ramorum and P. pseudosyringae) was tested for pathogenicity on tanoak stems (11.4 to 16.0 cm DBH) in the field. A 5-mm-diameter plug from the margin of a V8 agar culture was placed in a hole in the bark, covered with wet cheesecloth, and sealed with aluminum foil and duct tape. Each isolate was inoculated into five different stems. Each stem received three different isolates and an agar control. After 4 weeks, bark was removed to reveal lesion development. Lesions were measured (length by width), and pieces from four points on the lesion margin were plated in selective media to reisolate. P. cambivora, P. cinnamomi, P. gonapodyides, P. nemorosa, P. siskiyouensis and P. taxon "Pgchlamydo" all caused substantial lesions in inoculated tanoak trees (average area 11.5 to 18.6 cm). In all cases, the species used for inoculation was recovered on reisolation from lesion margins. Control inoculations caused necrotic areas averaging 0.2 cm. Isolations from these areas were clean. Prior to the recent SOD epidemic, no species of Phytophthora were known as pathogens of tanoak. The discovery of P. ramorum as a pathogen of tanoak in California was quickly followed by the discovery that P. nemorosa and P. pseudosyringae were also associated with tanoak cankers (2). Six years of diagnostic support for survey and detection of P. ramorum in tanoak forests of southwest Oregon has revealed the occurrence, at very low frequency, of at least five additional species of Phytophthora causing stem cankers in tanoak. References: (1) D. M. Rizzo et al. Ann. Rev. Phytopathol. 43:309, 2005. (2) A. C. Wickland et al. For. Pathol. Online publication. DOI:10.1111/j.1439-0329.2008.00552.x), 2008.
鞣皮栎(Lithocarpus densiflorus)是美国西部引起橡树猝死病(SOD)的病原菌——樟疫霉(Phytophthora ramorum)的主要寄主(1)。在俄勒冈州西南部进行的橡树猝死病调查过程中,发现其他疫霉属物种也会在鞣皮栎上引起茎溃疡,这些溃疡与樟疫霉引起的溃疡难以区分。在俄勒冈州,橡树猝死病需要进行检疫和根除。每年进行两次或更多次空中调查以定位有症状的鞣皮栎,然后从地面进行检查以确定死亡原因。尝试从所有具有典型疫霉茎溃疡的树木上在选择性培养基上进行分离培养。通过形态特征以及内部转录间隔区(ITS)或线粒体COX间隔区的DNA测序来鉴定疫霉属物种。将ITS序列与经过验证的GenBank记录进行比较,COX间隔区序列与俄勒冈州立大学(OSU)收藏中的已知参考分离株进行比较。从2001年到2006年,在采集的1057个鞣皮栎茎溃疡中,有482个分离出了疫霉属物种。从359个溃疡中分离出了樟疫霉,从102个溃疡中分离出了恶疫霉(P. nemorosa),从6个溃疡中分离出了异水霉(P. gonapodyides),从4个溃疡中分离出了食皮疫霉(P. cambivora,均为A1交配型),从4个溃疡中分离出了锡斯基尤疫霉(P. siskiyouensis),从2个溃疡中分离出了拟丁香疫霉(P. pseudosyringae),从1个溃疡中分离出了肉桂疫霉(P. cinnamomi,交配型A2),从1个溃疡中分离出了分类单元“Pgchlamydo”疫霉。有3个溃疡分离出的菌株未被鉴定,但根据ITS序列与拟丁香疫霉密切相关。其余溃疡未培养出疫霉属物种。对每个鉴定出的物种(除樟疫霉和拟丁香疫霉外)的一个分离株在野外对鞣皮栎茎(胸径11.4至16.0厘米)进行致病性测试。从V8琼脂培养基边缘切下一个直径5毫米的菌块,放入树皮的一个孔中,用湿粗棉布覆盖,并用铝箔和管道胶带密封。每个分离株接种到五个不同的茎中。每个茎接种三种不同的分离株和一个琼脂对照。4周后,去除树皮以观察病斑发展情况。测量病斑大小(长乘宽),并从病斑边缘的四个点取组织块接种在选择性培养基上进行再分离培养。食皮疫霉、肉桂疫霉、异水霉、恶疫霉、锡斯基尤疫霉和分类单元“Pgchlamydo”疫霉在接种的鞣皮栎树上均引起了明显的病斑(平均面积11.5至18.6平方厘米)。在所有情况下,接种的物种都能从病斑边缘再分离培养出来。对照接种引起的坏死区域平均为0.2平方厘米。从这些区域分离培养结果为无菌生长。在最近的橡树猝死病流行之前,没有疫霉属物种被认为是鞣皮栎的病原菌。在加利福尼亚发现樟疫霉是鞣皮栎的病原菌后,很快又发现恶疫霉和拟丁香疫霉也与鞣皮栎溃疡有关(2)。对俄勒冈州西南部鞣皮栎林中樟疫霉的调查和检测进行了六年的诊断支持,结果显示至少还有另外五种疫霉属物种以极低的频率在鞣皮栎上引起茎溃疡。参考文献:(1)D. M. Rizzo等人,《植物病理学年评》43:309,2005年。(2)A. C. Wickland等人,《森林病理学》在线出版物。DOI:10.1111/j.1439 - 0329.2008.00552.x,2008年。