Meng J, Wang Y C
Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, China.
Plant Dis. 2008 Sep;92(9):1365. doi: 10.1094/PDIS-92-9-1365B.
Phytophthora tentaculata causes root and stalk rot of Chrysanthemum spp., Delphinium ajacis, and Verbena spp. in nurseries in the Netherlands and Germany (2). In later years, P. tentaculata was isolated from Verbena hybrids (3) and lavender cotton (Santolina chamaecyparissus) in Spain (1). In August 2007, stalk rot symptoms were observed on Aucklandia lappa (Asteraceae), an economically important Chinese medicinal plant, in some fields in Yunnan Province of China. Small groups of infected plants were randomly distributed throughout the fields. Plants showing stalk rot and wilting died rapidly. Diseased tissues were cut into 10-mm pieces and plated onto Phytophthora selective medium, PARP (2), to obtain the pure cultures. Seven isolates were obtained, and five isolates were grown on solidified LBA (60 g of lima bean powder and 15 g of agar per 10,000 ml of distilled water) and 10% V8 juice liquid medium for examination of morphological and physiological characteristics (4). The colony surface texture was uniform and formed sparse, loosely branched mycelium on LBA medium. Radial growth rate was 2 to 3 mm per day at 24°C on LBA. In water, relatively small swellings were formed at hyphal branches. Sporangia were spherical or ovoid to obpyriform and some were distorted and papillate with a narrow exit pore. Approximately 10% of the sporangia were caducous with a short pedicel. Sporangial dimensions were 28 to 47 (35) × 21 to 36 (29) μm, length/breadth ratio 1.2. Chlamydospores formed on LBA after 1 week and were terminal, spherical, thin walled, and 21 to 31 (27) μm in diameter. The isolates were homothallic. Oogonia abundantly formed on LBA and were 25 to 36 (31) μm in diameter. One or two paragynous antheridia (15 × 10 μm) were attached to the oogonia. Oospores were spherical, hyaline, aplerotic, and 20 to 32 (25) μm in diameter. The minimum temperature for mycelium growth was 8°C and maximum temperature was 34°C. The internal transcribed spacer (ITS) region was amplified and sequenced and agreed 100% with sequences of four P. tentaculata isolates deposited in GenBank (Accession Nos. AJ854302, AY881001, DQ335634, and AF266775). Pathogenicity was assessed by flooding three potted A. lappa plants with a 10 ml zoospore suspension and incubating at 20 to 22°C. As controls, two potted A. lappa plants were flooded with deionized water. All three inoculated A. lappa plants exhibited stalk rot after 15 days, from which the pathogen was reisolated using selective medium, PARP. Controls remained healthy 15 days after inoculation, To our knowledge, this is the first report of P. tetaculata H. Kroeber & R. Marwitz infection of A. lappa in China. We speculate that the pathogen might have been introduced from other countries on seeds. References: (1) L. A. Álvarez et al. Plant Dis. 90:523, 2006. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (3) E. Moralejo et al. Plant Pathol. 53:806. 2004. (4) X. B. Zheng. Methods in Phytophthora. Chinese Agriculture Press, Beijing, China, 1995.
樟疫霉可引起荷兰和德国苗圃中菊花属植物、翠雀和马鞭草属植物的根腐病和茎腐病(2)。近年来,在西班牙从马鞭草杂交种(3)和绵杉菊(Santolina chamaecyparissus)中分离到了樟疫霉(1)。2007年8月,在中国云南省的一些田块中,发现一种具有重要经济价值的中药材木香(菊科)出现茎腐病症状。小块受感染植株随机分布于田间。表现出茎腐病和萎蔫症状的植株迅速死亡。将患病组织切成10毫米的片段,接种到樟疫霉选择性培养基PARP上(2),以获得纯培养物。获得了7个分离物,其中5个分离物在固化的LBA培养基(每10000毫升蒸馏水中含60克利马豆粉和15克琼脂)和10% V8果汁液体培养基上培养,用于形态和生理特征检测(4)。菌落表面质地均匀,在LBA培养基上形成稀疏、分枝松散的菌丝体。在LBA上24℃时的径向生长速率为每天2至3毫米。在水中,菌丝分支处形成相对较小的肿胀。孢子囊呈球形、卵形至倒梨形,有些变形且具乳头状突起,有狭窄的出口孔。约10%的孢子囊具短柄,脱落性。孢子囊大小为28至47(35)×21至36(29)微米,长宽比为1.2。在LBA上培养1周后形成厚垣孢子,厚垣孢子为顶生、球形、薄壁,直径为21至31(27)微米。这些分离物为同宗配合。在LBA上大量形成藏卵器,直径为25至36(31)微米。一个或两个侧生雄器(15×10微米)附着于藏卵器上。卵孢子呈球形、透明、无孔,直径为20至32(25)微米。菌丝生长的最低温度为8℃,最高温度为34℃。扩增并测序了内部转录间隔区(ITS),与GenBank中保存的4个樟疫霉分离物(登录号分别为AJ854302、AY881001、DQ335634和AF266775)的序列100%一致。通过用10毫升游动孢子悬浮液浇灌3株盆栽木香植株并在20至22℃下培养来评估致病性。作为对照,用去离子水浇灌2株盆栽木香植株。接种的3株木香植株在15天后均表现出茎腐病症状,从病组织中用选择性培养基PARP再次分离到病原菌。接种15天后对照植株仍保持健康。据我们所知,这是樟疫霉H. Kroeber & R. Marwitz侵染中国木香首次报道。我们推测该病原菌可能是通过种子从其他国家传入的。参考文献:(1)L. A. Álvarez等人,《植物病害》90:523,2(2)D. C. Erwin和O. K. Ribeiro,《世界疫霉病害》,美国植物病理学会,明尼苏达州圣保罗,1996。(3)E. Moralejo等人,《植物病理学》53:806,2004。(4)X. B. Zheng,《疫霉研究方法》,中国农业出版社,中国北京,1995。