Stevanović M, Stanković I, Vučurović A, Dolovac N, Pfaf-Dolovac E, Krstić B, Bulajić A
Institute for Plant Protection and Environment, Department of Plant Pathology, Teodora Drajzera 9, 11000 Belgrade, Serbia.
Institute of Plant Protection, Department of Phytopathology, University of Belgrade-Faculty of Agriculture, Nemanjina 6, 11080 Belgrade, Serbia.
Plant Dis. 2012 Jun;96(6):912. doi: 10.1094/PDIS-02-12-0179-PDN.
In September 2011, tomato (Solanum lycopersicum L. 'Big Beef') plants showing typical symptoms of powdery mildew were collected in a greenhouse in the vicinity of Padinska Skela (District of City of Belgrade) in Serbia. Numerous circular, white colonies of powdery mildew were observed predominantly on the adaxial surface of the leaves, the petioles, and the stems. The foliage of infected plants turned yellow and necrotic, which was followed by rapid defoliation. Disease incidence was estimated by counting plants with powdery mildew symptoms in a random batch of 100 plants in four replicates and estimated to be extremely high, approaching 90%. Tomato plants ('Novosadski Jabučar') were inoculated with conidia released from diseased tomato leaves positioned above the tomato leaves and maintained at 25°C with a 14-h photoperiod. Healthy tomato plants from the same lot, which were not exposed to the conidia shower, were used as negative control. The first white fungal colonies appeared on the leaves of the inoculated plants within 4 to 7 days after inoculation, while no fungal growth was observed in the control plants. To determine the morphological characteristics of the pathogen, surface mycelium was removed with small strips of clear adhesive tape and examined using light microscopy. Microscopic observation revealed mycelium with lobed appressoria and hyaline, ellipsoid-ovoid or doliform conidia (32.5 to 47.5 × 17.5 to 25 μm) with no distinct firosin bodies and which produced sub-terminal germ tubes. Conidia were produced on the unbranched, erect conidiophores (82.5 to 150 μm) consisting of a cylindrical foot-cell followed by one to three short cells. No chasmothecia were found. On the basis of morphological characteristics, the pathogen was identified as Oidium neolycopersici (4), which was confirmed by internal transcribed spacer (ITS) sequence analysis. Total DNA was extracted directly from the whitish spots of superficial mycelium on the leaves with a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) following the manufacturer's instructions. PCR amplification and sequencing were performed with primers ITS1F and ITS4 (1). The nucleotide sequence of the representative isolate 809-11 (Accession No. JQ619840) shared 100% identity with 16 O. neolycopersici isolates deposited in GenBank from different parts of the world. Tomato powdery mildew caused by O. neolycopersici is present in many European (4) and other countries around the world (3) and is becoming economically very important as majority of the tomato cultivars have shown to be susceptible (2). To our knowledge, this is the first report of O. neolycopersici in Serbia. Because tomato is a very popular and widely grown vegetable in Serbia, the presence of a new and potentially harmful disease could endanger greenhouse as well as open field tomato production. References: (1) J. H. Cunnington et al. Australas. Plant Pathol. 32:421, 2003. (2) T. Jankovics et al. Phytopathology 98:529, 2008. (3) H. Jones et al. Mol. Plant Pathol. 2:303, 2001. (4) L. Kiss et al. Mycol. Res. 105:684, 2001.
2011年9月,在塞尔维亚帕丁斯卡·斯凯拉(贝尔格莱德市辖区)附近的一个温室里,采集到了表现出白粉病典型症状的番茄(Solanum lycopersicum L. 'Big Beef')植株。在叶片的正面、叶柄和茎上,观察到大量圆形的白色白粉菌菌落。受感染植株的叶子变黄并坏死,随后迅速落叶。通过在四组随机抽取的100株植株中统计有白粉病症状的植株来估计发病率,结果估计极高,接近90%。将患病番茄叶片释放的分生孢子接种到番茄植株('Novosadski Jabučar')上,接种位置在番茄叶片上方,并将植株置于25°C、14小时光周期的环境中。同一批次未接触分生孢子喷淋的健康番茄植株用作阴性对照。接种后4至7天,接种植株的叶片上出现了第一批白色真菌菌落,而对照植株未观察到真菌生长。为了确定病原菌的形态特征,用小条透明胶带去除表面菌丝体,并用光学显微镜进行观察。显微镜观察显示,菌丝体带有叶状附着胞,分生孢子透明,呈椭圆形 - 卵形或桶形(32.5至47.5×17.5至25μm),没有明显的嗜铁体,且产生亚末端芽管。分生孢子在不分枝的直立分生孢子梗(82.5至150μm)上产生,分生孢子梗由一个圆柱形基部细胞和一至三个短细胞组成。未发现闭囊壳。根据形态特征,该病原菌被鉴定为新番茄粉孢(Oidium neolycopersici)(4),通过内转录间隔区(ITS)序列分析得以证实。按照制造商的说明,使用DNeasy植物微量提取试剂盒(Qiagen,德国希尔德)直接从叶片表面菌丝体的白色斑点中提取总DNA。用引物ITS1F和ITS4进行PCR扩增和测序(1)。代表性分离株809 - 11(登录号JQ619840)的核苷酸序列与GenBank中来自世界不同地区的16个新番茄粉孢分离株的序列100%相同。由新番茄粉孢引起的番茄白粉病在许多欧洲国家(4)和世界其他国家(3)都有发生,并且由于大多数番茄品种已被证明易感,其经济重要性日益凸显(2)。据我们所知,这是新番茄粉孢在塞尔维亚的首次报道。由于番茄在塞尔维亚是一种非常受欢迎且广泛种植的蔬菜,一种新的潜在有害病害的出现可能会危及温室以及露地番茄生产。参考文献:(1) J. H. Cunnington等人,Australas. Plant Pathol. 32:421, 2003。(2) T. Jankovics等人,Phytopathology 98:529, 2008。(3) H. Jones等人,Mol. Plant Pathol. 2:303, 2001。(4) L. Kiss等人,Mycol. Res. 105:684, 2001。