Petróczy M, Palkovics L
Department of Plant Pathology, Corvinus University of Budapest, H-1118 Budapest, Villányi Road 29-43, Hungary.
Plant Dis. 2006 Mar;90(3):375. doi: 10.1094/PD-90-0375B.
Brown rot caused by Monilinia fructicola is one of the most important diseases of peach. The pathogen is included in the EPPO A2 list of quarantine organisms for Europe (2). M. laxa and M. fructigena are common in Hungary, but M. fructicola has never been reported in orchards, in trade, or in markets. In early October 2005, brown rot was observed on imported peaches from Italy and Spain at a vegetable market and some supermarkets in Budapest. The variety of peach was identified as 'Michellini' by colleagues in the Department of Pomology of Corvinus University. The pathogen was identified as M. fructicola on the basis of morphological and molecular characteristics. Symptoms began with a small, circular brown spot, and the rot spread rapidly. At the same time, numerous small, grayish stromata developed. Finally, the whole surface of the fruit was covered with conidial tufts. The conidia were one-celled, lemon-shaped, hyaline, 15.7 × 10.3 μm, and produced in branched monilioid chains. Conidia from infected fruit were transferred to potato dextrose agar. Fungal mycelium grew at a linear rate of 10.7 mm per 24 h at 22°C in the dark. The color of the colony was grayish, and the sporulation showing concentric rings was abundant (sporulation is sparse in M. laxa or M. fructigena). The colony was not rosetted and the margin was not lobed, in contrast with M. laxa. Pathogenicity was tested by inoculating surface-sterilized, mature peach fruits with conidia. Inoculated and control fruits were placed in a sterilized glass container at room temperature. After 5 days of incubation, typical brown rot symptoms developed on inoculated fruits while control fruits remained healthy. M. fructicola was reisolated from the inoculated fruits. PCR was used to identify the fungus (1). Species-specific internal transcribed spacer (ITS) primers for M. fructicola, M. laxa, and M. fructigena were used to amplify the DNA of isolates. Three type-cultures were used as the positive control. Following the removal of the mycelia from the agar, total DNA was extracted using a cetyltrimethylammoniumbromide extraction. The nucleic acid-containing pellet was resuspended in RNase containing Tris-EDTA buffer. DNA quality was assessed by gel electrophoresis on 1% agarose gel stained with ethidium bromide. The molecular genetic identification method confirmed the results of morphological identification. To our knowledge, this is the first report of M. fructicola on peaches in Hungary or in eastern Europe. References: (1) R. Ioos and P. Frey. Eur. J. Plant Pathol. 106:373, 2000. (2) OEPP/EPPO. List of A2pests regulated as quarantine pests in the EPPO region. Version 2005-09. Online publication, 2005.
由果生链核盘菌引起的褐腐病是桃树最重要的病害之一。该病原菌被列入欧洲植物保护组织(EPPO)A2类检疫性有害生物名单(2)。核果链核盘菌和果产链核盘菌在匈牙利很常见,但果生链核盘菌从未在果园、贸易或市场中被报道过。2005年10月初,在布达佩斯的一个蔬菜市场和一些超市中,从意大利和西班牙进口的桃子上发现了褐腐病。科维努斯大学果树学系的同事将桃子品种鉴定为“米歇利尼”。基于形态学和分子特征,病原菌被鉴定为果生链核盘菌。症状始于一个小的圆形褐色斑点,腐烂迅速蔓延。同时,形成了许多小的灰色子座。最后,果实整个表面布满分生孢子丛。分生孢子单细胞,柠檬形,无色透明,15.7×10.3μm,呈分支的念珠状链产生。将感染果实上的分生孢子转移到马铃薯葡萄糖琼脂上。在22℃黑暗条件下,真菌菌丝体以每24小时10.7毫米的线性速度生长。菌落颜色为灰色,产孢丰富,呈现同心环(核果链核盘菌或果产链核盘菌产孢稀疏)。与核果链核盘菌不同,该菌落不呈莲座状,边缘不呈叶状。通过用分生孢子接种表面消毒的成熟桃子果实来测试致病性。将接种和对照果实置于室温下的消毒玻璃容器中。培养5天后,接种果实上出现典型的褐腐病症状,而对照果实保持健康。从接种果实中重新分离出果生链核盘菌。使用PCR鉴定真菌(1)。使用果生链核盘菌、核果链核盘菌和果产链核盘菌的物种特异性内转录间隔区(ITS)引物扩增分离株的DNA。使用三种模式菌株作为阳性对照。从琼脂中去除菌丝体后,使用十六烷基三甲基溴化铵提取法提取总DNA。将含核酸的沉淀重悬于含有核糖核酸酶的Tris-EDTA缓冲液中。通过在含有溴化乙锭的1%琼脂糖凝胶上进行凝胶电泳评估DNA质量。分子遗传鉴定方法证实了形态学鉴定结果。据我们所知,这是果生链核盘菌在匈牙利或东欧桃树上的首次报道。参考文献:(1)R. Ioos和P. Frey。《欧洲植物病理学杂志》106:373,2000。(2)OEPP/EPPO。EPPO地区被列为检疫性有害生物的A2类有害生物清单。2005年9月版。在线出版物,2005。