Liu X-B, Shi T, Li C-P, Cai J-M, Huang G-X
Environment and Plant Protection Institute, CATAS, Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture of China, Danzhou, Hainan, 71737, P. R. China.
Plant Dis. 2010 Jul;94(7):916. doi: 10.1094/PDIS-94-7-0916A.
Cassava (Manihot esculenta) is an important economic crop in the tropical area of China. During a survey of diseases in July and September of 2009, leaf spots were observed on cassava plants at three separate plantations in Guangxi (Yunfu and Wuming) and Hainan (Baisha) provinces. Circular or irregular-shaped leaf spots were present on more than one-third of the plants. Spots were dark brown or had white papery centers delimited by dark brown rims and surrounded by a yellow halo. Usually, the main vein or small veinlets adjacent to the spots were dark. Some defoliation of plants was evident at the Wuming location. A fungus was isolated from symptomatic leaves from each of the three locations and designated CCCGX01, CCCGX02, and CCCHN01. Single-spore cultures of these isolates were incubated on potato dextrose agar (PDA) for 7 days with a 12-h light/dark cycle at a temperature of 28 ± 1°C. Conidiophores were straight to slightly curved, unbranched, and pale to light brown. Conidia were formed singly or in chains, obclavate to cylindrical, straight or curved, subhyaline-to-pale olivaceous brown, 19.6 to 150.3 μm long and 5.5 to 10.7 μm wide at the base, with 4 to 13 pseudosepta. Morphological characteristics of the specimen and their conidia were similar to the descriptions for Corynespora cassiicola (2). The isolate CCCGX01 was selected as a representative for molecular identification. Genomic DNA was extracted by the cetyltrimethylammoniumbromide protocol (3) from mycelia and used as a template for amplification of the internal transcribed spacer (ITS) region of rDNA with primer pair ITS1/ITS4. The sequence (GenBank Accession No. GU138988) exactly matched several sequences (e.g., GenBank Accession Nos. FJ852715, EF198117, and AY238606) of C. cassiicola (1). Young, healthy, and fully expanded green leaves of cassava cv. SC205 were surface sterilized. Ten leaves were inoculated with 10-μl drops of 10 ml suspension of conidia and five leaves were inoculated with the same volume of sterile water to serve as controls. After inoculation, leaves were placed in a dew and dark chamber for 36 h at 25°C and subsequently transferred to the light for 5 days. All inoculated leaves with isolates showed symptoms similar to those observed in natural conditions, whereas the controls remained symptom free. The morphological characteristics of reisolated conidia that formed on the diseased parts were identical with the nature isolates. To our knowledge, this is the first report of leaf spot caused by C. cassiicola on cassava in China. References: (1) L. J. Dixon et al. Phytopathology 99:1015, 2009. (2) M. B. Ellis et al. Corynespora cassiicola. No. 303 in: CMI Description of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, UK 1971. (3) J. R. Xu et al. Genetics 143:175, 1996.
木薯(Manihot esculenta)是中国热带地区一种重要的经济作物。在2009年7月和9月的病害调查期间,在广西(云浮和武鸣)和海南(白沙)三省的三个不同种植园的木薯植株上观察到叶斑病。超过三分之一的植株上出现了圆形或不规则形状的叶斑。病斑呈深褐色,或有白色纸质中心,由深褐色边缘界定,并被黄色晕圈包围。通常,与病斑相邻的主脉或小脉呈深色。在武鸣种植地,一些植株出现了明显的落叶现象。从这三个地点的病叶中分离出一种真菌,分别命名为CCCGX01、CCCGX02和CCCHN01。将这些分离株的单孢培养物在马铃薯葡萄糖琼脂(PDA)上于28±1°C、12小时光照/黑暗周期下培养7天。分生孢子梗直或微弯,不分枝,浅褐色至浅棕色。分生孢子单个或成链形成,倒棍棒形至圆柱形,直或弯,近无色至浅橄榄褐色,长19.6至150.3μm,基部宽5.5至10.7μm,有4至13个假隔膜。该标本及其分生孢子的形态特征与尖孢炭疽菌(Corynespora cassiicola)的描述相似(2)。选择分离株CCCGX01作为分子鉴定的代表。采用十六烷基三甲基溴化铵法(3)从菌丝体中提取基因组DNA,并用作引物对ITS1/ITS4扩增rDNA内部转录间隔区(ITS)的模板。该序列(GenBank登录号GU138988)与尖孢炭疽菌的几个序列(如GenBank登录号FJ852715、EF198117和AY238606)完全匹配(1)。对木薯品种SC205的幼嫩、健康且完全展开的绿色叶片进行表面消毒。用10μl 10 ml分生孢子悬浮液滴接种10片叶子,并用相同体积的无菌水滴接种5片叶子作为对照。接种后,将叶片置于25°C的结露黑暗室中36小时,随后转移至光照下培养5天。所有接种分离株的叶片均表现出与自然条件下观察到的症状相似的症状,而对照叶片则无症状。在患病部位形成的再分离分生孢子的形态特征与自然分离株相同。据我们所知,这是中国首次报道由尖孢炭疽菌引起的木薯叶斑病。参考文献:(1)L. J. Dixon等人,《植物病理学》99:1015,2009年。(2)M. B. Ellis等人,《尖孢炭疽菌》。载于《英联邦真菌研究所病原真菌和细菌描述》第303号。英国邱园英联邦真菌研究所,1971年。(3)J. R. Xu等人,《遗传学》143:175,1996年。