Wu M D, Li G Q, Jiang D H
The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
The State Key Laboratory of Agricultural Microbiology and The Key Laboratory of Plant Pathology of Hubei Province, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
Plant Dis. 2009 Jun;93(6):667. doi: 10.1094/PDIS-93-6-0667A.
Pink reineckia (Reineckea carnea (Andrews) Kunth) is an evergreen herbaceous perennial plant widely grown as groundcover or for medical purposes in southern China. In 2006 and 2007, severe leaf blight was observed on pink reineckia in Wuhan, China. On newly formed pink reineckia leaves, symptoms were first noted in early May as grayish to dark brown, oval or irregular-shaped lesions, 1.5 to 0.2 × 0.5 to 0.1 cm (n = 50), on the leaf margin or leaf tip. A yellowish halo surrounded each lesion. Lesions enlarged and coalesced and diseased leaves became blighted during the fall and winter. In severely infected plots, most plants became straw-colored and had to be replaced with healthy seedlings. A fungus was isolated from surface-disinfested lesions on potato dextrose agar (PDA) at a frequency of 85.7%. One of 30 isolates, designated C2, was characterized further. The fungus growing on PDA at 20°C for 14 days formed zonate white colonies and black acervular conidiomata. Conidia of the fungus aggregated on acervuli as droplets. Conidia were fusiform and 20.7 to 32.2 × 5.8 to 9.8 μm (n = 50). Each conidium had one hyaline apical cell, one hyaline basal cell, and three dark brown median cells. There were two to four hyaline filamentous appendages 8.1 to 20.4 μm long attached to each apical cell and one hyaline appendage 2.4 to 7.1 μm long attached to each basal cell. The cultural and morphological characteristics of isolate C2 matched the description for Pestalotiopsis microspora (Speg.) Batista & Peres (1,2). The internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1-5.8S-ITS2) was PCR-amplified and sequenced. The ITS sequence (606 bp) for isolate C2 (GenBank Accession No. EU935587) was 100% similar to P. microspora isolates TA-57 (GenBank Accession No. AY924267) and LK32 (GenBank Accession No. DQ001002). Pathogenicity of isolate C2 was tested with the method described by Keith et al. (2). Four detached leaves were wound inoculated or inoculated without wounding with mycelia on agar plugs (4 mm in diameter; three plugs per leaf) or conidial suspensions (10 conidia per ml; 20 μl on each of three sites per leaf). Control leaves were wound inoculated with PDA or sterile water. All inoculated leaves were maintained in a moist enamel tray under fluorescent light for 7 days at 20°C. The test was performed twice. After 4 days of incubation, necrotic leaf lesions resembling symptoms that occurred in the field were observed on the wound-inoculated leaves, whereas the control leaves and C2-inoculated leaves without wounding remained healthy. Therefore, wounding was necessary for symptom development (2). A fungus was reisolated from the C2-induced leaf lesions and the morphology of colonies and conidia were identical to that for isolate C2 of P. microspora. On the basis of the results of isolations, inoculations, and fungal identification, P. microspora was determined to be the causal agent for leaf blight of pink reineckia occurring in Wuhan, China. This fungus previously has been reported as the causal agent of scab disease of Psidium guajava in Hawaii (2), decline of Torreya taxifolia in Florida (3), and leaf blight of Lindera obtusiloba in Korea (1). To our knowledge, this is the first report of the occurrence of P. microspora on R. carnea. References: (1) Y. H. Jeon et al. Plant Pathol. 56:349, 2007. (2) L. M. Keith et al. Plant Dis. 90:16, 2006. (3) M. W. Schwartz et al. Plant Dis. 80:600, 1996.
粉叶沿阶草(Reineckea carnea (Andrews) Kunth)是一种多年生常绿草本植物,在中国南方广泛用作地被植物或用于医学用途。2006年和2007年,在中国武汉的粉叶沿阶草上观察到严重的叶枯病。在新形成的粉叶沿阶草叶片上,症状最初于5月初在叶缘或叶尖出现,表现为浅灰色至深褐色、椭圆形或不规则形病斑,大小为1.5至0.2×0.5至0.1厘米(n = 50)。每个病斑周围有一个淡黄色晕圈。病斑扩大并融合,患病叶片在秋冬季节枯萎。在严重感染的地块中,大多数植株变成稻草色,不得不被健康的幼苗取代。从经表面消毒的病斑上在马铃薯葡萄糖琼脂(PDA)上分离出一种真菌,分离频率为85.7%。对30个分离物中的一个,命名为C2,进行了进一步鉴定。该真菌在20°C下于PDA上生长14天,形成具轮纹的白色菌落和黑色分生孢子盘。真菌的分生孢子在分生孢子盘上聚集成滴状。分生孢子呈梭形,大小为20.7至32.2×5.8至9.8微米(n = 50)。每个分生孢子有一个透明的顶端细胞、一个透明的基部细胞和三个深褐色的中间细胞。每个顶端细胞附有2至4条长8.1至20.4微米的透明丝状附属物,每个基部细胞附有一条长2.4至7.1微米的透明附属物。分离物C2的培养和形态特征与微小拟盘多毛孢(Pestalotiopsis microspora (Speg.) Batista & Peres)的描述相符(1,2)。对核糖体DNA的内部转录间隔区(ITS)区域(ITS1 - 5.8S - ITS2)进行PCR扩增并测序。分离物C2的ITS序列(606 bp)(GenBank登录号EU935587)与微小拟盘多毛孢分离物TA - 57(GenBank登录号AY924267)和LK32(GenBank登录号DQ001002)100%相似。按照Keith等人(2)描述的方法对分离物C2的致病性进行了测试。用直径4毫米的琼脂块上的菌丝体(每片叶子3个琼脂块)或分生孢子悬浮液(每毫升10个分生孢子;每片叶子的三个部位各20微升)对四片离体叶片进行创伤接种或非创伤接种。对照叶片用PDA或无菌水进行创伤接种。所有接种的叶片在20°C下置于荧光灯下的潮湿搪瓷盘中保持7天。该试验进行了两次。培养4天后,在创伤接种的叶片上观察到类似田间出现症状的坏死叶斑,而对照叶片和未创伤接种C2的叶片保持健康。因此,创伤是症状发展所必需的(2)。从C2诱导的叶斑中重新分离出一种真菌,其菌落和分生孢子的形态与微小拟盘多毛孢的分离物C2相同。根据分离、接种和真菌鉴定的结果,确定微小拟盘多毛孢是中国武汉粉叶沿阶草叶枯病的病原菌。此前该真菌已被报道为夏威夷番石榴疮痂病(2)、佛罗里达榧树衰退病(3)以及韩国钝叶钓樟叶枯病(1)的病原菌。据我们所知,这是微小拟盘多毛孢在粉叶沿阶草上发生的首次报道。参考文献:(1)Y. H. Jeon等人,《植物病理学》56:349,2007年。(2)L. M. Keith等人,《植物病害》90:16,2006年。(3)M. W. Schwartz等人,《植物病害》80:600,1996年。