Wang X X, Chen J, Wang B, Liu L J, Huang X, Ye S T, Peng D X
College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, P.R. China.
Plant Dis. 2011 Oct;95(10):1318. doi: 10.1094/PDIS-04-11-0356.
Ramie (Boehmeria nivea (L.) Gaud) anthracnose is regarded as one of the most widely spread and devastating diseases of ramie. This disease is most severe during warm and humid conditions. In China, ramie anthracnose is found in approximately 10,000 ha, with yield losses averaging 20% and ranging as high as 55% in some fields (3). In September 2010, typical anthracnose symptoms were observed in cultivated fields near Xianning, HuBei Province, China. Lesions on diseased leaves were initially small, scattered, bluish white, and water soaked. As the disease progressed, irregular spots developed on the leaves and the spots turned gray in the center with a brown margin. The diameter of the lesions was approximately 1 to 3 mm. Initially, lesions on the stems were fusiform and then expanded, causing the stem to break. Leaf and stem tissue adjacent to and including lesions were surface disinfected in 0.1% sodium hypochlorite and then planted on potato dextrose agar (PDA) plus oxalic acid to inhibit bacterial growth. The plates were incubated at 25°C for 3 to 5 days until the appearance of pink spore masses with numerous dense clusters of black setae. On PDA, the fungus initially produced gray colonies with an orange conidial mass and then the colonies turned black after 5 days. Spores were single celled, colorless, straight, oval, obtuse at both ends, 10.0 to 20.0 × 3.0 to 5.0 μm with an average size of 15.8 ± 2.4 × 4.6 ± 0.4 μm, and a length/width ratio of 3.47 ± 0.62. The setae were dark brown, 1 to 3 septa. These morphological traits corresponded to Colletotrichum higginsianum Sacc (1). The ITS1, 5.8S, and ITS2 sequences with primers ITS1 and ITS4 of this fungus (GenBank Accession No. JF830783) were 99% similar to sequences of multiple isolates of C. higginsianum (GenBank Accession Nos. GU935872 and AB042303). In pathogenicity tests, both sides of ramie plant leaves from 10-day-old, 30-cm high plants were sprayed with conidial suspensions (1 × 10 conidia/ml) of a representative fungal isolate. This experiment was repeated three times. Inoculated plants were incubated in an artificial climate chamber with a 12-h photoperiod at 25 to 28°C and a relative humidity of 90%. Three days after inoculation, brown spots and water-soaked lesions were observed on all inoculated leaves, but no symptoms were seen on water-treated control plants. Koch's postulates were fulfilled by the reisolation of C. higginsianum from diseased leaves. C. higginsianum is known to cause anthracnose leaf spot disease on many cultivars of Brassica and Raphanus spp., but there have been no reports on it causing ramie anthracnose. C. boehmeriae Sawada and C. gloeosporioides Penz are known to be the agents of ramie anthracnose (2,3). To our knowledge, this is the first report of C. higginsianum causing anthracnose of ramie in China and in the world. References: (1) A. J. Caesar et al. Plant Dis. 94:1166, 2010 (2) R. M. Li and H. G. Ma. J. Plant Prot. 20:83, 1993. (3) X. X. Wang et al. Plant Dis. 94:1508, 2010.
苎麻炭疽病被认为是苎麻中传播最广、危害最大的病害之一。这种病害在温暖潮湿的条件下最为严重。在中国,苎麻炭疽病发病面积约10000公顷,产量损失平均为20%,在一些田块高达55%(3)。2010年9月,在中国湖北省咸宁市附近的种植田中观察到典型的炭疽病症状。病叶上的病斑最初较小,分散,蓝白色,水渍状。随着病情发展,叶片上出现不规则斑点,斑点中心变为灰色,边缘褐色。病斑直径约1至3毫米。最初,茎上的病斑为梭形,然后扩展,导致茎折断。将与病斑相邻并包括病斑的叶和茎组织在0.1%次氯酸钠中进行表面消毒,然后接种在添加草酸的马铃薯葡萄糖琼脂(PDA)上以抑制细菌生长。平板在25°C下培养3至5天,直到出现带有大量密集黑色刚毛簇的粉红色孢子团。在PDA上,该真菌最初产生带有橙色分生孢子团的灰色菌落,5天后菌落变为黑色。孢子单细胞,无色,直形,椭圆形,两端钝圆,大小为10.0至20.0×3.0至5.0微米,平均大小为15.8±2.4×4.6±0.4微米,长宽比为3.47±0.62。刚毛深褐色,1至3个隔膜。这些形态特征与菜豆炭疽菌(Colletotrichum higginsianum Sacc)相符(1)。该真菌(GenBank登录号JF830783)的ITS1、5.8S和ITS2序列与菜豆炭疽菌多个分离株(GenBank登录号GU935872和AB042303)的序列相似度为99%。在致病性测试中,用代表性真菌分离株的分生孢子悬浮液(1×10个分生孢子/毫升)喷洒10日龄、30厘米高的苎麻植株叶片两面。该实验重复三次。接种的植株在人工气候箱中培养,光照周期为12小时,温度为25至28°C,相对湿度为90%。接种三天后,所有接种叶片上均观察到褐色斑点和水渍状病斑,但水处理对照植株上未出现症状。从病叶上重新分离出菜豆炭疽菌,从而证实了柯赫氏法则。已知菜豆炭疽菌会在许多甘蓝和萝卜品种上引起炭疽叶斑病,但尚未有其引起苎麻炭疽病的报道。已知苎麻炭疽病菌为苎麻炭疽盘长孢(C. boehmeriae Sawada)和盘长孢状炭疽菌(C. gloeosporioides Penz)(2,3)。据我们所知,这是菜豆炭疽菌引起中国乃至世界苎麻炭疽病的首次报道。参考文献:(1)A. J. Caesar等人,《植物病害》94:1166,2010年(2)R. M. Li和H. G. Ma,《植物保护学报》20:83,1993年。(3)X. X. Wang等人,《植物病害》94:1508,2010年。