Gai Y, Pan R, Xu D, Ji C, Deng M, Chen W
Laboratory of Bacteria and Fungicides, South China Agricultural University, Guangzhou 510642, China.
Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.
Plant Dis. 2011 Aug;95(8):1031. doi: 10.1094/PDIS-04-11-0287.
During October 2009, the occurrence of a disease on soybean (Glycine max) was observed in several fields in Boluo County and Zengcheng City, Guangdong Province. Top leaves of infected plants initially turned yellow and plants eventually were defoliated, while stems and roots became black and rotted. The stem lesions sometimes extended 10 to 15 cm upward from the soil surface. Orange-to-brown spherical fruiting bodies, which were very similar with those of the soybean red crown rot pathogen, scattered or congregated on the stem lesions. Plants with symptoms were sampled from fields. Fruiting bodies were excised from diseased tissues. Microscopic examination revealed that they were perithecia, globose to pyriform, ostiolate with a short neck, and measured 160 to 298 × 151 to 235 μm. Under gentle pressure, asci and ascospores were exuded from these perithecia. Asci were eight spored, narrowly cylindrical to clavate, thin walled, with a short stalk, and measured 58 to 124 μm long and 8 to 15 μm in diameter. Ascospores were uniseriately arranged, globose to ellipsoid, thick walled, one celled, hyaline to pale, and measured 14 to 17 × 8 to 12 μm. Isolation was made from stem tissues at the edge of disease lesions on potato dextrose agar (PDA) amended with streptomycin sulfate and incubated at 25°C. Mycelia were white and floccose. Conidia were cylindrical to oblong-ellipsoidal, hyaline, one celled, and measured 6 to 22 × 2 to 5 μm, aggregating in a slimy mass on the apex of the conidiogenous cell. Abundant orange-to-brown spherical perithecia were produced on the colony. Ascospores had walls with a rugose ornamentation that could be clearly seen under a scanning electron microscope. The fungus was identified as Neocosmospora vasinfecta var. vasinfecta (anamorph Acremonium sp.) (1). The internal transcribed spacer (ITS) region of rDNA of two isolates were amplified with universal primers ITS1/ITS4 and sequenced (GenBank Accession No. JF705861 and JF705862), and comparisons with GenBank accessions showed 99% similarity with N. vasinfecta strain Pec070 (Accession No. FJ940902) and strain NRRL22497 (Accession No. AY381142). Pathogenicity tests were conducted. Five, 3-week-old seedlings of soybean cv. Huaxia No. 3 planted in plastic pots (20 cm in diameter) were wounded with a needle at the base of the stem below the soil line and near the root system, and then inoculated by drenching the soil with a conidial suspension (10 per ml). Control plants were inoculated with sterile water. There were six replicates for each treatment. The treated plants were incubated at 25 ± 2°C in a greenhouse. All inoculated plants exhibited symptoms of leaf yellowing and black rot of stems and roots 3 weeks after inoculation. N. vasinfecta var. vasinfecta was reisolated from the diseased plants. All control plants remained healthy. To our knowledge, this is the first observation of Neocosmospora stem rot of soybean in China. The pathogen could pose a threat to soybean, which is a major crop in China. This disease has been previously reported in the United States though the anamorph of the pathogen has either not been identified or has been identified as a Cylindrocarpon sp. (2,4). This fungus is also associated with human infections (3). References: (1) P. F. Cannon and D. L. Hawksworth. Trans. Br. Mycol. Soc. 82:673, 1984. (2) F. A. Gray et al. Plant Dis. 64:321, 1980. (3) P. Manikandan et al. Med. Mycol. 46:279, 2008. (4) D. V. Phillips. Phytopathology 62:612, 1972.
2009年10月期间,在广东省博罗县和增城市的多个田块中,观察到大豆(Glycine max)出现一种病害。染病植株的顶部叶片最初变黄,最终植株落叶,而茎和根变黑并腐烂。茎部病斑有时从土壤表面向上延伸10至15厘米。橙色至褐色的球形子实体,与大豆赤冠腐病菌的子实体非常相似,散生或聚集在茎部病斑上。从田间采集有症状的植株。从患病组织上切下子实体。显微镜检查表明,它们是子囊壳,球形至梨形,有孔口和短颈,大小为160至298×151至235微米。在轻轻挤压下,子囊和子囊孢子从这些子囊壳中挤出。子囊有8个孢子,狭圆柱形至棒形,薄壁,有短柄,长58至124微米,直径8至15微米。子囊孢子单列排列,球形至椭圆形,厚壁,单细胞,透明至浅色,大小为14至17×8至12微米。从病斑边缘的茎组织在添加硫酸链霉素的马铃薯葡萄糖琼脂(PDA)上进行分离,并在25°C下培养。菌丝体白色,絮状。分生孢子圆柱形至长椭圆形,透明,单细胞,大小为6至22×2至5微米,聚集在分生孢子梗顶端的黏液团中。菌落上产生大量橙色至褐色的球形子囊壳。子囊孢子的壁有皱纹状纹饰,在扫描电子显微镜下可以清楚地看到。该真菌被鉴定为大豆新小赤壳病菌大豆变种(Neocosmospora vasinfecta var. vasinfecta)(无性型为顶孢霉属Acremonium sp.)(1)。用通用引物ITS1/ITS4扩增两个分离株的核糖体DNA的内部转录间隔区(ITS)区域并测序(GenBank登录号JF705861和JF705862),与GenBank登录序列比较显示与大豆新小赤壳病菌菌株Pec070(登录号FJ940902)和菌株NRRL22497(登录号AY381142)有99%的相似性。进行了致病性测试。将5株3周龄的大豆品种华夏3号幼苗种植在直径20厘米的塑料盆中,在土壤线以下靠近根系的茎基部用针刺伤,然后用分生孢子悬浮液(每毫升10个)浇灌土壤进行接种。对照植株接种无菌水。每个处理有6个重复。处理后的植株在温室中25±2°C下培养。接种后3周,所有接种的植株都表现出叶片变黄以及茎和根黑腐的症状。从患病植株上重新分离出大豆新小赤壳病菌大豆变种。所有对照植株保持健康。据我们所知,这是中国首次观察到大豆新小赤壳茎腐病。该病原菌可能对中国的主要作物大豆构成威胁。这种病害此前在美国有报道,不过该病原菌的无性型要么未被鉴定,要么被鉴定为柱孢属(Cylindrocarpon sp.)(2,4)。这种真菌也与人类感染有关(3)。参考文献:(1)P. F. Cannon和D. L. Hawksworth。《英国真菌学会会刊》82:673,1984年。(2)F. A. Gray等人。《植物病害》64:321,1980年。(3)P. Manikandan等人。《医学真菌学》46:279,2008年。(4)D. V. Phillips。《植物病理学》62:612,1972年。