Nicoli A, Zambolim L, Nasu E G C, Pinho D B, Pereira O L, Cabral P G C, Zambolim E M
Departamento de Fitopatologia, Universidade Federal de Viçosa, Viçosa, Minas Gerais, 36570-000, Brazil.
Plant Dis. 2011 Sep;95(9):1194. doi: 10.1094/PDIS-02-11-0081.
In Brazil, Capsicum chinense Jacq. is the predominant species of commercial hot peppers because of its popular citrus-like aroma and adaptability to different soils and climates (4). In June 2010, 30 samples of C. chinense with severe leaf spot were collected from a field in the city of Viçosa, state of Minas Gerais, Brazil. Symptoms were observed on leaves, calyxes, fruits, and stems on most of the plants found in the area. On leaves, symptoms included amphigenous lesions that were initially circular to ellipsoid, 1 to 5 mm in diameter, whitish to tan in the center, and surrounded by a dark brown or reddish purple border. Lesions coalesce and turned necrotic with age. A fungus isolated from the lesions matched well with the description of Cercospora apii Fresen. It formed erumpent stromata that were dark brown and spherical to irregular; fascicule conidiophores were clear brown or pale, straight or curved, unbranched, geniculate, 22.5 to 80 × 5 to 7.5 μm, 0 to 3 septate, subtruncate apex; and conidia were solitary, hyaline to subhyaline, filiform, base truncate, tip acute, straight to curved, 12.5 to 140 × 3.5 to 5 μm, and 0 to 11 septate (1,2). A sample was deposited in the herbarium of the Universidade Federal de Viçosa, Minas Gerais, Brazil (VIC 31415). Identity was confirmed by amplifying part of the calmodulin gene with species-specific primers CercoCal-apii and CercoCal-R (3) of fungal DNA from a single-spore culture. In amplification reaction, initial denaturation step was done at 94°C for 5 min, followed by 40 cycles of denaturation at 94°C (30 s), annealing at 56°C (30 s), and elongation at 72°C (30 s). Primers CercoCal-apii and CercoCal-R amplified a single DNA product of 176 bp, and coupled with the morphological characteristics, confirmed the identity of the fungus as Cercospora apii. To check pathogenicity, a 6-mm-diameter plug of the isolate was removed from the expanding edge of a 21-day-old culture grown on potato dextrose agar (PDA) and placed in contact with the adaxial face of the leaves of 8-week-old C. chinense grown in 2-liter plastic pots with soil substrate. Six plants, one per pot, were inoculated with the isolate and six plants were inoculated with the fungus-free PDA plug. Inoculated plants were maintained in a moist chamber for 24 h and then subsequently kept in a greenhouse at 26°C. Leaf spot was observed in all inoculated plants 15 days after inoculation and symptoms were similar to those expressed in the field. The fungus was reisolated from the inoculated plants and matched well with the description of Cercospora apii. All fungus-free PDA inoculated plants remained healthy. Cercospora apii comprises a complex of 281 morphologically indistinguishable species that can infect an extremely wide host range (2). To our knowledge, this pathogen has the potential to cause significant damage to the hot pepper industry of Brazil. References: (1) C. Chupp. A Monograph of the Fungus Cercospora. Cornell University Press, Ithaca, NY, 1954. (2) P. W. Crous and U. Braun. CBS Biodivers. Ser. 1:1, 2003. (3) M. Groenewald et al. Phytopathology 95:951, 2005. (4) S. D. Lannes et al. Sci. Hortic. 112:266, 2007.
在巴西,中国辣椒(Capsicum chinense Jacq.)是商业辣椒的主要品种,因其具有受欢迎的类似柑橘的香气,且能适应不同的土壤和气候(4)。2010年6月,从巴西米纳斯吉拉斯州维索萨市的一块田地中采集了30份患有严重叶斑病的中国辣椒样本。在该区域发现的大多数植株的叶片、花萼、果实和茎上均观察到症状。叶片上的症状包括两面生的病斑,最初为圆形至椭圆形,直径1至5毫米,中心为白色至棕褐色,周围有深褐色或红紫色边缘。病斑随着时间推移会合并并坏死。从病斑中分离出的一种真菌与芹菜尾孢(Cercospora apii Fresen.)的描述非常吻合。它形成了突出的子座,呈深褐色,球形至不规则形;束生分生孢子梗为透明褐色或浅色,直或弯曲,不分枝,膝状,22.5至80×5至7.5微米,0至3个隔膜,顶端近截形;分生孢子单个,透明至近透明,丝状,基部截形,顶端尖锐,直或弯曲,12.5至140×3.5至5微米,0至11个隔膜(1,2)。一个样本保存在巴西米纳斯吉拉斯州维索萨联邦大学的植物标本馆(VIC 31415)。通过使用真菌DNA单孢子培养物的物种特异性引物CercoCal-apii和CercoCal-R扩增钙调蛋白基因的部分片段来确认其身份(3)。在扩增反应中,初始变性步骤在94°C下进行5分钟,随后进行40个循环,94°C变性(30秒),56°C退火(30秒),72°C延伸(30秒)。引物CercoCal-apii和CercoCal-R扩增出一个176 bp的单一DNA产物,并结合形态特征,确认该真菌为芹菜尾孢。为了检查致病性,从在马铃薯葡萄糖琼脂(PDA)上生长21天的培养物的扩展边缘切下一个直径6毫米的分离物菌块,使其与在装有土壤基质的2升塑料盆中生长8周的中国辣椒叶片的正面接触。六株植物,每盆一株,接种该分离物,另外六株植物接种无真菌的PDA菌块。接种后的植物在保湿箱中保持24小时,然后随后置于26°C的温室中。接种后15天,在所有接种的植物中均观察到叶斑病,症状与田间表现相似。从接种的植物中重新分离出该真菌,与芹菜尾孢的描述非常吻合。所有接种无真菌PDA的植物均保持健康。芹菜尾孢包括281个形态上难以区分的物种的复合体,可感染极其广泛的寄主范围((2)。据我们所知,这种病原菌有可能对巴西的辣椒产业造成重大损害。参考文献:(1)C. Chupp. A Monograph of the Fungus Cercospora. Cornell University Press, Ithaca, NY, 1954.(2)P. W. Crous和U. Braun. CBS Biodivers. Ser. 1:1, 2003.(3)M. Groenewald等人. Phytopathology 95:951, 2005.(4)S. D. Lannes等人. Sci. Hortic., 112:266, 2007.