Fernández-Ortuño D, Li X, Chai W, Schnabel G
Department of Entomology, Soils, and Plant Sciences, Clemson University, Clemson, SC.
Plant Dis. 2011 Nov;95(11):1482. doi: 10.1094/PDIS-06-11-0529.
Gray mold caused by Botrytis spp. is one of the most economically important diseases of cultivated strawberry (Fragaria × ananassa) worldwide. From April to June 2011, strawberries with symptoms resembling gray mold disease were collected from different locations (Chesnee, Florence, Lexington, McBee, Monetta, and North Augusta) in South Carolina. Fruit infections began as small, firm, light brown lesions that enlarged quickly, becoming covered with a gray, fuzzy mass of spores followed by a soft rot. To isolate the causal agent, spores from symptomatic fruit were suspended in 1% Tween 20, streaked onto the surface of potato dextrose agar plates, and incubated at 22°C. Fungal colonies from single spores were at first colorless and later became gray to brown when the conidiphores and conidia developed. Conidia were identified by their morphological characteristics: an average size of 14 × 9 μm, ellipsoid to rounded without internal structure, and with a scar on the point of union to the conidiophore (1). Sclerotia produced in culture were hard, dark, irregular shaped, and formed after 2 weeks. The pathogen was identified as Botrytis cinerea Pers.: on the basis of morphology and confirmed by a restriction digest with ApoI of the 413-kb PCR amplification product obtained with BA2f/BA1r primers (2). Koch's postulates were conducted by inoculating 10 surface-sterilized strawberries with a conidial suspension (10 spores/ml) of a randomly chosen B. cinerea isolate previously characterized; 10 control fruit received sterile water without conidia. The inoculated fruit were incubated for 3 days at room temperature in air-tight plastic bags. Inoculated fruit developed typical gray mold symptoms with gray sporulating lesions. The developing spores on inoculated fruit were confirmed to be B. cinerea. All control fruit remained healthy. For many Botrytis spp., the internal transcribed spacer region does not reveal nucleotide variations and thus is useless for species identification. We used additional, more appropriate genetic markers for molecular-based species identification and verified that strawberries in South Carolina are affected by gray mold disease caused by B. cinerea. To our knowledge, this is the first scientific report of B. cinerea causing gray mold of strawberry in South Carolina. References: (1) W. R. Jarvis. Botryotinia and Botrytis Species: Taxonomy, Physiology and Pathogenicity. A Guide to the Literature. Monograph no. 15. Canada Department of Agriculture, Research Branch, Ottawa, 1977. (2) K. Nielsen et al. Plant Dis. 86:682, 2002.
由葡萄孢属(Botrytis spp.)引起的灰霉病是全球栽培草莓(Fragaria × ananassa)最具经济重要性的病害之一。2011年4月至6月,从南卡罗来纳州不同地点(切斯尼、佛罗伦萨、列克星敦、麦克比、莫内塔和北奥古斯塔)采集了有类似灰霉病症状的草莓。果实感染开始时为小而硬的浅褐色病斑,病斑迅速扩大,随后被灰色、毛茸茸的孢子团覆盖,接着出现软腐。为分离病原菌,将有症状果实上的孢子悬浮于1%吐温20中,划线接种到马铃薯葡萄糖琼脂平板表面,在22°C下培养。单个孢子形成的真菌菌落最初无色,当分生孢子梗和分生孢子形成后,菌落颜色变为灰色至褐色。分生孢子通过其形态特征进行鉴定:平均大小为14×9μm,椭圆形至圆形,无内部结构,在与分生孢子梗结合处有疤痕(1)。培养产生的菌核坚硬、深色、形状不规则,在2周后形成。根据形态学鉴定该病原菌为灰葡萄孢(Botrytis cinerea Pers.),并用BA2f/BA1r引物对获得的413-kb PCR扩增产物进行ApoI酶切消化进行确认(2)。通过用先前鉴定的随机选择的灰葡萄孢分离株的分生孢子悬浮液(10个孢子/ml)接种10个表面消毒的草莓来进行柯赫氏法则验证;另外10个对照果实接种无菌水而无分生孢子。接种的果实在室温下置于密封塑料袋中培养3天。接种的果实出现典型的灰霉病症状,有灰色产孢病斑。接种果实上发育中的孢子经确认是灰葡萄孢。所有对照果实保持健康。对于许多葡萄孢属物种,内部转录间隔区未显示核苷酸变异,因此对于物种鉴定无用。我们使用了其他更合适的遗传标记进行基于分子的物种鉴定,并验证了南卡罗来纳州的草莓受到灰葡萄孢引起的灰霉病影响。据我们所知,这是关于灰葡萄孢在南卡罗来纳州引起草莓灰霉病的第一份科学报告。参考文献:(1)W. R. Jarvis。葡萄孢盘菌属和葡萄孢属物种:分类学(Taxonomy)、生理学和致病性。文献指南。专论第15号。加拿大农业部,研究处,渥太华,1977年。(2)K. Nielsen等人。植物病害(Plant Dis.)86:682,2002年。