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中国西北陕西省油菜上核盘菌田间分离株对菌核净抗性的首次报道

First Report of Dimethachlon Resistance in Field Isolates of Sclerotinia sclerotiorum on Oilseed Rape in Shaanxi Province of Northwestern China.

作者信息

Zhou F, Zhu F X, Zhang X L, Zhang A S

机构信息

College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.

Plant Protection Research Institute, Shandong Academy of Agricultural Sciences, Jinan, China.

出版信息

Plant Dis. 2014 Apr;98(4):568. doi: 10.1094/PDIS-07-13-0730-PDN.

Abstract

Sclerotinia sclerotiorum (Lib.) de Bary is a necrotrophic fungal pathogen causing diseases in a wide range of plants, including oilseed rape (3). Substantial economic losses caused by S. sclerotiorum have been reported in the United States, Canada, Brazil, South Africa, Hungary, India, Nepal, and Japan (1). Application of fungicides is the principal tool for controlling S. sclerotiorum because of lack of high level of host resistance. Dicarboximide fungicides such as dimethachlon have been widely used to control S. sclerotiorum in recent years in China and field isolates with reduced sensitivity to dimethachlon have been reported in Jiangsu Province of eastern China (2). In order to understand the current status of dimethachlon resistance in S. sclerotiorum isolates of northwestern China, 196 and 344 isolates of S. sclerotiorum collected from oilseed rape fields in 10 counties throughout Shaanxi Province in 2011 and 2012, respectively, were assayed for sensitivity to dimethachlon using 5 μg ml dimethachlon as a discriminatory dose. Mycelial plugs (6 mm in diameter) cut from the margin of a 48-h-old colony were placed in the center of petri dishes containing potato dextrose agar (PDA) amended with 5 μg ml dimethachlon; PDA without fungicide served as the control. Cultures were incubated at 26°C and colony growth was measured after 72 h of incubation. Isolates that showed growth on PDA amended with fungicide were tentatively considered resistant to dimethachlon, whereas the completely inhibited isolates were considered sensitive. Results showed that 1.02% or 2 isolates of the 196 isolates collected in 2011 and 3.78% or 13 isolates of the 344 isolates collected in 2012 were resistant to dimethachlon. For all the isolates considered resistant and 42 randomly selected sensitive isolates, 50% effective concentrations (EC) were determined on PDA amended with a series of dimethachlon concentrations. The average EC value of dimethachlon for sensitive isolates was 0.29 ± 0.02 μg ml Resistance ratios (EC of resistant isolate / average EC of sensitive isolates) for the two resistant isolates detected in 2011 were 10.28 and 23.83, respectively, whereas resistance ratios for the 13 resistant isolates detected in 2012 ranged from 24.90 to 101.97. The average EC value of dimethachlon for the 13 resistant isolates detected in 2012 was 19.05 μg ml, and EC values for the two resistant isolates detected in 2011 were 2.98 and 6.91 μg ml, respectively. These results indicated that both resistance frequency and resistance level increased from 2011 to 2012. Bioassay results of three resistant isolates indicated that there was positive cross-resistance between dimethachlon and other dicarboximide fungicides such as iprodione and procymidone. To our knowledge, this is the first report of dimethachlon resistance in S. sclerotiorum in Shaanxi Province of northwestern China. The molecular mechanism of dimethachlon resistance in field isolates of S. sclerotiorum remains to be studied. Although resistance frequency is low at present, dimethachlon resistance should be kept in mind and fungicide resistance management tactics such as use of biological control agents, fungicide tank-mixing, or alternating dimethachlon with other fungicides having different modes of action is recommended in controlling S. sclerotiorum. References: (1) M. D. Bolton et al. Mol Plant Pathol. 7:1, 2006. (2) H. X. Ma et al. Plant Dis. 93:36, 2009. (3) L. H. Prudy. Phytopathology 69:875, 1979.

摘要

核盘菌(Sclerotinia sclerotiorum (Lib.) de Bary)是一种坏死营养型真菌病原体,可在包括油菜在内的多种植物上引发病害(3)。在美国、加拿大、巴西、南非、匈牙利、印度、尼泊尔和日本,均有报道称核盘菌造成了巨大的经济损失(1)。由于缺乏高水平的寄主抗性,使用杀菌剂是防治核盘菌的主要手段。近年来,在中国,诸如二甲菌核利等二甲酰亚胺类杀菌剂已被广泛用于防治核盘菌,并且在中国东部的江苏省已报道了对二甲菌核利敏感性降低的田间分离株(2)。为了解中国西北地区核盘菌分离株对二甲菌核利的抗性现状,分别于2011年和2012年从陕西省10个县的油菜田中采集了196株和344株核盘菌分离株,以5 μg/ml二甲菌核利作为鉴别剂量测定其对二甲菌核利的敏感性。从48小时龄菌落边缘切取直径为6 mm的菌丝块,置于含有添加了5 μg/ml二甲菌核利的马铃薯葡萄糖琼脂(PDA)的培养皿中央;不含杀菌剂的PDA作为对照。培养物在26°C下培养,培养72小时后测量菌落生长情况。在添加了杀菌剂的PDA上生长的分离株被初步认为对二甲菌核利具有抗性,而完全被抑制的分离株被认为是敏感的。结果表明,2011年采集的196株分离株中有1.02%(即2株)对二甲菌核利具有抗性,2012年采集的344株分离株中有3.78%(即13株)对二甲菌核利具有抗性。对于所有被认为具有抗性的分离株以及42株随机选择的敏感分离株,在添加了一系列二甲菌核利浓度的PDA上测定50%有效浓度(EC)。敏感分离株的二甲菌核利平均EC值为0.29±0.02 μg/ml。2011年检测到的两株抗性分离株的抗性倍数(抗性分离株的EC/敏感分离株的平均EC)分别为10.28和23.83,而2012年检测到的13株抗性分离株的抗性倍数范围为24.90至101.97。2012年检测到的13株抗性分离株的二甲菌核利平均EC值为19.05 μg/ml,2011年检测到的两株抗性分离株的EC值分别为2.98和6.91 μg/ml。这些结果表明,从2011年到2012年,抗性频率和抗性水平均有所增加。三株抗性分离株的生物测定结果表明,二甲菌核利与其他二甲酰亚胺类杀菌剂如异菌脲和腐霉利之间存在正向交互抗性。据我们所知,这是中国西北部陕西省核盘菌对二甲菌核利抗性的首次报道。核盘菌田间分离株对二甲菌核利抗性的分子机制仍有待研究。尽管目前抗性频率较低,但在防治核盘菌时应牢记对二甲菌核利的抗性问题,并建议采取诸如使用生物防治剂、杀菌剂桶混或二甲菌核利与其他具有不同作用方式的杀菌剂交替使用等杀菌剂抗性管理策略。参考文献:(1)M. D. Bolton等人,《分子植物病理学》7:1,2006年。(2)H. X. Ma等人,《植物病害》93:36,2009年。(3)L. H. Prudy,《植物病理学》69:875,1979年。

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