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加拿大新斯科舍省感染储存马铃薯的抗噻苯达唑接骨木镰刀菌分离株的首次报告

First Report of Thiabendazole-Resistant Isolates of Fusarium sambucinum Infecting Stored Potatoes in Nova Scotia, Canada.

作者信息

Peters R D, Macdonald I K, MacIsaac K A, Woodworth S

机构信息

Agriculture and Agri-Food Canada, Crops and Livestock Research Centre, P.O. Box 1210, Charlottetown, PE, Canada C1A 7M8.

Cavendish Agri Services, P.O. Box 247, Kentville, NS, Canada B4N 3W4.

出版信息

Plant Dis. 2001 Sep;85(9):1030. doi: 10.1094/PDIS.2001.85.9.1030A.

Abstract

Fusarium dry rot is a significant postharvest disease of potato (Solanum tuberosum L.) and is often controlled by applying thiabendazole to tubers prior to storage. However, thiabendazole-resistant isolates of Fusarium spp. have been reported from Europe (2), the United States (1), and Canada (1,4). To address concerns, samples of potato tubers showing symptoms of dry rot caused by Fusariumspp. were collected from three storage bays in a commercial storage facility in Nova Scotia, Canada, in February 2001. All tubers had been treated with thiabendazole after harvest and prior to storage. Tubers were cut longitudinally, and small tissue samples (10 × 5 × 3 mm) were taken from the margins of internal necrotic regions with a sterile scalpel, surface-sterilized in 0.6% sodium hypochlorite for 15 s, rinsed twice in sterile distilled water (SDW), and blotted dry on sterile filter paper. Tissue pieces were plated on 0.5-strength potato dextrose agar (PDA) amended with tetracycline (0.05 g/liter) and streptomycin sulfate (0.1 g/liter). Petri dishes were incubated in the dark at 22°C for 4 to 7 days. After incubation, hyphal tips from the margins of actively growing isolates were removed with a sterile probe and plated on 0.5-strength PDA to generate pure cultures. Of 35 potato tubers examined, 10 (29%) yielded Fusarium isolates for further study. All 10 isolates were identified as F. sambucinum Fuckel according to Nelson et al. (3). Agar plugs (5 mm diameter) taken from the margins of 7- to 10-day-old cultures of F. sambucinum isolates were transferred to petri dishes containing 0.5-strength PDA amended with thiabendazole at 0, 1, 5, 10, 20, 50, or 100 mg/liter. Thiabendazole was prepared as a stock solution in SDW and added to molten agar after autoclaving. Cultures were grown in the dark for 7 days at 22°C, after which mycelial growth diameter was measured using digital calipers. Two measurements, along orthogonal diameters, were taken from each of three replicate plates for a total of six measurements per thiabendazole concentration. Means were calculated, and the diameter of the inoculation plug was subtracted from each mean. Calculated EC values (thiabendazole concentration inhibiting pathogen growth by 50%) were obtained by regression of the log of the chemical concentration against the corresponding probit of percent fungal inhibition. All isolates of F. sambucinum were resistant to thiabendazole, with EC values ranging from 7 to 82 mg/liter. Six isolates had EC values between 40 and 82 mg/liter. Control isolates of F. sambucinum, F. avenaceum, F. solani, and F. oxysporum were sensitive to thiabendazole, with EC values of <1 mg/liter. Although isolates of F. sambucinum resistant to thiabendazole have been recovered from eastern Canada (1,4), this is the first report of thiabendazole resistance in F. sambucinum isolates from tubers in commercial storage in the Annapolis Valley of Nova Scotia, Canada, a production region that concentrates on growing processing potatoes for the potato chip industry and is several hundred kilometers from other potato-growing regions of Prince Edward Island and New Brunswick. References: (1) A. E. Desjardins. Am. Potato J. 72:145, 1995. (2) G. A. Hide et al. Plant Pathol. 41:745, 1992. (3) P. E. Nelson et al. 1983. Fusarium Species: An Illustrated Manual for Identification . Pennsylvania State University Press, University Park, PA. (4) H. W. Platt. Phytoprotection 78:1, 1997.

摘要

镰刀菌干腐病是马铃薯(Solanum tuberosum L.)一种重要的采后病害,通常通过在贮藏前对块茎施用噻苯达唑来控制。然而,欧洲(2)、美国(1)和加拿大(1,4)均已报道了对噻苯达唑具有抗性的镰刀菌分离株。为解决相关问题,2001年2月从加拿大新斯科舍省一个商业贮藏设施的三个贮藏隔间中采集了表现出镰刀菌引起干腐症状的马铃薯块茎样本。所有块茎在收获后和贮藏前均已用噻苯达唑处理。将块茎纵向切开,用无菌手术刀从内部坏死区域边缘取小组织样本(10×5×3毫米),在0.6%次氯酸钠中进行表面消毒15秒,在无菌蒸馏水中冲洗两次,然后在无菌滤纸上吸干。将组织块接种在添加了四环素(0.05克/升)和硫酸链霉素(0.1克/升)的0.5强度马铃薯葡萄糖琼脂(PDA)上。将培养皿在22°C黑暗条件下培养4至7天。培养后,用无菌探针从活跃生长的分离株边缘取菌丝尖端,接种在0.5强度PDA上以获得纯培养物。在检查的35个马铃薯块茎中,有10个(29%)产生了镰刀菌分离株以供进一步研究。根据纳尔逊等人(3)的方法,所有10个分离株均被鉴定为接骨木镰刀菌(F. sambucinum Fuckel)。从接骨木镰刀菌分离株7至10天龄培养物边缘取直径5毫米的琼脂块,转移至含有添加了0、1、5、10、20、50或100毫克/升噻苯达唑的0.5强度PDA的培养皿中。噻苯达唑在无菌蒸馏水中配制成储备液,在高压灭菌后加入融化的琼脂中。培养物在22°C黑暗条件下生长7天,之后用数字卡尺测量菌丝生长直径。在每个噻苯达唑浓度下,从三个重复平板中的每一个平板沿着正交直径进行两次测量,共进行六次测量。计算平均值,并从每个平均值中减去接种块的直径。通过将化学物质浓度的对数与相应的真菌抑制百分比的概率单位进行回归分析,获得计算得出的EC值(抑制病原体生长50%的噻苯达唑浓度)。所有接骨木镰刀菌分离株均对噻苯达唑具有抗性,EC值范围为7至82毫克/升。六个分离株的EC值在40至82毫克/升之间。接骨木镰刀菌、燕麦镰刀菌、茄病镰刀菌和尖孢镰刀菌的对照分离株对噻苯达唑敏感,EC值<1毫克/升。尽管在加拿大东部已分离出对噻苯达唑具有抗性的接骨木镰刀菌(1,4),但这是加拿大新斯科舍省安纳波利斯谷商业贮藏块茎中接骨木镰刀菌分离株对噻苯达唑抗性的首次报道,该产区主要种植用于薯片产业的加工马铃薯,距离爱德华王子岛和新不伦瑞克省的其他马铃薯种植区有数百公里。参考文献:(1)A. E. Desjardins。《美国马铃薯杂志》72:145,1995年。(2)G. A. Hide等人。《植物病理学》41:745,1992年。(3)P. E. Nelson等人。1983年。《镰刀菌属:鉴定图谱手册》。宾夕法尼亚州立大学出版社,宾夕法尼亚州大学公园。(4)H. W. Platt。《植物保护》78:1,1997年。

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