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尖孢镰刀菌引起的辣椒果实腐烂在中国的首次报道。

First Report of Pepper Fruit Rot Caused by Fusarium concentricum in China.

作者信息

Wang J H, Feng Z H, Han Z, Song S Q, Lin S H, Wu A B

机构信息

Institute for Agri-Food Standards and Testing Technology, Laboratory of Quality and Safety Risk Assessment for Agro-Products (Shanghai), Ministry of Agriculture, Shanghai Academy of Agricultural Sciences, 1000 Jinqi Road, Shanghai 201403, P. R. China. Funding provided by the Shanghai Agriculture Commission (2011NO. 4-3).

出版信息

Plant Dis. 2013 Dec;97(12):1657. doi: 10.1094/PDIS-03-13-0325-PDN.

Abstract

Pepper (Capsicum annuum L.) is an important vegetable crop worldwide. Some Fusarium species can cause pepper fruit rot, leading to significant yield losses of pepper production and, for some Fusarium species, potential risk of mycotoxin contamination. A total of 106 diseased pepper fruit samples were collected from various pepper cultivars from seven provinces (Gansu, Hainan, Heilongjiang, Hunan, Shandong, Shanghai, and Zhejiang) in China during the 2012 growing season, where pepper production occurs on approximately 25,000 ha. Pepper fruit rot symptom incidence ranged from 5 to 20% in individual fields. Symptomatic fruit tissue was surface-sterilized in 0.1% HgCl for 1 min, dipped in 70% ethanol for 30 s, then rinsed in sterilized distilled water three times, dried, and plated in 90 mm diameter petri dishes containing potato dextrose agar (PDA). After incubation for 5 days at 28°C in the dark, putative Fusarium colonies were purified by single-sporing. Forty-three Fusarium strains were isolated and identified to species as described previously (1,2). Morphological characteristics of one strain were identical to those of F. concentricum. Aerial mycelium was reddish-white with an average growth rate of 4.2 to 4.3 mm/day at 25°C in the dark on PDA. Pigments in the agar were formed in alternating red and orange concentric rings. Microconidia were 0- to 1-septate, mostly 0-septate, and oval, obovoid to allantoid. Macroconidia were relatively slender with no significant curvature, 3- to 5-septate, with a beaked apical cell and a foot-shaped basal cell. To confirm the species identity, the partial TEF gene sequence (646 bp) was amplified and sequenced (GenBank Accession No. KC816735). A BLASTn search with TEF gene sequences in NCBI and the Fusarium ID databases revealed 99.7 and 100% sequence identity, respectively, to known TEF sequences of F. concentricum. Thus, both morphological and molecular criteria supported identification of the strain as F. concentricum. This strain was deposited as Accession MUCL 54697 (http://bccm.belspo.be/about/mucl.php). Pathogenicity of the strain was confirmed by inoculating 10 wounded, mature pepper fruits that had been harvested 70 days after planting the cultivar Zhongjiao-5 with a conidial suspension (1 × 10 spores/ml), as described previously (3). A control treatment consisted of inoculating 10 pepper fruits of the same cultivar with sterilized distilled water. The fruit were incubated at 25°C in a moist chamber, and the experiment was repeated independently in triplicate. Initially, green to dark brown lesions were observed on the outer surface of inoculated fruit. Typical soft-rot symptoms and lesions were observed on the inner wall when the fruit were cut open 10 days post-inoculation. Some infected seeds in the fruits were grayish-black and covered by mycelium, similar to the original fruit symptoms observed at the sampling sites. The control fruit remained healthy after 10 days of incubation. The same fungus was isolated from the inoculated infected fruit using the method described above, but no fungal growth was observed from the control fruit. To our knowledge, this is the first report of F. concentricum causing a pepper fruit rot. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (2) K. O'Donnell et al. Proc. Nat. Acad. Sci. USA 95:2044, 1998. (3) Y. Yang et al. 2011. Int. J. Food Microbiol. 151:150, 2011.

摘要

辣椒(Capsicum annuum L.)是全球重要的蔬菜作物。一些镰刀菌属物种可导致辣椒果实腐烂,造成辣椒产量大幅损失,并且对于某些镰刀菌属物种而言,还存在霉菌毒素污染的潜在风险。在2012年生长季,从中国七个省份(甘肃、海南、黑龙江、湖南、山东、上海和浙江)的不同辣椒品种中总共采集了106份患病辣椒果实样本,这些省份的辣椒种植面积约为25000公顷。单个田块中辣椒果实腐烂症状的发生率在5%至20%之间。有症状的果实组织先用0.1% HgCl进行表面消毒1分钟,再浸入70%乙醇中30秒,然后用无菌蒸馏水冲洗三次,晾干,接种到含有马铃薯葡萄糖琼脂(PDA)的90毫米直径培养皿中。在28°C黑暗条件下培养5天后,通过单孢分离法纯化疑似镰刀菌菌落。按照之前描述的方法(1,2)分离并鉴定出43株镰刀菌菌株到种。其中一个菌株的形态特征与同心镰刀菌一致。在PDA上,气生菌丝呈红白色,在25°C黑暗条件下平均生长速率为4.2至4.3毫米/天。琼脂中的色素形成交替的红色和橙色同心环。小型分生孢子0至1分隔,大多无分隔,呈椭圆形、倒卵形至腊肠形。大型分生孢子相对细长,无明显弯曲,3至5分隔,顶端细胞呈喙状,基部细胞呈足形。为确认物种身份,扩增并测序了部分TEF基因序列(646 bp)(GenBank登录号KC816735)。在NCBI和镰刀菌鉴定数据库中对TEF基因序列进行BLASTn搜索,结果显示与同心镰刀菌已知TEF序列的序列同一性分别为99.7%和100%。因此,形态学和分子标准均支持将该菌株鉴定为同心镰刀菌。该菌株保藏号为MUCL 54697(http://bccm.belspo.be/about/mucl.php)。按照之前描述的方法(3),用分生孢子悬浮液(1×10孢子/毫升)接种10个种植品种中椒5号70天后收获的受伤成熟辣椒果实,以确认该菌株的致病性。对照处理为用无菌蒸馏水接种10个相同品种的辣椒果实。果实置于25°C的保湿箱中培养,该实验独立重复三次。最初,在接种果实的外表面观察到从绿色到深褐色的病斑。接种10天后切开果实,在内壁观察到典型的软腐症状和病斑。果实中的一些受感染种子呈灰黑色,被菌丝体覆盖,类似于在采样点观察到的原始果实症状。对照果实培养10天后未观察到真菌生长。据我们所知,这是同心镰刀菌引起辣椒果实腐烂的首次报道。参考文献:(1)J. F. Leslie和B. A. Summerell。《镰刀菌实验室手册》。Blackwell Publishing,艾姆斯,爱荷华州,2006年。(2)K. O'Donnell等人。《美国国家科学院院刊》95:2044,1998年。(3)Y. Yang等人。2011年。《国际食品微生物学杂志》151:150,2011年。

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