山东省辣椒炭疽病病原菌的鉴定及高效防治药剂的筛选

doi:10.3864/j.issn.0578-1752.2017.08.009

摘要/Abstract

摘要： 【目的】明确山东省露地辣椒主产区炭疽病致病菌的种类，评价不同药剂对致病菌的毒力和活体接种防效，筛选高效防治药剂。【方法】从山东省菏泽、济宁等5个辣椒主产区采集感炭疽病的辣椒果实，经分离、纯化培养后，观察菌落及分生孢子的形态、大小和病原菌的致病性，利用真菌通用引物ITS4和ITS5对4个典型代表菌株的rDNA-ITS进行扩增，对扩增产物进行回收和测序，并利用MEGA 5.1软件进行基于病原菌的rDNA-ITS序列和GenBank中相关炭疽菌序列的系统发育树构建，确定病原菌的种类。采用菌丝生长速率法和孢子萌发法测定11种杀菌剂对辣椒炭疽病菌的毒力，通过活体接种试验验证药剂对辣椒炭疽病的防效。【结果】分离得到的27个菌株形态特征相同，气生菌丝较发达，初呈白色，而后逐渐变为浅灰色，分生孢子无色单胞，椭圆形，一端稍尖，大小为（7.48—14.69）μm×（2.52—5.64）μm。将所分离得到的菌株接种于刺伤果实或无伤口果实，两者均可发病，但刺伤果实上病斑发展快。病菌既侵染未成熟的辣椒果实，也侵染成熟的果实，病斑呈椭圆形凹陷，表面有橘黄色的分生孢子团。所选取的4个代表性菌株的rDNA-ITS序列的长度为562、541、557和553 bp，通过系统进化树分析，4个代表菌株与炭疽菌属尖孢炭疽菌（Colletotrichum acutatum）聚在同一分支上，亲缘关系置信度为100%。室内毒力测定试验表明，吡唑醚菌酯、咯菌腈和啶菌唑对尖孢炭疽菌的菌丝生长和孢子萌发均具有较高的抑制活性。活体接种试验表明，吡唑醚菌酯、咯菌腈和啶菌唑对辣椒炭疽病具有较好的预防和治疗作用，在浓度为400 μg·mL-1时，对辣椒炭疽病的预防效果和治疗效果均大于60%，高于对照药剂嘧菌酯的防效。【结论】引起山东省露地辣椒主产区辣椒炭疽病的致病菌为尖孢炭疽菌。吡唑醚菌酯、咯菌腈和啶菌唑对该菌具有较高的毒力和活体防效，在辣椒炭疽病的田间防治中具有较大的应用潜力。

关键词: 辣椒炭疽病, 尖孢炭疽菌, 杀菌剂, 室内毒力, 防治效果

Abstract: 【Objective】The objective of this study is to identify the pathogen species of pepper anthracnose in major pepper growing areas in Shandong Province, and to screen the highly effective fungicides by inhibitory activity test in vitro and relative control efficacy trials on detached pepper fruits inoculated with conidial suspension. 【Method】 The isolates of pepper anthracnose were collected from the major pepper producing areas in Jining, Heze, Weifang, Taian and Dezhou. After isolation and purification, the isolates were identified by using the methods of morphological characteristics analysis, its pathogenicity test on detached pepper fruits and molecular diagnostics. Morphology parameters mainly include the colony and conidial morphology, conidial size. The internal transcribed spacer (ITS) rDNA region of the four typical isolates was amplified by using the fungal universal primers ITS4 and ITS5, and then the amplified product was recycled and sequenced. MEGA 5.1 was used to generate phylogenetic tree to confirm the pathogen species. The inhibitory activity in vitro of 11 fungicides to Colletotrichum acutatum isolate was determined by the inhibition of the 11 fungicides against mycelial growth and spore germination of the pathogen. Control efficacies of the selected fungicides against pepper anthracnose were determined on the detached pepper fruits.【Result】All of the twenty-seven isolates exhibited similar morphology, which produced white to pale grey colonies. The oval conidia were single-celled with a sub-cute end, (7.48-14.69) μm × (2.52-5.64) μm in size. Both of the mature and immature pepper fruits with or without wound could be infected by this pathogen, and disease spot expanded rapidly on wounded pepper fruits. Typical symptom of pepper anthracnose was sunken and necrotic lesion that covered with massive orange conidia. The rDNA-ITS sequence length of the four representative isolates was 562, 541, 557 and 553 bp (GenBank Accession No KX830854-KX830857), respectively. Phylogenetic tree analysis indicated that the four representative isolates and C. acutatum were divided into one group, and the bootstrap value was 100%. The inhibitory activity test in vitro indicated that pyraclostrobin, fludioxonil, and pyrisoxazole exhibited strong inhibitory activity against mycelial growth and spore germination of C. acutatum. Moreover, pyraclostrobin, fludioxonil and pyrisoxazole at the dose of 400 μg·mL^-1 also showed high preventive and curative activity against C. acutatum on detached pepper fruits, with the efficacy greater than 60%, which showed significantly higher efficacy than the preventive and curative activity of azoxystrobin.【Conclusion】 The major pathogenic fungus of pepper anthracnose in Shandong Province was identified as C. acutatum. Pyraclostrobin, fludioxonil and pyrisoxazole showed excellent inhibitory activity against C. acutatum both in vitro and in vivo, make these fungicides become potential candidates for the control of pepper anthracnose in the field.

Key words: pepper anthracnose, Colletotrichum acutatum, fungicides, inhibitory activity in vitro, control efficacy

高杨杨，禾丽菲，李北兴，林琎，慕卫，刘峰. 山东省辣椒炭疽病病原菌的鉴定及高效防治药剂的筛选[J]. 中国农业科学, 2017, 50(8): 1452-1464.

GAO YangYang, HE LiFei, LI BeiXing, LIN Jin, MU Wei, LIU Feng. Identification of the Pathogen Causing Pepper Anthracnose in Shandong Province and Screening of Highly Effective Fungicides[J]. Scientia Agricultura Sinica, 2017, 50(8): 1452-1464.

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