浙江农业学报 ›› 2025, Vol. 37 ›› Issue (2): 456-465.DOI: 10.3969/j.issn.1004-1524.20240313
四种食用菌采后主要病原菌的分离与鉴定
巩鑫鑫1,2(
), 刘瑞玲2, 韩延超2, 孟祥红1, 郜海燕2,*(), 陈杭君2,*()
- 1.中国海洋大学 食品科学与工程学院,山东 青岛 266100
2.浙江省农业科学院 食品科学研究所,农业农村部蔬菜采后保鲜与加工重点实验室(省部共建),农业农村部果品采后处理重点实验室,全省生鲜食品智慧物流与加工重点实验室,中国轻工业果蔬保鲜与加工重点实验室,浙江 杭州 310021
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收稿日期:2024-04-03出版日期:2025-02-25发布日期:2025-03-20 -
作者简介:陈杭君,E-mail: spshangjun@sina.com
郜海燕,E-mail: spsghy@163.com;
巩鑫鑫(1998—),女,河南许昌人,硕士研究生,研究方向为果蔬采后保鲜技术研究。E-mail:gongxx88@qq.com -
通讯作者:郜海燕,陈杭君 -
基金资助:浙江省“三农九方”科技协作计划项目(2022SNJF045);国家自然科学基金(32072294)
Isolation and identification of main postharvest pathogens of four edible fungi
GONG Xinxin1,2(), LIU Ruiling2, HAN Yanchao2, MENG Xianghong1, GAO Haiyan2,*(), CHEN Hangjun2,*()
- 1. College of Food Science and Engineering, Ocean University of China, Qingdao 266100, Shandong, China
2. Key Laboratory of Postharvest Vegetable Preservation and Processing of the Ministry of Agriculture and Rural Affairs (Co-Constructed by the Ministry and Province), Key Laboratory of Post-Harvest Handling of Fruits, Ministry of Agriculture and Rural Affairs, Zhejiang Key Laboratory of Intelligent Food Logistic and Processing, Key Laboratory of Postharvest Preservation and Processing of Fruits and Vegetables, China National Light Industry, Institute of Food Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
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Received:2024-04-03Online:2025-02-25Published:2025-03-20 -
Contact:GAO Haiyan,CHEN Hangjun
摘要：
为明确浙江食用菌采后主要病原菌的种类,文章选取香菇、双孢蘑菇、黑木耳和秀珍菇为试验材料,采用组织分离法和平板划线法对病原菌进行分离纯化,并进行形态学及分子生物学鉴定。序列比对和系统发育树分析结果表明,4种食用菌的主要病原菌分别为哈茨木霉(Trichoderma harzianum)、绿色木霉(Trichoderma virens)、塔宾曲霉(Aspergillus tubingensis)和恶臭假单胞菌(Pseudomonas putida)。食用菌采后主要病原菌的确定可为其现阶段的保鲜防治措施提供依据,从而及时保障食用菌产品的安全。
中图分类号:
引用本文
巩鑫鑫, 刘瑞玲, 韩延超, 孟祥红, 郜海燕, 陈杭君. 四种食用菌采后主要病原菌的分离与鉴定[J]. 浙江农业学报, 2025, 37(2): 456-465.
GONG Xinxin, LIU Ruiling, HAN Yanchao, MENG Xianghong, GAO Haiyan, CHEN Hangjun. Isolation and identification of main postharvest pathogens of four edible fungi[J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 456-465.
图1 病原菌分离与回接发病症状 a,XG-1在香菇上的发病状态;b,SB-1在双孢蘑菇上的发病状态;c,ME-1在黑木耳上的发病状态;d,XZ-1在秀珍菇上的发病状态。
Fig.1 Isolation of strains of the causative organisms and the disease of the splice back a, XG-1 on Lentinula edodes; b, SB-1 on Agaricus bisporus; c, ME-1 on Auricularia auricula; d, XZ-1 on Pleurotus pulmonarius.
图1 病原菌分离与回接发病症状 a,XG-1在香菇上的发病状态;b,SB-1在双孢蘑菇上的发病状态;c,ME-1在黑木耳上的发病状态;d,XZ-1在秀珍菇上的发病状态。
Fig.1 Isolation of strains of the causative organisms and the disease of the splice back a, XG-1 on Lentinula edodes; b, SB-1 on Agaricus bisporus; c, ME-1 on Auricularia auricula; d, XZ-1 on Pleurotus pulmonarius.
图2 病原菌在培养基25 ℃培养2 d的菌落特征 a,XG-1在PDA平板上的形态特征;b,SB-1在PDA平板上的形态特征;c,ME-1在PDA平板上的形态特征;d,XZ-1在LB平板上的形态特征。
Fig.2 Colony characteristics of strains of pathogenic microbes grown on medium for 2 d at 25 ℃ a, Morphology of XG-1 on PDA plate; b, Morphology of SB-1 on PDA plate; c, Morphology of ME-1 on PDA plate; d, Morphology of XZ-1 on LB plate.
图2 病原菌在培养基25 ℃培养2 d的菌落特征 a,XG-1在PDA平板上的形态特征;b,SB-1在PDA平板上的形态特征;c,ME-1在PDA平板上的形态特征;d,XZ-1在LB平板上的形态特征。
Fig.2 Colony characteristics of strains of pathogenic microbes grown on medium for 2 d at 25 ℃ a, Morphology of XG-1 on PDA plate; b, Morphology of SB-1 on PDA plate; c, Morphology of ME-1 on PDA plate; d, Morphology of XZ-1 on LB plate.
图3 病原菌在培养基25 ℃培养2 d的显微形态观察结果 a,XG-1分离株的孢子及孢子梗形态;b,SB-1分离株的孢子及孢子梗形态;c,ME-1分离株的孢子及孢子梗形态;d,革兰氏染色后的菌体XZ-1(1 000×)。
Fig.3 Results of morphological characteristics of pathogenic microbes cultured on plate for 2 d at 25 ℃ a, Spore and peduncle morphology of XG-1 isolate; b, Spore and peduncle morphology of SB-1 isolate; c, Spore and peduncle morphology of ME-1 isolate; d, Gram-stained organism XZ-1 (1 000×).
图3 病原菌在培养基25 ℃培养2 d的显微形态观察结果 a,XG-1分离株的孢子及孢子梗形态;b,SB-1分离株的孢子及孢子梗形态;c,ME-1分离株的孢子及孢子梗形态;d,革兰氏染色后的菌体XZ-1(1 000×)。
Fig.3 Results of morphological characteristics of pathogenic microbes cultured on plate for 2 d at 25 ℃ a, Spore and peduncle morphology of XG-1 isolate; b, Spore and peduncle morphology of SB-1 isolate; c, Spore and peduncle morphology of ME-1 isolate; d, Gram-stained organism XZ-1 (1 000×).
表1 食用菌病原菌序列检测结果
Table 1 Sequence detection results of edible fungi pathogenic microbes
| 名称 Name | 序列 Sequence |
|---|---|
| XG-1 | TCCGTAGGTGAACCTGCGGAGGGATCATTACCGAGTTTACAACTCCCAAACCCAATGTGAACGTTACCAAACTGTTGCCTCGGCGGG- ATCTCTGCCCCGGGTGCGTCGCAGCCCCGGACCAAGGCGCCCGCCGGAGGACCAACCAAAACTCTTTTTGTATACCCCCTCGCGGGT- TTTTTATAATCTGAGCCTTCTCGGCGCCTCTCGTAGGCGTTTCGAAAATGAATCAAAACTTTCAACAACGGATCTCTTGGTTCTGGCAT- CGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCGC- CAGTATTCTGGCGGGCATGCCTGTCCGAGCGTCATTTCAACCCTCGAACCCCTCCGGGGGGTCGGCGTTGGGGATCGGCCCTGCCTTT- GGCGGTGGCCGTCTCCGAAATACAGTGGCGGTCTCGCCGCAGCCTCTCCTGCGCAGTAGTTTGCACACTCGCATCGGGAGCGCGGCG- CGTCCACAGCCGTTAAACACCCAACTTCTGAAATGTTGACCTCGGATCAGGTAGGAATACCCGCTGAACTTAAGCATATCAATAAGC- GGAGGAA |
| SB-1 | TGGAAGTTAAAAAAACGAAACAAGGTCTCCGTTGGTGAACCAGCGGAGGGATCATTACCGAGTTTACAACTCCCAAACCCAATGTG- AACGTTACCAAACTGTTGCCTCGGCGGGATCTCTGCCCCGGGTGCGTCGCAGCCCCGGACCAAGGCGCCCGCCGGAGGACCAACCA- AAACTCTTATTGTATACCCCCTCGCGGGTTTTTTACTATCTGAGCCATCTCGGCGCCCCTCGTGGGCGTTTCGAAAATGAATCAAAAC- TTTCAACAACGGATCTCTTGGTTCTGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATC- ATCGAATCTTTGAACGCACATTGCGCCCGCCAGTATTCTGGCGGGCATGCCTGTCCGAGCGTCATTTCAACCCTCGAACCCCTCCGG- GGGGTCGGCGTTGGGGATCGGCCCTTTACGGGGCCGGCCCCGAAATACAGTGGCGGTCTCGCCGCAGCCTCTCCTGCGCAGTAGTTT- GCACACTCGCATCGGGAGCGCGGCGCGTCCACAGCCGTTAAACACCCCAAACTTCTGAAATGTGACCTCGGATCAGGTAGGAATTC- CCAG |
| ME-1 | CTTCCGTAGGTGAACCTGCGGAAGGATCATTACCGAGTGCGGGTCCTTTGGGCCCAACCTCCCATCCGTGTCTATTATACCCTGTTGC- TTCGGCGGGCCCGCCGCTTGTCGGCCGCCGGGGGGGCGCCTTTGCCCCCCGGGCCCGTGCCCGCCGGAGACCCCAACACGAACACT- GTCTGAAAGCGTGCAGTCTGAGTTGATTGAATGCAATCAGTTAAAACTTTCAACAATGGATCTCTTGGTTCCGGCATCGATGAAGAA- CGCAGCGAAATGCGATAACTAATGTGAATTGCAGAATTCAGTGAATCATCGAGTCTTTGAACGCACATTGCGCCCCCTGGTATTCCG- GGGGGCATGCCTGTCCGAGCGTCATTGCTGCCCTCAAGCCCGGCTTGTGTGTTGGGTCGCCGTCCCCCTCTCCGGGGGGACGGGCCC- GAAAGGCAGCGGCGGCACCGCGTCCGATCCTCGAGCGTATGGGGCTTTGTCACATGCTCTGTAGGATTGGCCGGCGCCTGCCGACGT- TTTCCAACCATTTTTTCCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCAATAAGCGGAGGAA |
| XZ-1 | GAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGATGACGGGAGCTTGCTCCTTGATT- CAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTGGTAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGTC- CTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGGGGTAATGGCTCAC- CAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAG- TGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTT- GGGAGGAAGGGCAGTAAGCTAATACCTTGCTGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGT- AATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTCGTTAAGTTGGATGTGAAAGCCCCGGG- CTCAACCTGGGAACTGCATCCAAAACTGGCGAGCTAGAGTACGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAG- ATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGAT- TAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGAATCCTTGAGATTTTAGTGGCGCAGCTAACGCATTAAGT- TGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATT- CGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCTGACACAG- GTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACG- TTATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTG- GGCTACACACGTGCTACAATGGTCGGTACAGAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCTCACAAAACCGATCGTAGTCCGGA- TCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGCCTTGT- ACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGCTAGTCTAACCTTCGGGAGGACGGTTACCACGGTGTGATTC- ATGACTGGGGTGAAG |
表1 食用菌病原菌序列检测结果
Table 1 Sequence detection results of edible fungi pathogenic microbes
| 名称 Name | 序列 Sequence |
|---|---|
| XG-1 | TCCGTAGGTGAACCTGCGGAGGGATCATTACCGAGTTTACAACTCCCAAACCCAATGTGAACGTTACCAAACTGTTGCCTCGGCGGG- ATCTCTGCCCCGGGTGCGTCGCAGCCCCGGACCAAGGCGCCCGCCGGAGGACCAACCAAAACTCTTTTTGTATACCCCCTCGCGGGT- TTTTTATAATCTGAGCCTTCTCGGCGCCTCTCGTAGGCGTTTCGAAAATGAATCAAAACTTTCAACAACGGATCTCTTGGTTCTGGCAT- CGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCGC- CAGTATTCTGGCGGGCATGCCTGTCCGAGCGTCATTTCAACCCTCGAACCCCTCCGGGGGGTCGGCGTTGGGGATCGGCCCTGCCTTT- GGCGGTGGCCGTCTCCGAAATACAGTGGCGGTCTCGCCGCAGCCTCTCCTGCGCAGTAGTTTGCACACTCGCATCGGGAGCGCGGCG- CGTCCACAGCCGTTAAACACCCAACTTCTGAAATGTTGACCTCGGATCAGGTAGGAATACCCGCTGAACTTAAGCATATCAATAAGC- GGAGGAA |
| SB-1 | TGGAAGTTAAAAAAACGAAACAAGGTCTCCGTTGGTGAACCAGCGGAGGGATCATTACCGAGTTTACAACTCCCAAACCCAATGTG- AACGTTACCAAACTGTTGCCTCGGCGGGATCTCTGCCCCGGGTGCGTCGCAGCCCCGGACCAAGGCGCCCGCCGGAGGACCAACCA- AAACTCTTATTGTATACCCCCTCGCGGGTTTTTTACTATCTGAGCCATCTCGGCGCCCCTCGTGGGCGTTTCGAAAATGAATCAAAAC- TTTCAACAACGGATCTCTTGGTTCTGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATC- ATCGAATCTTTGAACGCACATTGCGCCCGCCAGTATTCTGGCGGGCATGCCTGTCCGAGCGTCATTTCAACCCTCGAACCCCTCCGG- GGGGTCGGCGTTGGGGATCGGCCCTTTACGGGGCCGGCCCCGAAATACAGTGGCGGTCTCGCCGCAGCCTCTCCTGCGCAGTAGTTT- GCACACTCGCATCGGGAGCGCGGCGCGTCCACAGCCGTTAAACACCCCAAACTTCTGAAATGTGACCTCGGATCAGGTAGGAATTC- CCAG |
| ME-1 | CTTCCGTAGGTGAACCTGCGGAAGGATCATTACCGAGTGCGGGTCCTTTGGGCCCAACCTCCCATCCGTGTCTATTATACCCTGTTGC- TTCGGCGGGCCCGCCGCTTGTCGGCCGCCGGGGGGGCGCCTTTGCCCCCCGGGCCCGTGCCCGCCGGAGACCCCAACACGAACACT- GTCTGAAAGCGTGCAGTCTGAGTTGATTGAATGCAATCAGTTAAAACTTTCAACAATGGATCTCTTGGTTCCGGCATCGATGAAGAA- CGCAGCGAAATGCGATAACTAATGTGAATTGCAGAATTCAGTGAATCATCGAGTCTTTGAACGCACATTGCGCCCCCTGGTATTCCG- GGGGGCATGCCTGTCCGAGCGTCATTGCTGCCCTCAAGCCCGGCTTGTGTGTTGGGTCGCCGTCCCCCTCTCCGGGGGGACGGGCCC- GAAAGGCAGCGGCGGCACCGCGTCCGATCCTCGAGCGTATGGGGCTTTGTCACATGCTCTGTAGGATTGGCCGGCGCCTGCCGACGT- TTTCCAACCATTTTTTCCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCAATAAGCGGAGGAA |
| XZ-1 | GAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGATGACGGGAGCTTGCTCCTTGATT- CAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTGGTAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGTC- CTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGGGGTAATGGCTCAC- CAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAG- TGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTT- GGGAGGAAGGGCAGTAAGCTAATACCTTGCTGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGT- AATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTCGTTAAGTTGGATGTGAAAGCCCCGGG- CTCAACCTGGGAACTGCATCCAAAACTGGCGAGCTAGAGTACGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAG- ATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGAT- TAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGAATCCTTGAGATTTTAGTGGCGCAGCTAACGCATTAAGT- TGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATT- CGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCTGACACAG- GTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACG- TTATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTG- GGCTACACACGTGCTACAATGGTCGGTACAGAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCTCACAAAACCGATCGTAGTCCGGA- TCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGCCTTGT- ACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGCTAGTCTAACCTTCGGGAGGACGGTTACCACGGTGTGATTC- ATGACTGGGGTGAAG |
图4 食用菌病原菌rDNA的PCR扩增结果
Fig.4 PCR products of rDNA sequence of edible fungi pathogenic microbes
图4 食用菌病原菌rDNA的PCR扩增结果
Fig.4 PCR products of rDNA sequence of edible fungi pathogenic microbes
图5 食用菌病原菌XG-1(a)、SB-1(b)、ME-1(c)、XZ-1(d)的系统进化树
Fig.5 Phylogenetic tree of pathogens XG-1 (a), SB-1 (b), ME-1 (c) and XZ-1 (d)
图5 食用菌病原菌XG-1(a)、SB-1(b)、ME-1(c)、XZ-1(d)的系统进化树
Fig.5 Phylogenetic tree of pathogens XG-1 (a), SB-1 (b), ME-1 (c) and XZ-1 (d)
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