秀珍菇黄菇病病原菌鉴定及其致病相关的转录组分析

doi:10.3969/j.issn.1004-1524.20240034

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (2): 426-437.DOI: 10.3969/j.issn.1004-1524.20240034

秀珍菇黄菇病病原菌鉴定及其致病相关的转录组分析

陆娜(), 王伟科(), 林佳瑶, 周祖法

杭州市农业科学研究院,浙江杭州 310024

收稿日期:2024-01-04 出版日期:2025-02-25 发布日期:2025-03-20
作者简介:*王伟科,E-mail:akeok@126.com
陆娜(1983—),女,江苏太仓人,学士,高级农艺师,主要从事食药用菌育种与栽培研究。E-mail:13738068366@163.com
通讯作者: 王伟科
基金资助:
浙江省农业新品种选育重大科技专项(2021C02073);国家食用菌产业技术体系(CARS-20)

Identification of yellow rot disease of Pleurotus pulmonarius and analysis of its pathogenicity-related transcriptome

LU Na(), WANG Weike(), LIN Jiayao, ZHOU Zufa

Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China

Received:2024-01-04 Online:2025-02-25 Published:2025-03-20
Contact: WANG Weike

摘要/Abstract

摘要： 在秀珍菇的工厂化生产中,黄菇病严重影响秀珍菇的产量和质量。为此,采集患病样本并经分离、纯化获得病原菌,采用柯赫氏法则,结合显微特征和16S rRNA等信息鉴定病原菌,并通过菌丝生长速度和转录组分析来揭示其特性。结果显示,从患病样本分离得到的CAJ01-D、CAJ06-C、TXX01和LAD07菌株均能引起黄菇病,这些菌株为假单胞菌属(Pseudomonas spp.)的不同种。不同菌株对秀珍菇菌丝的生长速度有影响,但菌丝长势无异。病原菌侵染下的相关基因表达水平存在差异,其中,LAD07和CAJ06-C菌株侵染下的差异表达基因的表达水平多数下调。GO、KEGG功能的聚类和富集显示,LAD07菌株引起细胞色素c还原酶活性下调,线粒体呼吸作用减弱;CAJ06-C菌株引起酵解或糖异生、酪氨酸代谢和苯丙氨酸代谢出现特有的显著下调。据此推测,LAD07和CAJ06-C菌株对秀珍菇子实体生长发育的阻碍作用更强,与其侵染下菇体发黄、出现水渍和停止发育等现象更重的结果较为一致。

关键词: 秀珍菇(Pleurotus pulmonarius), 黄菇病, 假单胞菌属(Pseudomonas), 分子鉴定

Abstract:

Yellow rot disease has a serious impact on the yield and quality of Pleurotus pulmonarius in the factory production mode. In the present study, pathogenic bacteria were isolated and purified from diseased samples. The pathogenicity of pathogenic bacteria were determinated by Koch postulates. Morphological characteristics and 16S rRNA sequences were used for pathogen identification, and their characteristic properties were analyzed by mycelium growth rate and transcriptome. It was shown that the isolated strains CAJ01-D, CAJ06-C, TXX01 and LAD07 could cause yellow rot disease, which were different species of Pseudomonas spp. The treatment using different Pseudomonas strains impacted the growth rate of P. pulmonarius mycelia, but there was no significant difference in mycelium growth vigor. There was difference in the expression of pathogenicity-related genes when P. pulmonarius was infected by Pseudomonas strains, but most of the differentially expressed genes under the infection of strains LAD07 and CAJ06-C were down-regulated. The clustering and enrichment of GO and KEGG functions showed that the cytochrome c reductase was down-regulated and mitochondrial respiration was weakened by the infection of LAD07 strain. The CAJ06-C strain caused down-regulation of glycolysis or gluconeogenesis, tyrosine metabolism and phenylalanine metabolism. It was speculated that the LAD07 and CAJ06-C strains hindered the growth and development of the fruiting body of P. pulmonarius, which was consistent with the observed severe disease symptoms, including yellowing, water stain and developmental arrest of the fruiting body under infection.

Key words: Pleurotus pulmonarius, yellow rot disease, Pseudomonas, molecular identification

中图分类号:

S432.42
S646

陆娜, 王伟科, 林佳瑶, 周祖法. 秀珍菇黄菇病病原菌鉴定及其致病相关的转录组分析[J]. 浙江农业学报, 2025, 37(2): 426-437.

LU Na, WANG Weike, LIN Jiayao, ZHOU Zufa. Identification of yellow rot disease of Pleurotus pulmonarius and analysis of its pathogenicity-related transcriptome[J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 426-437.

图/表 9

图1 样品的症状特征 A,原基期;B,菇蕾期;C,生长期;D,成熟期。

Fig.1 Disease characteristics of the collected samples A, Primordia; B, Early fruiting bodies; C, Growing fruiting bodies; D, Fruiting bodies at harvest time.

图2 接种病原菌后的症状特征 A,原基期;B,菇蕾期;C,生长期;D,成熟期。

Fig.2 Characteristics of disease symptoms after inoculation of strains A, Primordia; B, Early fruiting bodies; C, Growing fruiting bodies; D, Fruiting bodies at harvest time

图3 病原菌的菌落特征与细胞学超微结构

Fig.3 Colony characteristics and cytological and ultrastructural micrographs of bacterial pathogens 1,CAJ01-D;2,CAJ06-C;3,LAD07;4,TXX01。

图4 基于16S rRNA构建的病原菌系统发育树

Fig.4 Phylogenetic tree of the isolated bacterial pathogens based on 16S rRNA

表1 不同处理对秀珍菇菌丝生长的影响

Table 1 Effect of treatments on mycelia growth of Pleurotus pulmonarius

t/d	不同病原菌处理下秀珍菇菌丝的生长速度Growth rate of mycelia under different pathogens/(cm·d^-1)
t/d	CK	CAJ01-D	CAJ06-C	LAD07	TXX01
1	0.525±0.108	0.637±0.119	0.550±0.112	0.563±0.041	0.675±0.082
2	0.485±0.043	0.375±0.043	0.475±0.147	0.450±0.050	0.450±0.050
3	0.463±0.054	0.350±0.050	0.513±0.143	0.525±0.043	0.375±0.083
4	0.528±0.079	0.350±0.031	0.520±0.097	0.510±0.038	0.453±0.084
5	0.542±0.075	0.345±0.026	0.518±0.103	0.540±0.034	0.450±0.085
6	0.567±0.074	0.636±0.033	0.518±0.096	0.512±0.037	0.452±0.078

图5 秀珍菇菌株与病原菌对峙培养5 d后的菌落形态从左到右依次为CK(不接种病原菌)和接种CAJ01-D、TXX01、CAJ06-C、LAD07的结果。

Fig.5 Colony morphological characteristics after dual cultures of Pleurotus pulmonarius and pathogen for 5 d Photos from left to right show the results of CK (without pathogen inoculation) and treatments after inoculation of CAJ01-D, TXX01, CAJ06-C, LAD07, respectively.

图6 不同病原菌处理下差异基因表达量的火山图 -lg(FDR),错误发现率常用对数值的相反数;log2(FC),差异表达倍数以2为底的对数值。

Fig.6 Volcano map of differential expressed genes under treatments -lg(FDR), The opposite number of common logarithm value of false discovery rate. log2(FC), The logarithm value of fold change based on 2.

图7 不同病原菌处理下差异基因表达量的维恩图

Fig.7 Venn diagram of differential expressed genes under treatments

图8 GO功能的聚类分析图

Fig.8 Cluster analysis diagram of GO function

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秀珍菇黄菇病病原菌鉴定及其致病相关的转录组分析

Identification of yellow rot disease of Pleurotus pulmonarius and analysis of its pathogenicity-related transcriptome

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