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中国引起马铃薯黑胫病的机会致病菌的鉴定与特性分析

Identification and characterization of opportunistic pathogen causing potato blackleg in China.

作者信息

Han Wanxin, Wang Jinhui, Pirhonen Minna, Pan Yang, Qin Jingxin, Zhang Shangqing, Zhu Jiehua, Yang Zhihui

机构信息

College of Plant Protection, Hebei Agricultural University, Baoding, China.

Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland.

出版信息

Front Plant Sci. 2023 Mar 3;14:1097741. doi: 10.3389/fpls.2023.1097741. eCollection 2023.

Abstract

Blackleg and aerial stem rot of potato ( L.), caused by soft rot enterobacteria of the genera and , has recently increased years in Hebei Province, China. Field surveys were performed during the 2021 potato growing season in Hebei to identify and characterize bacterial pathogens. Sixteen potato plants showing blackleg or aerial stem rot were collected from three potato-producing areas, and ten representative pectinolytic bacteria were isolated from symptomatic plants. 16S rDNA sequencing and multilocus sequence analysis were performed to determine the taxonomic position of the bacterial isolates. The isolates belonged to the genus , including , , , and . The exceptions were isolates BY21311 and BY21312, which belonged to a new species of previously found in groundwater. The taxonomy of isolate BY21311 was confirmed using whole genome-based analysis. has only been identified in potato plants on one farm in Baoding region in China. Isolates BY21311 and BY21312 displayed similar physiological and biochemical traits to the type strain DPMP315. Artificial inoculation assays revealed that isolate BY21311 fulfilled Koch's postulates for potato blackleg. These findings represent the first time , a water-associated species may be the cause of blackleg in the field. Interestingly, BY21311 has reduced ability to macerate potato tubers when compared to , , , and , which is more virulent in tubers than the type strain DPMP315. The host range of isolate BY21311 was determined by injection method, which can impregnate five plants. Although the genome of isolate BY21311 harbors gene clusters encoding a type III secretion system, it did not elicit a hypersensitive response (HR) in or leaves. T3SS effector AvrE and T4SS effector PilN were obtained by predicting isolate BY21311 genome. appears to show significant variations in gene content between two genomes, and gene content varies between isolates BY21311 and DPMP315, with strain specific-genes involved in many aspects, including lipopolysaccharide biosynthesis, substrate translocation, T4SS and T6SS among others, suggesting that isolates BY21311 and DPMP315 might represent distinct clades within the species.

摘要

马铃薯黑胫病和茎腐病由欧文氏菌属和芽孢杆菌属的软腐肠杆菌引起,近年来在中国河北省呈上升趋势。2021年马铃薯生长季期间在河北进行了田间调查,以鉴定和表征细菌病原体。从三个马铃薯产区采集了16株表现出黑胫病或茎腐病的马铃薯植株,并从有症状的植株中分离出10株具有代表性的果胶分解细菌。进行了16S rDNA测序和多位点序列分析,以确定细菌分离株的分类地位。这些分离株属于芽孢杆菌属,包括芽孢杆菌、嗜麦芽窄食单胞菌、解淀粉芽孢杆菌和枯草芽孢杆菌。例外的是分离株BY21311和BY21312,它们属于先前在地下水中发现的一个芽孢杆菌新物种。使用基于全基因组的分析确认了分离株BY21311的分类。在中国保定地区的一个农场,仅在马铃薯植株中鉴定出芽孢杆菌。分离株BY21311和BY21312表现出与模式菌株DPMP315相似的生理和生化特性。人工接种试验表明,分离株BY21311符合马铃薯黑胫病的科赫法则。这些发现首次表明,一种与水相关的芽孢杆菌物种可能是田间黑胫病病因。有趣的是,与芽孢杆菌、嗜麦芽窄食单胞菌、解淀粉芽孢杆菌和枯草芽孢杆菌相比,BY21311使马铃薯块茎浸解的能力降低,这些菌在块茎中的毒性比模式菌株DPMP315更强。通过注射法确定分离株BY21311的寄主范围,该方法可侵染五种植物。尽管分离株BY21311的基因组含有编码III型分泌系统的基因簇,但它在烟草或本氏烟草叶片中未引发过敏反应(HR)。通过预测分离株BY21311的基因组获得了III型分泌系统效应蛋白AvrE和IV型分泌系统效应蛋白PilN。芽孢杆菌在两个基因组之间似乎显示出基因含量的显著差异,并且分离株BY21311和DPMP315之间的基因含量也有所不同,有许多方面涉及菌株特异性基因,包括脂多糖生物合成、底物转运、IV型分泌系统和VI型分泌系统等,这表明分离株BY21311和DPMP315可能代表该物种内不同的进化枝。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a427/10020715/9b474f18d290/fpls-14-1097741-g001.jpg

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