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台湾健康葱叶内生真菌和细菌的多样性及生物防治潜力

Diversity and Biocontrol Potential of Endophytic Fungi and Bacteria Associated with Healthy Welsh Onion Leaves in Taiwan.

作者信息

Wang Jian-Yuan, Jayasinghe Himanshi, Cho Yi-Tun, Tsai Yi-Chen, Chen Chao-Ying, Doan Hung Kim, Ariyawansa Hiran A

机构信息

Department of Plant Pathology and Microbiology, College of Bio-Resources and Agriculture, National Taiwan University, Taipei 106319, Taiwan.

Hualien District Agricultural Research and Extension Station, Hualien 973044, Taiwan.

出版信息

Microorganisms. 2023 Jul 13;11(7):1801. doi: 10.3390/microorganisms11071801.

Abstract

Foliar diseases caused by and species are among the major biotic factors limiting Welsh onion production in Taiwan. Owing to concerns about the environment and the development of pathogen resistance to existing fungicides, biological control using endophytes is emerging as an eco-friendly alternative to chemical control. The aim of the present study was to isolate endophytes from healthy Welsh onion leaves and investigate their antagonistic potential against the major phytopathogenic fungi associated with Welsh onion plants in Taiwan. A total of 109 bacterial and 31 fungal strains were isolated from healthy Welsh onion leaves and assigned to 16 bacterial and nine fungal genera using morphological and molecular characterization based on DNA sequence data obtained from nuclear internal transcribed spacer (nrITS) (fungi) and 16S rRNA (bacteria). Evaluation of these endophytic isolates for biocontrol activity against leaf blight pathogens strain SX15-2 and strain SX20-2 by dual culture assay and greenhouse experiments resulted in the identification of two bacterial isolates (GFB08 and LFB28) and two fungal isolates (GFF06 and GFF08) as promising antagonists to leaf blight pathogens. Among the four selected isolates, strain GFB08 exhibited the highest disease control in the greenhouse study. Therefore, strain GFB08 was further evaluated to understand the mechanism underlying its biocontrol efficacy. A phylogenetic analysis based on six genes identified strain GFB08 as . The presence of antimicrobial peptide genes (, , , , , , , and ) and the secretion of several cell wall degrading enzymes (CWDEs), including cellulase and protease, confirmed the antifungal nature of strain GFB08. Leaf blight disease suppression by preventive and curative assays indicated that strain GFB08 has preventive efficacy on strain SX15-2 and both preventive and curative efficacy on strain SX20-2. Overall, the current study revealed that healthy Welsh onion leaves harbour diverse bacterial and fungal endophytes, among which the endophytic bacterial strain, strain GFB08, could potentially be used as a biocontrol agent to manage the leaf blight diseases of Welsh onion in Taiwan.

摘要

由[具体病原菌名称1]和[具体病原菌名称2]引起的叶部病害是限制台湾地区大葱生产的主要生物因素之一。由于对环境的关注以及病原菌对现有杀菌剂产生抗性,利用内生菌进行生物防治正成为一种替代化学防治的环保方法。本研究的目的是从健康大葱叶片中分离内生菌,并研究它们对与台湾大葱植株相关的主要植物病原真菌的拮抗潜力。从健康大葱叶片中总共分离出109株细菌和31株真菌菌株,并根据从核糖体DNA内转录间隔区(nrITS)(真菌)和16S rRNA(细菌)获得的DNA序列数据,通过形态学和分子特征鉴定,将它们归为16个细菌属和9个真菌属。通过对峙培养试验和温室试验评估这些内生菌分离株对叶枯病病原菌[病原菌名称1]菌株SX15 - 2和[病原菌名称2]菌株SX20 - 2的生防活性,结果鉴定出两株细菌分离株(GFB08和LFB28)和两株真菌分离株(GFF06和GFF08)是叶枯病病原菌有潜力的拮抗剂。在这四株选定的分离株中,GFB08菌株在温室研究中表现出最高的病害防治效果。因此,对GFB08菌株进行了进一步评估,以了解其生防功效的潜在机制。基于六个基因的系统发育分析将GFB08菌株鉴定为[具体细菌种类]。抗菌肽基因([基因名称1]、[基因名称2]、[基因名称3]、[基因名称4]、[基因名称5]、[基因名称6]、[基因名称7]和[基因名称8])的存在以及包括纤维素酶和蛋白酶在内的几种细胞壁降解酶(CWDEs)的分泌,证实了GFB08菌株的抗真菌特性。通过预防和治疗试验对叶枯病的抑制作用表明,GFB08菌株对[病原菌名称1]菌株SX15 - 2有预防效果,对[病原菌名称2]菌株SX20 - 2有预防和治疗效果。总体而言,当前研究表明健康大葱叶片中存在多种细菌和真菌内生菌,其中内生细菌菌株GFB08有潜力作为生物防治剂来管理台湾地区大葱的叶枯病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/60a2/10386586/14c3b23cf669/microorganisms-11-01801-g001.jpg

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