Rapid and Easy Modified Plate-based Screening Methods for Quantitative and Qualitative Detection of Protease Production by Fungi

K. Karimi, A. Narmani, I. Pertot, M. Arzanlou
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引用次数: 1

Abstract

Proteases constitute a significant part of cell wall-degrading enzymes (CWDEs) produced by fungal biocontrol agents and particularly crucial in mycoparasitism of fungal phytopathogens. Plate-based screening methods are routinely used for screening protease-producing microorganisms including fungi. Skim milk agar (SMA) is one of the most popular media for the detection of protease producing bacteria. However, SMA is not efficient to test fast growing fungi, because it does not give an estimation of the actual amount of secreted protease produced by fungal inocula. In the current study, the efficacy of two modified plate-screening methods, including split-SMA (SSMA) and minimal medium supplemented with skim milk (MSMW) was assessed for detection of protease production by three representative fungal strains including Trichoderma longibrachiatum strain N, Beauveria bassiana strain B and Purpureocillium lilacinum strain PL. Protease production was revealed on the three tested media by the three strains. However, the halo diameter of the fungal strains (a proxy for protease production) was the smallest on SMA. Furthermore, protease production could not be detected for T. longibrachiatum strain N on SMA due to its fast growth; while it showed the highest protease activity on both modified media compared with the other strains. According to the result of this study, the SSMA medium is an easy and more accurate method compared with the two other different methods as it displays the actual amount of protease produced by fungal strains and therefore this method is recommended for quantitative and qualitative detection of protease production by slow and fast growing fungi.
快速简便的改良板筛选方法用于真菌蛋白酶生产的定量和定性检测
蛋白酶是真菌生物防治剂产生的细胞壁降解酶(CWDE)的重要组成部分,在真菌植物病原体的真菌寄生中尤为重要。基于平板的筛选方法通常用于筛选产生蛋白酶的微生物,包括真菌。脱脂乳琼脂(SMA)是检测蛋白酶产生菌最常用的培养基之一。然而,SMA不能有效地测试快速生长的真菌,因为它不能估计真菌接种物产生的蛋白酶的实际分泌量。在本研究中,评估了两种改良平板筛选方法,包括裂解SMA(SSMA)和添加脱脂乳的最小培养基(MSMW),对三种具有代表性的真菌菌株(包括长臂木霉菌株N、球孢白僵菌菌株B和淡紫色紫浆菌菌株PL)产蛋白酶的检测效果。三种菌株在三种测试培养基上显示了蛋白酶的产生。然而,SMA上真菌菌株的晕圈直径(蛋白酶产生的指标)最小。此外,由于长臂T.N菌株生长迅速,在SMA上无法检测到蛋白酶的产生;而与其他菌株相比,它在两种修饰的培养基上都表现出最高的蛋白酶活性。根据这项研究的结果,与其他两种不同的方法相比,SSMA培养基是一种简单、准确的方法,因为它显示了真菌菌株产生的蛋白酶的实际量,因此推荐该方法用于定量和定性检测生长缓慢和快速的真菌产生的蛋白酶。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Acta Phytopathologica Et Entomologica Hungarica
Acta Phytopathologica Et Entomologica Hungarica Agricultural and Biological Sciences-Plant Science
CiteScore
1.10
自引率
0.00%
发文量
7
期刊介绍: The journal publishes papers on the infectious diseases of plants, damages caused by insects and deals with the basic aspects of chemical and biological protection. Within its field of interest additional topics such as resistance against plant diseases, and physiological, biochemical and molecular questions of plant resistance and susceptibility are also discussed. Publishes book reviews and advertisements.
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