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从乳制品中分离的戊糖片球菌对禾谷镰刀菌的抗真菌及玉米赤霉烯酮抑制活性

Antifungal and Zearalenone Inhibitory Activity of Pediococcus pentosaceus Isolated from Dairy Products on Fusarium graminearum.

作者信息

Sellamani Muthulakshmi, Kalagatur Naveen K, Siddaiah Chandranayaka, Mudili Venkataramana, Krishna Kadirvelu, Natarajan Gopalan, Rao Putcha Venkata L

机构信息

Toxicology and Immunology Division, DRDO-BU-Centre for Life sciences, Bharathiar University Campus, Coimbatore India.

Food Microbiology Division, Defence Food Research Laboratory Mysore, India.

出版信息

Front Microbiol. 2016 Jun 13;7:890. doi: 10.3389/fmicb.2016.00890. eCollection 2016.

Abstract

The present study was aimed to evaluate the bio-control efficacy of Pediococcus pentosaceus isolated from traditional fermented dairy products originated from India, against the growth and zearalenone (ZEA) production of Fusarium graminearum. The cell-free supernatants of P. pentosaceus (PPCS) were prepared and chemical profiling was carried out by GC-MS and MALDI-TOF analysis. Chemical profiling of PPCS evidenced that, the presence of phenolic antioxidants, which are responsible for the antifungal activity. Another hand, MALDI-TOF analysis also indicated the presence of antimicrobial peptides. To know the antioxidant potential of PPCS, DPPH free radical scavenging assay was carried out and IC50 value was determined as 32 ± 1.89 μL/mL. The antifungal activity of P. pentosaceus was determined by dual culture overlay technique and zone of inhibition was recorded as 47 ± 2.81%, and antifungal activity of PPCS on F. graminearum was determined by micro-well dilution and scanning electron microscopic techniques. The minimum inhibitory concentration (MIC) of PPCS was determined as 66 ± 2.18 μL/mL in the present study. Also a clear variation in the micromorphology of mycelia treated with MIC value of PPCS compared to untreated control was documented. Further, the mechanism of growth inhibition was revealed by ergosterol analysis and determination of reactive oxygen species (ROS) in PPCS treated samples. The effects of PPCS on mycelial biomass and ZEA production were observed in a dose-dependent manner. The mechanism behind the suppression of ZEA production was studied by reverse transcriptase qPCR analysis of ZEA metabolic pathway genes (PKS4 and PKS13), and results showed that there is a dose dependent down-regulation of target gene expression in PPCS treated samples. The results of the present study were collectively proved that, the antifungal and ZEA inhibitory activity of PPCS against F. graminearum and it may find a potential application in agriculture and food industry as a natural bio-controlling agent.

摘要

本研究旨在评估从印度传统发酵乳制品中分离出的戊糖片球菌对禾谷镰刀菌生长及玉米赤霉烯酮(ZEA)产生的生物防治效果。制备了戊糖片球菌的无细胞上清液(PPCS),并通过气相色谱 - 质谱联用(GC-MS)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)分析进行化学谱图分析。PPCS的化学谱图分析表明,存在负责抗真菌活性的酚类抗氧化剂。另一方面,MALDI-TOF分析也表明存在抗菌肽。为了解PPCS的抗氧化潜力,进行了二苯基苦味酰基自由基(DPPH)清除试验,测定IC50值为32±1.89μL/mL。通过双培养覆盖技术测定戊糖片球菌的抗真菌活性,抑菌圈记录为47±2.81%,并通过微孔稀释和扫描电子显微镜技术测定PPCS对禾谷镰刀菌的抗真菌活性。在本研究中,PPCS的最低抑菌浓度(MIC)测定为66±2.18μL/mL。此外,记录了与未处理对照相比,用PPCS的MIC值处理的菌丝体微观形态有明显变化。进一步通过麦角固醇分析和测定PPCS处理样品中的活性氧(ROS)揭示了生长抑制机制。观察到PPCS对菌丝体生物量和ZEA产生的影响呈剂量依赖性。通过对ZEA代谢途径基因(PKS4和PKS13)的逆转录酶定量聚合酶链反应(qPCR)分析研究了ZEA产生受抑制的机制,结果表明在PPCS处理的样品中靶基因表达呈剂量依赖性下调。本研究结果共同证明,PPCS对禾谷镰刀菌具有抗真菌和ZEA抑制活性,它可能作为一种天然生物防治剂在农业和食品工业中找到潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/befb/4904835/0a3d2ecc0073/fmicb-07-00890-g001.jpg

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