Sundar Ranjitha Dhevi V, Arunachalam Sathiavelu
Laboratory of Microbiology, Department of Biotechnology, School of Biosciences and Technology, Vellore Institute of Technology, Vellore, India.
Laboratory of Microbiology, Department of Agriculture Microbiology, VIT School of Agricultural Innovations and Advanced Learning, Vellore Institute of Technology, Vellore, India.
Front Microbiol. 2024 Feb 16;15:1327190. doi: 10.3389/fmicb.2024.1327190. eCollection 2024.
Endophytes refer to microorganisms residing within the endosphere of plants, particularly perennials, without inflicting noticeable injury or inducing obvious morphological variations to their host plant or host organism. Endophytic fungi, although often overlooked microorganisms, have garnered interest due to their significant biological diversity and ability to produce novel pharmacological substances.
In this study, fourteen endophytic fungi retrieved were from the stem of the perennial plant of the Asparagaceae family. These fungal crude metabolites were tested for antagonistic susceptibility to Multi-Drug Resistant (MDR) pathogens using agar well diffusion, Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC) assays. The chequerboard test was used to assess the synergistic impact of active extract.
In early antibacterial screening using the Agar plug diffusion test, three of fourteen endophytes demonstrated antagonism against Methicillin-resistant (MRSA) and Vancomycin-resistant (VRE). Three isolates were grown in liquid medium and their secondary metabolites were recovered using various organic solvents. Eight extracts from three endophytic fungi displayed antagonism against one or more human pathogens with diameters ranging from 11 to 24 mm. The highest antagonistic effect was obtained in ethyl acetate extract for PTS8 isolate against two MRSA (ATCC 43300, 700699) with 20 ± 0.27 and 22 ± 0.47 mm zones of inhibition, respectively, among different solvent extracts. The extract had MICs of 3.12 ± 0.05 and 1.56 ± 0.05 μg/mL, and MBCs of 50 ± 0.01 and 12.5 ± 0.04 μg/mL, respectively. Antagonism against VRE was 18 ± 0.23 mm Zone of Inhibition (ZOI) with MIC and MBC of 6.25 ± 0.25 and 25 ± 0.01 μg/mL. When ethyl acetate extract was coupled with antibiotics, the chequerboard assay demonstrated a synergistic impact against MDR bacteria. In an antioxidant test, it had an inhibitory impact of 87 ± 0.5% and 88.5 ± 0.5% in 2,2-Diphenyl-1-Picrylhydrazyl and reducing power assay, respectively, at 150 μg/mL concentration. PTS8 was identified as a strain by 18S rRNA internal transcribed spacer (ITS) sequencing. To our insight, it is the foremost study to demonstrate the presence of an endophyte in the stem of P. tuberosa and the first report to study the antibacterial efficacy of which might serve as a powerful antibacterial source against antibiotic-resistant human infections.
内生菌是指存在于植物(尤其是多年生植物)内皮层中的微生物,它们不会对宿主植物或宿主生物体造成明显伤害或引起明显的形态变化。内生真菌虽然常常被忽视,但因其显著的生物多样性和产生新型药理物质的能力而受到关注。
在本研究中,从天门冬科多年生植物的茎中分离出14株内生真菌。使用琼脂孔扩散法、最低抑菌浓度(MIC)测定法和最低杀菌浓度(MBC)测定法,对这些真菌的粗代谢产物进行了对多重耐药(MDR)病原体的拮抗敏感性测试。采用棋盘试验评估活性提取物的协同作用。
在早期使用琼脂块扩散试验进行的抗菌筛选中,14株内生菌中有3株对耐甲氧西林金黄色葡萄球菌(MRSA)和耐万古霉素肠球菌(VRE)表现出拮抗作用。将3株分离菌在液体培养基中培养,并用各种有机溶剂回收其次级代谢产物。来自3株内生真菌的8种提取物对一种或多种人类病原体表现出拮抗作用,抑菌圈直径为11至24毫米。在不同溶剂提取物中,PTS8分离株的乙酸乙酯提取物对两种MRSA(ATCC 43300、700699)的拮抗作用最强,抑菌圈直径分别为20±0.27毫米和22±0.47毫米。该提取物的MIC分别为3.12±0.05微克/毫升和1.56±0.05微克/毫升,MBC分别为50±0.01微克/毫升和12.5±0.04微克/毫升。对VRE的拮抗作用表现为抑菌圈直径18±0.23毫米,MIC和MBC分别为6.25±0.25微克/毫升和25±0.01微克/毫升。当乙酸乙酯提取物与抗生素联合使用时,棋盘试验显示对MDR细菌有协同作用。在抗氧化试验中,在150微克/毫升浓度下,其在2,2-二苯基-1-苦基肼自由基清除试验和还原力试验中的抑制率分别为87±0.5%和88.5±0.5%。通过18S rRNA内部转录间隔区(ITS)测序将PTS8鉴定为一个菌株。据我们所知,这是首次证明在块茎石豆兰茎中存在内生菌的研究,也是首次报道其抗菌功效的研究,其可能成为对抗耐抗生素人类感染的强大抗菌来源。