Rana Shiwali, Singh Sanjay Kumar, Dufossé Laurent
National Fungal Culture Collection of India, Biodiversity and Palaeobiology Group, MACS' Agharkar Research Institute, G.G. Agarkar Road, Pune 411004, India.
Faculty of Science, Savitribai Phule Pune University, Ganeshkhind Road, Ganeshkhind, Pune 411007, India.
J Fungi (Basel). 2022 Jun 24;8(7):662. doi: 10.3390/jof8070662.
The taxonomy of the genus has been in a because of ambiguous circumscription of species-level identification based on morphotaxonomic criteria. In this study, multigene phylogeny was conducted to resolve the evolutionary relationships of 88 Indian isolates based on the internal transcribed spacer region, 28S large subunit, translation elongation factor 1-alpha, RNA polymerase second largest subunit, beta-tubulin and calmodulin gene regions. species are well known to produce metabolites such as beauvericin (BEA) and enniatins. These identified isolates were subjected to fermentation in -defined media for BEA production and tested using TLC, HPLC and HRMS. Among 88 isolates studied, 50 were capable of producing BEA, which varied from 0.01 to 15.82 mg/g of biomass. NFCCI 5201 showed maximum BEA production (15.82 mg/g of biomass). The extract of . NFCCI 5201 showed promising antibacterial activity against MLS16 MTCC 2940 and MTCC 2470 with MIC of 62.5 and 15.63 µg/mL, respectively. Similarly, the . NFCCI 5201 extract in potato dextrose agar (40 µg/mL) exhibited antifungal activity in the food poison technique against plant pathogenic and other fungi, NFCCI 4327, NFCCI 4263, NFCCI 3744 and sp. NFCCI 3482, showing % inhibition of 84.31, 49.76, 38.22 and 35.13, respectively. The antibiotic effect was found to synergize when extract and amphotericin B (20 µg/mL each in potato dextrose agar) were used in combination against sp. NFCCI 2108, NFCCI 4263, NFCCI 4690 and sp. NFCCI 2716, showing % inhibition of 50.35, 79.37, 48.07 and 76.72, respectively. The extract also showed satisfactory dose-dependent DPPH radical scavenging activity with an IC value of 0.675 mg/mL. This study reveals the correct identity of the Indian isolates based on multigene phylogeny and also throws light on BEA production potential, suggesting their possible applicability in the medicine, agriculture and industry.
由于基于形态分类标准的物种水平鉴定的界定不明确，该属的分类一直处于混乱状态。在本研究中，基于内转录间隔区、28S大亚基、翻译延伸因子1-α、RNA聚合酶第二大亚基、β-微管蛋白和钙调蛋白基因区域，进行了多基因系统发育分析以解析88株印度该菌分离株的进化关系。该菌物种以产生诸如白僵菌素（BEA）和恩镰孢菌素等代谢产物而闻名。这些鉴定出的分离株在限定培养基中进行发酵以生产BEA，并使用薄层色谱法（TLC）、高效液相色谱法（HPLC）和高分辨质谱法（HRMS）进行检测。在所研究的88株分离株中，有50株能够产生BEA，其产量在0.01至15.82 mg/g生物量之间变化。NFCCI 5201显示出最高的BEA产量（15.82 mg/g生物量）。NFCCI 5201的提取物对MLS16 MTCC 2940和MTCC 2470显示出有前景的抗菌活性，其最低抑菌浓度（MIC）分别为62.5和15.63 μg/mL。同样，NFCCI 5201提取物在马铃薯葡萄糖琼脂（40 μg/mL）中采用食物中毒技术对植物病原真菌和其他真菌，即NFCCI 4327、NFCCI 4263、NFCCI 3744和NFCCI 3482种真菌，分别显示出84.31%、49.76%、38.22%和35.13%的抑制率。当NFCCI 5201提取物与两性霉素B（在马铃薯葡萄糖琼脂中各为20 μg/mL）联合用于对抗NFCCI 2108种、NFCCI 4263种、NFCCI 4690种和NFCCI 2716种真菌时，发现抗生素效果具有协同作用，分别显示出50.35%、79.37%、48.07%和76.72%的抑制率。该提取物还显示出令人满意的剂量依赖性1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除活性，其半数抑制浓度（IC）值为0.675 mg/mL。本研究基于多基因系统发育解析了印度该菌分离株的正确身份，也揭示了BEA的生产潜力，表明它们在医药、农业和工业中可能具有的适用性。
