Arias-Aravena Matias, Altimira Fabiola, Gutiérrez Daniela, Ling Jian, Tapia Eduardo
Laboratorio de Entomología y Biotecnología, Instituto de Investigaciones Agropecuarias, INIA La Platina, Santiago 8831314, Chile.
Facultad de Medicina Veterinaria y Agronomía, Universidad de Las Américas, Santiago 8370040, Chile.
J Fungi (Basel). 2022 Oct 14;8(10):1083. doi: 10.3390/jof8101083.
RGM 2184 has shown 80% maximum efficacy against the pest in the autumn and winter seasons. This suggests that the strain possesses an interesting battery of enzymes that are cold-adapted to penetrate the thick and hydrophobic cocoon of . In this study, screening of the proteolytic, lipolytic, and chitinolytic activity of enzyme extracts secreted by the RGM 2184 strain was carried out in various culture media. The enzyme extracts with the highest activity were subjected to zymography and mass spectrometry. These analyses allowed the identification of two proteases, two lipases, and three chitinases. Comparative analysis indicated that the degree of similarity between these enzymes was substantially reduced when the highest degree of taxonomic relatedness between RGM 2184 and the entomopathogenic fungus strain was at the family level. These results suggest that there is a wide variety of exoenzymes in entomopathogenic fungi species belonging to the order Hypocreales. On the other hand, exoenzyme extract exposure of cocoons and pupae of provoked damage at 10 °C. Additionally, an analysis of the amino acid composition of the RGM 2184 exoenzyme grouped them close to the cold-adapted protein cluster. These results support the use of this strain to control pests in autumn and winter. Additionally, these antecedents can form a scaffold for the future characterization of these exoenzymes along with the optimization of the strain's biocontrol ability by overexpressing them.
RGM 2184在秋冬季节对该害虫显示出80%的最大防治效果。这表明该菌株拥有一系列有趣的酶,这些酶适应低温,能够穿透[害虫名称]厚厚的疏水茧。在本研究中,在各种培养基中对RGM 2184菌株分泌的酶提取物的蛋白水解、脂肪水解和几丁质分解活性进行了筛选。对活性最高的酶提取物进行了酶谱分析和质谱分析。这些分析鉴定出了两种蛋白酶、两种脂肪酶和三种几丁质酶。比较分析表明,当RGM 2184与昆虫病原真菌菌株之间的最高分类学相关性处于科级水平时,这些酶之间的相似程度大幅降低。这些结果表明,肉座菌目昆虫病原真菌物种中存在多种胞外酶。另一方面,在10℃下,用胞外酶提取物处理[害虫名称]的茧和蛹会造成损害。此外,对RGM 2184胞外酶的氨基酸组成分析将它们归类为接近冷适应蛋白簇。这些结果支持使用该菌株在秋冬季节防治害虫。此外,这些前期研究结果可为未来这些胞外酶的特性表征以及通过过表达优化该菌株的生物防治能力提供一个框架。