Phytopathogenic fungi devastate agricultural crops worldwide. The biological agents, such as Trichoderma spp., antagonize phytopathogenic fungi by secreting various cell wall‐degrading enzymes, for example, endochitinase and β‐1,3‐glucanase that target glycosidic linkages in β‐glucan and chitin polymers of fungal cell walls, thus inhibiting pathogen growth. In this study, two antifungal genes endochitinase and β‐1,3‐glucanase cloned from local Trichoderma spp. were ligated in pET28a+ expression vector individually to generate two recombinant vectors. The vectors were mobilized into Escherichia coli host strain Rosetta‐gami 2 for protein expression, and the 6xHis‐tagged recombinant proteins were purified through Ni‐NTA affinity chromatography. The purified proteins were individually confronted in vitro with pure cultures of Phytophthora parasitica (destructive pathogen affecting several hundred plant species worldwide) for analyzing their effect on pathogen growth. In vitro confrontation assay revealed P. parasitica growth inhibition by purified β‐1,3‐glucanase. The pathogen growth inhibition was due to hyphal morphological distortions, such as breakages, swelling, and holes evinced through electron micrography confirming direct role of β‐1,3‐glucanase in pathogen structural degradation.

中文翻译：

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木霉菌的抗真菌作用。寄生疫霉菌上的β-1,3-葡聚糖酶：菌丝形态变异

植物致病真菌破坏了全世界的农作物。诸如木霉属（Trichoderma spp。）之类的生物制剂可通过分泌各种细胞壁降解酶来拮抗植物病原真菌，例如，内切壳聚糖酶和β-1,3-葡聚糖酶以真菌细胞壁的β-葡聚糖和几丁质聚合物中的糖苷键为目标，从而抑制病原体的生长。在这项研究中，从当地木霉属物种中克隆了两个抗真菌基因内切几丁质酶和β-1,3-葡聚糖酶。将其分别连接到pET28a +表达载体中以产生两个重组载体。载体动员到大肠杆菌中宿主菌株Rosetta-gami 2进行蛋白表达，并通过Ni-NTA亲和层析纯化6xHis标记的重组蛋白。纯化的蛋白质分别在体外与纯疫霉菌（破坏性病原体影响全球数百种植物）的纯培养物相面对，以分析其对病原体生长的影响。体外对抗试验显示，纯化的β-1,3-葡聚糖酶可抑制寄生性疟原虫的生长。病原体的生长抑制是由于菌丝形态畸变，例如破损，肿胀和电子显微镜显示的孔洞，从而证实了β-1,3-葡聚糖酶在病原体结构降解中的直接作用。