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从食虫蝙蝠粪便中分离的产几丁质酶细菌的细胞外抗真菌活性。

Extracellular Antifungal Activity of Chitinase-Producing Bacteria Isolated From Guano of Insectivorous Bats.

机构信息

INQUISAL, CONICET. FQByF, UNSL, Chacabuco 917 (D5700BWS), 5700, San Luis, Argentina.

PROIMI, CONICET, Av. Belgrano Y Pje. Caseros, 4000, San Miguel de Tucumán, Argentina.

出版信息

Curr Microbiol. 2021 Jul;78(7):2787-2798. doi: 10.1007/s00284-021-02555-0. Epub 2021 Jun 4.

Abstract

A reduction in the use of agrochemicals requires the development of either alternatives or complementary control methods in order to limit their dangerousness. An alternative is the biological control of fungi by bacteria. The fungal cell wall is a unique structure of the fungi, composed of glucan, chitin, and glycoproteins. Therefore, bacteria producing mycolytic enzymes, like chitinases, are of great interest to degrade fungal cell-wall components. The objectives of this work were to isolate chitinolytic bacteria from the guano of insectivorous bats (Tadarida brasiliensis) and to verify the presence of antifungal activities against phytopathogenic fungi. From the guano samples, 28 bacterial isolates were obtained, 70% of which presented chitinolytic activity. Four isolates were selected since they showed the highest values of chitinase activity, and they were characterized as belonging to Bacillus genus, by analyzing the 16S ribosomal RNA gene sequence. Cell-free supernatants of bacterial cultures were used in inhibition tests on 16 fungi: Alternaria and Colletotrichum acutatum were the most affected. Chitinase and antifungal activities were observed in the cell-free supernatant regardless of the culture medium used. Both activities were stable to heat and proteinase K treatments. Finally, when the culture medium was supplemented with 1 ml of cell-free supernatants (0.33%) and incubated for 120 h, the inhibition of hyphae formation and germination spores of reporter fungus were observed under light microscopy. These results suggest the feasibility of using cell-free supernatants as eco-friendly fungicides. The use of them may contribute to reducing the dose of toxic chemicals.

摘要

减少农用化学品的使用需要开发替代品或补充控制方法,以限制其危害性。一种替代品是利用细菌对真菌进行生物防治。真菌细胞壁是真菌的独特结构,由葡聚糖、几丁质和糖蛋白组成。因此,产生几丁质酶(如几丁质酶)的细菌对于降解真菌细胞壁成分非常有兴趣。这项工作的目的是从食虫蝙蝠(Tadarida brasiliensis)的粪便中分离出具有几丁质分解活性的细菌,并验证其对植物病原菌真菌的抗真菌活性。从粪便样本中获得了 28 个细菌分离物,其中 70%具有几丁质分解活性。选择了 4 个分离物,因为它们表现出最高的几丁质酶活性,并且通过分析 16S 核糖体 RNA 基因序列,它们被表征为属于芽孢杆菌属。细菌培养物的无细胞上清液用于对 16 种真菌进行抑制试验:链格孢属和炭疽菌属受到的影响最大。无论使用何种培养基,无细胞上清液中都观察到几丁质酶和抗真菌活性。两种活性都对热和蛋白酶 K 处理稳定。最后,当培养基中补充 1ml 无细胞上清液(0.33%)并孵育 120 小时时,在光镜下观察到报告真菌的菌丝形成和孢子萌发受到抑制。这些结果表明,使用无细胞上清液作为环保型杀真菌剂是可行的。它们的使用可能有助于减少有毒化学物质的剂量。

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