Gene Cloning, Expression, and Antifungal Activities of Permatin from Naked Oat (Avena nuda)
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Thaumatin-like proteins (TLPs) are the products of a large, highly complex gene family involved in host defense. TLPs also belong to the pathogenesis-related family 5 (PR-5) of plant defense proteins. Most TLPs exhibit potential antifungal activities, and their accumulation in the plant is related to many physiological processes. In this study, a gene encoding TLP named permatin with an open reading frame of 678 bp encoding a protein of 225 amino acids with a calculated molecular mass of 23.5 kDa was cloned from naked oat leaves. Phylogenetic analysis revealed that permatin shares high homology with a number of other TLPs among diverse taxa. Model of structure by homology modeling showed that permatin consists of an acidic cleft region consistent with most TLPs. Recombinant NusA-permatin was overexpressed in Escherichia coli strain BL21 and purified by Heparin column combined with Sephacryl S-200 column. The protein exhibited antifungal activity to Fusarium oxysporum (half maximal inhibitory concentration, IC50 = 21.42 μM). Morphological observation showed that NusA-permatin can induce mycelium deformation of F. oxysporum, the cell membrane is blurred, and the diaphragm is not obvious. NusA-permatin also causes membrane permeabilization and reactive oxygen species accumulation in the mycelium of F. oxysporum. Permatin may play an important role in the disease resistance responses of plants against pathogen attacks through its antifungal activity.
KeywordsPermatin Fusarium oxysporum Gene cloning Recombinant expression Antifungal activity Naked oat
We acknowledge that this project is supported by Key Research and Development Program of Shanxi Province of China (no.201603D211104), Special Funds of the Natural Science Foundation of TaiYuan, Shanxi Province (grant no. 11014908), and the fund for Shanxi “1331 project” collaborative innovation center and Research Project supported by Shanxi Scholarship Council of China (no.2017019).
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Conflict of Interest
The authors declare that they have no conflict of interest.
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