Transgenic expression of plant-specific insert of potato aspartic proteases (StAP-PSI) confers enhanced resistance to Botrytis cinerea in Arabidopsis thaliana
Abstract The plant-specific insert of Solanum tuberosum aspartic proteases ( St AP-PSI) has high structural similarity with NK-lysin and granulysin, two saposin-like proteins (SAPLIPs) with antimicrobial activity. Recombinant St AP-PSI and some SAPLIPs show antimicrobial activity against pathogens that affect human and plants. In this work, we transformed Arabidopsis thaliana plants with St AP-PSI encoding sequence with its corresponding signal peptide under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Results obtained show that St AP-PSI significantly enhances Arabidopsis resistance against Botrytis cinerea infection. St AP-PSI is secreted into the leaf apoplast and acts directly against pathogens; thereby complementing plant innate immune responses. Data obtained from real-time PCR assays show that the constitutive expression of St AP-PSI induces the expression of genes that regulate jasmonic acid signalling pathway, such as PDF1.2, in response to infection due to necrotrophic pathogens. On the other hand, according to the data described for other antimicrobial peptides, the presence of the St AP-PSI protein in the apoplast of A. thaliana leaves is responsible for the expression of salicylic acid-associated genes, such as PR-1, irrespective of infection with B. cinerea . These results indicate that the increased resistance demonstrated by A. thaliana plants that constitutively express St AP-PSI owing to B. cinerea infection compared to the wild-type plants is a consequence of two factors, i.e., the antifungal activity of St AP-PSI and the overexpression of A. thaliana defense genes induced by the constitutive expression of St AP-PSI. We suggest that the use of this protein would help in minimizing the ecological and health risks that arise from the use of pesticides. We suggest that the use of this protein would help in minimizing the ecological and health risks that arise from the spreading of resistance of agriculturally important pathogens. Highlights The constitutive expression of St AP-PSI induces defense genes in Arabidopsis thaliana. The St AP-PSI domain exerts cytotoxic activity toward Botrytis cinerea. The constitutive expression of St AP-PSI reduces B. cinerea infection in A. thaliana. The constitutive expression of St AP-PSI increases growth in A. thaliana. Graphical abstract [DISPLAY OMISSION]
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