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Rubber tree ( Hevea brasiliensis  Muell. Arg) is an important economic crop in Thailand. Leaf fall and black stripe diseases caused by the aggressive oomycete pathogen  Phytophthora palmivora , cause deleterious damage on rubber tree growth leading to decrease of latex production. To gain insights into the molecular function of  H .  brasiliensis  subtilisin-like serine proteases, the  HbSPA ,  HbSPB , and  HbSPC  genes were transiently expressed in  Nicotiana benthamiana  via agroinfiltration. A functional protease encoded by  HbSPA  was successfully expressed in the apoplast of  N .  benthamiana  leaves. Transient expression of  HbSPA  in  N .  benthamiana  leaves enhanced resistance to  P .  palmivora , suggesting that HbSPA plays an important role in plant defense.  P .  palmivora  Kazal-like extracellular protease inhibitor 10 (PpEPI10), an apoplastic effector, has been implicated in pathogenicity through the suppression of  H .  brasiliensis  protease. Semi-quantitative RT-PCR revealed that the  PpEPI10  gene was significantly up-regulated during colonization of rubber tree by  P .  palmivora . Concurrently, the  HbSPA  gene was highly expressed during infection. To investigate a possible interaction between HbSPA and PpEPI10, the recombinant PpEPI10 protein (rPpEPI10) was expressed in  Escherichia coli  and purified using affinity chromatography. In-gel zymogram and co-immunoprecipitation (co-IP) assays demonstrated that rPpEPI10 specifically inhibited and interacted with HbSPA. The targeting of HbSPA by PpEPI10 revealed a defense-counterdefense mechanism, which is mediated by plant protease and pathogen protease inhibitor, in  H .  brasiliensis - P .  palmivora  interactions.

plos one

The plant defense and pathogen counterdefense mediated by Hevea brasiliensis serine protease HbSPA and Phytophthora palmivora extracellular protease inhibitor PpEPI10