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Summary  Small signalling peptides, generated from larger protein precursors, are important components to orchestrate various plant processes such as development and immune responses. However, small signalling peptides involved in plant immunity remain largely unknown. Here, we developed a pipeline using transcriptomics‐ and proteomics‐based screening to identify putative precursors of small signalling peptides: small secreted proteins ( SSP s) in rice, induced by rice blast fungus  Magnaporthe oryzae  and its elicitor, chitin. We identified 236  SSP s including members of two known small signalling peptide families, namely rapid alkalinization factors and phytosulfokines, as well as many other protein families that are known to be involved in immunity, such as proteinase inhibitors and pathogenesis‐related protein families. We also isolated 52 unannotated  SSP s and among them, we found one gene which we named  immune response peptide  (  IRP  ) that appeared to encode the precursor of a small signalling peptide regulating rice immunity. In rice suspension cells, the expression of   IRP   was induced by bacterial peptidoglycan and fungal chitin. Overexpression of   IRP   enhanced the expression of a defence gene,   PAL 1  and induced the activation of the  MAPK s in rice suspension cells. Moreover, the  IRP  protein level increased in suspension cell medium after chitin treatment. Collectively, we established a simple and efficient pipeline to discover  SSP  candidates that probably play important roles in rice immunity and identified 52 unannotated  SSP s that may be useful for further elucidation of rice immunity. Our method can be applied to identify  SSP s that are involved not only in immunity but also in other plant functions.

Identification of endogenous small peptides involved in rice immunity through transcriptomics‐ and proteomics‐based screening