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Topping damage can induce the nicotine synthesis in tobacco roots, which involves the activation of JA and auxin signal transduction. It remains unclear how these hormone signals are integrated to regulate nicotine synthesis. Here we isolated a transcription factor  NtWRKY-R1  from the group IIe of WRKY family and it had strong negative correlation with the expression of  putrescine N-methyltransferase , the key enzyme of nicotine synthesis pathway.  NtWRKY-R1  was specifically and highly expressed in tobacco roots, and it contains two transcriptional activity domains in the N- and C-terminal. The promoter region of  NtWRKY-R1  contains two cis-elements which are responding to JA and auxin signals, respectively. Deletion of  NtWRKY-R1  promoter showed that JA and auxin signals were subdued by  NtWRKY-R1 , and the expression of  NtWRKY-R1  was more sensitive to auxin than JA. Furthermore, Yeast two-hybrid experiment demonstrated that NtWRKY-R1 can interact with the actin-binding protein. Our data showed that the intensity of JA and auxin signals can be translated into the expression of  NtWRKY-R1 , which regulates the balance of actin polymerization and depolymerization through binding actin-binding protein, and then regulates the expression of genes related to nicotine synthesis. The results will help us better understand the function of the WRKY-IIe family in the signaling crosstalk of JA and auxin under damage stress.

NtWRKY-R1, a Novel Transcription Factor, Integrates IAA and JA Signal Pathway under Topping Damage Stress in Nicotiana tabacum