Screening and Identification of Interacting Proteins of CsMYC2 and CsMPK6 in Citrus Based on Pull-Down/MS Technique
The purpose of this study is to explore the interaction network of CsMPK6-CsMYC2 module and identify the proteins involved in the citrus peel coloring process mediated by jasmonic acid through the interaction screening with CsMPK6-CsMYC2 module.To this purpose,Pull-Down with mass spectrometry technique(Pull-Down/MS)was performed to screen and identify the CsMPK6-CsMYC2 interacting proteins using the total proteins isolated from MeJA-treated citrus fruit.The interactive proteins were involved in secondary metabolites biosynthesis,plant mitogen-activated protein kinase signaling pathway,ribosome pathway,stress response,hormone response,abiotic stress responses and other biological processes according to bioinformatics analysis.Among them,trihelix transcription factor CsGT2 and germin-like protein CsGLP play important roles in fruit development.Therefore,the interactions of CsGT2 with CsMYC2,CsGLP with CsMPK6 and CsGLP with CsMYC2 were further confirmed.The promoters of CsGT2 and CsGLP contained multiple cis-acting elements involving in light,hormone and stress-response.This work provides a reference for species that are difficult to screen interacting proteins by using yeast two-hybrid system and IP-MS technology.Meanwhile,this work provides gene pool for further study regarding the regulatory network for the jasmonate-induced coloration in citrus fruit.
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