Cloning And Function Analysis Of14-3-3Family And NPR1-like Genes In Melon

NPR1(nonexpressor of PR gene1), involved in regulating plant broad-spectrum resistance, plays important roles in plant system resistance.14-3-3proteins widely exist in eukaryotic with highly conservative, involved in the regulation of basic metabolism, signal transduction, cell division, photoperiod, biotic stress and abiotic stress, through interaction of many target proteins, In our study, We cloned NPR1-like genes and the14-3-3family members in melon, analyzed their expression patterns by qRT-PCR, and studied the function of CmNPR3and CmNPR4in disease resistance through their transiently overexpression in Nicotiana benthamiana.Cloning and subcellular localization of NPR1-like genes and the14-3-3family members in melon. A total of9members of the14-3-3family and2NPR1-like genes (CmNPR3/CmNPR4) in melon were cloned by RT-PCR, and subcellular localization assays showed that the9proteins of the14-3-3family were localized on the plasma membrane and cytoplasm, whereas CmNPR3/CmNPR4were localized in the nucleus.Interaction of melonl4-3-3family members and2NPR1-like proteins. To determine the interaction of the9members of the14-3-3family by BiFC assays,14-3-3gene fusion constructs were generated and transiently expressed in Nicotiana benthamiana. The results observed by laser confocal microscopy showed that the914-3-3members can interact by themselves and to each other. Also, BiFC assays and yeast two-hybrid system confirmed that CmNPR3and CmNPR4can form both homo-and heterodimer in a SA-independent manner.Expression patterns of NPR1-like genes and the14-3-3family members in melon. We studied the expression patterns of NPR1-like genes and the14-3-3family members in melon in hormones (SA/JA/ACC) and abiotic stress (cold, heat, salt, drought) treatments. qRT-PCR results showed that CmNPR3/CmNPR4were induced by SA, and partial members of14-3-3family were reduced in drought and heat stress.CmNPR3and CmNPR4positively regulated the resistance to pathogens. B. cinerea and Pst DC3000were inoculated to the N.benthamianain which CmNPR3andCmNPR4were transiently overexpressed. Results showed that CmNPR3andCmNPR4transiently overexpressed plants had positive roles in resistance to Pst DC3000, but had no obvious function on B. cinerea resistance.