关键词: 月季, RhNAC31, 酵母双杂交, 互作蛋白

Abstract: To further understand the molecular mechanism of RhNAC31 in stress response，we used the RhNAC31 protein as bait through cut rose‘Samantha’water deficit stress yeast two-hybrid（Y2H）cDNA libraries and screened its interacting proteins. RhNAC31 transactivation domain was located in its C-terminal（157–286 aa）. Two different deletion regions，including RhNAC31-C1（157–250 aa）and RhNAC31-C2（251–286 aa），were constructed to pGBKT7 vector，respectively. Self-activation activity and toxicity were also examined in Y2H Gold yeast cells. The results showed that both pGBKT7-RhNAC31-C1 and pGBKT7-RhNAC31-C2 had no toxicity in Y2H Gold cells and could stimulate Aureobasidin A（AbA）reporter gene with auto-activation activity. The activities were obviously inhibited when supplemented with 400 ng ? mL-1 AbA under SD deficient medium. The SD deficient medium plus 400 ng ? mL-1 AbA was used as Y2H screening library medium. Twenty-one positive colonies were screened by Y2H，and 16 of them may be candidate interacting proteins of RhNAC31 which were further sequenced. In addition，these interacting proteins were also functional associations based on gene ontology through Uniprot website. RhNAC31 may interact with RZFP34，MIEL1，RUB E3 ubiquitin ligase，SGT1，DnaJ and other proteins. These different proteins may participate in glycolipid biosynthesis，protein ubiquitination，plant defense，chlorophyll biosynthesis，pattern recognition receptor signaling pathway，protein folding，ethylene biosynthesis，stress response and other biological processes.

Key words: rose, RhNAC31, yeast two-hybrid, interacting protein