Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (23): 4754-4762.doi: 10.3864/j.issn.0578-1752.2014.23.020

• RESEARCH NOTES • Previous Articles     Next Articles

Regional Analysis of Transcription Activity and Screening of Interaction Proteins of AtMYB73 Transcription Factor in Arabidopsis

FAN Jin-tao1, JIA Jiao2, JIANG Chen-xi1, WANG Guan-yu1, ZHANG Jing1, XING Ji-hong1, DONG Jin-gao1
  

  1. 1The Laboratory of Mycotoxin and Molecular Plant Pathology, Agricultural University of Hebei, Baoding 071001, Hebei 
    2Institute of Plant Protection, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, Jilin
  • Received:2014-07-11 Revised:2014-08-26 Online:2014-12-01 Published:2014-12-01

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Abstract: 【Objective】 The objective of this study is to analyze the transcription activity region and screen interaction proteins of Arabidopsis resistance related transcription factor AtMYB73 and the study will lay a foundation for clarifying the regulation mechanism of the AtMYB73 gene in Arabidopsis resistance in the future. 【Method】 The bait vector pAS1-AtMYB73 was constructed and introduced into yeast Y190 by PEG/LiAC mediated transformation method. The self-activation and cytotoxicity of pAS1-AtMYB73 were detected in this paper. The transcription activity domain of the AtMYB73 was analyzed through detecting the transcription activity of the N-terminus and C-terminus of the AtMYB73. The AtMYB73 was used as bait to screen Arabidopsis cDNA library by the yeast two-hybrid system. The positive clones were screened on SD/-Ade/-His/-Leu/-Trp plates, identified by PCR and sequenced, and function of the interaction proteins were analyzed using TAIR database. The pGBDT7-AtMYB73 and pGADT7-F12F1.4 vectors were constructed and co-transformed into yeast AH109. The interaction relationship between AtMYB73 and F12F1.4 was analyzed by yeast two-hybrid system.【Result】The bait vector of the AtMYB73, pAS1-AtMYB73, was successfully constructed and transformed into yeast Y190. The pAS1-AtMYB73 yeast could grow on SD/-His/-Trp/ and SD/-Ade/-Trp plates with different concentrations of 3-AT, suggesting that AtMYB73 has higher self-activation activity. The OD600 of the pAS1-AtMYB73 yeast cultured for 24 h in SD/-Trp/Amp liquid media was greater than 0.8, showing that the bait vector has no cytotoxicity. Vectors of pAS1-AtMYB73-N and pAS1-AtMYB73-C were successfully constructed and transformed into yeast Y190, respectively. The pAS1-AtMYB73-N yeast was colorless, but the pAS1-AtMYB73-C yeast was blue. These results indicated that the C-terminus of AtMYB73 has obvious self-activation activity and the N-terminus of AtMYB73 has no self-activation activity. Eight candidate interacting proteins of AtMYB73 were obtained by screening Arabidopsis cDNA library using yeast two-hybrid system. Function annotation showed that these candidate interacting proteins were related to photosynthesis, defense reaction and resistance. The interaction relationship between AtMYB73 and F12F1.4 was determined by yeast two hybrid system. 【Conclusion】 Transcription factor AtMYB73 has higher self-activation activity and the transcription activity region of AtMYB73 was localized in its C-terminus. Eight interaction proteins related to photosynthesis, defense reaction and resistance were obtained by screening Arabidopsis cDNA library. Transcription factor AtMYB73 interacting with F12F1.4 was determined by yeast two hybrid system.

Key words: Arabidopsis thaliana, transcription factor AtMYB73, transcription activity, yeast two-hybrid, interaction protein

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