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| Gene Cloning, Expression Analysis and Identification of Interacting Proteins of Transcription Factor WRKY72 in Oilseed Rape (Brassica napus) |
| LI Qi*, LI Cui*, WANG, Chen SUN Yun J, IANG Yuan-Qing**, YANG Bo** |
| College of Life Sciences, Northwest A&F University, Yangling 712100, China |
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Abstract WRKY transcription factors are a class of plant-specific proteins involved in the regulation of growth and development, secondary metabolism, senescence and response to biotic and abiotic stresses.The present study firstly cloned the cDNA sequence of WRKY72 (GenBank No.KC246585) from oilseed rape (Brassica napus) by the method of reverse transcription PCR (RT-PCR).The encoded protein of BnaWRKY72 included a conserved WRKY domain and a C2H2-type zinc finger motif.Subcellular localization study using green fluorescence protein revealed that BnaWRKY72 transcription factor localized in the nucleus.Dual luciferase reporter assay detected that BnaWRKY72 had trans-activation activity towards firefly luciferase gene driven by tandem repeat of W-box element.The expression of BnaWRKY72 was examined by qRT-PCR under various hormones, biotic and abiotic stresses.It was found that BnaWRKY72 responded to cold stress and Sclerotinia sclerotiorum infection.Yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays were used to screen and confirm the interacting proteins of WRKY72 which included homologous BnaWRKY31 and BnaWRKY42 in the same subclade and itself.It was also found that BnaWRKY72 could interact with small ubiquitin-like modifier (SUMO) 3, suggesting that SUMO modification might participate in regulating of BnaWRKY72 activity.The present study explored the molecular characteristics of BnaWRKY72 for the first time, which provides a reference for further study of its stress resistance function and related molecular mechanism.
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Received: 10 November 2018
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Corresponding Authors:
yangwl@nwafu.edu.cn;jiangyq@nwafu.edu.cn
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