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| Analysis of Transactivation Activity and Expression Characteristics of RhMYB96 in Rosa hybrida |
| DING Ai-Qin, LI Shao-Cui, LIU Qing-Chao, WANG Kui-Ling, LIU Qing-Hua, JIANG Xin-Qiang* |
| College of Landscape Architecture and Forestry, Qingdao Agriculture University, Qingdao 266109, China |
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Abstract External environment and internal gene regulation could influence the germplasm of Rosa hybrida genetic characteristics, and transcription regulation plays vital roles in these biological processes. To determine the biological function of R2R3-MYB (v-myb avian myeloblastosis viral oncogene homolog) transcription factors in rose, a new MYB transcription factor gene named as RhMYB96 (GenBank No. MF185658) was cloned from cut rose 'Samantha' based on the expressed sequence tags from transcriptome assembly and rapid amplification of cDNA ends (RACE) methods. The RhMYB96 ORF was 1 065 bp, which encoded 354 amino acids. Bioinformatics analysis showed that the molecular weight and theoretical isoelectric point of RhMYB96 was 39.49 kD and 6.89, respectively, and the formula was C1693H2655N519O548S14. Multiply protein alignment revealed that RhMYB96 contained 2 highly conserved domains R2-MYB and R3-MYB in its N terminus regions. The secondary structure of RhMYB96 was typical helix-turn-helix mode. In addition, the C terminal region of RhMYB96 and other plant MYB proteins were used to construct phylogenic tree. RhMYB96 had a close relationship with Arabidopsis AtMYB96, AtMYB94 and AtMYB30, which belonged to the R2R3-MYB proteins of subgroup 1. qRT-PCR results showed that the expression of RhMYB96 in rose petals was the highest when compared with other organs. In addition, salt stress and abscisic acid (ABA) treatment significantly increased the expression of RhMYB96. Meanwhile, salt stress plus ABA treatment significantly increased the expression of RhMYB96. Moreover, 6 recombinant plasmids in yeast cells, which contained different fragments of RhMYB96 were constructed. The transactivation analysis of RhMYB96 was also examined by yeast one hybrid-assays. The results showed that RhMYB96 was a transcriptional activator, and an activation domain located at the C terminus. The N terminal DNA-binding domain lost transcription activity. Above results showed that transactivator RhMYB96 might participate in salt stress response via ABA dependent pathway. The study provides a foundation for predicting the roles of RhMYB96 in transcription regulatory networks and inferring functions in molecular breeding of rose plant species.
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Received: 30 August 2018
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Corresponding Authors:
* jiangxinqiang8@163.com
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