Identification and Spatiotemporal Expression Analysis of R2R3-MYB Transcription Factor in Crocus sativus
WU Rui1,2, LIN Ding1,2, LUO Dong1,2, ZHOU Yi-Ru1,2, ZHANG Ling-Yi1,2, SHAO Qing-Song1,2, XING Bing-Cong1,2,*
1 Zhejiang Provincial Key Laboratory of Traditional Chinese Medicine Resources Protection and Innovative Utilization, Hangzhou 311300, China; 2 College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China
Abstract R2R3-MYB transcription factor is one of the largest transcription factor family in plants, and it is widely involved in the regulation of plant secondary metabolism. Based on the sequencing data of the full-length transcriptome of Crocus sativus, explored the temporal and spatial expression characteristics of R2R3-MYB transcription factors and screend the CsR2R3-MYB transcription factors related to the secondary metabolism of C. sativus, bioinformatics methods were used to identify and systematically analyzed the CsR2R3-MYB transcription factor, and the temporal and spatial expression pattern of the CsR2R3-MYB transcription factor was explored by qRT-PCR. Totally 42 R2R3-MYB transcription factors were screened from the C. sativus transcriptome. Physical and chemical properties analysis of the proteins showed that the CsR2R3-MYB proteins were all unstable hydrophilic proteins. Subcellular localization prediction showed that most of the CsR2R3-MYB were present in the nucleus. The phylogenetic tree was constructed together with Arabidopsis thaliana R2R3-MYB, and the results showed that the CsR2R3-MYB transcription factors could be divided into 26 subclasses. According to the function conserved properties of homologous proteins, CsMYB9, CsMYB16, CsMYB17, CsMYB18, CsMYB20, CsMYB21, which were homologous to the S7, S19, and S20 subclasses, were predicted to participate in the secondary metabolism regulation of active substances in C. sativus by responding to jasmonic acid (JA) signals. The results of temporal and spatial expression analysis showed that CsMYB5, CsMYB6, CsMYB7, CsMYB9, CsMYB10, CsMYB13, CsMYB17, CsMYB19, CsMYB21, CsMYB22, CsMYB23, CsMYB26, CsMYB28, CsMYB30, CsMYB31 and CsMYB32 showed relatively high expression levels in the stigma tissues and responds to JA signals. The previous studies of this research group showed that JA could promote flavonoids biosynthesis. According to the prediction results of the phylogenetic tree and the temporal and spatial expression characteristics, screened CsMYB9, CsMYB17, and CsMYB21 regulate the secondary metabolism of C. sativus, by responding to JA signals. This research provides a reference for the further exploration of the functions of CsR2R3-MYB. And it also provides ideas for the study of transcription regulation of C. sativus secondary metabolism.
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