Cloning and Expression Pattern Analysis of OlMYB35 Gene from Opisthopappus longilobus
ZHANG Ya-Ru, WANG Yan-Ru, JING Ru-Yue, SU Jiang-Shuo, SONG Ai-Ping, WANG Hai-Bin, JIANG Jia-Fu*
State Key Laboratory of Crop Genetic & Germplasm Enhancement and Utilization/Key Laboratory of Landscape Agriculture, Ministry of Agriculture and Rural Affairs/College of Horticulture, Nanjing Agricultural University, Nanjing 211800, China
Abstract：MYB transcription factors constitute one of the largest transcription factor families in plants and are extensively involved in various biological processes, including growth and development, secondary metabolism, and stress responses. Opisthopappus longilobus, a wild species closely related to chrysanthemum and endemic to China, has long thrived in arid, nutrient-poor cliff habitats characterized by substantial water fluctuations, exhibiting strong environmental adaptability. In this study, the OlMYB35 gene was cloned from O. longilobus. Sequence analysis revealed that the ORF was 888 bp in length, encoding a protein of 295 amino acids with a predicted molecular mass of approximately 33.12 kD and a theoretical isoelectric point (pI) of 8.59. OlMYB35 belonged to the R2R3-MYB transcription factor family. Secondary structure prediction indicated that the protein was mainly composed of random coils, α-helices, and extended strands. Phylogenetic analysis showed that OlMYB35 was most closely related to homologous proteins from Chrysanthemum species and was evolutionarily conserved. A p35S::OlMYB35-GFP fusion expression vector was constructed, and subcellular localization analysis revealed that the protein was localized in both the nucleus and the cytoplasm. Yeast transcriptional activation assays demonstrated that OlMYB35 possessed transcriptional activation activity. Expression pattern analysis under water stress conditions showed that the OlMYB35 gene was rapidly induced by polyethylene glycol 6000 (PEG6000)-simulated drought stress and significantly upregulated under waterlogging stress, suggesting its potential involvement in regulating the response of O. longilobus to water stress. This study provides a theoretical basis for further functional characterization of the transcription factor OlMYB35 in O. longilobus and for the application of chrysanthemum molecular breeding for stress tolerance.
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