Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (5): 205-216.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0812

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Functional Study and Validation of Transcriptional Coactivator PwMBF1c in Picea wilsonii

LIU Kui(), LI Xing-fen, YANG Pei-xin, ZHONG Zhao-chen, CAO Yi-bo(), ZHANG Ling-yun()   

  1. School of Forestry, State Key Laboratory of Forest Cultivation and Protection of Ministry of Education, Beijing Forestry University, Beijing 100083
  • Received:2022-07-02 Online:2023-05-26 Published:2023-06-08
  • Contact: CAO Yi-bo, ZHANG Ling-yun E-mail:742174371@qq.com;caoyibo@bjfu.edu.cn;lyzhang@bjfu.edu.cn

Abstract:

We cloned PwMBF1c, a candidate gene in response to drought stress based on the transcriptomes of Picea wilsonii under drought stress and verified its biological function in order to explore the drought tolerance mechanism of P. wilsonii and to provide genetic resources for breeding excellent resistant forest varieties. RT-qPCR was used to analyze the expression profile of PwMBF1c to drought, high temperature, low temperature and hormone treatments. Transiently transformed tobacco leaf and onion epidermal cells were used to detect the subcellular localization of PwMBF1c. The transcriptional activity of PwMBF1c was verified by yeast one-hybrid. The function of PwMBF1c was verified in Arabidopsis thaliana and potato systems. PwMBF1c was mainly expressed in mature leaves, and its transcription level was strongly up-regulated by drought, high temperature and salicylic acid(SA)treatment. The C-terminus of PwMBF1c showed transcriptional activation activity, but not for the N-terminus or full-length of PwMBF1c. PwMBF1c was localized in cell membrane, cytoplasm and nucleus, and overexpression of PwMBF1c significantly increased drought tolerance in both Arabidopsis and potato. Under drought stress, compared with the control(Col-0 and VC), PwMBF1c-L1 and PwMBF1c-L2 lines showed higher survival rate, higher chlorophyll content and lower MDA content. After PEG treatment, PwMBF1c significantly increased seedling height compared with wild-type potato(Y5). The expression of PwMBF1c significantly increased by multiple abiotic stresses such as drought. PwMBF1c is mainly localized in cell membrane, cytoplasm and nucleus, and the C-terminus of PwMBF1c showed transcriptional activation activity. PwMBF1c significantly enhanced the drought tolerance of A. thaliana and potato.

Key words: Picea wilsonii, MBF1c coactivator, stress response, gene expression, drought tolerance