Abstract
Main conclusion
We performed genome-wide and heterologous expression analysis of the safflower cysteine protease family and found that inhibition of CtCP1 expression enhanced plant cold resistance.
Abstract
Cysteine protease (CP) is mainly involved in plant senescence and stress responses. However, the molecular mechanism of endogenous cysteine protease inhibition in plant stress tolerance is yet unknown. Here, we report the discovery and functional characterization of a candidate CP1 gene from safflower. The conserved structural topology of CtCPs revealed important insights into their possible roles in plant growth and stress responses. The qRT-PCR results implied that most of CtCP genes were highly expressed at fading stage suggesting that they are most likely involved in senescence process. The CtCP1 expression was significantly induced at different time points under cold, NaCl, H2O2 and PEG stress, respectively. The in-vitro activity of heterologously expressed CtCP1 protein showed highest protease activity for casein and azocasein substrates. The expression and phenotypic data together with antioxidant activity and physiological indicators revealed that transgenic plants inhibited by CtCP1-anti showed higher tolerance to low temperature than WT and CtCP1-OE plants. Our findings demonstrated the discovery of a new Cysteine protease 1 gene that exerted a detrimental effect on transgenic Arabidopsis under low-temperature stress.
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Availability of data and materials
All the data associated with manuscript is included in this paper and its supplementary materials. Vectors and Arabidopsis seeds can be requested from the corresponding authors. The vector pEASY-T1 is also available via Miaoling (https:// miaolingbio.com). The whole genome sequence of Carthamus tinctorius is derived from NCBI (https://www.ncbi.nlm.nih.gov), accession number PRJNA399628. The data of Arabidopsis thaliana was obtained from http://www.Arabidopsis.org/.
Abbreviations
- CAT:
-
Catalase
- CP:
-
Cysteine protease
- MDA:
-
Malondialdehyde
- PCD:
-
Programmed cell death
- POD:
-
Peroxidase
- SOD:
-
Superoxide dismutase
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Acknowledgements
We are grateful to Professor Li Haiyan (Hainan University) for providing pCAMBIA3301.
Funding
This work was funded by grants from the National Natural Science Foundation of China (Nos. 31771868, 31501366), the Science and Technology Development Project of Jilin Province (Nos. 20190201172JC, 20190201175JC, JJKH20210347KJ), Student innovation and entrepreneurship training program of Jilin Agricultural University.
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Yingqi, H., Lv, Y., Zhang, J. et al. Identification and functional characterization of safflower cysteine protease 1 as negative regulator in response to low-temperature stress in transgenic Arabidopsis. Planta 255, 106 (2022). https://doi.org/10.1007/s00425-022-03875-6
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DOI: https://doi.org/10.1007/s00425-022-03875-6