College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, 130118, China.
Institute of Crop Germplasm Resources (Institute of Biotechnology), Shandong Academy of Agricultural Sciences; Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan, 250100, People's Republic of China.
Planta. 2022 Apr 21;255(5):106. doi: 10.1007/s00425-022-03875-6.
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. 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, HO 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.
我们对红花半胱氨酸蛋白酶家族进行了全基因组和异源表达分析，发现抑制 CtCP1 表达可增强植物的抗寒性。半胱氨酸蛋白酶（CP）主要参与植物衰老和应激反应。然而，内源性半胱氨酸蛋白酶抑制在植物应激耐受中的分子机制尚不清楚。在这里，我们报道了红花候选 CP1 基因的发现和功能特征。CtCPs 的保守结构拓扑结构揭示了它们在植物生长和应激反应中可能的作用的重要见解。qRT-PCR 结果表明，大多数 CtCP 基因在褪色期高度表达，这表明它们很可能参与衰老过程。CtCP1 的表达在冷、NaCl、HO 和 PEG 胁迫下的不同时间点均显著诱导。异源表达的 CtCP1 蛋白的体外活性对酪蛋白和偶氮酪蛋白底物表现出最高的蛋白酶活性。表达和表型数据以及抗氧化活性和生理指标表明，CtCP1-anti 抑制的转基因植物对低温的耐受性高于 WT 和 CtCP1-OE 植物。我们的研究结果表明，在低温胁迫下，拟南芥中的一种新的半胱氨酸蛋白酶 1 基因被发现会产生有害影响。
