Antifreeze protein from Ammopiptanthus nanus

In this research resercher discuss about the Temperature stress restricts in plant growth and development. Antifreeze protein (AFP) can improve plants antifreeze ability. In our previous study, the AnAFP gene cloned from Ammopiptanthus nanus was confirmed to be an excellent candidate enhancing plant cold resistance. But, AnAFP protein shared similar structures with KnS type dehydrins including K, N and S domains except ice crystal binding domain A. Here, we generatedAnAFPΔA, AnAFPΔK, AnAFPΔN and AnAFPΔS, and transformed them into ordinary and cold sensitive strains of E. coli, and Arabidopsis KS type dehydrin mutant to evaluate their function. Expression of AnAFPΔAdecreases cold and heat tolerance in E. coli, meanwhile, AnAFP enhances heat tolerance in Arabidopsis, suggesting that domain A is a thermal stable functional domain. AnAFP, AnAFPΔA and AnAFPΔS localize in whole cell, but AnAFPΔK and AnAFPΔN only localizes in nucleus and cytoplasm, respectively, exhibiting that K and N domains control localization of AnAFP.


FIG: Phenotype of cold sensitive BX40 strain of E. coli with AnAFP, AnAFPΔA,AnAFPΔK, AnAFPΔN and AnAFPΔS gene under low temperature stress (17 °C) for 9 days. (a) Colony growth of different dilution times. (b) Monoclone of transformed strains. (c) Average survival rate of transformed strains. Lower case letter indicates the significant difference at p < 0.05 level in student’s t-test. The experiment was performed with three replicates. The data were presented as the mean values ± SD.

Likewise, K domain blocks interaction between AnAFP and AnICE1. The result of RT-qPCR showed that expression ofAnAFP, AnICE1 and AnCBF genes was significantly induced by high-temperature, indicating that the AnAFP is likely regulated by ICE1-CBF-COR signal pathway. Taken together, the study provides insights into understanding the mechanism of AnAFP in response to temperature stress and gene resource to improve heat or cold tolerance of plants in transgenic engineering.

Yu, H., Zheng, H., Liu, Y. et al. Antifreeze protein fromAmmopiptanthus nanus functions in temperature-stress through domain A. Sci Rep 11, 8458 (2021).

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