[Objective]CIM(cytokinin induce messager)belongs to the Expansin protein family,which plays a critical role in plant growth and development.To analyze the structure,expression,subcellular localization,and function of MtCIM during Medicago truncatula growth and development.[Method]Bioinformatics tools were employed to conduct a comprehensive study of molecular weight,theoretical isoelectric point,instability index,and protein structure prediction and analysis.Meanwhile,RT-qPCR method was used to measure the gene expressions under different hormone treatments(IAA,ABA and GA3).Fusion protein transient expression and yeast expression methods were used to identify its subcellular localization and transcriptional self-activation,respectively.Analysis of MtCIM-transgenic plants was performed in terms of their adaptation to a low phosphorus environment.[Result]Bioinformatics analysis showed that the CIM protein had a relative molecular weight of 29.563 kD,theoretical isoelectric point of 5.54,and instability index of 41.62,indicating its protein property were unstable.The secondary and tertiary structure predictions showed that the protein was mainly composed of irregular curls.Exogenous IAA significantly increased the expression of MtCIM,while the expression of MtCIM showed a trend of rising and then decreasing after exogenous ABA and GA3.The subcellular localization results showed that it was located in the cytoplasm.Upon analysis of transcriptional self-activation,it was found that there existed no self-activation activity in MtCIM.Analysis of the 20 obtained transgenic plants revealed that the cell walls of the leaves from these transgenic lines were thicker.Moreover,the transgenic plants demonstrated superior adaptability to low-phosphorus environments.[Conclusion]A comprehensive analysis of the MtCIM gene in M.truncatula revealed its key role as a member of the Expansin protein family in plant growth and development.The phenotype of the transgenic plants demonstrated that MtCIM enhanced the adaptability of plants in low-phosphorus environments.These findings provide new insights for further understanding the role of the Expansin protein family in plant growth and development.
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