Characterization of LIM gene family from Tetragonia tetragonoides(Pall.)Kuntze and cloning and functional analysis of TtWLIM1b
[Objective]The study aimed to explore the possible biological roles for development and abiotic stress responses of TtLIM gene family from Tetragonia tetragonoides(Pall.)Kuntze,mainly based on the RNA-seq expression patterns and the phenotype analysis of heter-ologous expression in yeast.[Method]In the study,we screened the cDNA library of T.tetragonoides(Pall.)Kuntze,and cloned a novel LIM gene from this species,named TtWLIM1b.The ectopic expression of TtWLIM1b in yeast was performed.Combined with the whole ge-nome sequencing data,a total of 16 TtLIM genes were identified from T.tetragonoides genome.The chromosomal localization,physicochemi-cal properties of the encoded proteins,phylogenetic evolution,and conserved motifs of these family members were comprehensively analyzed.A phylogenetic tree was constructed to compare the LIM protein family members between T.tetragonoides and Arabidopsis thaliana(L.)Heynh,Oryza sativa L,and Medicago sativa.Finally,RNA-seq analysis was performed to investigate the relative expression levels of TtLIM gene family members in different tissues and under various stress treatments.[Result]The heterologous expression of some TtLIM1b members could improve the heavy metals and heat tolerance of yeast.TtLIM proteins consisted of one or two LIM domains,and LIM domain was cyste-ine-rich zinc finger sequence,therefore TtLIM proteins also belonged to cysteine rich protein(CRP).The sequence analysis indicated that TtLIMs were highly conserved and their genes'reduplicating amplification was prominent during evolution.The RNA-seq results showed that TtLIMs were widely expressed with specific patterns in different organs in T.tetragonoides plant.[Conclusion]TtLIMs might play important roles in regulation of T.tetragonoides development and stress responses.
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