Abstract:
Based on the transcriptome data of Anoectochilus roxburghii leaves, an LBD transcription factor gene, named ArLBD1,was cloned by RT-PCR.The function of the gene was analyzed by bioinformatics, prokaryotic expression, Western blot, subcellular localization and expression patterns of different organs, in order to provide reference for further research on the gene′s function.The results showed that the CDS length of ArLBD1 gene was 864 bp, encoding 287 amino acids.The molecular weight of ArLBD1 protein was 30.62 kD,theoretical isoelectric point was 6.25,and instability coefficient was 55.55.ArLBD1 protein belonged to unstable protein.ArLBD1 protein contained a LOB superfamily conserved domain, its amino acid 7-28 was C-block, amino acid 36-84 was GAS-block, and amino acid 89-107 was Leucine-zipper-like block.It had a complete Leucine-zipper-like-block and belonged to Class I.Phylogenetic analysis showed that ArLBD1 was most closely related to Arabidopsis thaliana AtLBD36,followed by AtLBD6.SDS-PAGE and Western blot results showed that ArLBD1 gene was successfully induced and expressed in Escherichia coli.Subcellular localization analysis showed that ArLBD1 protein was in the nucleus.The results of qPCR analysis showed that ArLBD1 gene could be detected in leaves,stem and roots of Anoectochilus roxburghii,with the highest expression level in leaves,followed by roots,and the lowest expression level in stems.It was speculated that the ArLBD1 gene mainly played a role in the development of Anoectochilus roxburghii leaves.