Abstract:To understand the function of CRABS CLAW (CRC) subgene of the YABBY gene family of Anoectochilus roxburghii in flower and leaf development, the ArCRC gene was cloned by RT-PCR based on the full-length transcriptome data of A. roxburghii. The bioinformatics, prokaryotic expression and subcellular localization of encoded ArCRC protein were analyzed, and the expression pattern of ArCRC gene was analyzed by qPCR. The results showed that the CDS length of ArCRC gene was 576 bp (GenBank accession No.: OR394646), coding 191 amino acids. ArCRC contains YABBY superfamily and HMG-box_SF superfamily conserved domain, with molecular weight 21.514 kD, theoretical isoelectric point 9.16, instability coefficient 41.12, belonging to the unstable protein. Phylogenetic analysis showed that ArCRC was clustered with OsDL of rice (Oryza sativa) and AtCRC of Arabidopsis thaliana, belonging to the CRC subfamily and localized in the nucleus. SDS-PAGE electrophoresis and Western blot results showed that ArCRC gene could be successfully induced in Escherichia coli. qRT-PCR analysis showed that the expression of ArCRC gene was the highest in flowers, followed by leaves, and the expression in the midrib of leaves was significantly higher than that in the margin of leaves. Therefore, it was speculated that ArCRC gene mainly played a role in the development of flower organs and was also involved in regulating the development of the midrib of leaves.
