Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia

doi:10.7525/j.issn.1673-5102.2021.06.006

Abstract

Abstract:

In order to clarify the mechanism of MADS-box transcription factor gene family member in flowering and floral organ development in Macadamia integrifolia， aMiMADS-box gene was cloned from the leaves of M. integrifolia “GUIRE1” cultivar by PCR technique and analyzed its structure and function by Bioinformatics method. The cloned cDNA sequence of MiMADS-box gene was 1 179 bp in length， 729 bp open reading frame （ORF）， encoded 242 amino acids. MiMADS-box， an unstable hydrophilic protein that had no transmembrane structure， no signal peptide and localized in the nucleus， had MADS superfamily and K-box superfamily conserved domains belonging to the MIKCc-type family. Phylogenetic analysis clustered MiMADS-box and AGL7/AP1， AGL10/CAL1， AGL8/FUL， AGL79 into FUL-AP1（SQUA） subfamily. The transcriptome data were used to analyze the expression patterns of MiMADS-box gene in branch， flower and leave of M. integrifolia “GUIRE1” and “695” cultivar. Results indicated that MiMADS-box had the lowest expression in the flower of “GUIRE1”， but the expression level of “695” was the highest. The green fluorescent protein fusion vector pGREEN-MiMADS-box-ORF-GFP was constructed by homologous recombination. It was speculated that MiMADS-box was closely related to flowering and floral organ development. The study provided a theoretical and technical guide for clarification the role MiMADS-box in flowering regulation mechanism of M. integrifolia.

Key words: Macadamia integrifolia, MiMADS-box, gene cloning, vector construction

CLC Number:

S664.9

Qiu-Jin TAN, Tao ZHANG, Yuan-Rong WEI, Shu-Fang ZHENG, Xiu-Hua TANG, Wen-Lin WANG. Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia[J]. Bulletin of Botanical Research, 2021, 41(6): 888-895.

Figures/Tables 6

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