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Bulletin of Botanical Research ›› 2021, Vol. 41 ›› Issue (6): 888-895.doi: 10.7525/j.issn.1673-5102.2021.06.006

• Research Report • Previous Articles     Next Articles

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

Qiu-Jin TAN, Tao ZHANG, Yuan-Rong WEI, Shu-Fang ZHENG, Xiu-Hua TANG, Wen-Lin WANG()   

  1. Guangxi South Subtropical Agricultural Science Research Institute,Longzhou 532400
  • Received:2021-03-04 Online:2021-11-20 Published:2021-10-29
  • Contact: Wen-Lin WANG E-mail:wwlin3949@163.com
  • About author:TAN Qiu-Jin(1980—),famale,senior agronomist,mainly engaged in the genetic breeding of tropical fruit trees.
  • Supported by:
    Guangxi Academy of Agricultural Sciences Science and Technology Development Fund(Guinongke 2020YM141,Guinongke 2020YM46,Guinongke 2019ZX2015,2019ZX17);Major Science and Technology Projects of Guangxi(Guike AA17204058-4);National Key Research and Development Project(2016YFC0502406)


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

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