Molecular Cloning and Characterization of the GoMADS-box2 Gene from Guzmania Ruiz & Pav  

Jianxin Liu , Weiyong Wang , Fuquan Shen , Huaqiao Ding , Fei Zhang , Xiaojing Liu
Flower Research and Development Center, Zhejiang Academy of Agricultural Sciences, No. 79 Tangwan Village, Beigan Sub-District, Xiaoshan District, Hangzhou, 311202, P.R.China
Author    Correspondence author
Molecular Plant Breeding, 2013, Vol. 4, No. 2   doi: 10.5376/mpb.2013.04.0002
Received: 21 Nov., 2012    Accepted: 07 Dec., 2012    Published: 24 Dec., 2012
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This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Liu J.X., et al., 2012, Molecular Cloning and Characterization of the GoMADS-box2 Gene from Guzmania Ruiz & Pav, Molecular Plant Breeding, Vol.4, No.2 15-23 (doi: 10.5376/mpb.2013.04.0002)

Abstract

The MADS-box gene plays a crucial role in the formation of the floral meristem, development of floral organs and florescence regulation in plants. Based on a full-length cDNA library of floral organs and bracts in ostara, a Guzmania cultivar, four EST sequences from the same contig that included the MADS-box gene were obtained by large-scale random sequencing. Using the primer walking sequencing method, a 985 bp full-length cDNA sequence of the MADS-box gene was identified, which was named GoMADS-box2 gene (GenBank accession No.:JN936044). It has a 675 bp open reading frame that encodes 225 amino acid residuals, and its theoretical molecular weight and isoelectric point were predicted to be 26.3 kD and 9.36, respectively, by using a ProtParam analysis. The putative GoMADS-box2 protein contains a MIKC domain, which only exists in plant MADS-box proteins. The SPOMA program predicted that the secondary structure of the protein was composed of alpha helices (56.00%), random coils (24.44%), extended strands (13.78%) and beta turns (5.78%). Based on the 3D structure of the 1TQE P chain, a tertiary structure model was predicted by the ESyPred 3D program. Molecular phylogenetic tree analysis revealed that the cloned gene belongs to the AP3-type family of genes as well as B-class genes, which might control the development of the petals and stamens. Finally, real-time quantitative PCR analysis indicated that the GoMADS-box2 gene could be expressed in all of the tested tissues, and the amount expressed in the flower was much higher than that of other three tissues; while the expression levels of the scape and leaf were nearly equal and the expression level in the bract was the lowest that was close to zero.

Keywords
Guzmania Ruiz & Pav; GoMADS-box2; real-time quantitative PCR
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