Molecular Cloning and Characterization of the GoMADS-box2 Gene from Guzmania Ruiz & Pav
Flower Research and Development Center, Zhejiang Academy of Agricultural Sciences, No. 79 Tangwan Village, Beigan Sub-District, Xiaoshan District, Hangzhou, 311202, P.R.China
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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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)
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.
Guzmania Ruiz & Pav; GoMADS-box2; real-time quantitative PCR