Molecular characterization and expression analysis of phosphate transporter gene <italic>TaPT2-1</italic> in wheat (<italic>Triticum aestivum</italic> L.)

doi:10.1007/s11703-011-1101-7

Front Agric Chin

2011, Vol. 5

Issue (3) : 274-283 https://doi.org/10.1007/s11703-011-1101-7

RESEARCH ARTICLE

Molecular characterization and expression analysis of phosphate transporter gene TaPT2-1 in wheat (Triticum aestivum L.)

Xirong CUI¹, Yongsheng ZHANG¹, Fanghua ZHAO¹, Chengjin GUO¹, Juntao GU², Wenjing LU², Xiaojuan LI², Kai XIAO¹(

)

1. College of Agronomy, Agricultural University of Hebei, Baoding 071001, China; 2. College of Life Sciences, Agricultural University of Hebei, Baoding 071001, China

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Abstract

A transcript-derived fragment (TDF) showing up-regulated expression under low Pi stress and being identical to an uncharacterized phosphate transporter gene TaPT2-1 was cloned in wheat. TaPT2-1 was 2075 bp in length and encoded a 568-aa polypeptide. Transmembrane prediction analysis suggested that TaPT2-1 had 13 conserved transmembrane domains. TaPT2-1 shared much higher similarities to other four homologs from Arabidopsis thaliana, Solanum tuberosum, Capsicum frutescens, and Solanum melongena. The expression of TaPT2-1 was root specific and low Pi inducible, suggesting that it plays roles in roots and is involved in the Pi acquisition under Pi-starved condition. The promoter region of TaPT2-1 was cloned based on genome walk analysis. Several types of cis-regulatory elements, such as low Pi responding and tissue specific, were identified in TaPT2-1 promoter. The transgenic tobacco plants with the integrated TaPT2-1 promoter GUS were generated, and GUS histochemical staining analysis in the roots and leaves of the transgenic plants was performed. The results of GUS staining in roots and leaves under various Pi supply conditions were in accordance with the TaPT2-1 transcripts detected based on RT-PCR analysis. Taken together, the distinct expression of low Pi-induced and root-specific TaPT2-1 suggested that it could be used as the potential gene resource on generation of elite crop germplasms with high Pi use efficiency in the future.

Keywords wheat (Triticum aestivum L.), phosphate transporter, expression cis-regulatory element

Corresponding Author(s): XIAO Kai,Email:xiaokai@hebau.edu.cn

Issue Date: 05 September 2011

Cite this article:

Xirong CUI,Yongsheng ZHANG,Fanghua ZHAO, et al. Molecular characterization and expression analysis of phosphate transporter gene TaPT2-1 in wheat (Triticum aestivum L.)[J]. Front Agric Chin, 2011, 5(3): 274-283.

URL:

https://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1101-7
https://academic.hep.com.cn/fag/EN/Y2011/V5/I3/274

Fig.1 Identification of a TDF responding to the starved Pi stress. A: A TDF band in a polyacrylamide gel by an arrow responds to starved Pi condition of 24 and 48 h. B: The sequence of the TDF. The restriction digestion site of I and I, two restriction endonucleases used for digestion of the cDNAs of roots of CK, 24 h, and 48 h of starved Pi treatment, are shaded.

Fig.2 The cDNA sequence and the corresponding translated polypeptide of .

Fig.3 The diagram of transmembrane domains in TaPT2-1.

Fig.4 Phylogenetic analysis of and its homologs in plant species.

Fig.5 Expression patterns of in roots and leaves under various Pi supply conditions.

Fig.6 Cloning of promoter region based on genome walking. SP1, SP2, and SP3 are the three rounds of specific PCR products of promoter.

Fig.7 The promoter sequence. The translation start codon (ATG) of was boxed. The -regulatory elements PIBS and PHO-like putatively involved in regulation of gene expression to be low Pi responding are shaded and underlined, respectively. The primers used for the promoter integration into binary vector pCAMBIA3301 are underlined and labeled concomitantly with forward arrow and reverse arrow, respectively. The three specific primers, SP1, SP2, and SP3, used for cloning of promoter in genome walking, are shown by reverse arrow.

Tab.1 The putative important -regulatory elements identified in promoter

Fig.8 Molecular identification of the transgenic tobacco lines that were integrated promoter. A: The genome DNA extracted from CK and the transgenic tobacco lines. B: PCR results in which the genome DNA extracted from the CK and the transgenic tobacco lines was used as the template.

Fig.9 GUS staining analysis of the roots and leaves of control (transformed the empty binary vector) and representative transgenic plants. A: The histochemical staining of roots and leaves in which the empty binary plasmid integrated. B and C: The GUS activities of roots and leaves in which the reporter gene was driven by promoter under normal Pi supply condition (2 mM) and starved Pi condition (20 μM), respectively.

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