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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (01): 76-83.doi: 10.3724/SP.J.1006.2013.00076

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Isolation and Expression Analysis of Gm TIP1;1 in Soybean (Glycine max L.)

ZHANG Da-Yong1,HU Guo-Min3,YI Jin-Xin1,*,XU Ling1,Ali ZULFIQAR4,LIU Xiao-Qing1,YUAN Ling-Ling2,XU Zhao-Long1,2, HE Xiao-Lan1, HUANG Yi-Hong1,MA Hong-Xiang1   

  1. 1 Provincial Key Laboratory of Agrobiology, Institute of Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210094, China; 2 College of Agriculture, Nanjing Agricultural University, Nanjing 210095, China; 3College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, China; 4 Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad 38040, Pakistan
  • Received:2012-02-21 Revised:2012-09-05 Online:2013-01-12 Published:2012-11-14
  • Contact: 易金鑫, E-mail: yij@jaas.ac.cn, Tel: 025-84391105

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Abstract:

The full length open reading frame (ORF) of gene named Glyma03g34310.1 was amplified from the soybean root tissues by reverse transcriptase polymerase chain reaction (RT-PCR) method. Sequencing, Blast and homology analyses showed that the amino acids encoded by this gene had the higher similarity with the TIP1;1 from other species, so designated Gm TIP1;1 (GenBank accession number: AK285481), its ORF was 753 bp, encoded a polypeptide with 250 amino acids, and contained a 94 bp intron at the site of the 381th nucleotide complying with the ↓GT--AG↓ mode of splicing. Gm TIP1;1 had two copies in the soybean genome, with the other one of Glyma19g37000.1. Multiple ali MEGA5.05gnment using protein indicated that GmTIP1;1 contained six conserved transmembrane domains and two higher conserved NPA motifs. Phylogenetic tree analysis showed that Gm TIP1;1 was indeed grouped into the legumes clade and several different clades which belonged to the different plant coleus were regularly separated based on the TIP1;1 protein sequences, which implied that TIP1;1 sequence probably could be regard as a proof of plant taxonomy. Semi-quantity RT-PCR analysis demonstrated that the Gm TIP1;1 gene constitutively expressed in soybean organs including root, stem, leaf, flower and pod, and the expression levels were no obvious difference in different tissues at the different developmental stages, which implied Gm TIP1;1 gene plays important roles in plant growth. The expression of Gm TIP1;1 appeared a declined  trend at the different time points under the treatment of salt solution (200 mmol L–1 NaCl), although still showing the abundant transcript. In addition, the recombinant plasmid pYES2-Gm TIP1;1 was constructed by inserting the Gm TIP1;1 gene into the yeast expression vector pYES2. The recombinant plasmid pYES2-Gm TIP1;1 was transformed into yeast Saccharomyces cerevisiae INVScl, then treated with salt and drought stresses, respectively. The results showed that the survival rate of the recombinant yeast INVScl (pYES2-Gm TIP1;1) was higher than that of the control strain under the salinity condition, but no difference under the drought. These results indicated that the heterologous expression of Gm TIP1;1 could effectively improve the tolerance of yeast to salinity stress.

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