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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (10): 1801-1808.doi: 10.3724/SP.J.1006.2011.01801

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

Molecular Cloning and Expression Analysis of C4 Phosphoenolpyruvate Carboxylase Gene from A. hypochondriacus L.

FENG Rui-Yun1,2,BAI Yun-Feng2,**,LI Ping3,ZHANG Wei-Feng2,*,WANG Yuan-Yuan1,YANG Wu-De1,*   

  1. 1 College of Agronomy, Shanxi Agricultural University, Taigu 030801, China; 2 Institute of Crop Sciences, Shanxi Academy of Agricultural Sciences, Taiyuan 030032, China; 3 Bio-engineering School of Shanxi University, Taiyuan 030006, China
  • Received:2011-03-23 Revised:2011-06-25 Online:2011-10-12 Published:2011-07-28
  • Contact: 张维锋, E-mail: zhwfen@sina.com; 杨武德, E-mail: sxauywd@126.com E-mail:byfok@126.com

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Abstract: Phosphoenolpyruvate carboxylase (PEPC) plays a crucial role in primarily fixing atmospheric CO2 in C4 and CAM plants. This paper aimed at cloning both cDNA and genomic DNA of PEPC gene from Amarnthus hypochondriacus in order to provide candidate genes for C4 genetic engineering. The cDNA was cloned by RT-PCR, and the corresponding gDNA was cloned by LA-PCR. The sequence characteristics were analyzed using bioinformatics softwares including Blast, ClustalX (1.8), DNAMAN, PlantCARE. The gene expression pattern was analyzed by using semi-quantitative RT-PCR. The results indicated that the cDNA with the complete coding region was 2 895 bp in length which encoded 964 putative amino acids. The amino acid at site 774 was the serine that is an invariant residue in all C4 phosphoenolpyruvate carboxylases regardless of their taxonomic. The corresponding gDNA was 6 785 bp in length which harbored ten exoned and nine introns. The longest intron was the first intron with 1 662 bp in length. A computer scan disclosed that the first intron harbored 15 light-induced elements, nine hormone-induced elements, 29 TAAT motifs and 72 TATA motifs.In comparisons of PEPC genes between different species the exons are usually conserved but the introns have often diverged in both nucleotide sequences and lengths. The GC ratios of exons are positively correlated with those of introns in PEPC genes from different species. And the GC ratios of PEPC gene in dicotyledon A. hypochondriacus, potato, A. thaliana and ice plant are lower than those in monocotyledon maize, rice and wheat. The semi-quantitative RT-PCR analysis revealed that the cloned PEPC gene was light-inducible leaf-specifically expressed and classified as C4-type sub-family groups.

Key words: Amarnthus hypochondriacus, C4 photosynthetic type, PEPC, gDNA, Leaf-specific, Light-inducible expression

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