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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (10): 1856-1863.doi: 0.3724/SP.J.1006.2012.01856

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

Cloning and Bioinformatic Analysis of γ-Tocopherol Methyltransferase Gene (γ-TMT) in Peanut

LI Shuan-Zhu,WAN Yong-Shan*,LIU Feng-Zhen*   

  1. State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, Shandong Agricultural University, Taian 271018, China
  • Received:2012-02-15 Revised:2012-06-10 Online:2012-10-12 Published:2012-07-27
  • Contact: 万勇善, E-mail: yswan@sdau.edu.cn, Tel: 0538-8241540; 刘风珍, E-mail: liufz@sdau.edu.cn, Tel: 0538-8241540

Abstract:

γ-tocopherol methyltransferase(γ-TMT) is one of the critical enzymes that determine the composition and activity of vitamin E in plant. γ-TMT catalyzes the biological reaction from γ-tocopherol to α-tocopherol, is the most bioactive component of vitamin E. In this study, DNA sequence of γ-TMT in peanut (designated as AhgTMT ) was cloned by using in silico cloning combined with PCR from six cultivars and four diploid wild species that belong to Arachis section. cDNA sequence of γ-TMT (designated as AhrTMT) was cloned from Fenghua 2. Nucleotide sequencesof AhgTMT are identical with AhrTMT, which means no intron in this gene. AhrTMT has a 1 059 bp open reading frame encoding a protein (AhTMT) of 352 amino acid residues with a molecular weight of 39.094 kD. The isoelectric points of AhTMT is 6.72 and its grand average of hydropathy is –0.12. Secondary structure prediction of AhTMT indicated that 55.40% of the protein sequence is alpha helixes, 10.51% beta turn and 34.09% random coil. AhTMT has a conserved SAM domain and is localized in chloroplasts. The similarity between AhTMT and γ-TMTs reported in other species is from 61.75% to 72.80%. Alignment analysis of AhgTMT nucleotide sequences indicated that the homology of AhgTMT from cultivars and those from A. ipaensis (BB), A. batizocoi (BB), A. dura nensis (AA) and A. kuhlmannii (AA) was 100%, 99.91%, 99.74%, and 95.63%, respectively.

Key words: Peanut (Arachis hypogaea L.), Vitamin E, &gamma, -tocopherol methyltransferase, Gene cloning, Sequence analysis

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