Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (9): 1685-1695.doi: 10.3864/j.issn.0578-1752.2012.09.003


Cloning and Polymorphism of 2-Methyl-6-Phytyl-1, 4-Benzoquinol Methyltransferase Gene (VTE3) in Arachis

 GUO  An-Qiang, WAN  Yong-Shan, LIU  Feng-Zhen   

  1. 山东农业大学农学院/作物生物学国家重点实验室/山东省作物生物学重点实验室,山东泰安 271018
  • Received:2011-04-08 Online:2012-05-01 Published:2012-04-16

Abstract: 【Objective】 The objective of this study is to isolate VTE3 (encoding MPBQ MT) in peanut, and to reveal the molecular characteristics and investigate polymorphism of the genes.【Method】A putative VTE3 of peanut was obtained via EST sequence splicing. cDNAs of VTE3 were cloned from cultivated varieties of A. hypogaea by RT-PCR, genomic DNAs of VTE3 were cloned from cultivated varieties and two wild species in Arachis by PCR. Polymorphism analysis of VTE3 from cultivated varieties and two wild species were performed, and phylogenetic tree was constructed by PHYLIP software. 【Result】 Two cDNA sequences of VTE3 (designated as rVTE3-1 andrVTE3-2) were isolated from each of the three cultivated varieties (Arachis hypogaea L.). rVTE3-1 and rVTE3-2 both had a DNA sequence of 1 059 bp in length, with the homology of 97.8% between the two sequences, and eight single-nucleotide polymorphisms (SNPs) exist within the sequences. The two cDNA sequences both encode 351 amino acids, and the homology of the two proteins was 98.6% with five amino acid differences. The two DNA sequences of VTE3 from each cultivar were designated as gVTE3-1 and gVTE3-2. The sequences of gVTE3-1 from the thirteen cultivars share a homology of 99.9% in nucleotide acid level and the thirteen sequences of gVTE3-2 are identical. gVTE3-1 of cultivated peanut samples from Fenghua 2 had a length of 2 710 bp, with three introns located at 44-163, 772-1 295 and 1 603-2 437 bp, and gVTE3-2 from Fenghua 2 had a length of 2 706 bp with three introns located at 44-169, 778-1 291 and 1 599-2 433 bp. Thirty-six SNPs and three variation sites of endonuclease recognition were identified between the intrones of the two sequences. Meanwhile, the VTE3 DNA (designated as gVTE3-A and gVTE3-B) was isolated from wild species A. duranensis (A-genome) and A. ipaensis (B-genome), respectively. The sequences of gVTE3-1 (Fenghua 2), gVTE3-2 (Fenghua 2), gVTE3-A and gVTE3-B were aligned and the phylogenetic tree was constructed. The tree demonstrated that FhgVTE3-1 and gVTE3-A clusters form a group and FhgVTE3-2 and gVTE3-B clusters form as an additional group. Sequence alignment analysis revealed that the homology of FhgVTE3-1 and gVTE3-A was 98.7% and the homology of FhgVTE3-2 and gVTE3-B was 100%. The amino acid homologies of MPBQ MT among different species shared higher similarities.【Conclusion】Full-length cDNA and DNA sequences in VTE3 of peanut were cloned. The present study demonstrated that abundant polymorphisms were observed in VTE3 from A-genome and B-genome of A. hypogaea, respectively. Additionally, polymorphisms were observed in the gVTE3-1 alleles of the thirteen cultivars and wild species of A. duranensis. However, no variation of gVTE3-2 was found in the gVTE3-2 alleles of the thirteen cultivars and wild species of A. ipaensis.

Key words: Arachis hypogaea L., vitamin E, 2-methyl-6-phytyl-1, 4-benzoquinone methyltransferase (MPBQ MT), gene cloning

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