Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (9): 1857-1866.doi: 10.3864/j.issn.0578-1752.2013.09.013

• HORTICULTURE • Previous Articles     Next Articles

Cloning, Sequence and Expression Analysis of NADP-Malic Enzyme Genes in Apple

 DONG  Qing-Long, WANG  Hai-Rong, AN  Miao, YU  Xian-Mei, WANG  Chang-Jun   

  1. Shandong Institute of Pomology, Tai’an 271000，Shandong
  • Received:2012-10-23 Online:2013-05-01 Published:2013-03-01

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Abstract: 【Objective】This study is aimed to characterize the genes of MdNADP-MEs involved in malic acid metabolism in Malus×domestica B. by gene cloning, sequence and expression of MdNADP-MEs in different tissues.【Method】 Three full-length cDNA sequences of MdNADP-ME1,2,3 were isolated by RT-PCR. The obtained cDNA sequences and the deduced amino acid sequences were analyzed with bioinformatics methods. qRT-PCR was used to assess the expression of MdNADP-MEs in different tissues.【Result】The sequencing results showed that three cDNAs (designated as MdNADP-ME1, 2 and 3; GenBank Accession No. JX971883, JX971884 and JX971885) were 1 512 bp, 1 782 bp and 1 926 bp. They contained an open reading frame (ORF) of 1 512 bp, 1 782 bp and 1 926 bp, respectively, and their ORFs encoded an protein with 503, 593 and 641 amino acids, respectively. Amino acid sequence and structure analysis indicated that MdNADP-MEs contained five conservative amino acid areas (motif I-V) and two functional structure domains: malic and NAD_bind_1_malic_enz. The result of phylogenetic analysis showed that MdNADP-ME1 and 2 belonged to cytosolic dicot NADP-ME (group I), MdNADP-ME3 belonged to plastidic dicot NADP-ME. qRT-PCR result showed that MdNADP-MEs were constitutively expressed in all examined tissues, but showed different expression levels.【Conclusion】MdNADP-ME1,2,3 belong to NADP-ME family, possess highly conserved structures, and show different expression patterns in different tissues of apple.

Key words: apple , malic enzyme , cloning , sequence analysis , expression analysis

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