Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (23): 4584-4592.doi: 10.3864/j.issn.0578-1752.2016.23.011

• HORTICULTURE • Previous Articles     Next Articles

Isolation and Expression Analysis of a Vacuolar Glucose Transporter Gene MdVGT1 in Apple

XU Hai-feng, LIU Jing-xuan, WANG Yi-cheng, ZUO Wei-fang, QU Chang-zhi, WANG De-yun, ZHANG Jing, JIANG Sheng-hui, WANG Nan, CHEN Xue-sen   

  1. College of Horticulture Science and Engineering, Shandong Agricultural University/State Key Laboratory of Crop Biology, Tai’an 271018, Shandong
  • Received:2016-04-26 Online:2016-12-01 Published:2016-12-01

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Abstract: 【Objective】 In order to develop the theory and breeding technology of functional apple, the bioinformatics, the expression level and the function in the sugar metabolism several aspects of vacuolar glucose transporter gene MdVGT1 in ‘Hongcui No.1’ of Malus sieversii f. neidzwetzkyana F1 population were studied. 【Method】 The MdVGT1 in ‘Hongcui No.1’ was cloned and the bioinformation of it was analyzed. The expression level of MdVGT1 in different tissues and different development stages was studied by the qRT-PCR and the expression of MdVGT1 induced by glucose in tissue culture seedlings of ‘gala’ was also analyzed. Meanwhile, the interaction between MdVGT1 and MdTMT1 was verified by yeast two-hybrid system and the recombinant protein by the prokaryotic induction technology was obtained. 【Result】 The full length of MdVGT1 in ‘Hongcui No.1’ was cloned, and the gene was 1 506 bp which encoded 501 amino acids. It was predicted that the molecular mass of this protein was 53.16 kD, and pI was 5.92. Furthermore, it was inferred that the gene includes 14 exons and 13 introns, and is located on the chromosome 1 of the apple genome. A phylogenetic tree indicated that MdVGT1, AtVGT1, VvVGT1 and CsVGT1 are located in the same evolutionary branch. Analysis of functional domain showed that MdVGT1 contains 12 transmembrane regions. MdVGT1 has the higher expression level in flowers, leaves and young fruits of apples, and its expression has a significant correlation with the content of glucose during the development stage of fruits. The promoter of MdVGT1 contains several typical cis-acting elements, including defense responsive elements, sugar signaling responsive elements and phytohormone responsive elements, and the expression of MdVGT1 in tissue culture seedlings of ‘gala’ was significantly increased after glucose treatment a week later. The yeast two hybrid experiments showed that the MdVGT1 could interact with MdTMT1 in vitro. In addition, the recombinant protein of MdVGT1 was obtained by the prokaryotic induction technology. 【Conclusion】The vacuolar glucose transporter gene MdVGT1 in ‘Hongcui No.1’ apple was cloned, and it was found that it could interact with MdTMT1 to transport glucose into the vacuole membrane.

Key words: apple, vacuolar glucose transporter, VGT1, gene clone, yeast two hybrid, prokaryotic expression

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