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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (02): 201-209.doi: 10.3724/SP.J.1006.2017.00201

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

Cloning, Molecular Characterization, and Functional Analysis of Wheat TaVIP1 Genes

ZHAO Pei1,**,TENG Li-Jie2,**,WANG Ke1,DU Li-Pu1,REN Xian2,SHE Mao-Yun3,*,YE Xing-Guo1,*   

  1. 1Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement, Beijing 100081; 2 College of Biology and Engineering, Beifang University of Nationalities, Yinchuan 750002, China; 3Crop Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230031, China
  • Received:2016-03-25 Revised:2016-09-18 Online:2017-02-12 Published:2016-09-28
  • Contact: 佘茂云, E-mail: ahxiaoshe@126.com; 叶兴国, E-mail: yexingguo@caas.cn
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31401380, 31401376).

Abstract:

Plant VIP1 (VirE2 interacting protein 1) is involved in the transportation of T-DNA in plant cells after Agrobacterium infection, affecting plant transformation efficiency. However, the function of TaVIP1 is unknown. Using in silico technique, a family of TaVIP1 genes were successfully cloned from common wheat in this study. The gene family encompasses four exons and shares high similarity among its members while only 50.7%–51.4% similarity to the AtVIP1 in Arabidopsis. Southern blotting assay showed that the TaVIP1 had three allelic copies in wheat genome. The three copies of TaVIP1 were further assigned to wheat chromosomes 4AL, 4BS, and 4DS according to Blastn in the IWGSC (International Wheat Genome Sequencing Consortium) database, and experimentally confirmed by PCR using specific primers to each TaVIP1 allele based on their DNA sequences and Langdon durum disomic substitution lines. Subcellular localization analysis showed that TaVIP1 proteins were located in cell membrane, cytoplasm and nucleus. Over-expression of TaVIP1 in tobacco obviously reduced Agrobacterium-mediated transformation efficiency. The amino acid sequence encoded by TaVIP1 only had less similarity to the corresponding amino acid sequence encoded by AtVIP1 in Arabidopsis or NtVIP1 in tobacco. This might be a reason of the low transformation efficiency of wheat mediated by Agrobacterium.

Key words: Wheat, TaVIP1, Tobacco, Agrobacterium-mediated transformation

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