Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (8): 1505-1512.doi: 10.3864/j.issn.0578-1752.2012.08.006

• PLANT PROTECTION • Previous Articles     Next Articles

Cloning and Analysis of StLTPa7 from Solanum torvum

 XIE  Chao, YANG  Qing, SHI  Ce, JUE  Deng-Wei, ZHU  Yan-Ping, LIU  Shui-Ping   

  1. 1.南京农业大学生命科学学院，南京 210095
  • Received:2011-06-20 Online:2012-04-15 Published:2011-10-31

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Abstract: 【Objective】The objective of this study is to clone non-specific lipid transfer protein gene StLTPa7 from Solanum torvum and to analyze the function of the gene. 【Method】 Primers were designed according to homologous cloning method. In order to clone StLTPa7, the gene cDNA was amplified by RT-PCR. Over-expression transgenic tobacco plants were generated via Agrobacterium-mediated transformation. Analysis on the inhibitory activity of proteins extracted from transgenic tobacco plants was done using mycelia growth rate method.【Result】StLTPa7 cDNA contained an ORF of 345 bp long and encoded a putative protein of 114 amino acids with a molecular weight of 11.42 kD and a theoretical pI of 9.01. StLTPa7 was induced to express by both salicylic acid (SA) and Verticillium dahliae and the level of transcript was the highest 24 to 48 h after treatment. To analyze the resistance of StLTPa7 to V. dahliae, the plants over-expressing StLTPa7 were generated by Agrobacterium-mediated transformation and 4 transgenic lines were identified by PCR. Quantitative RT-PCR indicated that the gene over-expressed in transgenic lines L5 and L7. Anti-fungal assay revealed that the inhibitory rate of the proteins extracted from the transgenic line L5 to V. dahliae was 2.5 times compared to the control.【Conclusion】StLTPa7 exhibits associated to inhibitory effect of V. dahliae growth, suggesting that it may be involved in plant defense against V. dahliae.

Key words: wild eggplant, StLTPa7, gene cloning, genetic transformation, function analysis

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