Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (4): 815-822.doi: 10.3864/j.issn.0578-1752.2012.04.024

• RESEARCH NOTES • Previous Articles    

Cloning and Analysis of Three Ethylene Response Factor Genes in Alfalfa（Medicago sativa L.）

 CHEN  Ting-Ting, YANG  Qing-Chuan, KANG  Jun-Mei, DING  Wang, ZHANG  Tie-Jun, ZHANG  Xin-Quan   

  1. 1.中国农业科学院北京畜牧兽医研究所，北京 100193
    2.四川农业大学动物科技学院草业科学系，四川雅安 625014
  • Received:2011-09-15 Online:2012-02-15 Published:2011-11-16

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Abstract: 【Objective】 Cloning and characterization of three ethylene response factor genes in alfalfa to provide a foundation for further study of their function.【Method】Three ethylene response factor genes, MsERF5, MsERF8, and MsERF11 were isolated from alfalfa by PCR, and their expression patterns under different treatments and in different tissues were analyzed using semi-quantitative RT-PCR. The expression vectors of these three genes, was constructed, and then were introduced into tobacco by Agrobacterium mediated transformation system and the salt-tolerance of trangenic plants was analyzed.【Result】 MsERF5, MsERF8, and MsERF11 had no intron. The expression patterns of these three genes were significantly different in different tissues, the expression level of MsERF5 in roots, leaves and flower buds was higher than in other tissues. The expression of MsERF8 was higher in roots and leaves, and the expression of MsERF11 in leaves was higher than in other tissues, the expression of MsERF5, MsERF8, and MsERF11 were both induced by salt, drought, Al3+ and diverse hormones. The expression vectors of these three genes were constructed, and they were introduced into tobacco by Agrobacterium mediated transformation system. The calluses transformed MsERF5, MsERF8, and MsERF11 were changed into adventitious bud differ from the non-transformed calluses. The chlorophyll and soluble sugars contents of the transgenic plants were significantly higher than wild-type plants. The electrolyte leakage was significantly lower than wild-type plants. 【Conclusion】MsERF5, MsERF8, and MsERF11 had no intron, the expression patterns of these three genes were significantly different in different tissues, and both of them could be induced by abiotic stresses and diverse hormones treatments. All of them could change the calluses into adventitious buds on the saline medium and produce transgenic plants, and the salt-tolerance of transgenic plants was higher than wild-type plants.      

Key words: Medicago sativa, ethylene response factor, expression analysis, tobacco transformation, salt stress

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