Research on Transformation of DHN Gene from Ammopiptanthus mongolicus into Alfalfa(Medicago sativa L. )

doi:10.3969/j.issn.1000-7091.2012.03.019

Abstract

Abstract: In order to obtain transgenic plants of resistant drought，by comparative analysis on the different explant tissues，concentrations of Agrobacterium and kanamycin，time of inoculation and coculture，etc，an Agrobacterium-mediated transformation for an alfalfa variety Zhongmu No. 2 was established. On the basis of the transformation，?dehydrin of Ammopiptanthus mongolicus was transformated into genome of Zhongmu No. 2 by A. tumefaciens-mediated. The results showed that cotyledon and hypocotyl was the best explant for callus induction and transformation.?The optimal kanamycin concentration for selection of resistant callus and transgenic plant was 25 mg/L in the selection medium. The explants and bacteria was cocultured for 5 d. One hundred and twenty-six independent resistant?plants were obtained，30 plants were confirmed to be transgenic by PCR amplification of AmDHN gene and 35S?gene.

Key words: Alfalfa, Genetic transformation, Dehydrin

CLC Number:

S541.9

NIE Li-zhen, GUO Jiu-feng, SUN Jie, SUN Guo-qin, LIU Yong-zhi. Research on Transformation of DHN Gene from Ammopiptanthus mongolicus into Alfalfa(Medicago sativa L. )[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(3): 96-101. doi: 10.3969/j.issn.1000-7091.2012.03.019.

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