Studies on Construction of Regeneration System and Genetic Transformation of Puccinellia chinampoensis
2. Asian Natural Environmental Science Center(ANESC), University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 188-0002, Japan
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Bioscience Methods, 2011, Vol. 2, No. 5 doi: 10.5376/bm.2011.02.0005
Received: 02 Jun., 2011 Accepted: 20 Jun., 2011 Published: 23 Jun., 2011
Wang et al., 2011, Studies on Construction of Regeneration System and Genetic Transformation of Puccinellia chinampoensis, Bioscience Methods, doi:10.5376/bm.2011.02.0005
The regeneration system was established and the genetic transformation was processed, using the mature seeds of Puccinellia chinampoensis as explants. The results showed that the callus induction rate could reach to 39.75% after 19days of induction with the altered mediumâ… (adding 4 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine) (pH 5.8~6.0). When using the modified medium S (adding 2 mg/L 2,4-D, 1 mg/L ABA, 500 mg/L proline and 600 mg/L caseinhydrolysate) (pH 5.8~6.0) as the subculture medium, we can get much more embryogenic callus. The differentiation rate could reach to 53.73% using the differentiation medium (MS as the basic medium, adding 0.4 mg/L ABA, 0.04 mg/L IAA, 500 mg/L proline and 600 mg/L caseinhydrolysate) (pH 5.8~6.0). On the genetic transformation, the transformation efficiency was best when the infect time lasted 30 min, making using of the Agrobacterium tumefaciens mediated pBI121-GUS transformation when OD600 reached to 0.8~1.0.
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