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Bulletin of Botanical Research ›› 2009, Vol. 29 ›› Issue (6): 721-727.doi: 10.7525/j.issn.1673-5102.2009.06.014

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Establishment and Optimization of Agrobacterium-mediated Transformation of Suspension Embryogenic Callus of Alfalfa

SHEN Yu-Hua;LI Wang-Feng;JIN Tai-Cheng;CHANG Qing;YIN Dong-Xu;WANG De-Li;LIU Li-Xia*   

  1. (1.School of Life Sciences,Northeast Normal University,Changchun 130024) (2.Inner Mongolia Chifeng College,Chifeng 024000)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-11-20 Published:2009-11-20
  • Contact: LIU Li-Xia
  • Supported by:

Abstract: Genotype is a key restrictive factor for genetic transformation in alfalfa. A high frequency-regeneration potential alfalfa cultivar named Gongnong No.1 was screened from 7 cultivars by embryonic callus inducing test, and was successfully used in establishing a high efficiency genetic transformation system. Additionally, we analyzed several factors affecting the genetic transformation and optimized the suspension culture conditions, and established sonication-assisted Agrobacterium-mediated transformation system in alfalfa primarily. The suspension callus from alfalfa hypocotyls were taken as explants, as concentration was determined at 100 μmol·L-1, sonication treatment time was 8 s, kanamycin concentration was 30 mg·L-1 and co-culture time was 4 days. Conclusively, large numbers of transgenic plants were obtained by the transformation system. Molecular analysis confirmed that exogenous vacuolar Na+/H+ antiporter gene from Suaeda corniculata was integrated into the genome of alfalfa.

Key words: Alfalfa, suspension embryogenic callus, NHX1 gene, transformation

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