Mungbean transformation

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  1. Prepare binary vector ex) pMDC 43
  2. Vector are inserted into Agrobacterium competent cell by heat shock method
  3. Spreading them on LB agar media with antibiotics for selecting cells (until colonies were visible)
  4. Seed planting in vitro, dark condition (during 2 days)
  5. Single colony picking into 2ml LB liquid media with antibiotics (over 16h incubate)
  6. Prep and confirm inserting DNA using PCR
  7. 20ul bacteria suspension + 20ml LB media with antibiotics (over 16h incubate)
  8. Harvest cells on 4000 RPM for 10 min
  9. Resuspending cells in liquid B5 plant culture media at a density of 109 cells per ml(OD560=1)
  10. Immerse plant(Primary leaves with petiole or Cotyledonary nodes with proximal halves of cotyledon or hypocotyl) with suspension and gently shaking
  11. Explants were blotted on sterile filter paper and co-cultivated on B5 basal medium for 3 days under dark conditions at 25±2℃
  12. Wash the explants 3~4 times with T.D.W and blot
  13. Cultivating on B5 medium containing 5 * 10-6 M BAP and 2.5 * 10-6 M each of 2,4-D and NAA(also antibiotics)
  14. The explants along with newly initiated callus were transferred every 2 weeks culture period for 8-10 weeks onto fresh medium


  1. Agrobacterium tumefaciens-mediated genetic transformation of mungbean (Vigna radiata L. wilczek) - a recalcitrant grain legume, Pawan et al, 2001, Plant Science