Stable Transformation in Lotus japonicus

Materials and Reagents

1. Germination plate (1% agar in sterilized water)
   
2. B5 salt (Wako Pure Chemical Industries, catalog number: 399-00621 )
   
3. Gamborg’s vitamin solution
   
4. BAP (Wako Pure Chemical Industries, catalog number: 020-07621 )
   
5. 1-Naphthaleneacetic acid (NAA)
   
6. Acetosyringone
   
7. MES
8. (NH₄)₂SO₄
   
9. Phytagel (Sigma-Aldrich, catalog number: P8169 )
   
10. Meropen (Dainippon Sumitomo Pharma)
    
11. Hygromycin B
    
12. LB medium (nacalai tesque, catalog number: 20066-95 )
13. Co-cultivation medium (see Recipes)
    
14. YMB medium (see Recipes)
    
15. Callus medium (see Recipes)
    
16. Shoot elongation medium (see Recipes)
    
17. Root induction medium (see Recipes)
    
18. Root elongation medium (see Recipes)

Equipment

1. Clean bench
   
2. L. japonicus growth facility (we use several types of Biotrons such as LH-410S purchased from NIPPON MEDICAL & CHEMICAL INSTRUMENTS CO., LTD.)
   
3. Surgical knife
4. Sterilized dish ( 9 cm in diameter x 2 cm)
5. Sterilized magenta box
6. Sterilized filter paper (7 cm in diameter)
7. Sterilized filter paper (6 x 6 cm)
8. Vermiculite (Hakugen)
   

Procedure