From: Transformation and gene editing in the bioenergy grass Miscanthus
Media | Msa & Msi | Mxg |
---|---|---|
Callus induction media (CIM) | CIM-1: 2 mg 2,4-D + 0.25 mg zeatin | CIM-2: 3 mg 2,4-D + 0.1 mg BA1 |
Callus maintenance media (CMM) | CMM-1: 2 mg 2,4-D + 0.25 mg zeatin; 3.64 g mannitol and sorbitol | same as CIM-2 |
Callus osmotic medium (COM); biolistics | COM-1: CIM-1 with 36.4Â g mannitol and sorbitol2 | N/A |
Coculture media (CCM); A. tumefaciens | CCM-1: 2 mg 2,4-D + 0.25 mg zeatin; 0.1X MS salts, 0.1X B5 vitamins, 30 g maltose, 10 g glucose, 100 mg L-cysteine, pH 5.4; 39.24 mg acetosyringone (200 μM) | CCM-2: CIM-2 plus 58.86 mg acetosyringone (300 μM) |
Recovery media (RCM); A. tumefaciens | RCM-1: CIM-1 plus 100Â mg timentin | RCM-2: CIM-2 plus 300Â mg cefotaxime |
Callus selection media (CSM) | CSM-1: CIM-1 with paromomycin (200Â mg Msac, 100Â mg Msin); CSM-1Â T: recipe above plus 100Â mg timentin | CSM-2: CIM-2 plus 300Â mg cefotaxime and 100Â mg paromomycin |
Regeneration selection media (RSM); no selection = RM | RSM-1: 2 mg BA + 0.1 mg NAA; 20 g sucrose, 10 g maltose3; paromomycin (200 mg Msac, 100 mg Msin); RSM-1 T: recipe above plus 100 mg timentin | RSM-2: 5 mg BA + 0.24 mg NAA; 20 g sucrose, no MgCl2 or L-proline1; 300 mg cefotaxime and 50 mg paromomycin |
Rooting selection media (RtSM); no selection = RtM | RtSM-1: 0.25 mg IBA; 0.5X MS salts, 1X B5 vitamins, 15 g sucrose, 0.1 mg myo-inositol; 100 mg paromomycin | RtM-2: 0.5X for MS salts, vitamins and sucrose; pH 5.8; no MgCl2 or L-proline |