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Table 1 Additives to media used for culture initiation, transformation and plant regeneration in three Miscanthus species

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 36.4 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 100 mg timentin

RCM-2: CIM-2 plus 300 mg cefotaxime

Callus selection media (CSM)

CSM-1: CIM-1 with paromomycin (200 mg Msac, 100 mg Msin); CSM-1 T: recipe above plus 100 mg timentin

CSM-2: CIM-2 plus 300 mg cefotaxime and 100 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

  1. The media sequence when starting with immature inflorescences. Plant growth regulators and non-standard additives included with amounts given per liter; further details provided in text. Msa and Msi media designations followed by -1; Mxg media followed by -2
  2. Msa M. sacchariflorus, Msi M. sinensis, Mxg M. x giganteus
  3. 1Reference [61]
  4. 2Sugar alcohols based on [19]
  5. 3Reference [26]