Evaluation of parameters affecting switchgrass tissue culture: toward a consolidated procedure for Agrobacterium-mediated transformation of switchgrass (Panicum virgatum)

Table 2 Tissue culture medium composition used in this protocol

Purpose	Medium name	Composition
Callus induction	CIM	NB₀ medium (pH 5.8) (3 mg l⁻¹ 2,4-d + 0.2 mg l⁻¹ BAP + 500 mg l⁻¹ l-proline)
Callus resting	CRM	MS medium (pH 5.8) (5 mg l⁻¹ 2,4-d + 1 mg l⁻¹ BAP + 2 g l⁻¹ l-proline + 200 mg l⁻¹ Timentin)
Callus selection	CSM	MS medium (pH 5.8) (5 mg l⁻¹ 2,4-d + 1 mg l⁻¹ BAP + 2 g l⁻¹ l-proline + 200 mg l⁻¹ Timentin) (50–100 mg l⁻¹ hygromycin B)
Regeneration	REG	MS medium (pH 5.8) (2 mg l⁻¹ BAP + 1 mg l⁻¹ IAA + 1 mg l⁻¹ kinetin + 1 mg l⁻¹ NAA + 500 mg l⁻¹ l-proline + 200 mg l⁻¹ Timentin) (20 mg l⁻¹ hygromycin B)
Rooting	RM	1/2 MS salts + 30 g l⁻¹ maltose (pH 5.8) (200 mg l⁻¹ Timentin + 50 mg l⁻¹ hygromycin B)

NB₀ medium, NB Basal Medium + 30 g l⁻¹ maltose + 300 mg l⁻¹ casein enzymatic hydrolysate (CH) + 6.5 g l⁻¹ agar MS medium, MS Basal medium with vitamins + 30 g l⁻¹ maltose + 6.5 g l⁻¹ agar or 3 g l⁻¹ phytagel (in REG)

ISSN: 1746-4811