Optimized protocols for protoplast isolation, transfection, and regeneration in the Solanum genus for the CRISPR/Cas-mediated transgene-free genome editing

Yang, So Hee; Kim, Suk Weon; Lee, Sujin; Koo, Yeonjong

doi:10.1186/s13765-024-00870-1

Applied Biological Chemistry

Table 2 List of the protoplast regeneration media used in previous studies

From: Optimized protocols for protoplast isolation, transfection, and regeneration in the Solanum genus for the CRISPR/Cas-mediated transgene-free genome editing

Species	Protoplast culture medium	Callus Induction medium	Callus growth medium	Shoot Induction medium	Rooting Induction medium	Reference
Tomato (solanum lycopersicum)	Microcalli proliferation medium (3.62 g/L KM medium, 3% Sucrose, 9% Mannitol), 2 mg/L 2,4-D, 0.5 mg/L BAP, 10% KM Vitamin Solution	4.4 g/L MS(Murashige & Skoog) medium containing vitamins, Myo-Inositol 60 g/L, 2 mg/L 2.4-D, 0.5 mg/L 6-BAP, 30 g/L Sucrose, 8 g/L Plant Agar	4.4 g/L MS(Murashige & Skoog) medium containing vitamins, 2 mg/L 2.4-D, 0.5 mg/L 6-BAP, 30 g/L Sucrose, 8 g/L Plant Agar	4.4 g/L MS(Murashige & Skoog) medium containing vitamins, 0.5 mg/L a-NAA, 2 mg/L 6-BAP, 30 g/L Sucrose, 30 g/L Plant Agar	4.4 g/L MS(Murashige & Skoog) medium containing vitamins, 1 mg/L IBA, 30 g/L Sucrose, 8 g/L Plant Agar
Tomato (Solanum peruvianum)	Microcalli not transferred	None	None	4.405 g/L MS medium with vitamins, 30 g/L sucrose, 6 g/L phyto agar, e 0.1 mg/L IAA and 0.75 mg/L zeatin	None	Lin et al. [10]
Tomato (Solanum esculentum)	MS salts without NH₄NO₃, 1900 mg/L KNO₃, 1000 mg/L myo-Inositol, 20 mg/L Casein hydrolysate, 2 mg/L Thiamine-HCl, 2 mg/L Giycine, 0.5 mg/L Nicotinic acid, 1 mg/L 2.4-D, 0.5 mg/L IAA, 0.5 mg/L 6-BAP, 10 g/L Sucrose, 80 g/L Mannitol, 5000 mg/L Glucose	MS salts without NH₄NO₃, 1900 mg/L KNO₃, 1000 mg/L myo-Inositol, 20 mg/L Casein hydrolysate, 2 mg/L Thiamine-HCl, 2 mg/L Giycine, 0.5 mg/L Nicotinic acid, 1 mg/L 2.4-D, 0.5 mg/L IAA, 0.5 mg/L 6-BAP, 10 g/L Sucrose, 80 g/L Mannitol, 5000 mg/L Glucose	MS(Murashige & Skoog) medium containing vitamins, 1 mg/L IAA, NAA, 0.4% Bacto Agar,	MS(Murashige & Skoog) medium containing vitamins, 2 mg/L Zeatin, 30 g/L Sucrose	MS(Murashige & Skoog) medium containing vitamins, 10 g/L sucrose, 7 g/L Bacto agar	Randall et al. [39]
Potato (Solanum tuberosum)	Microcalli not transferred	MS(Murashige & Skoog) medium containing vitamins, 20 g/L Sucrose, 0.05 mg/L 6-BAP, 10 g/L Bacto agar	Macro A and micro A stocks, iron elements A, carbohydrate A, vitamins A, 1 g/L BSA, 1.0 mg/L NAA, 0.4 mg/L BAP, and other organics,	30 g/L sucrose, 0.01 mg/L NAA, 2.0 mg/L zeatin, 0.1 mg/L Gibberellic acid (GA3), 4.0 g/L plant agar, 10 g/L sucrose, 0.01 mg/L NAA, 2.0 mg/L zeatin, 0.1 mg/L GA3, 2.5 g/L gelrite	None	Moon et al. [67]
Eggplant (Solanum melongena L.)	KM medium, 0.5 mg/L Zeatin, 0.2 mg/L 2.4-D, 1 mg/L NAA, 0.05% MES, 6.3% Glucose	MS medium, 7 g/L Agar, 20 g/L Sucrose, vitamin, 1–4 mg/L 6-BAP, 0.2 mg/L IAA, 1–4 mg/L Zeatikn	MS medium, 7 g/L Agar, 20 g/L Sucrose, vitamin, 1–4 mg/L 6-BAP, 0.2 mg/L IAA, 2 mg/L Zeatikn, 0.5 mg/L P, 0.5 mg/L NAA	None	None	Sihachakr et al. [68]
Eggplant (Solanum melongena L ,Solanum insanum L.)	1/2 MS salts (200 mg/L NH₄NO₃), 1 mg/L NAA, 1 mg/L 2.4-D, 1 mg/L Kinetin	MS salts (200 mg/L NH4NO3), 1 mg/L NAA, 1 mg/L 2.4-D, 0.2 M mannitol, 3% Sucrose, 0.8% Agar	MS salts (200 mg/L NH4NO3), 1 mg/L NAA, 1 mg/L 2.4-D, 0.2 M mannitol, 3% Sucrose, 0.8% Agar	1/3 MS, 1% Sucrose	MS medium, 1 mg/L NAA	Nishio et al. [37]
Eggplant (Solanum melongena L.)	KM medium, 0.5 mg/L Zeatin, 0.2 mg/L 2.4-D, 1 mg/L NAA	KM medium, 0.5 mg/L Zeatin, 0.2 mg/L 2.4-D, 1 mg/L NAA	KM medium, 2 mg/L 6-BAP, 0.1 mg/L 2.4-D	MS solid medium, 20 g/L Sucrose, vitamins, 2 mg/L Zeatin, 0.1 mg/L IAA	None	Fournier et al. [20]

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