Table 1 Enzyme solution for protoplast isolation in previous studies

Tomato

(solanum lycopersicum)

Leaves

Sliced

2% cellulase R-10, 0.5% macerozyme R-10

2 mM Ca(NO₃)₂, 1 mM MgSO₄, 0.5 mM KH₂PO₄, 0.5 mM Na₂HPO₄, 0.5 mM KCl, 0.4 M sucrose-K3 solution, micronutrients

12 h, overnight, 27 °C, dark

575,000 ± 153,292.7 (86.91%)

Horvath [29]

Tomato

(solanum lycopersicum, Solanum pennellii, Solanum pimpinellifolium)

Leaves

Sliced

1.5% Cellulase RS, 0.75% Macerozyme R-10

0.6 M mannitol, 10 mM MES, 10 mM CaCl₂, 0.1% BSA

16 h, 25 ℃, dark,

1.0 × 10⁶ protoplasts/ml

Nicolia et al. [11], Liu et al. [3]

Tomato

(Solanum peruvianum)

Leaves, Stems

Sliced

1.5% Cellulase RS, 0.75% Macerozyme R-10

0.6 M mannitol, 10 mM MES, 10 mM CaCl₂, 0.1% BSA

4–5 h, dark, 60-80 rpm

4.8 × 10⁶

Lin et al. [16]

Tomato

(Solanum esculentum)

Roots

Sliced

5% Myrothecium cellulase, 0.02 M phosphate buffer

0.6 M Sucrose

2 h, 27 ℃

Not disclosed

Cocking et al. [34]

Leaves

Sliced

0.75% Cellulase, 0.1% macerozyme

0.3 M mannitol, 170 mg/ L KH₂PO₄, 440 mg/L CaCl₂·2H₂O, 1500 mg/L KNO₃, MgSO₄·7H₂O, 0.3 M sucrose, vitamins, 1.0% polyvinylpyrolidine, 5 uM MES

4–6 h, 28 ℃, 60 rpm

6.6 × 10⁶/g

Shahin [46]

Leaves

Sliced

3% Meicelase-P, 0.1% Macerase

13% mannitol, 9% Sorbitol, CPW salt

12–14 h, 27 °C, 45 rpm

3 × 10⁴/ml

Randall et al. [39]

Roots

Sliced

2% cellulase, 2% macerozyme, 0.1% pectolyase

500 mM sorbitol, 10 mM CaCl_2, 0.5% BSA, 5 mM DTT, 0.5% PVP, 5 mM MES

2 h, 27 ℃, 60 rpm

1 × 10⁵ Cell·ml⁻¹

Shin et al. [40]

Tomato

(Lycopersicon hirsutum,

Solanum esculentum)

Leaves

Sliced

0.6% cellulase, 0.1% pectinase

9% Mannitol, 1.0 mM NaCl, 0.2 mM KCl, 0.2 mM CaSO₄, 3 mM MES

8 h, dark

Not disclosed

Rueda et al. [42]

Potato

(Solanum tuberosum)

Tubers

Sliced

0.2% Pectolyase, 0.5% Rhozyme, 1% Cellulase R10

0.5% BSA, 10% mannitol

4–5 h, dark

7.5 × 10⁴ to 5 × 10⁵/g

Jones et al. [45, 58]

Tubers

Sliced

4% cellulose R-10, 0.8% macerozyme R-10, 1% hemicellulase

0.55 M mannitol, 2 mM CaCl2, 1 mM KH₂ PO₄, 10 mM MES,

1 mM MgCl_2, 50 mM Tris buffer

18–20 h, 29 ℃, dark, 180 rpm

1.6 × 10⁵ protoplasts/g fresh weigh

Laimbeer et al. [43]

Leaves

Sliced

1% Cellulase R-10, 0.5% Macerozyme R-10

0.5 M mannitol, 20 mM KCl, 10 mM CaCl₂, 20 mM MES, 15% sucrose, 0.1% BSA

6 h, dark, 40 rpm

1.6 × 10⁶ protoplasts/g fresh weigh

Moon et al. [67]

Leaves

Sliced

0.5% Cellulase R-10, 0.5% Macerase R-10

5 mM CaCl₂, 0.01 M MES, 0.5 M sucrose

15–16 h, 28–30 ℃,

2.4 × 10⁶ to 4.6 × 10⁶

Konovalova et al. [44]

Eggplant

(Solanum melongena L. and Solanum insanum L.)

Leaves

Sliced

0.067% Macerozyme R-10, 0.33% Meicelase

0.5 M mannitol, 1/2 MS salts (200 mg/L NH₄NO₃), 1% sucrose, 1/2 MS vitamins

16 h, 25 ℃

1 × 10⁴ per hypocotyl

NISHIO et al. [37]

Eggplant

(Solanum melongena L.)

Stems

Sliced

1.5% Cellulase R-10, 0.75% Macerozyme R-10

6 mM CaCl₂, 1 mM KH₂PO₄, 0.5 M Sucrose, 0.05% MES

3 h, 27 ℃, dark

4 to 5 × 10⁴ protoplasts per ml

Sihachakr et al. [68]

Leaves

Sliced

1.25% cellulose, 0.4% macerozyme

0.5 M mannitol, 10 mM CaCl₂, 20 mM MES, 1.5 mM MgCl₂, 20 mM KCl, 1.5 mM KH₂PO₄

3 h, 26 ℃

1.2 × 10⁷/g fresh weight (FW)

Wang et al. [48]

Leaves

Sliced

0.3% cellulase R10, 0.65% pectinase

0.3 M mannitol, 2% PVP, 0.2% BSA, 3 mM MES, 1/10 RA salts

Overnight, 25 ℃, dark

2 × 10⁵ ml⁻¹

Yu et al. [18]

Leaves

Sliced

1.5% Cellulase R-10, 0.5% Macerozyme R-10

6 mM CaCl2, 1 mM KH2PO4, 0.5 M Sucrose, 0.05% MES

Overnight, dark

5 × 10⁴ protoplasts per ml

Fournier et al. [20]