Table 1 Comparative analysis of protoplast isolation protocol of different woody plants as per literature

Jasminum sambac and J. mesnyi

Callus

1.5% cellulase, 0.4% macerozyme, 0.8% pectinase

4 h

26–30 °C in dark, 50 rpm

21% sucrose,

100×g for 3 min at 4 °C

23.8 ± 4.3 × 10⁶

88

[32]

Leaves

4 h

5.6 ± 3.9 × 10⁶

89.6

Flowers

10 h

3.9 ± 5.0 × 10⁶

83

Stems

6 h

3.3 ± 0.5 × 10⁶

72.5

Holm Oak (Quercus ilex L.)

Leaf tissue

2% cellulase, 1% macerozyme

25 °C for 4 h in dark, 50 rpm

11% mannitol, 1000×g for 5 min at RT

61.5 ± 9.7 × 10⁶

-

[42]

Rubber tree

5–7 day old etiolated

leaves

1.5% cellulase, 0.6% macerozyme

4–5 h at 26–28 °C in dark, 60 rpm

900 rpm for 3 min at 4 °C

-

-

[43]

Peach

Mesocarp tissue (Fruits)

2% cellulase

and 0.2% macerozyme

30 °C

for 4 h

300×g for 10 min at 4 °C

-

-

[27]

Asian white birch

Young microculture

0.5% cellulase, 0.1% maceroenzyme

16-18 h, 50 rpm

350×g for 10 min

100 ± 0.3 × 10⁴

95

[44]

‘Boule de Neige’ Rhododendron

260.0 ± 0.3 × 10⁴

99

‘Gibraltar’ Azalea

2% cellulase, 0.5% maceroenzyme

4–6 h, 50 rpm

52 ± 0.6 × 10⁴

62

Apricot

Leaves from in vitro shoots

1% cellulase, 0.1% pectolyase,

1% hemicellulase

13–16 h

21% sucrose

75×g at RT

20.97 × 10⁶ (Plasmolysis of the leaves in a 13% sorbitol solution for 90 min)

83

[28]

Magnolia

Young leaves

3% cellulase, 0.8% macerozyme, 0.04% pectinase

6 h, 25 °C, 60 rpm

100×g for

10 min

1.89 × 10⁵

-

[45]

Ginkgo biloba L.

Leaves

2% cellulase, 0.2% pectolyase, 1.5% macerozyme

5 h in dark at 25 °C, 50 rpm

50×g for 3 min

5.39 × 10⁶

80.23

[31]

Albizia julibrissin

Leaves from in-vitro seedlings

1.5% cellulase, 1% pectolyase

6 h in dark at 25 ± 2 °C, 40 rpm

100×g for 5 min

6.31 × 10⁵

87

[46]

Callus

2% cellulase, 1% pectolyase

16 h in dark at 25 ± 2 °C, 40 rpm

5.53 × 10⁵

85

Platycladus orientalis

Young and fresh scale leaves

1.5% cellulase, 0.4% macerozyme, 0.4% pectolyase, 1.0% ligninase

16 h in dark at 25 °C, 40 rpm

-

9.60 × 10³

52.4

[47]

Areca catechu

Leaf peels

2% cellulase, 0.5% macerozyme

12 h dark at 25 ℃, 40 rpm

100×g for 3 min

2.5 × 10⁷

86.6

[33]

Camellia oleifera

Leaves

1.5% cellulase, 0.5% macerozyme, 0.25% Snailase

28 ℃ in dark, 40 rpm

15×g for 4 min

3.5 × 10⁷

90.9%

[40]

C. sinensis var. sinensis cv. ‘shuchazao’

Leaf

1.5% cellulase, 0.5% macerozyme, 0.7% snailase

25 °C for 4 h in the dark

100×g for 2 min with a swinging bucket rotor

3.5–6.6 × 10⁶

73-82%

[39]

Camellia sinensis (L.) O. Kuntze

Cultivars used:

Zijuan, Shuchazao, Huang shanbaicha and Huangkui

Tender leaves

1.5% cellulase, 0.4–0.6% macerozyme

10 h

65% iodixanol

200×g for 3 min

3.27 × 10⁶

92.94

[38]

Mature leaves

1.48 × 10⁶

83.23

Unlignified branches

1.20 × 10⁶

80.97

3.20 × 10⁶

89

Roots

Camellia sinensis (L.) O. Kuntze

Cultivars used:

Him Sphurti (China type), TV 23 (Cambod type), Upasi 9 (Assam type), and Kangra Asha (China type)

Tissue cultured

0.6% macerozyme, 3% cellulase, 1% hemicellulase

12 h in dark at 28 ºC, 60 rpm

25% sucrose 100×g for 5 min with a swinging bucket rotor

4.5 × 10⁷

95%

This study

Field grown

1.5 × 10⁷

80%

Hydroponically grown

3.9 × 10⁷

89%