Table 5 Summary of electroporation-mediated transformation protocols for different fungal species
From: Methods for genetic transformation of filamentous fungi
Strain | Waveform | Instrument | Electroporation parameters | Raw material | Number of cell | Name and size of vectors | Amount of DNA | Selective markers | Transformation efficiency (per μg DNA) | References |
|---|---|---|---|---|---|---|---|---|---|---|
Rhizopus oryzae AS 3.819 | NA | Multiporator (Eppendorf) | 12 kV/cm | Protoplasts | 1 × 107 | pBS-hygro-ldhA (8533 bp) | 5 μg | hygB | 10.2 transformants | [36] |
R. oryzae AS 3.819 | NA | Multiporator (Eppendorf) | 15 kV/cm | Germinated spores | 1 × 107 | pBS-hygro-ldhA (8533 bp) | 5 μg | hygB | 8.8 transformants | [36] |
A. niger ATCC 20739 | Exponential decay waveform | Electro Gene Transfer Unit | 6 kV/cm and 3 ms time constant | Germinated spores | 1 × 107/0.4 mL | pXbal92 (5.7 kb) and pBXba2 (6.0 kb) | 1–10 μg | argB − | 1.2 transformants for integrative vector and 100 colonies for plasmid DNA. | [52] |
Penicillium urticae NRRL 2159A | Exponential decay waveform | Gene Pulser (Bio-Rad) Multiporatr (Eppendorf) | 12.5 kV/cm, 8.6 ms time constant and 25 μF | Germinated conidia | 8 × 106 | pCSN44(NA) | 1–5 μg | hygB | 1.8 × 103 transformants | [64] |
A. oryzae ATCC 14895 | Exponential decay waveform | Gene Pulser (Bio-Rad) Multiporatr (Eppendorf) | 11–12.5 kV/cm, 4.6–4.8 ms time constant and 25 μF | Germinated conidia | 2.5 × 106 | pBEN(NA) | 1–5 μg | BenomylR | 2.6 × 103 transformants | [64] |
Leptosphaeria maculans “Virulent” | Exponential decay waveform | Gene Pulser (Bio-Rad) Multiporatr (Eppendorf) | 12.5 kV/cm, 4.8 ms time constant and 25 μF | Germinated conidia | 1.2 × 106 | pCSN44(NA) | 1-5 μg | hygB | 1–6 × 102 transformants | [64] |
N. Crassa R-206A | Exponential decay waveform | Gene Pulser (Bio-Rad) Multiporatr (Eppendorf) | 12.5 kV/cm, 5 ms time constant and 25 μF | Germinated conidia | 3–6 × 106 | Bsqa(NA) | 1–5 μg | hygB | 5.7 × 103 transformants | [64] |