Table 3 Summary of Agrobacterium-mediated transformation protocols for different fungal species
From: Methods for genetic transformation of filamentous fungi
Fungus | Agrobacterium | Starting material | The number of fungal cells | Pre-culturing of Agrobacterium | The conditions of co-culturing | Name of vector (size) | Selective marker | Transformation efficiency | References |
|---|---|---|---|---|---|---|---|---|---|
Aspergillus awamori | A. tumefaciens LBA1100 | Conidiospores | 1 × 106 | Incubate at 28 °C 4–5 h with 0.2 M AS and 100 rpm to OD600 nm of 0.8 | Incubate the plates for 3 days at 22.5 °C | NA | hygB | 200–250 transformants per 106 conidiospores | [45] |
Aspergillus fumigatus B-5233 | A. tumefaciens EHA105 | Conidia | 1 × 107 | Further incubation at 28 °C for 24 h to a ratio of 1:10 (conidia to bacteria) | Incubated at 37 °C for 3 days | pDHt/hph (NA) | hygB | 100 transformants per 107 conidia | [46] |
Trichoderma reesei QM 9414 | A. tumefaciens AGL-1 | Protoplasts and conidia | 1 × 107 | Further incubation for 6 h at 28 °C to OD600 nm of 0.15 | Incubation at 25 °C for 48 h | pBI-hph (15846 bp) | hygB | Start from protoplasts: 2000 to 9000 transformants per 107 protoplasts. Start from conidia: 200–500 transformants per 107 condia | [47] |
Mucor circinelloides ATCC1216b | A. tumefaciens AGL-1 | Spores | 1 × 105 spores per mL and | Incubate for 4 h at 20 °C to OD600 nm of 0.6 | Incubation at 15 °C for 6 days | pTiBo542 (NA) | hygB | 2000–9000 transformants per 107 sporangiospores | [48] |
Aspergillus giganteus IfGB15/0903 | A. tumefaciens LBA1100 | Germinated conidia | 5 × 107 | Incubate for 6 h at 30 °C with 200 μM AS to OD 600 nm of 2.5 | Incubation at 24 °C for 3 days | pUR5750 (NA) | hygB | 500 to 7900 transformants per 108 conidia | [23] |