Table 1. Use of Wus2 and Bbm to Enhance Transformation of Sorghum, Rice, and Sugarcane.
| Species | Vector | Developmental Gene Expression Cassettes | Marker Expression Cassettes | No. Explants Treated | No. Txn Events | Txn Freq. (%) |
|---|---|---|---|---|---|---|
| Sorghum bicolor TX430 | pPHP32371 | Ubipro:Bbm:pinII + nospro:Wus2:pinII | Ubipro:CYAN:pinII + Ubipro:moPAT:pinII | 393 immature embryos | 72 | 18 |
| Sorghum bicolor TX430 | pPHP32269 | None | Ubipro:moPAT-YFP: pinII + Ubipro:PMI:pinII | 376 immature embryos | 7 | 2 |
| Oryza sativa (ssp indica IRV95) | pPHP57324 | Ubipro:OsBbm:pinII + nospro:Wus2:pinII | CaMV 35Spro:YFP-HYG:pinII | 300 calli | 130 | 43 |
| Oryza sativa (ssp indica IRV95) | pPHP66801 | Ubipro:Bbm:pinII + nospro:Wus2:pinII | CaMV 35Spro:YFP-HYG:pinII | 300 calli | 80 | 27 |
| Oryza sativa (ssp indica IRV95) | pPHP48195 | None | CaMV 35Spro:HYG:nos3′ + Ubipro:CYAN:pinII | 300 calli | 8 | 3 |
| Oryza sativa (ssp indica IRV95) | pPHP56800 | Ubipro:Bbm:pinII + nospro:Wus2:pinII | Ubipro:CYAN:pinII + Ubipro:PMI:pinII | 300 calli | 45 | 15 |
| Oryza sativa (ssp indica IRV95) | pPHP49754 | None | Ubipro:CYAN:pinII + Ubipro:PMI:pinII | 300 calli | 0 | 0 |
| Saccharum officinarum (v. CP01-1372) | pPHP54561 | Ubipro:Bbm:pinII + nospro:Wus2:pinII | Ubipro:YFP:pinII + Ubipro:moPAT:pinII | 20 calli | 177 | 885 |
| Saccharum officinarum (v. CP01-1372) | pPHP35648 | Ubipro:Bbm:pinII + nospro:Wus2:pinII | Ubipro:CFP:pinII + Ubipro:moPAT:pinII | 40 calli | 109 | 273 |
| Saccharum officinarum (v. CP01-1372) | pPHP24600 | None | CaMV 35Spro:PAT:35S 3′ + Ubipro:DsRED:pinII | 48 calli | 1 | 2 |
Transformation of sorghum immature embryos, indica rice established callus explants, and sugarcane established callus explants using Agrobacterium strain AGL1 in sugarcane and LBA4404 in rice and sorghum that delivered either (1) T-DNAs containing Ubipro:Bbm and nospro:Wus2 or (2) T-DNAs with marker genes but no Bbm or Wus2 (control vectors). Sorghum and rice transformation frequencies based on phenotype (both resistance to a selectable marker and fluorescence) and qPCR, while sugarcane frequencies were based on fluorescence. For rice transformation experiments, one experiment was performed with the rice ZmBbm ortholog OsBbm + the Zea mays Wus2 (pPHP57324). For all other experiments, the Zea mays Bbm and Wus2 genes were used. Transformation is abbreviated Txn in the column labels.