Table 1. Use of Wus2 and Bbm to Enhance Transformation of Sorghum, Rice, and Sugarcane
SpeciesVectorDevelopmental Gene Expression CassettesMarker Expression CassettesNo. Explants TreatedNo. Txn EventsTxn Freq. (%)
Sorghum bicolor TX430pPHP32371Ubipro:Bbm:pinII + nospro:Wus2:pinIIUbipro:CYAN:pinII + Ubipro:moPAT:pinII393 immature embryos7218
Sorghum bicolor TX430pPHP32269NoneUbipro:moPAT-YFP: pinII + Ubipro:PMI:pinII376 immature embryos72
Oryza sativa (ssp indica IRV95)pPHP57324Ubipro:OsBbm:pinII + nospro:Wus2:pinIICaMV 35Spro:YFP-HYG:pinII300 calli13043
Oryza sativa (ssp indica IRV95)pPHP66801Ubipro:Bbm:pinII + nospro:Wus2:pinIICaMV 35Spro:YFP-HYG:pinII300 calli8027
Oryza sativa (ssp indica IRV95)pPHP48195NoneCaMV 35Spro:HYG:nos3′ + Ubipro:CYAN:pinII300 calli83
Oryza sativa (ssp indica IRV95)pPHP56800Ubipro:Bbm:pinII + nospro:Wus2:pinIIUbipro:CYAN:pinII + Ubipro:PMI:pinII300 calli4515
Oryza sativa (ssp indica IRV95)pPHP49754NoneUbipro:CYAN:pinII + Ubipro:PMI:pinII300 calli00
Saccharum officinarum (v. CP01-1372)pPHP54561Ubipro:Bbm:pinII + nospro:Wus2:pinIIUbipro:YFP:pinII + Ubipro:moPAT:pinII20 calli177885
Saccharum officinarum (v. CP01-1372)pPHP35648Ubipro:Bbm:pinII + nospro:Wus2:pinIIUbipro:CFP:pinII + Ubipro:moPAT:pinII40 calli109273
Saccharum officinarum (v. CP01-1372)pPHP24600NoneCaMV 35Spro:PAT:35S 3′ + Ubipro:DsRED:pinII48 calli12
  • 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.