Agrobacterium tumefaciens-mediated transformation of maize endosperm as a tool to study endosperm cell biology.
Developing maize (Zea mays) endosperms can be excised from the maternal tissues and undergo tissue/cell-type differentiation under in vitro conditions. We have developed a method to transform in vitro-grown endosperms using Agrobacterium tumefaciens and standard binary vectors. We show that both aleurone and starchy endosperm cells can be successfully transformed using a short cocultivation with A. tumefaciens cells. The highest transformation rates were obtained with the A. tumefaciens EHA101 strain and the pTF101.1 binary vector. The percentage of aleurone cells transformed following this method varied between 10% and 22% whereas up to the eighth layer of starchy endosperm cells underneath the aleurone layer showed transformed cells. Cultured endosperms undergo normal cell type (aleurone and starchy endosperm) differentiation and storage protein accumulation, making them suitable for cell biology and biochemical studies. In addition, transgenic cultured endosperms are able to express and accumulate epitope-tagged storage proteins that can be isolated for biochemical assays or used for immunolabeling techniques.
Reyes
Francisca C
FC
Department of Botany, University of Wisconsin, Madison, Wisconsin 53706, USA.
Sun
Beimeng
B
Guo
Hena
H
Gruis
Darren Fred
DF
Otegui
Marisa S
MS
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D056625:Endosperm / Q000166:cytology / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics; D010957:Plasmids; D055314:Seed Storage Proteins / Q000378:metabolism; D014170:Transformation, Genetic; D003313:Zea mays / Q000235:genetics*
D055314:Seed Storage Proteins; D049452:Green Fluorescent Proteins