Agrobacterium tumefaciens-mediated transient transformation of Arabidopsis thaliana leaves.
Transient assays provide a convenient alternative to stable transformation. Compared to the generation of stably transformed plants, agroinfiltration is more rapid, and samples can be analyzed a few days after inoculation. Nevertheless, at difference of tobacco and other plant species, Arabidopsis thaliana remains recalcitrant to routine transient assays. In this chapter, we describe a transient expression assay using simple infiltration of intact Arabidopsis leaves with Agrobacterium tumefaciens carrying a plasmid expressing a reporter fluorescent protein. In this protocol, Agrobacterium aggressiveness was increased by a prolonged treatment in an induction medium deficient in nutrients and containing acetosyringone. Besides, Arabidopsis plants were cultivated in intermediate photoperiod (12 h light-12 h dark) to promote leaf growth.
Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA)-CCT-La Plata-CONICET, Universidad de La Plata, La Plata, Argentina.
Silvina
S
Mangano
Cintia Daniela
CD
Gonzalez
Silvana
S
Petruccelli
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D046508:Culture Techniques; D018274:Electroporation; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018515:Plant Leaves / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D014170:Transformation, Genetic; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein