Analysis of a LEA gene promoter via Agrobacterium-mediated transformation of the desiccation tolerant plant Lindernia brevidens.
An Agrobacterium tumefaciens-based transformation procedure was developed for the desiccation tolerant species Lindernia brevidens. Leaf explants were infected with A. tumefaciens strain GV3101 harbouring a binary vector that carried the hygromycin resistance gene and an eGFP reporter gene under the control of a native dehydration responsive LEA promoter (Lb2745pro). PCR analysis of the selected hygromycin-resistant plants revealed that the transformation rates were high (14/14) and seeds were obtained from 13/14 of the transgenic lines. A combination of RNA gel blot and microscopic analyses demonstrated that eGFP expression was induced upon dehydration and ABA treatment. Comparison with existing procedures used to transform the well studied resurrection plant and close relative, Craterostigma plantagineum, revealed that the transformation process is both rapid and leads to the production of viable seed thus making L. brevidens a candidate species for functional genomics approaches to determine the genetic basis of desiccation tolerance.
Smith-Espinoza
Claudia
C
Institute of Molecular Physiology and Biotechnology of Plants, University of Bonn, Kirschallee 1, 53115 Bonn, Germany.
Bartels
Dorothea
D
Phillips
Jonathan
J
D032181:Craterostigma / Q000235:genetics*; D018744:DNA, Plant / Q000302:isolation & purification; D003890:Desiccation; D017343:Genes, Plant; D018745:Genome, Plant; D049452:Green Fluorescent Proteins / Q000378:metabolism; D019441:Plant Epidermis / Q000166:cytology; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D021381:Protein Transport; D012231:Rhizobium / Q000502:physiology*; D012639:Seeds / Q000166:cytology; D012641:Selection, Genetic; D014170:Transformation, Genetic
D018744:DNA, Plant; D010940:Plant Proteins; C485184:enhanced green fluorescent protein; C066246:late embryogenesis abundant protein, plant; D049452:Green Fluorescent Proteins