Agroinjection of tomato fruits. A tool for rapid functional analysis of transgenes directly in fruit.
Transient expression of foreign genes in plant tissues is a valuable tool for plant biotechnology. To shorten the time for gene functional analysis in fruits, we developed a transient methodology that could be applied to tomato (Solanum lycopersicum cv Micro Tom) fruits. It was found that injection of Agrobacterium cultures through the fruit stylar apex resulted in complete fruit infiltration. This infiltration method, named fruit agroinjection, rendered high levels of 35S Cauliflower mosaic virus-driven beta-glucuronidase and yellow fluorescence protein transient expression in the fruit, with higher expression levels around the placenta and moderate levels in the pericarp. Usefulness of fruit agroinjection was assayed in three case studies: (1) the heat shock regulation of an Arabidopsis (Arabidopsis thaliana) promoter, (2) the production of recombinant IgA antibodies as an example of molecular farming, and (3) the virus-induced gene silencing of the carotene biosynthesis pathway. In all three instances, this technology was shown to be efficient as a tool for fast transgene expression in fruits.
Orzaez
Diego
D
Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas, Universidad Politécnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain.
Mirabel
Sophie
S
Wieland
Willemien H
WH
Granell
Antonio
A
D000910:Antibodies, Heterophile / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D017796:Caulimovirus / Q000235:genetics; D005638:Fruit / Q000235:genetics / Q000382:microbiology; D020868:Gene Silencing; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D007267:Injections; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018551:Solanum lycopersicum / Q000033:anatomy & histology / Q000235:genetics* / Q000382:microbiology; D018613:Microscopy, Confocal; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D012231:Rhizobium / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic; D019076:Transgenes / Q000502:physiology*
D000910:Antibodies, Heterophile; D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D010088:Oxidoreductases; C022398:phytoene dehydrogenase; D005966:Glucuronidase