Transient GUS Gene Expression in Date Palm Fruit Using Agroinjection Transformation Technique

Methods Mol Biol. 2017:1637:295-305. doi: 10.1007/978-1-4939-7156-5_24.

Abstract

Transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy of transformation methods. In this work, we present, for the first time, the utilization of agroinjection as an efficient transformation system for gene delivery in date palm fruit. The research utilized Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pRI201-AN-GUS carrying the beta-glucuronidase (GUS) gene, under the control of a CaMV 35S and kanamycin (NPTII) as an antibiotic gene under the control of a NOS promoter. Based on histochemical assay of agroinjected fruit for the GUS gene expressions, this protocol has proved to be an efficient and reliable tool for transgene expression in date palm. PCR for plasmid DNA, extracted from the transformed Agrobacterium, demonstrated the generation of the expected amplicon, corresponding to the GUS gene using GUS primers.

Keywords: Agrobacterium; Agroinjection; Beta-glucuronidase GUS gene; Genetic transformation.

MeSH terms

  • Agrobacterium tumefaciens / genetics
  • Agrobacterium tumefaciens / physiology*
  • Gene Expression
  • Gene Expression Regulation, Plant
  • Gene Transfer Techniques
  • Glucuronidase / genetics*
  • Phoeniceae / genetics
  • Phoeniceae / microbiology*
  • Plants, Genetically Modified
  • Transformation, Genetic

Substances

  • Glucuronidase