Transient expression of uida reporter gene in regenerable callus tissues of Anthurium andraeanum lind. by agrobacterium mediated transformation

Abstract

A tissue culture scheme for plant regeneration via callus, and a protocol for gene transfer to regenerable callus are described for Anthurium andraeanum variety A vo Nette. Callus was induced from shoot bases and from leaf blades of Anthurium andraeanum on modified MS media containing 0.3 mg t' 2,4-D and 0.5 mg t' BAP. Shoot bases, leaf blades with a single cut at the base, and sectioned leaf blades produced callus at frequencies of 90%, 20% and 8%, respectively. Plant regeneration was obtained at a high frequency in all callus types, irrespective of their origin, in modified MS media containing 0.5 mg t' BAP. Callus was tested for gene transfer efficiency by 2 Agrobacterium tumefaciens strains, C58 andLB4404, carrying binary vector p CAMBIAi } oi containing hpt and uidA reporter genes under plant expression signals. Transient expression of uidA gene monitored by GUS histochemical assay was observed only in callus inoculated with Agrobacterium strain LB4404 at a frequency of 3.33%.