Testing protocols for transformation of Phalaenopsis amabilis l. by biolistic and Agrobacterium tumefaciens

Abstract

We have succeeded in testing protocols for producing transgenic plant using Agrobacterium tumefaciens and microprojectile bombardment for PLB laminae of Phalaenopsis amabilis L. cultured in stationary liquid medium. In mediated transformation, A.tumefaciens C58pGV2260 containing p35SGUS-INT with the uidA gene and nptII gene was used, and maximum level of gus expression was observed 4 days ater co-cultivation with acetosyringone concentration was 50µM. In direct gene transformation, we used Biolistic^TM PDS-1000/He system to transfer  pBAR-GUS (containing uidA gene and nptII gene) into plant cells. The result showed that PBL had the highest GUS activity at 6 cm fire distance with 500 μg tungsten and 0,5 μg DNA mix.