Development of transgenic Phalaenopsis resistant to two viral infections

Abstract

Cymbididum mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV) are the two of the most prevalent plant viruses in orchids. To generate CymMV and ORSV-resistant Phalaenopsis amabilis overexpressing either the sense or anti-sense strand of the CymMV and ORSV coat protein (CP) genes, the gene comprising the capsid proteins of CymMV and the ORSV has been constructed, optimum conditions for particle bombardment have been investigated. Resistant calli and transgenic plantlets have been screened on selection medium. The selection medium was optimized for shoot induction with L-methionine sulfoximine (MSO) added as selection agent. Selection was done in both solid and liquid medium. It was found that liquid media and experiment showed that selection in liquid medium was faster and more efficient. PCR amplification of the inserted CP genes were used for genotyping and PCR Southern assay confirm the insertion of sense and anti-sense strand of CymMV and ORSV CP genes into P. amabilis transformants. Northern blot and Western blot have also been done for the putative transformed plants. Our results showed that the transgenic P. amabilis has been successfully created, and hopefully, the transgenic orchids will be resistant against the two viral infection.