Genetic Engineering of ORSV-resistant Phalaenopsis
Odontoglossum ringspot virus (ORSV) is one of the most prevalent plant viruses in orchids. The aim of this research is to generate ORSV-resistant Phalaenopsis equestris van alba overexpressing either the sense or anti-sense strand of the ORSV coat protein (CP) genes using particle bombardment In this research, calli have been induced from different explants of P. equestris var. alba. Plant regeneration system has been established from callus. The gene comprising the capsid proteins of 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 anti-sense strand of ORSV CP genes into several lines of P. equestris var. alba transformants. Northern blot and Western blot have also been done for the putative transformed plants, and viral inoculation and detection will performed on the transgenic Phalaenopsis when they grow to be big. Our results showed that the transgenic P. equestris var. alba has been successfully created, and hopefully, the transgenic orchids will be resistant against viral infection.
Odontoglossum ringspot virus (ORSV) Phalaenopsis equestris var. alba coat protein
Shuguo Fan
Department of Chemistry and Life Science, Chuxiong Normal University, Chuxiong 675000 Institute for Bio-resources Development and Utilization in Central Yunnan Plateau, Chuxiong 675000
国际会议
上海
英文
1446-1449
2011-10-15（万方平台首次上网日期，不代表论文的发表时间）