Production of Genetically Stable and Odontoglossum Ringspot Virus-Free Cymbidium Orchid ‘New True’ Plants via Meristem-Derived Protocorm-Like Body (Plb) Subcultures

Authors

Jova Riza Campol
Aung Htay Naing
Hay Mon Aung
Su Bin Cho
Hyunhee Kang
Mi Young Chung
Chang Kil Kim

Publication Date

9-19-2024

Journal

Plant Methods

Abstract

BACKGROUND: This study aimed to produce Odontoglossum ringspot virus (ORSV)-free Cymbidium orchid 'New True' plants from ORSV-infected mother plants by culturing their meristems and successively repeating subcultures of protocorm-like bodies (PLBs) derived from the meristems.

RESULTS: Initially, ORSV was confirmed as the causative agent of viral symptoms in orchid leaves via reverse transcription-polymerase chain reaction (RT-PCR) analysis. Meristems from infected plants were cultured to generate PLBs, which in sequence were repeatedly subcultured up to four times. RT-PCR and quantitative RT-PCR analyses revealed that while ORSV was undetectable in shoots derived from the first subculture, complete elimination of the virus required at least a second subculture. Genetic analysis using inter-simple sequence repeat markers indicated no somaclonal variation between regenerated plants and the mother plant, suggesting that genetic consistency was maintained.

CONCLUSION: Overall, our findings demonstrate that subculturing PLBs for a second time is ideal for producing genetically stable, ORSV-free Cymbidium orchids, thus offering a practical means of generating genetically stable, virus-free plants and enhancing plant health and quality in the orchid industry.

Keywords

Cymbidium orchid, Genetic stability, Meristem culture, ISSR marker, qRT-PCR analysis, Protocorm-like bodies (PLBs)

Comments

Supplementary Materials