Campol Jova Riza, Naing Aung Htay, Aung Hay Mon, Cho Su Bin, Kang Hyunhee, Chung Mi Young, Kim Chang Kil
Department of Horticultural Science, Kyungpook National University, Daegu, South Korea.
Department of Agricultural Education, Sunchon National University, Suncheon, South Korea.
Plant Methods. 2024 Sep 19;20(1):145. doi: 10.1186/s13007-024-01269-1.
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.
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.
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.
本研究旨在通过培养感染齿舌兰环斑病毒（ORSV）的母株的分生组织，并对源自分生组织的原球茎状体（PLB）进行连续继代培养，从而培育出无ORSV的大花蕙兰“新真”植株。
最初，通过逆转录-聚合酶链反应（RT-PCR）分析确认ORSV是兰花叶片病毒症状的致病因子。对受感染植株的分生组织进行培养以产生PLB，这些PLB依次进行了多达四次的继代培养。RT-PCR和定量RT-PCR分析表明，虽然在第一次继代培养获得的芽中未检测到ORSV，但完全消除该病毒至少需要进行第二次继代培养。使用简单序列重复区间标记进行的遗传分析表明，再生植株与母株之间没有体细胞克隆变异，这表明遗传一致性得以保持。
总体而言，我们的研究结果表明，对PLB进行第二次继代培养是培育遗传稳定、无ORSV的大花蕙兰的理想方法，从而为培育遗传稳定、无病毒的植株以及提高兰花产业中植株的健康状况和品质提供了一种切实可行的方法。
