Optimizing virus-induced gene silencing efficiency with Cymbidium mosaic virus in Phalaenopsis flower

Ming Hsien Hsieh, Hsiang Chia Lu, Zhao Jun Pan, Hsin Hung Yeh, Shyh Shyan Wang, Wen Huei Chen, Hong Hwa Chen

研究成果: Article同行評審

47 引文 斯高帕斯(Scopus)


Virus-induced gene silencing (VIGS) is a good way to study floral gene functions of orchids, especially those with a long life cycle. To explore the applicability and improve viral silencing efficiency for application of Cymbidium mosaic virus (CymMV)-induced gene silencing, we examined several variables, including the optimal length of the DNA fragment, the effect of developmental maturation status of inflorescence, and suitable inoculation sites. A CymMV-based VIGS system can be used with orchids to silence genes including PeUFGT3, PeMADS5 and PeMADS6 and induce prominent phenotypes with silencing efficiency up to 95.8% reduction. The DNA fragment size used for silencing can be as small as 78-85. bp and still reach 61.5-95.8% reduction. The effect of cDNA location as a target in VIGS varies among genes because of non-target gene influence when using the 5' terminus of the coding region of both PeMADS5 and PeMADS6. Use of VIGS to knock down a B-class MADS-box gene (PeMADS6) in orchids with different maturation status of inflorescence allowed for observing discernable knockdown phenotypes in flowers. Furthermore, silencing effects with Agro-infiltration did not differ with both leaf and inflorescence injections, but injection in the leaf saved time and produced less damage to plants. We propose an optimized approach for VIGS using CymMV as a silencing vector for floral functional genomics in Phalaenopsis orchid with Agro-infiltration: (1) DNA fragment length about 80. bp, (2) a more mature status of inflorescence and (3) leaf injection.

頁(從 - 到)25-41
期刊Plant Science
出版狀態Published - 2013 3月

All Science Journal Classification (ASJC) codes

  • 遺傳學
  • 農學與作物科學
  • 植物科學


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