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Bulletin of Botanical Research ›› 2021, Vol. 41 ›› Issue (4): 514-521.doi: 10.7525/j.issn.1673-5102.2021.04.006

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Elimination of Cymbidium mosaic virus from in vitro Shoot Tips of Phalaenopsis aphrodite by Droplet Vitrification

Ren-Rui WANG, Ling-Li TANG, Shi-Lin SU, Cheng-Cai LIU, Jie LI()   

  1. School of Life Science and Engineering,Southwest University of Science and Technology,Mianyang 621010
  • Received:2020-08-07 Online:2021-07-20 Published:2021-03-24
  • Contact: Jie LI E-mail:jay0224@sina.com
  • About author:WANG Ren-Rui(1981—),female,lecturer,doctor degree,majoring in cryopreservation and cryotherapy of horticultural plants.
  • Supported by:
    Project of the education department of Sichuan province‘Elimination of viruses from orchids of shoot tips by cryotherapy’(16zd1137);Science foundation of Southwest university of science and technology‘Study on elimination of viruses from orchids by cryotherapy’(15zx7117)


In vitro shoots of Phalaenopsis Aphrodite ‘Mantianhong’ which were infected by Cymbidium mosaic virus(CymMV) were used as initial materials in the study. Droplet vitrification protocol of shoot tips was established by screening sucrose concentrations in preculture medium, preculture time and time of exposure to PVS2 (Plant vitrification solution 2, PVS2). The regenerative shoot tips were induced to form protocorm-like-body(PLB) and differentiated to shoots. After detection by RT-PCR for CymMV, negative shoots were micropropagated and induced to form roots. The results showed that shoot tips being precultured on BM+0.6 mol·L-1 sucrose for 1-2 days was the optimal, resulting 70%-76.7% survival rate and 53.3%-56.7% regeneration rate of cryopreserved shoot tips. The optimal time duration of PVS2 was 60-90 minutes, resulting 73.3%-76.7% survival rate and 50.0%-56.7% regeneration rate of cryopreserved shoot tips. Being tested by RT-PCR, half of the regenerated shoots were negative. The CymMV eradication frequency was 50% by droplet vitrification protocol, which laid a technical and theoretical foundation for CymMV eradication of orchids.

Key words: Phalaenopsis Aphrodite, shoot tips, cryopreservation, droplet vitrification, Cymbidium mosaic virus

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