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

doi:10.7525/j.issn.1673-5102.2021.04.006

Abstract

Abstract:

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

CLC Number:

S632.5

Ren-Rui WANG, Ling-Li TANG, Shi-Lin SU, Cheng-Cai LIU, Jie LI. Elimination of Cymbidium mosaic virus from in vitro Shoot Tips of Phalaenopsis aphrodite by Droplet Vitrification[J]. Bulletin of Botanical Research, 2021, 41(4): 514-521.

Figures/Tables 5

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预培养的蔗糖浓度 Sucrose concentration	成活率Survival rate（%）		再生率Regrowth rate（%）
预培养的蔗糖浓度 Sucrose concentration	对照茎尖 -LN	超低温茎尖 +LN	对照茎尖 -LN	超低温茎尖 +LN
BM+0.3 mol·L^-1蔗糖	93.3±3.3x	43.3±3.3b	73.3±3.3x	10.0±5.8bc
BM+0.6 mol·L^-1蔗糖	86.7±6.7x	76.7±3.3a	66.7±6.7x	56.7±3.3a
BM+0.8 mol·L^-1蔗糖	83.3±3.3x	70.0±5.8a	46.7±3.3y	30.0±5.8b
BM+1.0 mol·L^-1蔗糖	56.7±3.3y	66.7±3.3a	33.3±3.3y	6.7±3.3c

预培养时间 Preculture duration（d）	成活率（%） Survival rate		再生率（%） Regrowth rate
预培养时间 Preculture duration（d）	对照茎尖 -LN	超低温茎尖 +LN	对照茎尖 -LN	超低温茎尖 +LN
1	86.7±6.7x	76.7±3.3a	66.7±6.7x	56.7±3.3a
2	80.0±5.8xy	70.0±5.8a	60.0±5.8x	53.3±3.3a
3	73.3±3.3xy	66.7±3.3a	53.3±6.7xy	30.0±5.8b
4	60.0±0y	20.0±5.8b	33.3±3.3y	6.7±3.3c

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