Eradication of CymMV and ORSV from Tissue Culture of Phalaenopsis aphrodita with Chemotherapy

doi:10.11924/j.issn.1000-6850.casb2020-0342

Abstract

Abstract:

The paper aims to explore the virus removal of Phalaenopsis with shoot tip culture and provide technical support for the industrialized virus-free seedling culture. Due to long-term industrial production, Phalaenopsis is often infected with various viruses, which have resulted in withered leaves, chlorosis, necrosis and so on, causing abnormal discoloration of flowers. The peduncles of plants detected with CymMV and ORSV were cultured to induce cespitose buds. The cespitose buds were multiplicated by sub-culture and test-tube plantlets were taken for virus eradicated treatment. Meristem-tips of vitro plantlets were cut for culturing. When the stem-tips were 1-6 mm in length, with the tip length increase, the survival rate of tips increased, and the virus-free rate of regenerated plantlets was significantly reduced. 2-3 mm length of meristem-tips were cut and immersed separately with 0, 10, 20, 30 and 40 mg/L of antiviral agent ribavirin for 15 min, and then cultured for 30 d in the medium with ribavirin of the corresponding concentration, followed by sub-culture, and the regenerated plantlets were obtained. The plants before virus eradicating and the regenerated plantlets after virus eradicating were detected for CymMV and ORSV by the way of ELISA, and whether there were viruses in the samples were judged by chromogenic reaction. The experiment results showed that the survival rate of the regenerated plantlets reduced as the concentration of ribavirin increased, under the treatment of 40 mg/L of ribavirin, the meristem of vitro plantlets was transparent and browned seriously, which could not achieve regenerated plantlets. Oppositely the virus-free rate of the regenerated plantlets rose with the increase of the ribavirin concentration, by adding 30 mg/L ribavirin to the culture medium, the virus-free rate increased by 22.3% compared with that of the purely shoot tip culture.

Key words: Phalaenopsis aphrodita, Cymbidium mosaic virus (CymMV), Odontogssum ringspot virus (ORSV), chemical treatment, shoot tip culture

CLC Number:

S682.31

Jing Jing, Li Jun, Liu Fengjun, Xu Jun, Jiang Hongwei, Li Qing. Eradication of CymMV and ORSV from Tissue Culture of Phalaenopsis aphrodita with Chemotherapy[J]. Chinese Agricultural Science Bulletin, 2021, 37(28): 115-120.

Figures/Tables 4

References 27

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茎尖大小/mm	茎尖成活数	茎尖总数	茎尖成活率/%
1~2	10	50	20Ee
2~3	38	50	76Dd
3~4	43	50	86Cc
4~5	46	50	92Bb
5~6	50	50	100Aa

茎尖大小/mm	茎尖成活数	茎尖总数	茎尖成活率/%
1~2	10	50	20Ee
2~3	38	50	76Dd
3~4	43	50	86Cc
4~5	46	50	92Bb
5~6	50	50	100Aa

茎尖大小/mm	ELISA		茎尖脱毒率/%
茎尖大小/mm	阴性反应的样品数	被检样品数	茎尖脱毒率/%
1~2	10	10	100Aa
2~3	26	38	68Bb
3~4	23	43	54Cc
4~5	12	46	26Dd
5~6	6	50	12Ee
阳性对照	8	8	0
阴性对照	0	8	100

茎尖大小/mm	ELISA		茎尖脱毒率/%
茎尖大小/mm	阴性反应的样品数	被检样品数	茎尖脱毒率/%
1~2	10	10	100Aa
2~3	26	38	68Bb
3~4	23	43	54Cc
4~5	12	46	26Dd
5~6	6	50	12Ee
阳性对照	8	8	0
阴性对照	0	8	100

处理浓度/(mg/L)	ELISA		茎尖脱毒率/%
处理浓度/(mg/L)	阴性反应样品数	被检样品数	茎尖脱毒率/%
0	26	38	68Dd
10	28	35	80Cc
20	30	33	91Bb
30	31	31	100Aa
40	—	—	褐变,未出苗
阳性对照	8	8	0
阴性对照	0	8	100