Method for improving detoxification efficiency of fructus momordicae seedlings

Abstract

A method for improving the detoxification efficiency of fructus momordicae seedlings includes: step 1, preparing a rapid fructus momordicae propagation culture medium containing a virus inhibitor; step 2, transferring the fructus momordicae seedlings into the culture medium of the step 1 to subculture at 25-30 DEG C; step 3, cultivating the fructus momordicae seedlings of the step 2 at 35-39 DEG C, and cutting off stem tips of the seedlings; and step 4, inoculating a common rapid propagation culture medium with the stem tips of the fructus momordicae seedlings of the step 3 to obtain virus-free fructus momordicae plants. The culture medium containing the virus inhibitor at least comprises one of 10-60 ppm of virazole, 100-500 ppm of moroxydine and 20-100 ppm of oseltamivir. The method hasthe advantages that the virus inhibitor and a stem-tip detoxification method are combined to perform secondary detoxification on the fructus momordicae seedlings, and accordingly, the detoxification efficiency of the fructus momordicae seedlings is remarkably improved; the detoxification culture method is simple, the operation process is simplified, the production cost is low, and the method is suitable for industrial production.

Description

technical field

[0001] The invention relates to the field of grosvenoria plant propagation, in particular to a method for improving the detoxification efficiency of grosvenoria seedlings. Background technique

[0002] At present, the detoxification methods of Luo Han Guo mainly contain two: 1) conventional shoot tip culture method: take the tissue cultured seedlings of Luo Han Guo as test material, strip off the shoot tip after high-temperature heat treatment for different days, and carry out the detoxification of Luo Han Guo mosaic virus. 2) Use detoxification medium for detoxification, such as:

[0003] Chinese patent CN106718922A discloses a tissue culture method for effective detoxification of Luo Han Guo, including: sterilizing the explants to obtain sterile explants; inoculating the obtained sterile explants into MS induction medium for induction culture to obtain sterile Bacteria test-tube plantlets; pre-cultivate the obtained sterile test-tube plantlets in MS pre-...

Examples

Embodiment 1

[0022] 1. Preparation of virus inhibitor culture medium:

[0023] Add plant growth regulator 6-BA and NAA in MS medium, make its final concentration reach 0.3ppm and 0.03ppm respectively. Add ribavirin 10 ppm after autoclaving.

[0024] 2. Detoxification pretreatment of Luo Han Guo susceptible seedlings:

[0025] Take the 1cm terminal buds of the virus seedlings and transfer them to the virus inhibitor medium for subculture once, for 10 days each time. The culture conditions are: temperature 25°C, light intensity 33umol / s / m 2 , The light time is 12 hours a day. Then transfer it to 35°C and take it out after 15 days for stripping off the shoot tip.

[0026] 3. Preparation and cultivation of shoot tips:

[0027] The virus seedlings treated with high temperature were taken out, and the shoot tip meristem of 0.3mm was stripped under a stereomicroscope, and immediately after the shoot tip was stripped, it was placed in a medium without virus inhibitors for cultivation. The cul...

Embodiment 2

[0031] 1. Preparation of virus inhibitor culture medium:

[0032] Add plant growth regulator 6-BA and NAA in MS medium, make its final concentration reach 0.7ppm and 0.07ppm respectively. After autoclaving, 100 ppm of morpholinidine was added.

[0033] 2. Detoxification pretreatment of Luo Han Guo susceptible seedlings:

[0034] Take the 1cm terminal buds of the virus seedlings and transfer them to the virus inhibitor medium for subculture 5 times, each time for 30 days. The culture conditions are: temperature 30°C, light intensity 33umol / s / m 2 , The light time is 12 hours a day. Then transfer it to 39°C and take it out after 5 days for stripping off the shoot tip.

[0035] 3. Preparation and cultivation of shoot tips:

[0036] The virus seedlings treated with high temperature were taken out, and the shoot tip meristem of 0.3mm was stripped under a stereomicroscope, and immediately after the shoot tip was stripped, it was placed in a medium without virus inhibitors for cul...

Embodiment 3

[0040] 1. Preparation of virus inhibitor culture medium:

[0041] Add plant growth regulator 6-BA and NAA in MS medium, make its final concentration reach 0.5ppm and 0.05ppm respectively. Add Tamiflu 20ppm after autoclaving.

[0042] 2. Detoxification pretreatment of Luo Han Guo susceptible seedlings:

[0043] Take the 1cm terminal buds of the virus seedlings and transfer them to the virus inhibitor medium for subculture for 3 times, each time for 20 days. The culture conditions are: temperature 30°C, light intensity 33umol / s / m 2 , The light time is 12 hours a day. Then transfer it to 38.5°C and take it out after 15 days for stripping off the shoot tip.

[0044] 3. Preparation and cultivation of shoot tips:

[0045] The virus seedlings treated with high temperature were taken out, and the shoot tip meristem of 0.2 mm was stripped under a stereomicroscope, and immediately after the shoot tip was stripped, it was placed in a medium without virus inhibitors for cultivation. ...

technical field

[0001] The invention relates to the field of grosvenoria plant propagation, in particular to a method for improving the detoxification efficiency of grosvenoria seedlings. Background technique

[0002] At present, the detoxification methods of Luo Han Guo mainly contain two: 1) conventional shoot tip culture method: take the tissue cultured seedlings of Luo Han Guo as test material, strip off the shoot tip after high-temperature heat treatment for different days, and carry out the detoxification of Luo Han Guo mosaic virus. 2) Use detoxification medium for detoxification, such as:

[0003] Chinese patent CN106718922A discloses a tissue culture method for effective detoxification of Luo Han Guo, including: sterilizing the explants to obtain sterile explants; inoculating the obtained sterile explants into MS induction medium for induction culture to obtain sterile Bacteria test-tube plantlets; pre-cultivate the obtained sterile test-tube plantlets in MS pre-...

Examples

Embodiment 1

[0022] 1. Preparation of virus inhibitor culture medium:

[0023] Add plant growth regulator 6-BA and NAA in MS medium, make its final concentration reach 0.3ppm and 0.03ppm respectively. Add ribavirin 10 ppm after autoclaving.

[0024] 2. Detoxification pretreatment of Luo Han Guo susceptible seedlings:

[0025] Take the 1cm terminal buds of the virus seedlings and transfer them to the virus inhibitor medium for subculture once, for 10 days each time. The culture conditions are: temperature 25°C, light intensity 33umol / s / m 2 , The light time is 12 hours a day. Then transfer it to 35°C and take it out after 15 days for stripping off the shoot tip.

[0026] 3. Preparation and cultivation of shoot tips:

[0027] The virus seedlings treated with high temperature were taken out, and the shoot tip meristem of 0.3mm was stripped under a stereomicroscope, and immediately after the shoot tip was stripped, it was placed in a medium without virus inhibitors for cultivation. The cul...

Embodiment 2

[0031] 1. Preparation of virus inhibitor culture medium:

[0032] Add plant growth regulator 6-BA and NAA in MS medium, make its final concentration reach 0.7ppm and 0.07ppm respectively. After autoclaving, 100 ppm of morpholinidine was added.

[0033] 2. Detoxification pretreatment of Luo Han Guo susceptible seedlings:

[0034] Take the 1cm terminal buds of the virus seedlings and transfer them to the virus inhibitor medium for subculture 5 times, each time for 30 days. The culture conditions are: temperature 30°C, light intensity 33umol / s / m 2 , The light time is 12 hours a day. Then transfer it to 39°C and take it out after 5 days for stripping off the shoot tip.

[0035] 3. Preparation and cultivation of shoot tips:

[0036] The virus seedlings treated with high temperature were taken out, and the shoot tip meristem of 0.3mm was stripped under a stereomicroscope, and immediately after the shoot tip was stripped, it was placed in a medium without virus inhibitors for cul...

Embodiment 3

[0040] 1. Preparation of virus inhibitor culture medium:

[0041] Add plant growth regulator 6-BA and NAA in MS medium, make its final concentration reach 0.5ppm and 0.05ppm respectively. Add Tamiflu 20ppm after autoclaving.

[0042] 2. Detoxification pretreatment of Luo Han Guo susceptible seedlings:

[0043] Take the 1cm terminal buds of the virus seedlings and transfer them to the virus inhibitor medium for subculture for 3 times, each time for 20 days. The culture conditions are: temperature 30°C, light intensity 33umol / s / m 2 , The light time is 12 hours a day. Then transfer it to 38.5°C and take it out after 15 days for stripping off the shoot tip.

[0044] 3. Preparation and cultivation of shoot tips:

[0045] The virus seedlings treated with high temperature were taken out, and the shoot tip meristem of 0.2 mm was stripped under a stereomicroscope, and immediately after the shoot tip was stripped, it was placed in a medium without virus inhibitors for cultivation. ...

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