CN110651709B - Method for improving detoxification efficiency of siraitia grosvenorii seedlings - Google Patents
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Abstract
A method for improving the detoxification efficiency of Momordica grosvenori seedlings, comprising the following steps: firstly, preparing a rapid propagation culture medium for siraitia grosvenorii seedlings containing virus inhibitors; step two, transferring the siraitia grosvenorii seedlings into the culture medium in the step one for subculture at 25-30 ℃; step three, culturing the Momordica grosvenori germchits of the step two at 35-39 ℃, and cutting stem tips of the germchits; step four, inoculating the stem tip of the momordica grosvenori seedling obtained in the step three to a conventional rapid propagation culture medium to be cultured for 30-45 days, so as to obtain a detoxified momordica grosvenori plant; wherein the virus inhibitor medium comprises at least one of: 10-60 ppm of ribavirin, 100-500 ppm of moroxydine and 20-100 ppm of tamiflu. The invention combines virus inhibitor and stem tip detoxification method to detoxify the Momordica grosvenori seedlings for the second time, thereby obviously improving the detoxification efficiency of the Momordica grosvenori seedlings. Meanwhile, the detoxification culture method is simple, simplifies the operation procedure, has low production cost and is suitable for industrial production.
Description
Technical Field
The invention relates to the field of fructus momordicae plant propagation, in particular to a method for improving the detoxification efficiency of fructus momordicae seedlings.
Background
The momordica grosvenori is a mature fruit of a plant of the genus momordica of the family cucurbitaceae, is mainly produced in counties such as Guangxi Yongfu, Lingui and Longsheng, is cool in nature, sweet in taste and non-toxic, has the effects of moistening lung to arrest cough, cooling blood and relaxing bowel, is one of traditional export commodities in China, and is popular in Hongkong and Australian regions, southeast Asia and European and American countries. In recent years, the Guangxi fructus momordicae industry is rapidly developed, the fructus momordicae planting area is continuously enlarged, more and more enterprises are added into the ranks of industrial production of fructus momordicae tissue culture seedlings, the tissue culture seedlings are uniform in specification, the number of seedlings can be large, and detoxification can be carried out technically, however, due to various reasons, viral diseases generally occur in the main production area of the Guangxi fructus momordicae, the disease rate of serious plots reaches 100%, the yield of damaged plants is reduced by 25% -45%, and the serious results reach more than 50%, so that the quality of the fructus momordicae is reduced, the secondary fruits are increased, and huge economic losses are caused. The virus causing the greatest harm to the grosvenor momordica is mosaic virus at present, and detoxification culture of grosvenor momordica seedlings is an effective measure for removing the mosaic virus.
At present, the detoxification method of the momordica grosvenori mainly comprises two methods: 1) conventional shoot tip culture: taking the tissue culture seedling of the momordica grosvenori as a test material, stripping the stem tip after different days of high-temperature heat treatment, and detoxifying the momordica grosvenori mosaic virus. 2) Detoxification was performed using detoxification media such as:
chinese patent CN106718922A discloses a tissue culture method for efficient detoxification of fructus momordicae, which comprises the following steps: sterilizing the explant to obtain a sterile explant; inoculating the obtained sterile explant into an MS induction culture medium for induction culture to obtain a sterile test-tube plantlet; pre-culturing the obtained sterile test-tube plantlet in an MS pre-culture medium to obtain a pre-cultured test-tube plantlet; inoculating the obtained pre-cultured test-tube plantlet into an MS virus-free culture medium for virus-free culture to obtain test-tube cluster buds; selecting strong test tube cluster buds, inoculating the test tube cluster buds to 1/2MS rooting culture medium for rooting culture to obtain rooted plants, and finally hardening and transplanting the rooted plants; wherein the MS induction culture medium at least comprises 0.5-1.0 mg/L of adamantanone, the MS pre-culture medium at least comprises 0.5-1.0 mg/L of adamantanone and 0.1-0.6 mg/L of silver nitrate, and the detoxification culture medium comprises: 0.2-1.0 mg/L TDZ 0.1-0.5 mg/L ZT 0.1-1.0 mg/L IBA 30g/L sucrose and 5g/L agar, and the pH value is 5.5-6.
However, the detoxification method has certain defects, and the detoxification method is only carried out once regardless of the conventional stem tip culture method or the detoxification method using the detoxification culture medium, so that the detoxification efficiency cannot be guaranteed; meanwhile, the method has the advantages of strong technical performance, complex virus-free culture method and high production cost, and is not suitable for industrial production.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a method for improving the detoxification efficiency of the siraitia grosvenori seedlings, remarkably improves the detoxification efficiency of the siraitia grosvenori plants with viruses, and reduces the industrial production cost.
The purpose of the invention is realized by the following scheme:
the invention provides a method for improving the detoxification efficiency of momordica grosvenori seedlings, which comprises the following steps: firstly, preparing a rapid propagation culture medium for siraitia grosvenorii seedlings containing virus inhibitors; step two, transferring the siraitia grosvenorii seedlings into the culture medium in the step one to culture at 25-30 ℃; step three, culturing the Momordica grosvenori germchits of the step two at 35-39 ℃, and cutting stem tips of the germchits; step four, inoculating the stem tip of the momordica grosvenori seedling obtained in the step three to a conventional rapid propagation culture medium to be cultured for 30-45 days, so as to obtain a detoxified momordica grosvenori plant; wherein the virus inhibitor medium comprises at least one of: 10-60 ppm of ribavirin, 100-500 ppm of moroxydine and 20-100 ppm of tamiflu;
preferably, the number of days of each subculture in the second step is 10-30 days;
preferably, the number of days for culturing in the third step is 5-15 days;
preferably, the temperature for culturing in the third step is 38-39 ℃;
preferably, the stem tip in the third step is 0.2-0.5 mm;
preferably, the detoxified momordica grosvenori plants in the fourth step are propagated on a rapid propagation medium;
preferably, the preparation method of the culture medium in the first step is as follows: adding plant growth regulator 6-BA and NAA into MS culture medium to make their final concentrations reach 0.3-0.7 ppm and 0.03-0.07 ppm respectively, and adding virus inhibitor after high-pressure sterilization.
The virus inhibitor is adopted to pretreat the siraitia grosvenorii seedlings, the virus density in the seedling cells is reduced, and then the stem tip culture method is used for detoxification, so that the secondary detoxification treatment is carried out on the siraitia grosvenorii tissue culture seedlings, and the detoxification effect of the siraitia grosvenorii seedlings is remarkably improved. Meanwhile, compared with a comparative example in which a virus inhibitor is used alone and a stem tip detoxification method is used, the detoxification rate of papaya ringspot virus (PRSV) is as high as 75%, the detoxification rate of courgette yellows mosaic virus (PRSV) is as high as 90%, which is obviously higher than 10% and 35% of the comparative example, and the unexpected effect is achieved by using the virus inhibitor and the stem tip detoxification method in combination.
The invention has the beneficial effects that:
1. the virus inhibitor and stem tip detoxification method are combined, so that the optimal conditions for virus inhibitor and stem tip detoxification in the combined use process of the virus inhibitor and the stem tip are optimized, secondary detoxification is carried out on the siraitia grosvenori seedlings, and the detoxification efficiency of the siraitia grosvenori seedlings is remarkably improved.
2. The detoxification culture method is simple, low in production cost and suitable for industrial production.
3. The invention carries out heat treatment by taking the tissue culture seedling as the material, avoids the defects of sterilization and high pollution rate when the field materials are obtained, saves the step of explant sterilization, simplifies the operation procedure and improves the working efficiency.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
Example one
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.3ppm and 0.03ppm, respectively. 10ppm ribavirin was added after autoclaving.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm of apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 1 time, 10 days each time, wherein the culture conditions are as follows: at a temperature of 25 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 35 ℃ for treatment for 15 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.3mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: at a temperature of 25 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
Example two
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.7ppm and 0.07ppm, respectively. 100ppm moroxydine was added after autoclaving.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
taking virus vaccine1cm of terminal bud is transferred to a virus inhibitor culture medium for subculture for 5 times, each time for 30 days, and the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 39 ℃ for treatment for 5 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.3mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: at a temperature of 25 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
EXAMPLE III
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively. After autoclaving, duffy 20ppm was added.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 3 times, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 38.5 ℃ for processing for 15 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.2mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
Example four
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.4ppm and 0.05ppm, respectively. 60ppm of ribavirin and 100ppm of tamiflu are added after autoclaving.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 3 times, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 38.5 ℃ for treatment for 12 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.5mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
EXAMPLE five
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.6ppm and 0.05ppm, respectively. After autoclaving, 100ppm moroxydine and 50ppm duffy were added.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 4 times, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 39 ℃ for treatment for 7 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.2mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
EXAMPLE six
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively. Sterilizing under high pressure, and adding ribavirin 30 ppm; moroxydine 150 ppm; 50ppm of tamiflu.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 3 times, 20 days each time, wherein the culture conditions are as follows: at a temperature of 28 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 38.5 ℃ for treatment for 10 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.4mm shoot apical meristem under a stereomicroscope, and immediately putting the shoot apical meristem into a culture medium without virus inhibitor for culture after stripping. The culture conditions were: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
EXAMPLE seven
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively. After autoclaving, 60ppm ribavirin, 500ppm moroxydine and 100ppm duffy are added.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm of apical bud of the virus vaccine to a virus inhibitor culture medium for subculture for 2 times, each time for 25 days, wherein the culture conditions are as follows: at a temperature of 29 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day. Then, the stem tip was removed after the stem was treated at 38 ℃ for 12 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.5mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: the temperature is 32 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
Example eight
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively. After autoclaving, 40ppm ribavirin and 65ppm duffy were added.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm of apical bud of the virus vaccine into a virus inhibitor culture medium for subculture for 1 time, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 38.5 ℃ for treatment for 12 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.5mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: at a temperature of 28 ℃ and a light intensity of 33umol/s/m2Illumination of lightThe time is 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
Example nine
Firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively. Sterilizing under high pressure, and adding ribavirin 30 ppm; moroxydine 160 ppm; 60ppm of tamiflu.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 3 times, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 38.5 ℃ for treatment for 10 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.5mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: at a temperature of 25 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
Comparative example 1 for evaluating the Effect of the viral inhibitors ribavirin, moroxydine, Tamiflu
Comparative example 1
Compared with the example 9, the ribavirin, moroxydine, duffine and the like are not added, and other conditions are the same as follows:
firstly, preparing a culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively.
II, detoxification pretreatment of the momordica grosvenori infected seedlings:
inoculating 1cm of apical bud of the virus vaccine to a culture medium for subculture for 3 times, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day. Then the stem tip is taken out for stripping after being transferred to 38.5 ℃ for treatment for 10 days.
Thirdly, preparing and culturing stem tips:
taking out the virus vaccine treated at high temperature, stripping 0.5mm stem tip meristem under a stereomicroscope, and immediately putting the stem tip stripped into a culture medium without virus inhibitor for culture. The culture conditions were: at a temperature of 25 ℃ and a light intensity of 33umol/s/m2The illumination time was 12 hours per day.
Fourthly, detecting the formed virus of the stem tip regeneration seedlings:
after approximately 45 days under the above culture conditions, the shoot tips were cultured to produce whole plants, and the leaves were taken for virus detection.
Comparative example 2 evaluation of Effect of Combined use of Virus inhibitor and Secondary detoxification by Stem tip detoxification method comparative example 2
Compared with example 9, the secondary detoxification of the stem tip is not used, and the optimal culture conditions are used at the same time, and other conditions are the same, and are as follows:
firstly, preparing a virus inhibitor culture medium:
plant growth regulators 6-BA and NAA were added to the MS medium to make the final concentrations 0.5ppm and 0.05ppm, respectively. Sterilizing under high pressure, and adding ribavirin 30 ppm; moroxydine 160 ppm; 60ppm of tamiflu.
II, detoxification treatment of the momordica grosvenori infected seedlings:
inoculating 1cm apical bud of the virus vaccine into a virus inhibitor culture medium to perform subculture for 6 times, 20 days each time, wherein the culture conditions are as follows: the temperature is 30 ℃, and the light intensity is 33umol/s/m2The illumination time was 12 hours per day.
And taking the plant leaves after 6 times of subculture under the culture condition for virus detection.
Results and analysis
The detoxification effects of papaya ringspot virus (PRSV) and courgette yellows mosaic virus (SMMV) of the leaves of the momordica grosvenori in the examples 1 to 9 and the momordica grosvenori in the comparative examples 1 to 2 are detected by an indirect ELISA method and an electron microscope observation method, and the results are shown in tables 1 to 2.
TABLE 1 papaya ringspot virus (PRSV) detoxification Effect of examples 1 to 9 and comparative examples 1 to 2
| Number of stem tips | Number of adult plants | Analysis of plant number | Number of detoxifications | Detoxification rate (%) | |
| Example 1 | 35 | 33 | 30 | 10 | 33 |
| Example 2 | 25 | 25 | 20 | 3 | 15 |
| Example 3 | 20 | 20 | 14 | 2 | 14 |
| Example 4 | 27 | 27 | 20 | 9 | 45 |
| Example 5 | 30 | 28 | 25 | 7 | 28 |
| Example 6 | 23 | 21 | 20 | 11 | 55 |
| Example 7 | 35 | 28 | 25 | 18 | 72 |
| Example 8 | 18 | 18 | 15 | 11 | 73 |
| Example 9 | 30 | 24 | 20 | 15 | 75 |
| Comparative example 1 | 30 | 28 | 20 | 2 | 10 |
| Comparative example 2 | 35 | 35 | 20 | 0 | 0 |
TABLE 2 detoxification effects of yellow mosaic Virus of Cucurbita pepo in examples 1 to 9 and comparative examples 1 to 2
| Number of stem tips | Number of adult plants | Analysis of plant number | Number of detoxifications | Detoxification rate (%) | |
| Example 1 | 35 | 33 | 30 | 14 | 47 |
| Example 2 | 25 | 25 | 20 | 8 | 40 |
| Example 3 | 20 | 20 | 14 | 6 | 43 |
| Example 4 | 27 | 27 | 20 | 13 | 65 |
| Example 5 | 30 | 28 | 25 | 13 | 52 |
| Example 6 | 23 | 21 | 20 | 15 | 75 |
| Example 7 | 35 | 28 | 25 | 21 | 84 |
| Example 8 | 18 | 18 | 15 | 13 | 87 |
| Example 9 | 30 | 24 | 20 | 18 | 90 |
| Comparative example 1 | 30 | 28 | 20 | 7 | 35 |
| Comparative example 2 | 35 | 35 | 20 | 0 | 0 |
The results of examples 1-3 show that compared with comparative example 1, the detoxification effect of the momordica grosvenori germchit is improved by treating the germchit with a single virus inhibitor. The results of examples 4-9 show that the combination of 2 or 3 inhibitors can further improve the detoxification rate, and is as effective as the two viruses. Meanwhile, the invention can treat different concentrations of ribavirin to Momordica grosvenori papaya ringspot virus (PRSV) and small zucchini yellow mosaic virus
(ZYMV) influence of detoxification Effect, the specific results are as follows:
TABLE 3 Effect of different concentrations of ribavirin on the detoxification of Momordica grosvenori papaya Ringspot Virus (PRSV)
| Ribavirin (mg/L) | Number of stem tips | Number of adult plants | Analysis of plant number | Number of detoxifications | Detoxification rate (%) |
| 0mg/L | 14 | 14 | 14 | 1 | 7.1％ |
| 15mg/L | 14 | 8 | 8 | 4 | 50％ |
| 30mg/L | 14 | 11 | 10 | 7 | 70％ |
TABLE 4 influence of ribavirin of different concentrations on detoxification of yellow mosaic virus of Momordica grosvenori Small zucchini
| Ribavirin (mg/L) | Number of stem tips | Number of adult plants | Analysis of plant number | Number of detoxifications | Detoxification rate (%) |
| 0mg/L | 20 | 20 | 20 | 9 | 45％ |
| 15mg/L | 18 | 16 | 15 | 11 | 73.3％ |
| 30mg/L | 18 | 18 | 18 | 13 | 72.2％ |
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (6)
1. A method for improving the detoxification efficiency of Momordica grosvenori seedlings is characterized by comprising the following steps: firstly, preparing a rapid propagation culture medium for siraitia grosvenorii seedlings containing virus inhibitors; step two, transferring the siraitia grosvenorii seedlings into the culture medium in the step one for subculture at 25-30 ℃; step three, culturing the Momordica grosvenori germchits of the step two at 35-39 ℃, and cutting stem tips of the germchits; step four, inoculating the stem tip of the momordica grosvenori seedling obtained in the step three to a conventional rapid propagation culture medium to be cultured for 30-45 days, so as to obtain a detoxified momordica grosvenori plant; wherein the virus inhibitor medium comprises at least two of: 10-60 ppm of ribavirin, 100-500 ppm of moroxydine and 20-100 ppm of tamiflu; the preparation method of the culture medium in the first step is as follows: adding plant growth regulators 6-BA and NAA into an MS culture medium to enable the final concentrations to reach 0.3-0.7 ppm and 0.03-0.07 ppm respectively; after autoclaving, the virus inhibitor is added after a filtration sterilization treatment.
2. The method for improving the detoxification efficiency of the seedlings of the siraitia grosvenorii as claimed in claim 1, wherein the number of days of each subculture in the second step is 10-30 days.
3. The method for improving the detoxification efficiency of the seedlings of the momordica grosvenori according to claim 1, wherein the number of days of cultivation in the third step is 5-15 days.
4. The method for improving the detoxification efficiency of the seedlings of the momordica grosvenori fruits according to claim 1, wherein the temperature for culturing in the third step is 38-39 ℃.
5. The method for improving the detoxification efficiency of the seedlings of the siraitia grosvenorii as claimed in claim 1, wherein the stem tip in the third step is 0.2-0.5 mm.
6. The method of claim 1, wherein the detoxified Luo Han Guo plant of step four is propagated on a rapid propagation medium.
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