CN112088775A - Method for raising seedlings of hispid fig in test tube - Google Patents
Method for raising seedlings of hispid fig in test tube Download PDFInfo
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- CN112088775A CN112088775A CN202010614840.2A CN202010614840A CN112088775A CN 112088775 A CN112088775 A CN 112088775A CN 202010614840 A CN202010614840 A CN 202010614840A CN 112088775 A CN112088775 A CN 112088775A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention provides a method for cultivating seedlings of hispid fig in a test tube, which comprises the steps of inducing an explant, selecting a tender bud of the hispid fig as the explant, sterilizing the explant, inoculating the sterilized explant into a sterile induction culture medium, and culturing under a specific culture condition; proliferating cluster buds, transferring well-grown buds into a proliferation culture medium, and culturing under a specific culture condition; selecting seedlings with the height of 2-3 cm, transferring the seedlings into a strong seedling culture medium, and culturing under a specific culture condition to obtain seedlings with the height of 4-6 cm, the diameter of 0.3-0.45 mm and the number of sections of 3-5; rooting culture, namely inoculating the seedlings into a rooting culture medium and culturing under a specific culture condition; and (4) greenhouse domestication, namely cleaning the rooted seedlings and planting the rooted seedlings in a domestication greenhouse. The quincuncial peach seedlings can be quickly cultivated without being limited by seasons; the cultivated hispid fig seedlings have more adventitious roots and high rooting rate, the yield of target products is improved, and 2.5 kilograms of roots can be produced after a single hispid fig grows for 18 months.
Description
Technical Field
The invention relates to a seedling raising method, in particular to a method for raising seedlings of hispid fig in a test tube.
Background
Radix fici simplicissimae is a common herb in Lingnan, is prepared from root of Ficus benjamina of Ficus of Moraceae, has mild nature, sweet and pungent taste, has effects of invigorating spleen, tonifying lung, removing dampness, and relaxing muscles and tendons, and can be used for treating spleen deficiency, edema, anorexia, weakness, cough due to pulmonary tuberculosis, night sweat, rheumatalgia, puerperal agalactia, etc. In addition, hispid fig is a plant with the same source of food and medicine and is used for cooking soup in Guangdong region. In recent years, the hispid fig, a precious plant resource, draws high attention of pharmaceutical workers, researches on chemical components, pharmacological activity and other aspects are deepened, and the psoralen is proved to be one of the main active components of the hispid fig and has the effects of resisting bacteria, viruses, blood coagulation, inhibiting tumors, regulating immunity and the like.
The hispid fig favors warm and humid environment with good illumination, and the growing soil of the hispid fig is required to be fertile, so the growth of the hispid fig is limited by seasonal environment and land environment, the rooting rate is low, and the four-season supply and demand of the hispid fig root cannot be met.
Disclosure of Invention
Aiming at the problems pointed out in the background technology, the invention provides a method for raising seedlings of hispid fig in a test tube.
The technical scheme of the invention is realized as follows:
a method for raising seedlings of hispid fig in test tubes is characterized by comprising the following steps:
s1, explant induction, namely selecting a tender bud as an explant, sterilizing the explant, and inoculating the sterilized explant into a sterile induction culture medium, wherein the sterile induction culture medium comprises 1/2MS, NAA with the concentration of 0.5mg/L, potato juice with the concentration of 50ml/L, white sugar with the concentration of 30g/L and agar powder with the concentration of 3.6-4.4 g/L, and is cultured for 50-60 days under the culture conditions that the temperature is 25-27 ℃, the illumination intensity is 1500-2000 Lx and the illumination time is 10 hours/day to form a bud with the height of about 0.5 cm;
s2, proliferating cluster buds, and transferring well-grown buds into a proliferation culture medium, wherein the proliferation culture medium comprises MS, 6-BA with the concentration of 0.3-0.5 mg/L, NAA with the concentration of 1.0mg/L, potato juice with the concentration of 100ml/L, white sugar with the concentration of 30g/L and agar powder with the concentration of 3.6g/L, and 3-5 strains of each cluster are subjected to proliferation culture for 50-60 days under the culture conditions of the temperature of 25-27 ℃, the illumination intensity of 1800-2000 Lx and the illumination time of 10-12 hours/day, and the proliferation rate is more than 3.20;
s3, strengthening seedlings, namely selecting seedlings 2-3 cm high in the step S2, transferring the seedlings into a seedling strengthening culture medium, wherein the seedling strengthening culture medium comprises 1/2MS, NAA with the concentration of 0.5-1.0 mg/L, potato juice with the concentration of 20ml/L, white sugar with the concentration of 20g/L and agar powder with the concentration of 3.6g/L, and culturing for 45-60 days under the culture conditions that the temperature is 24-28 ℃, the illumination intensity is 2000-3000 Lx, and the illumination time is 11-13 hours/day to obtain seedlings 4-6 cm high, 0.3-0.45 mm in diameter and 3-5 knots;
s4, rooting culture, namely inoculating the seedlings in the step S3 into a rooting culture medium, wherein the rooting culture medium comprises MS, 6-BA with the concentration of 1.0-2.0 mg/L, banana puree with the concentration of 5%, white sugar with the concentration of 30g/L and agar with the concentration of 3.6g/L, and after 30-40 days of culture, new roots of the plants grow to be about 1 cm long and can reach 2 cm after 50 days of culture under the culture conditions that the temperature is 24-28 ℃, the illumination intensity is 2000-3000 Lx and the illumination time is 10-12 hours/day;
s5, greenhouse domestication, namely cleaning the rooted seedlings in the step S4 to a cleaning degree of 90%, and planting the rooted seedlings in a domesticated greenhouse;
the invention is further configured to: the 1/2MS culture medium contains the following components in each liter and the weight of each component is as follows: 825mg of NH4NO3, 950mg of KNO3, 220mg of CaCl 2.2H 20, 185mg of MgSO 4.7H 20, 85mg of KH2PO4, 0.83mg of KI, 6.2mg of H3BO3, 16.9mg of MnS 04.H 2O, 8.6mg of ZnSO 4.7H 20, 0.25mg of Na2MoO 4.2H 20, 0.025mg of CuSO 4.5H 20, 0.025mg of CoCl 2.6H 20, 27.8mg of FeSO 4.7H 20, 37.3mg of Na2-EDTA, 100mg of inositol, 0.5mg of nicotinic acid, 0.5mg of pyridoxine hydrochloride, 0.1mg of thiamine hydrochloride and 2mg of glycine.
The invention is further configured to: the explant is tender stem of hispid fig.
The invention is further configured to: and the step S1, namely cleaning the selected tender stems by using detergent, soaking the tender stems for 10min by using 0.1% mercuric chloride, washing the tender stems for 5-8 times by using sterile water, cutting the disinfected tender stems, and inoculating the cut tender stems to the sterile induction culture medium in the step S1.
The invention is further configured to: in step S2, the value-added rate of the cluster buds reaches more than 5, and the plant height reaches 2-5 cm.
The invention is further configured to: when strong seedling rooting culture is performed in the step S4, the height of the hispid fig seedlings is 3-8 cm, each adventitious root is 3-5, and the length of the root is more than 1 cm.
The invention is further configured to: in the step S4, the rooting rate of strong seedlings reaches more than 98 percent.
The invention is further configured to: the survival rate of the rooted seedlings planted in the step S5 reaches more than 95%.
In conclusion, the beneficial effects of the invention are as follows: the method for the test tube seedling raising of the hispid fig can quickly and freely cultivate hispid fig seedlings without season limitation; the cultivated hispid fig seedlings have more adventitious roots and high rooting rate, the yield of target products is improved, and 2.5 kilograms of roots can be produced after a single hispid fig grows for 18 months.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
A method for raising seedlings of hispid fig in test tubes comprises the following steps:
s1, inducing an explant, and selecting a tender bud as the explant, wherein the explant is a tender stem of hispid fig, and in the embodiment, the wild hispid fig in the Wenlizian county of Guangxi Lingshan is selected from the variety of the hispid fig. The method comprises the following steps of disinfecting tender stems of hispid fig, wherein the selected tender stems are cleaned by detergent, soaked for 10min by 0.1% mercuric chloride, washed for 5-8 times by sterile water, and cut. And (3) sterilizing the explant, and inoculating the sterilized explant into a sterile induction culture medium, wherein the components of the sterile induction culture medium comprise 1/2MS, NAA with the concentration of 0.5mg/L, potato juice with the concentration of 50ml/L, white sugar with the concentration of 30g/L and agar powder with the concentration of 3.6-4.4 g/L. Culturing for 50-60 days under the culture conditions that the temperature is 25-27 ℃, the illumination intensity is 1500-2000 Lx and the illumination time is 10 hours/day, and forming buds with the height of about 0.5 cm.
S2, proliferating cluster buds, transferring well-grown buds into a proliferation culture medium, wherein the proliferation culture medium comprises MS, 6-BA with the concentration of 0.3-0.5 mg/L, NAA with the concentration of 1.0mg/L, potato juice with the concentration of 100ml/L, white sugar with the concentration of 30g/L and agar powder with the concentration of 3.6g/L, and performing proliferation culture on 3-5 strains of each cluster. Culturing for 50-60 days under the culture conditions that the temperature is 25-27 ℃, the illumination intensity is 1800-2000 Lx and the illumination time is 10-12 hours/day, and the proliferation rate reaches more than 3.20. And (3) growing the seedling of the cluster buds while proliferating the cluster buds, wherein the proliferation rate of the cluster buds is more than 5, and the plant height is 2-5 cm.
S3, strengthening seedlings, namely selecting seedlings with the height of 2-3 cm in the step S2, and transferring the seedlings into a seedling strengthening culture medium, wherein the seedling strengthening culture medium comprises 1/2MS, NAA with the concentration of 0.5-1.0 mg/L, potato juice with the concentration of 20ml/L, white sugar with the concentration of 20g/L and agar powder with the concentration of 3.6 g/L. Culturing for 45-60 days under the culture conditions that the temperature is 24-28 ℃, the illumination intensity is 2000-3000 Lx and the illumination time is 11-13 hours/day to obtain seedlings with the height of 4-6 cm, the diameter of 0.3-0.45 mm and the number of sections of 3-5;
s4, rooting culture, namely inoculating the seedlings in the step S3 into a rooting culture medium, wherein the rooting culture medium comprises MS, 6-BA with the concentration of 1.0-2.0 mg/L, banana puree with the concentration of 5%, white sugar with the concentration of 30g/L and agar with the concentration of 3.6g/L, and after 30-40 days of culture, the new roots of the plants are about 1 cm long after 50 days of culture under the culture conditions that the temperature is 24-28 ℃, the illumination intensity is 2000-3000 Lx and the illumination time is 10-12 hours/day, and the new roots of the plants can reach 2 cm and the rooting rate reaches more than 98%. The method is characterized in that when strong seedlings of the hispid fig root are cultured and grow, the height of each hispid fig is 3-8 cm, each adventitious root is 3-5, the root is longer than 1 cm, and the longer the culture time is, the longer the root is.
S5, greenhouse domestication, namely cleaning the rooted seedlings obtained in the step S4 to a cleaning degree of 90%, planting the rooted seedlings in a domestication greenhouse to obtain a planting survival rate of more than 95%.
In the process, each liter of 1/2MS culture medium contains the following components by weight: 825mg of NH4NO3, 950mg of KNO3, 220mg of CaCl 2.2H 20, 185mg of MgSO 4.7H 20, 85mg of KH2PO4, 0.83mg of KI, 6.2mg of H3BO3, 16.9mg of MnS 04.H 2O, 8.6mg of ZnSO 4.7H 20, 0.25mg of Na2MoO 4.2H 20, 0.025mg of CuSO 4.5H 20, 0.025mg of CoCl 2.6H 20, 27.8mg of FeSO 4.7H 20, 37.3mg of Na2-EDTA, 100mg of inositol, 0.5mg of nicotinic acid, 0.5mg of pyridoxine hydrochloride, 0.1mg of thiamine hydrochloride and 2mg of glycine.
The MS culture medium is prepared by doubling the number of the large number of elements in 1/2MS and keeping the number of elements unchanged. The MS culture medium has higher inorganic salt concentration, can ensure mineral nutrition required by tissue growth and can accelerate the growth of callus.
NAA is naphthylacetic acid, is a broad-spectrum plant growth regulator, and can promote cell division and enlargement.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. A method for raising seedlings of hispid fig in test tubes is characterized by comprising the following steps:
s1, explant induction, namely selecting a tender bud as an explant, sterilizing the explant, and inoculating the sterilized explant into a sterile induction culture medium, wherein the sterile induction culture medium comprises 1/2MS, NAA with the concentration of 0.5mg/L, potato juice with the concentration of 50ml/L, white sugar with the concentration of 30g/L and agar powder with the concentration of 3.6-4.4 g/L, and is cultured for 50-60 days under the culture conditions that the temperature is 25-27 ℃, the illumination intensity is 1500-2000 Lx and the illumination time is 10 hours/day to form a bud with the height of about 0.5 cm;
s2, proliferating cluster buds, and transferring well-grown buds into a proliferation culture medium, wherein the proliferation culture medium comprises MS, 6-BA with the concentration of 0.3-0.5 mg/L, NAA with the concentration of 1.0mg/L, potato juice with the concentration of 100ml/L, white sugar with the concentration of 30g/L and agar powder with the concentration of 3.6g/L, and 3-5 strains of each cluster are subjected to proliferation culture for 50-60 days under the culture conditions of the temperature of 25-27 ℃, the illumination intensity of 1800-2000 Lx and the illumination time of 10-12 hours/day, and the proliferation rate is more than 3.20;
s3, strengthening seedlings, namely selecting seedlings 2-3 cm high in the step S2, transferring the seedlings into a seedling strengthening culture medium, wherein the seedling strengthening culture medium comprises 1/2MS, NAA with the concentration of 0.5-1.0 mg/L, potato juice with the concentration of 20ml/L, white sugar with the concentration of 20g/L and agar powder with the concentration of 3.6g/L, and culturing for 45-60 days under the culture conditions that the temperature is 24-28 ℃, the illumination intensity is 2000-3000 Lx, and the illumination time is 11-13 hours/day to obtain seedlings 4-6 cm high, 0.3-0.45 mm in diameter and 3-5 knots;
s4, rooting culture, namely inoculating the seedlings in the step S3 into a rooting culture medium, wherein the rooting culture medium comprises MS, 6-BA with the concentration of 1.0-2.0 mg/L, banana puree with the concentration of 5%, white sugar with the concentration of 30g/L and agar with the concentration of 3.6g/L, and after 30-40 days of culture, new roots of the plants grow to be about 1 cm long and can reach 2 cm after 50 days of culture under the culture conditions that the temperature is 24-28 ℃, the illumination intensity is 2000-3000 Lx and the illumination time is 10-12 hours/day;
s5, greenhouse domestication, namely cleaning the rooted seedlings in the step S4 to reach 90 percent, and planting the rooted seedlings in a domesticated greenhouse.
2. The method for raising the seedling of the hispid fig in the test tube according to claim 1, wherein each liter of 1/2MS culture medium contains the following components by weight: 825mg of NH4NO3, 950mg of KNO3, 220mg of CaCl 2.2H 20, 185mg of MgSO 4.7H 20, 85mg of KH2PO4, 0.83mg of KI, 6.2mg of H3BO3, 16.9mg of MnS 04.H 2O, 8.6mg of ZnSO 4.7H 20, 0.25mg of Na2MoO 4.2H 20, 0.025mg of CuSO 4.5H 20, 0.025mg of CoCl 2.6H 20, 27.8mg of FeSO 4.7H 20, 37.3mg of Na2-EDTA, 100mg of inositol, 0.5mg of nicotinic acid, 0.5mg of pyridoxine hydrochloride, 0.1mg of thiamine hydrochloride and 2mg of glycine.
3. The method for raising the seedling of the hispid fig in the test tube according to claim 1, wherein the explant is a tender stem of the hispid fig.
4. The method for the test tube seedling culture of the hispid fig according to claim 3, wherein the explant sterilization process in the step S1 is as follows: and cleaning the selected tender stems with detergent, soaking the tender stems for 10min with 0.1% mercuric chloride, washing the tender stems with sterile water for 5-8 times, cutting the disinfected tender stems, and inoculating the cut tender stems to the sterile induction culture medium in the step S1.
5. The method for raising the seedling of the hispid fig in the test tube according to claim 1, wherein in step S2, the multiplication rate of the cluster buds is more than 5, and the plant height is 2-5 cm.
6. The method for tube seedling of hispid fig. according to claim 3, wherein in the step S4, when strong seedling and rooting culture is performed, the height of the hispid fig seedling is 3 to 8 cm, each adventitious root has 3 to 5 roots, and the root length is more than 1 cm.
7. The method for raising the seedling of the hispid fig in the test tube according to claim 1, wherein the rooting rate of strong seedlings in step S4 is more than 98%.
8. The method for raising the seedling of the hispid fig in the test tube according to claim 1, wherein the survival rate of the rooted seedling planted in the step S5 reaches more than 95%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111213588A (en) * | 2020-03-11 | 2020-06-02 | 钦州市林业科学研究所 | Simple and efficient tissue culture propagation method for hispid fig |
| CN115380825A (en) * | 2022-09-20 | 2022-11-25 | 广西壮族自治区药用植物园 | Hispid fig tissue culture method and psoralen preparation method |
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| WO2019104931A1 (en) * | 2017-11-30 | 2019-06-06 | 毛小达 | Traditional chinese medicine composition for treating leukaemia and preparation method therefor |
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Patent Citations (3)
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111213588A (en) * | 2020-03-11 | 2020-06-02 | 钦州市林业科学研究所 | Simple and efficient tissue culture propagation method for hispid fig |
| CN111213588B (en) * | 2020-03-11 | 2022-09-02 | 钦州市林业科学研究所 | Simple and efficient tissue culture propagation method for hispid fig |
| CN115380825A (en) * | 2022-09-20 | 2022-11-25 | 广西壮族自治区药用植物园 | Hispid fig tissue culture method and psoralen preparation method |
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Application publication date: 20201218 |