CN108617508B - Rooting culture medium and tissue culture method for tissue culture of sea buckthorn - Google Patents
Rooting culture medium and tissue culture method for tissue culture of sea buckthorn Download PDFInfo
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- 238000012136 culture method Methods 0.000 title description 12
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- 241000229143 Hippophae Species 0.000 claims abstract description 63
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- 239000003617 indole-3-acetic acid Substances 0.000 claims abstract description 32
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- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 24
- 229930006000 Sucrose Natural products 0.000 claims abstract description 14
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 14
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- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 claims abstract description 12
- 229910052564 epsomite Inorganic materials 0.000 claims abstract description 12
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims abstract description 12
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- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims abstract description 12
- 229910000162 sodium phosphate Inorganic materials 0.000 claims abstract description 12
- 241000196324 Embryophyta Species 0.000 claims description 19
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- 239000000463 material Substances 0.000 description 7
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- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 4
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
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Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Soil Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention provides a rooting culture medium for tissue culture of sea buckthorn, which is characterized in that: the rooting culture medium takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3230‑250mg/L，MgSO4·7H2O 30‑35mg/L，NaH2PO4·H215-20mg/L of O, 2-4mg/L of inositol, 0.1-0.2mg/L of glycine, 0.1mg/L of indole-3-acetic acid, 30g/L of sucrose and 6g/L of agar. The invention provides a rooting culture medium for sea-buckthorn tissue culture, which provides a rooting culture medium with balanced and reasonable nutrition for sea-buckthorn tissue culture rooted seedlings by adjusting the types and the concentrations of major elements, optimizing hormone regulation, optimizing illumination and the like on the basis of 1/2MS culture medium, and can promote the quick, efficient and robust growth of the sea-buckthorn rooted seedlings.
Description
Technical Field
The invention belongs to the technical field of plant regeneration through a tissue culture technology, and particularly relates to a rooting culture medium for tissue culture of sea buckthorn and a tissue culture method.
Background
Hippophae rhamnoides (Latin's name: Hippophe rhamnoides Linn.) is a perennial deciduous fruit tree, shrub or small tree. The fruit, pericarp, leaf, bark and their processed products contain more than 280 kinds of physiologically active substances, and have strong special effects of diminishing inflammation, sterilizing, relieving pain and promoting tissue regeneration. Many of the components show magical therapeutic effects in killing and inhibiting tumor cells, resisting radiation, resisting blood coagulation, lowering blood pressure, preventing vascular embolism, resisting aging and fatigue, enhancing body vitality and immunity and the like.
The sea-buckthorn has luxuriant branches and leaves, developed lateral roots and extremely strong root tillering property, can quickly bunch roots and self-tillering to form a dense colony, can be symbiotic with actinomycetes, bacteria, mycobacteria and the like to form a large number of root nodules, and has stronger nitrogen fixation capacity (180 kg of nitrogen can be fixed per hectare) than soybeans. In addition, the ecological fertilizer has the obvious functions of rapid growth, drought resistance, saline-alkali resistance and saline-alkali resistance, protection against severe cold and summer heat, maintenance of water and soil, wind prevention and sand fixation, soil improvement, strong adaptability and the like, and promotes ecological balance, and is not only a pioneer tree species for afforestation and greening in arid and windy sand areas, but also an important salary forest and feed forest tree species. Therefore, the planting of sea buckthorn has important nutritional value, ecological value and economic value in the current adjustment of the industrial structure of returning back to agriculture and forestry.
The characteristics of good varieties of sea-buckthorn are difficult to maintain in production by seed propagation of male and female sea-buckthorn plants; the root shoot propagation coefficient is low. The survival rate of the cutting propagation adopted in the prior production is not high, and the diseases are easy to spread. Compared with cutting seedlings, the tissue culture seedlings can better keep the excellent properties of parents and are more suitable for industrial seedling culture. However, the existing sea-buckthorn tissue culture rooting culture medium has the defects of few rooting numbers, low rooting rate, low root activity and the like.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a rooting culture medium for tissue culture of sea buckthorn and a tissue culture method.
The first purpose of the invention is to provide a rooting culture medium for sea-buckthorn tissue culture, which takes 1/2MS culture medium as basic culture medium and adds NH in the basic culture medium4NO3230-250mg/L，MgSO4·7H2O 30-35mg/L，NaH2PO4·H215-20mg/L of O, 2-4mg/L of inositol, 0.1-0.2mg/L of glycine, 0.1mg/L of indole-3-acetic acid, 30g/L of sucrose and 6g/L of agar.
Preferably, the rooting medium takes 1/2MS medium as a basic medium, and NH is added into the basic medium4NO3240mg/L，MgSO4·7H2O 32mg/L，NaH2PO4·H218mg/L of O, 3mg/L of inositol, 0.1mg/L of glycine, 0.1mg/L of indole-3-acetic acid, 30g/L of sucrose and 6g/L of agar.
Preferably, 0.2mg/L of indolebutyric acid is used instead of 0.1mg/L of indole-3-acetic acid.
The second purpose of the invention is to provide a method for tissue culture of sea-buckthorn by applying the rooting culture medium, which comprises the steps of cutting off individual plants of the sea-buckthorn clump seedlings, inoculating the cut individual plants of the sea-buckthorn clump seedlings into the rooting culture medium to induce rooting, and transferring test-tube seedlings into a sterilized substrate to be cultured when the roots grow to be more than 3.5cm, so as to obtain the sea-buckthorn tissue culture seedlings.
Preferably, the culture conditions for inducing rooting are as follows: temperature: 25. + -. 2 ℃ and light intensity: 2200-2600lx, the photoperiod is 16 h/d.
Preferably, the culture conditions for inducing rooting are as follows: temperature: 25 ℃, light intensity: 2400lx, the light period is 16 h/d.
Preferably, the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1: 2.
The invention provides a rooting culture medium for sea-buckthorn tissue culture, which provides a rooting culture medium with balanced and reasonable nutrition for sea-buckthorn tissue culture rooted seedlings by adjusting the types and the concentrations of major elements, optimizing hormone regulation, optimizing illumination and the like on the basis of 1/2MS culture medium, and can promote the quick, efficient and robust growth of the sea-buckthorn rooted seedlings.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples are commercially available unless otherwise specified.
The invention provides a rooting culture medium for sea-buckthorn tissue culture, which provides a rooting culture medium with balanced and reasonable nutrition for sea-buckthorn tissue culture rooted seedlings by adjusting the types and the concentrations of major elements, optimizing hormone regulation, optimizing illumination and the like on the basis of 1/2MS culture medium, and can promote the quick, efficient and robust growth of the sea-buckthorn rooted seedlings.
Specifically, the rooting culture medium for sea-buckthorn tissue culture takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3230-250mg/L，MgSO4·7H2O 30-35mg/L，NaH2PO4·H215-20mg/L of O, 2-4mg/L of inositol, 0.1-0.2mg/L of glycine, 0.1mg/L of indole-3-acetic acid (IAA), 30g/L of sucrose and 6g/L of agar.
The preparation method comprises the following steps: mixing the above materials except agar, heating with slow fire, stirring to dissolve, adjusting pH to 6.6-7.0 with acid such as diluted hydrochloric acid, adding agar, stirring, and performing wet heat sterilization.
MS culture medium and 1/2MS culture medium formulations are known to those skilled in the art, see Table 1 in particular.
TABLE 1
The tissue culture method of the sea buckthorn comprises the following steps:
cutting off single plants of the sea-buckthorn clustered seedlings, and inoculating the cut plants into a rooting culture medium to induce rooting. The culture conditions were: temperature: 25. + -. 2 ℃ and light intensity: 2200-2600lx with the photoperiod of 16h/d to obtain the test-tube plantlet. When the root grows to be more than 3.5cm, transferring the test-tube plantlet into a sterilized substrate for culturing, wherein the substrate can be any one of commercially available substrates, and is preferably prepared by mixing perlite and turfy soil according to the volume ratio of 1:2, so as to obtain the sea-buckthorn tissue culture plantlet.
Example 1
The rooting culture medium for sea-buckthorn tissue culture takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3240mg/L，MgSO4·7H2O 32mg/L，NaH2PO4·H218mg/L of O, 3mg/L of inositol, 0.1mg/L of glycine, 0.1mg/L of indole-3-acetic acid (IAA), 30g/L of sucrose and 6g/L of agar.
The preparation method comprises the following steps: mixing the above materials except agar, heating with slow fire, stirring to dissolve, adjusting pH to 6.8 with acid such as dilute hydrochloric acid, adding agar, stirring, and performing wet heat sterilization.
The tissue culture method of the sea buckthorn comprises the following steps:
cutting off single plants of the sea-buckthorn clustered seedlings, and inoculating the cut plants into a rooting culture medium to induce rooting. The culture conditions were: temperature: 25 ℃, light intensity: 2400lx, the light period is 16h/d, and test-tube plantlets are obtained. When the root grows to be more than 3.5cm, transferring the test-tube plantlet into a sterilized matrix for culture, wherein the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1:2, and obtaining the sea-buckthorn tissue culture plantlet.
Example 2
The rooting culture medium for sea-buckthorn tissue culture takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3230mg/L，MgSO4·7H2O 35mg/L，NaH2PO4·H2O15 mg/L, inositol 4mg/L, glycine 0.1mg/L, indole-3-acetic acid (IAA)0.1mg/L, sucrose 30g/L, agar 6 g/L.
The preparation method comprises the following steps: mixing the above materials except agar, heating with slow fire, stirring to dissolve, adjusting pH to 6.6 with acid such as dilute hydrochloric acid, adding agar, stirring, and performing wet heat sterilization.
The tissue culture method of the sea buckthorn comprises the following steps:
cutting off single plants of the sea-buckthorn clustered seedlings, and inoculating the cut plants into a rooting culture medium to induce rooting. The culture conditions were: temperature: 23 ℃, light intensity: 2600lx, the light period is 16h/d, and test-tube plantlets are obtained. When the root grows to be more than 3.5cm, transferring the test-tube plantlet into a sterilized matrix for culture, wherein the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1:2, and obtaining the sea-buckthorn tissue culture plantlet.
Example 3
The rooting culture medium for sea-buckthorn tissue culture takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3250mg/L，MgSO4·7H2O 30mg/L，NaH2PO4·H220mg/L of O, 2mg/L of inositol, 0.2mg/L of glycine, 0.1mg/L of indole-3-acetic acid (IAA), 30g/L of sucrose and 6g/L of agar.
The preparation method comprises the following steps: mixing the above materials except agar, heating with slow fire, stirring to dissolve, adjusting pH to 7.0 with acid such as diluted hydrochloric acid, adding agar, stirring, and performing wet heat sterilization.
The tissue culture method of the sea buckthorn comprises the following steps:
cutting off single plants of the sea-buckthorn clustered seedlings, and inoculating the cut plants into a rooting culture medium to induce rooting. The culture conditions were: temperature: 27 ℃, light intensity: 2200lx, the light cycle is 16h/d, and test-tube plantlets are obtained. When the root grows to be more than 3.5cm, transferring the test-tube plantlet into a sterilized matrix for culture, wherein the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1:2, and obtaining the sea-buckthorn tissue culture plantlet.
Example 4
The rooting culture medium for sea-buckthorn tissue culture takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3240mg/L，MgSO4·7H2O 30mg/L，NaH2PO4·H218mg/L of O, 4mg/L of inositol, 0.2mg/L of glycine, 0.1mg/L of indole-3-acetic acid (IAA), 30g/L of sucrose and 6g/L of agar.
The preparation method comprises the following steps: mixing the above materials except agar, heating with slow fire, stirring to dissolve, adjusting pH to 6.8 with acid such as dilute hydrochloric acid, adding agar, stirring, and performing wet heat sterilization.
The tissue culture method of the sea buckthorn comprises the following steps:
cutting off single plants of the sea-buckthorn clustered seedlings, and inoculating the cut plants into a rooting culture medium to induce rooting. The culture conditions were: temperature: 24 ℃, light intensity: 2300lx, the light period is 16h/d, and test-tube plantlets are obtained. When the root grows to be more than 3.5cm, transferring the test-tube plantlet into a sterilized matrix for culture, wherein the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1:2, and obtaining the sea-buckthorn tissue culture plantlet.
Example 5
The rooting culture medium for sea-buckthorn tissue culture takes 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3230mg/L，MgSO4·7H2O 33mg/L，NaH2PO4·H2O16 mg/L, inositol 3mg/L, glycine 0.1mg/L, indole-3-acetic acid (IAA)0.1mg/L, sucrose 30g/L, agar 6 g/L.
The preparation method comprises the following steps: mixing the above materials except agar, heating with slow fire, stirring to dissolve, adjusting pH to 7.0 with acid such as diluted hydrochloric acid, adding agar, stirring, and performing wet heat sterilization.
The tissue culture method of the sea buckthorn comprises the following steps:
cutting off single plants of the sea-buckthorn clustered seedlings, and inoculating the cut plants into a rooting culture medium to induce rooting. The culture conditions were: temperature: 26 ℃, light intensity: 2500lx, the light period is 16h/d, and test-tube plantlets are obtained. When the root grows to be more than 3.5cm, transferring the test-tube plantlet into a sterilized matrix for culture, wherein the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1:2, and obtaining the sea-buckthorn tissue culture plantlet.
Comparative example 1
The present embodiment is different from embodiment 1 in that: rooting culture medium for sea-buckthorn tissue culture, which takes 1/2MS culture medium as basic culture medium and is added with NH4NO3240mg/L, indole-3-acetic acid (IAA)0.1mg/L, sucrose 30g/L, agar 6 g/L.
The remaining steps and specific parameters were the same as in example 1.
Comparative example 2
The present embodiment is different from embodiment 1 in that: rooting culture medium for sea-buckthorn tissue culture, which takes 1/2MS culture medium as basic culture medium and adds MgSO4·7H232mg/L of O, 0.1mg/L of glycine, 0.1mg/L of indole-3-acetic acid (IAA), 30g/L of sucrose and 6g/L of agar.
The remaining steps and specific parameters were the same as in example 1.
Comparative example 3
The present embodiment is different from embodiment 1 in that: rooting culture medium for sea-buckthorn tissue culture, which takes 1/2MS culture medium as basic culture medium and adds MgSO4·7H2O 32mg/L，NaH2PO4·H218mg/L of O, 3mg/L of inositol, 0.1mg/L of indole-3-acetic acid (IAA), 30g/L of sucrose and 6g/L of agar.
The remaining steps and specific parameters were the same as in example 1.
Comparative example 4
The present embodiment is different from embodiment 1 in that: in a rooting culture medium for the tissue culture of sea buckthorn, 0.1mg/L of naphthylacetic acid (NAA) is used for replacing indole-3-acetic acid (IAA).
The remaining steps and specific parameters were the same as in example 1.
Comparative example 5
The present embodiment is different from embodiment 1 in that: indole Butyric Acid (IBA)0.2mg/L is used to replace indole-3-acetic acid (IAA) in rooting medium for sea buckthorn tissue culture.
The remaining steps and specific parameters were the same as in example 1.
Comparative example 6
The present embodiment is different from embodiment 1 in that: indole Butyric Acid (IBA)0.1mg/L and Naphthalene Acetic Acid (NAA)0.1mg/L are used in the rooting culture medium for sea buckthorn tissue culture to replace indole-3-acetic acid (IAA).
The remaining steps and specific parameters were the same as in example 1.
Comparative example 7
The present embodiment is different from embodiment 1 in that: in a rooting culture medium for the tissue culture of sea buckthorn, 0.1mg/L of 6-benzylamino adenine (6-BA) is used for replacing indole-3-acetic acid (IAA).
The remaining steps and specific parameters were the same as in example 1.
Comparative example 8
The present embodiment is different from embodiment 1 in that: in the sea-buckthorn tissue culture method, the culture conditions are as follows: temperature: 20. + -.2 ℃ and light intensity: 2000lx, photoperiod 12 h/d.
The remaining steps and specific parameters were the same as in example 1.
Comparative example 9
The present embodiment is different from embodiment 1 in that: in the sea-buckthorn tissue culture method, the culture conditions are as follows: temperature: 25. + -. 2 ℃ and light intensity: 3000lx, light period 18 h/d.
The remaining steps and specific parameters were the same as in example 1.
After rooting culture, the above examples and comparative examples were acclimatized and transplanted according to conventional acclimatization and transplantation methods for plant tissue culture, and the rooting conditions and the transplantation survival rates are shown in table 2.
TABLE 2 rooting and transplanting survival data for examples and comparative examples of the present invention
| Root number (root) | Root length (cm) | Plant height (cm) | Survival Rate of transplantation (%) | |
| Example 1 | 10-15 | 3.7-7.0 | 8.2-10.7 | 98 |
| Example 2 | 9-13 | 3.5-6.2 | 7.8-10.4 | 97 |
| Example 3 | 9-14 | 3.5-6.6 | 7.9-10.5 | 98 |
| Example 4 | 10-13 | 3.5-6.1 | 8.0-10.6 | 97 |
| Example 5 | 9-13 | 3.6-6.3 | 7.8-10.3 | 96 |
| Comparative example 1 | 4-7 | 2.1-4.1 | 5.8-8.4 | 82 |
| Comparative example 2 | 5-9 | 2.3-4.2 | 6.2-8.9 | 83 |
| Comparative example 3 | 5-8 | 2.0-4.3 | 5.9-8.6 | 85 |
| Comparative example 4 | 3-8 | 1.7-4.2 | 5.1-7.7 | 65 |
| Comparative example 5 | 8-11 | 3.2-5.9 | 7.2-9.5 | 72 |
| Comparative example 6 | 4-8 | 1.9-3.7 | 5.1-7.8 | 67 |
| Comparative example 7 | 3-8 | 1.5-3.6 | 5.4-7.6 | 71 |
| Comparative example 8 | 7-11 | 2.1-4.3 | 6.3-8.8 | 78 |
| Comparative example 9 | 6-10 | 2.0-4.2 | 5.4-7.8 | 81 |
Table 2 was analyzed: as can be seen from the data in comparative examples 1-3, when only a partial group was added to 1/2MS medium, the number of roots, root length, plant height and survival rate of transplantation were much lower than the medium formulation of the present application. As can be seen from the data in comparative examples 4-7, the effect of selecting one or more other growth regulators is much lower than the growth regulator in the present application, and when the growth regulator is indolebutyric acid (IBA), the rooting effect is inferior to the growth regulator used in the present application. As can be seen from the data in comparative examples 8 and 9, the effect is much lower in other lighting conditions than the lighting conditions used in this application.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (3)
1. The method for performing the tissue culture of the sea buckthorn by applying the tissue culture rooting culture medium of the sea buckthorn is characterized by comprising the following steps: cutting off individual plants of the seabuckthorn clump seedlings, inoculating the cut individual plants into a rooting culture medium to induce rooting, and transferring the test-tube seedlings into a sterilized matrix to be cultured when the roots grow to be more than 3.5cm to obtain seabuckthorn tissue culture seedlings;
the preparation method of the sea-buckthorn tissue culture rooting medium comprises the following steps: 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3230-250mg/L，MgSO4·7H2O 30-35mg/L，NaH2PO4·H215-20mg/L of O, 2-4mg/L of inositol, 0.1-0.2mg/L of glycine, 0.1mg/L of indole-3-acetic acid, 30g/L of sucrose and 6g/L of agar;
the culture conditions during rooting induction are as follows: temperature: 25. + -. 2 ℃ and light intensity: 2200-2600lx, the light period is 16 h/d;
the matrix is prepared by mixing perlite and turfy soil according to the volume ratio of 1: 2.
2. The method for tissue culture of seabuckthorn according to claim 1, wherein the method comprises the following steps: the preparation method of the sea-buckthorn tissue culture rooting medium comprises the following steps: 1/2MS culture medium as basic culture medium, and NH is added into the basic culture medium4NO3240mg/L，MgSO4·7H2O 32mg/L，NaH2PO4·H218mg/L of O, 3mg/L of inositol, 0.1mg/L of glycine, 0.1mg/L of indole-3-acetic acid, 30g/L of sucrose and 6g/L of agar.
3. The method according to claim 1 or 2, characterized in that: the culture conditions during rooting induction are as follows: temperature: 25 ℃, light intensity: 2400lx, the light period is 16 h/d.
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