CN116982559B - Test tube flowering induction culture method for primula sikkimensis - Google Patents
Test tube flowering induction culture method for primula sikkimensis Download PDFInfo
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- 230000006698 induction Effects 0.000 title claims abstract description 77
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 title claims abstract description 53
- 235000014947 Primula sikkimensis Nutrition 0.000 title claims abstract description 15
- 241000146405 Primula sikkimensis Species 0.000 title claims abstract description 15
- 238000012136 culture method Methods 0.000 title claims abstract description 11
- 238000005286 illumination Methods 0.000 claims abstract description 62
- 229920001817 Agar Polymers 0.000 claims abstract description 31
- 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 31
- 229930006000 Sucrose Natural products 0.000 claims abstract description 31
- 239000008272 agar Substances 0.000 claims abstract description 31
- 239000005720 sucrose Substances 0.000 claims abstract description 31
- 239000001963 growth medium Substances 0.000 claims abstract description 28
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 37
- 241000196324 Embryophyta Species 0.000 claims description 27
- 230000035755 proliferation Effects 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 14
- 238000005520 cutting process Methods 0.000 claims description 9
- 239000012883 rooting culture medium Substances 0.000 claims description 9
- 235000006044 Primula forbesii Nutrition 0.000 claims description 6
- 241000738919 Primula forbesii Species 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 3
- 241000245063 Primula Species 0.000 abstract description 35
- 235000000497 Primula Nutrition 0.000 abstract description 35
- 241000530105 Clerodendrum minahassae Species 0.000 abstract description 9
- 238000009776 industrial production Methods 0.000 abstract description 3
- 238000004161 plant tissue culture Methods 0.000 abstract description 3
- 230000001939 inductive effect Effects 0.000 abstract 1
- 239000000463 material Substances 0.000 description 21
- 238000000338 in vitro Methods 0.000 description 13
- 235000019092 Primula sieboldii Nutrition 0.000 description 8
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- 238000010413 gardening Methods 0.000 description 2
- 241001573881 Corolla Species 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
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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
- A01H4/002—Culture media for tissue culture
-
- 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
Abstract
The invention relates to a primula test tube flowering induction culture method, which belongs to the technical field of plant tissue culture, wherein the culture medium is 3g/L of Huabao No. 1, 0.5mg/L of NAA, 20g/L of sucrose, 1g/L of AC and 7g/L of agar, the pH is 5.8, the culture temperature is 15 ℃, the illumination intensity is 1600lx, and the illumination time is 12h/d. The culture medium and the culture method can be used for inducing the flowering of the primula sikkimensis in the test tube, have the advantages of simple experimental operation, short period, strong practicability and the like, and provide theoretical basis and technical support for the industrial production of the flowering test tube flowers.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to a test tube flowering induction culture method for primula.
Background
Primula (Primula sieboldii) belonging to Primula genus (Primula) belonging to Primula family (Primula), perennial herb, and is mainly distributed in the northeast three-province and the eastern portion of inner mongolia. She Jisheng the flower stand is in a shape of a long circle or a section, is in a shape of a lotus base, is upright, has an umbrella-shaped inflorescence, has a dense flower, has a corolla of purple red to light red, has a gorgeous color, is quite elegant and beautiful as a whole, is pavilion and jade, is commonly used for indoor potted plants, garden landscaping and fresh cut flowers, and has a higher gardening ornamental value.
The test tube flower is a technology for culturing plants in a miniature transparent closed container with better ornamental value by utilizing plant tissue culture technology under the sterile environment condition of manual control. Compared with the traditional gardening planting mode, the flower ornamental and cultivating mode is quite novel and unique, and has the advantages of being free from season limitation, avoiding diseases and natural disasters, being convenient to manage and the like. Meets the current young people's new and different hunting psychology and has wide development prospect. The test tube flowering refers to the process of flowering plants in a closed culture container by tissue culture technology, and is an important component in test tube flowers. In recent years, the main stream of the market mainly is test tube flowers mainly comprising leaves and plant types with long growth period and long playing time, the variety is monotonous, and the culture technology of the test tube flowers for flowers still has the problems in aspects of species selection, flowering induction, flowering period weathers and the like. The test tube flowering experiment system of primula provides a simple and controllable experiment platform for further researching physiological metabolism, gene differential expression, test tube hybridization germplasm innovation and the like in the transformation process from primula plant nutrition growth to reproductive growth, and also provides more possibility for test tube flower market diversification. At present, no report is made on the aspect of flowering induction of test tubes of primula sikkmensis.
Disclosure of Invention
In order to overcome the problems in the background art, the invention provides the flowering induction culture method of the primula fordii test tube, which can finish the induction of flowering in the primula fordii test tube by using the flowering induction culture medium, has the advantages of simple experimental operation, short period, strong practicability and the like, and provides theoretical basis and technical support for industrial production of flowering test tube flowers.
In order to achieve the above purpose, the invention is realized by the following technical scheme:
the primula forbesii test tube flowering induction culture method is used for culturing by the following steps:
1) Cutting leaf stalks of primula sikkimensis aseptic seedlings into 1cm lengths, inoculating the leaf stalks on an induction culture medium for callus induction, and differentiating adventitious buds after subculturing for 30d, wherein the induction culture medium comprises MS+BA0.5mg/L+NAA0.2mg/L+30 g/L of sucrose+6 g/L of agar, and the pH value is 5.8;
2) Inoculating the differentiated adventitious buds to a proliferation culture medium for adventitious bud proliferation, wherein the proliferation culture medium is MS+BA1.0mg/L+NAA0.2mg/L+sucrose 30 g/L+AC1g/L+agar 7g/L, and the pH is 5.8;
3) Inoculating the proliferated adventitious bud single plant to a rooting culture medium for rooting culture, wherein the rooting culture medium is 1/2MS+NAA0.5mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, and the pH value is 5.8;
4) Inoculating rooting seedling with 9-10 leaves to flowering induction culture medium for test tube flowering induction, wherein the flowering induction culture medium is flower bud No. 1, 3g/L, NAA0.5mg/L, sucrose 20g/L, AC 1g/L and agar 7g/L, the pH is 5.8, the culture temperature is 15 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, flower buds are formed after 60d of culture, and the flowers are continuously cultured for about 20d to bloom.
Preferably, the callus induction culture condition in the step 1) is that the temperature is 25+/-2 ℃, the illumination intensity is 1600lx, and the illumination time is 12h/d.
Preferably, the culture condition for proliferation of the adventitious buds in the step 2) is that the temperature is 25+/-2 ℃, the illumination intensity is 1600lx, and the illumination time is 12h/d.
Preferably, the culture condition of the rooting culture in the step 3) is that the temperature is 25+/-2 ℃, the illumination intensity is 1600lx, and the illumination time is 12 hours/d.
The invention has the beneficial effects that:
the invention can complete the induction of flowering in the test tube of the primula sikkmensis through the tissue culture technology and the reasonable allocation of culture medium components and plant growth regulators, solves the problems that the main flow of test tube flowers in the current market mainly comprises leaves and plant types with long growth period and long playing time, provides the flowering test tube flowering primula sikkmensis flowers through callus induction, adventitious bud proliferation, rooting culture and test tube flowering induction, has the advantages of simple experimental operation, short period, strong practicability and the like, and provides theoretical basis and technical support for the industrial production of the flowering test tube flowers.
Drawings
FIG. 1 shows the flowering induction process of the primula forbesii test tube;
in the figure, a represents primula sikkimensis test-tube plantlet, b represents petiole callus and induced differentiation, c represents adventitious bud proliferation, d represents rooting culture, e represents test-tube flowering plant, f represents test-tube flowering plant in a culture bottle with the height of 10cm, g represents test-tube flowering plant in a culture bottle with the height of 7cm, and h represents test-tube flowering plant in a culture bottle with the height of 6 cm.
Description of the embodiments
In order to make the objects, technical solutions and advantageous effects of the present invention more apparent, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, so as to facilitate understanding of the skilled person.
Test-tube plantlets (figure 1 a) of primula sikkimensis (Primula sieboldii) in the applied plant in-vitro library are used as materials, the materials are sterile clone established by primula sikkimensis seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the Chinese southwest wild organism germplasm resource library in the form of test-tube plantlets. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the primula sikkimensis aseptic seedling petiole obtained by the application into 1cm length, inoculating to an induction culture medium (MS+BA0.5 mg/L+NAA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 99.17%. After the callus was further subcultured for 30 days, the callus differentiated into adventitious buds (FIG. 1 b), and the induction rate was 79.66%.
The differentiated adventitious buds were inoculated onto proliferation medium (MS+BA1.0mg/L+NAA0.2mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, pH 5.8) to proliferate adventitious buds (FIG. 1 c). The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the proliferation coefficient after 30d is 3.17.
The propagated adventitious bud single plant is inoculated on a rooting culture medium (1/2MS+NAA0.5mg/L+30 g/L of sucrose+1 g/L of AC+7 g/L of agar, pH 5.8) for rooting culture (FIG. 1 d). The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical rooting rate reaches 100% after 30 d.
Rooting seedling with 9-10 leaves is inoculated to flowering induction culture medium (Huabao No. 1 3g/L+NAA0.5 mg/L+sucrose 20g/L+AC 1 g/L+agar 7g/L, pH 5.8) for test tube flowering induction, the diameter of the induced flowering pot is 7.0cm, the height is 11cm, the culture temperature is 15 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, flower buds are formed after 60d culture, the flowers are continuously cultured for about 20d to bloom (figure 1 e), the umbrella-shaped inflorescence and the test tube flowering induction rate reach 73.33%.
The test-tube plantlet of primula (Primula sieboldii) in the applied plant in-vitro library is used as a material, the material system is a sterile clone established by using primula seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the in-vitro plant library in the wild biomass resource library of southwest China in the form of the test-tube plantlet. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the primula sikkimensis aseptic seedling petiole obtained by the application into 1cm length, inoculating to an induction culture medium (MS+BA0.5 mg/L+NAA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 99.17%. After the callus is continuously subcultured for 30 days, the callus is differentiated into adventitious buds, and the induction rate is 79.66%.
The differentiated adventitious buds were inoculated onto a proliferation medium (MS+BA1.0mg/L+NAA0.2mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, pH 5.8) to proliferate the adventitious buds. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the proliferation coefficient after 30d is 3.17.
The proliferated adventitious bud single plant is inoculated on a rooting culture medium (1/2MS+NAA0.5mg/L+30 g/L of sucrose+1 g/L of AC+7 g/L of agar, pH is 5.8) for rooting culture. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical rooting rate reaches 100% after 30 d.
Rooting seedling with 9-10 leaves is inoculated on flowering induction culture medium (1/2MS+NAA0.5mg/L+1 g/L of active carbon+20 g/L of sucrose+6 g/L of agar and pH of 5.8) to perform test tube flowering induction, the diameter of the induced flowering pot is 7.0cm, the height is 11cm, the culture temperature is 15 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the plants are not flowering after 90d culture.
The test-tube plantlet of primula (Primula sieboldii) in the applied plant in-vitro library is used as a material, the material system is a sterile clone established by using primula seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the in-vitro plant library in the wild biomass resource library of southwest China in the form of the test-tube plantlet. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the primula sikkimensis aseptic seedling petiole obtained by the application into 1cm length, inoculating to an induction culture medium (MS+BA0.5 mg/L+NAA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 99.17%. After the callus is continuously subcultured for 30 days, the callus is differentiated into adventitious buds, and the induction rate is 79.66%.
The differentiated adventitious buds were inoculated onto a proliferation medium (MS+BA1.0mg/L+NAA0.2mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, pH 5.8) to proliferate the adventitious buds. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the proliferation coefficient after 30d is 3.17.
The proliferated adventitious bud single plant is inoculated on a rooting culture medium (1/2MS+NAA0.5mg/L+30 g/L of sucrose+1 g/L of AC+7 g/L of agar, pH is 5.8) for rooting culture. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical rooting rate reaches 100% after 30 d.
The rooting seedling with 9-10 leaves is inoculated on a flowering induction culture medium (MS+NAA0.5mg/L+1 g/L of active carbon+20 g/L of sucrose+6 g/L of agar and pH of 5.8) for test tube flowering induction, the diameter of an induced vase is 7.0cm, the height is 11cm, the culture temperature is 15 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the plant does not bloom after being cultured for 90 d.
The test-tube plantlet of primula (Primula sieboldii) in the applied plant in-vitro library is used as a material, the material system is a sterile clone established by using primula seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the in-vitro plant library in the wild biomass resource library of southwest China in the form of the test-tube plantlet. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the primula sikkimensis aseptic seedling petiole obtained by the application into 1cm length, inoculating to an induction culture medium (MS+BA0.5 mg/L+NAA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 99.17%. After the callus is continuously subcultured for 30 days, the callus is differentiated into adventitious buds, and the induction rate is 79.66%.
The differentiated adventitious buds were inoculated onto a proliferation medium (MS+BA1.0mg/L+NAA0.2mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, pH 5.8) to proliferate the adventitious buds. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the proliferation coefficient after 30d is 3.17.
The proliferated adventitious bud single plant is inoculated on a rooting culture medium (1/2MS+NAA0.5mg/L+30 g/L of sucrose+1 g/L of AC+7 g/L of agar, pH is 5.8) for rooting culture. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical rooting rate reaches 100% after 30 d.
The rooting seedling with 9-10 leaves is inoculated on a flowering induction culture medium (Huabao No. 1, 3g/L+NAA0.5 mg/L+sucrose 20g/L+AC 1 g/L+agar 7g/L, pH is 5.8) to perform test tube flowering induction, the diameter of the induced flowering pot is 7.0cm, the height is 11cm, the test tube flowering culture temperature is 25 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the plants are not flowering after being cultured for 90 d.
The test-tube plantlet of primula (Primula sieboldii) in the applied plant in-vitro library is used as a material, the material system is a sterile clone established by using primula seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the in-vitro plant library in the wild biomass resource library of southwest China in the form of the test-tube plantlet. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the primula sikkimensis aseptic seedling leaf obtained by the application into 1cm length, inoculating to an induction culture medium (MS+BA0.5 mg/L+NAA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 98.48%. After the callus is continuously subcultured for 30 days, the callus is differentiated into adventitious buds, and the induction rate is 28.73%. The callus induction rate was not significantly different from example 1, but the adventitious bud differentiation rate was extremely significantly lower than that of example 1.
The test-tube plantlet of primula (Primula sieboldii) in the applied plant in-vitro library is used as a material, the material system is a sterile clone established by using primula seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the in-vitro plant library in the wild biomass resource library of southwest China in the form of the test-tube plantlet. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the root tip of the primula sikkimensis aseptic seedling obtained by the application into 1cm length, inoculating the root tip to an induction culture medium (MS+BA0.5 mg/L+NA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 98.55%. After the callus is continuously subcultured for 30 days, the callus is differentiated into adventitious buds, and the induction rate is 34.33%. The callus induction rate was not significantly different from example 1, but the adventitious bud differentiation rate was extremely significantly lower than that of example 1.
The test-tube plantlet of primula (Primula sieboldii) in the applied plant in-vitro library is used as a material, the material system is a sterile clone established by using primula seed materials collected from Heilongjiang in 2017, and the sterile clone is stored in the in-vitro plant library in the wild biomass resource library of southwest China in the form of the test-tube plantlet. The specific method comprises the following steps: the primula seed is used as raw material, the surface of the seed is sterilized and aseptically germinated to obtain aseptic seedlings, and an asepsis clone is established.
Cutting the primula sikkimensis aseptic seedling petiole obtained by the application into 1cm length, inoculating to an induction culture medium (MS+BA0.5 mg/L+NAA0.2 mg/L+sucrose 30 g/L+agar 6g/L, pH of 5.8) for callus induction, wherein the culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical callus induction rate after 30d reaches 99.17%. After the callus is continuously subcultured for 30 days, the callus is differentiated into adventitious buds, and the induction rate is 79.66%.
The differentiated adventitious buds were inoculated onto a proliferation medium (MS+BA1.0mg/L+NAA0.2mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, pH 5.8) to proliferate the adventitious buds. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the proliferation coefficient after 30d is 3.17.
The proliferated adventitious bud single plant is inoculated on a rooting culture medium (1/2MS+NAA0.5mg/L+30 g/L of sucrose+1 g/L of AC+7 g/L of agar, pH is 5.8) for rooting culture. The culture temperature is 25+/-2 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, and the statistical rooting rate reaches 100% after 30 d.
Rooting seedling with 9-10 leaves is inoculated on flowering induction culture medium (Huabao No. 1 3g/L+NAA0.5 mg/L+sucrose 20g/L+AC 1 g/L+agar 7g/L, pH 5.8) to perform test tube flowering induction, the diameter of the flowering culture flask is 4.5cm, the heights are respectively 10cm, 7cm and 6cm, outer leaves are cut off, the culture temperature is 15 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, flower buds are formed after 60d, flowers are continuously cultured for about 20d, two flowers of the inflorescence of the culture flask with the height of 10cm (figure 1 f) and single flowers of the inflorescences of the culture flask with the height of 6cm and 7cm (figure 1 g-h).
And (3) injection: the different lowercase letters indicate that the same column between treatments was significantly different at a level of 0.05, and the uppercase letters indicate that the difference at a level of 0.01 was extremely significant, as follows.
TABLE 1 Effect of different flowering induction Medium on flowering induction in primula test tubes
TABLE 2 comparison of callus induction and differentiation of different explants
TABLE 3 Effect of different types of flasks on flowering induction in primula test tubes
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (4)
1. A primula forbesii test tube flowering induction culture method is characterized in that: the cultivation is carried out by the following method, which specifically comprises the following steps:
1) Cutting leaf stalks of primula sikkimensis aseptic seedlings into 1cm lengths, inoculating the leaf stalks on an induction culture medium for callus induction, and differentiating adventitious buds after subculturing for 30d, wherein the induction culture medium comprises MS+BA0.5mg/L+NAA0.2mg/L+30 g/L of sucrose+6 g/L of agar, and the pH value is 5.8;
2) Inoculating the differentiated adventitious buds to a proliferation culture medium for adventitious bud proliferation, wherein the proliferation culture medium is MS+BA1.0mg/L+NAA0.2mg/L+sucrose 30 g/L+AC1g/L+agar 7g/L, and the pH is 5.8;
3) Inoculating the proliferated adventitious bud single plant to a rooting culture medium for rooting culture, wherein the rooting culture medium is 1/2MS+NAA0.5mg/L+30 g/L of sucrose+AC1g/L+7 g/L of agar, and the pH value is 5.8;
4) Inoculating rooting seedling with 9-10 leaves to flowering induction culture medium for test tube flowering induction, wherein the flowering induction culture medium is flower bud No. 1, 3g/L, NAA0.5mg/L, sucrose 20g/L, AC 1g/L and agar 7g/L, the pH is 5.8, the culture temperature is 15 ℃, the illumination intensity is 1600lx, the illumination time is 12h/d, flower buds are formed after 60d of culture, and the flowers are continuously cultured for about 20d to bloom.
2. The primula forbesii test tube flowering induction culture method according to claim 1, wherein the method comprises the following steps of: the callus induction culture condition in the step 1) is that the temperature is 25+/-2 ℃, the illumination intensity is 1600lx, and the illumination time is 12h/d.
3. The primula forbesii test tube flowering induction culture method according to claim 1, wherein the method comprises the following steps of: the proliferation culture condition of the adventitious buds in the step 2) is that the temperature is 25+/-2 ℃, the illumination intensity is 1600lx, and the illumination time is 12h/d.
4. The primula forbesii test tube flowering induction culture method according to claim 1, wherein the method comprises the following steps of: and 3) culturing conditions of rooting culture are that the temperature is 25+/-2 ℃, the illumination intensity is 1600lx, and the illumination time is 12h/d.
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