Rapid seedling raising method for euphorbia filiformis

Abstract

The invention belongs to the technical field of euphorbia filiformis seedling culture, and particularly relates to a rapid seedling culture method of euphorbia filiformis, which comprises the following steps: explant sterilization treatment, callus induction, callus differentiation, strong seedling culture, acclimatization and transplantation and transplantation management. According to the method, tender leaves of euphorbia filiformis are selected as explants, rapid seedling culture of euphorbia filiformis is carried out on the explants through the steps of callus induction, callus differentiation, strong seedling culture, seedling hardening and the like, and an induction culture medium, a differential culture medium, a strong seedling culture medium and the like in the seedling culture process are optimized and improved. The induction effect, growth coefficient and rooting rate of calluses can be improved, cultivated seedlings can be robust and grow well, the purposes of effectively controlling the production cost and rapidly producing seedlings are achieved, and a foundation can be laid for large-scale production of euphorbia filiformis.

Description

Technical Field

[0001] The invention belongs to the technical field of Euphorbia fischeri seedling cultivation, and particularly relates to a rapid seedling cultivation method for Euphorbia fischeri. Background Art

[0002] Euphorbia francoisii Leandri, also known as Euphorbia francoisii or Colorful Kylin, is a succulent plant of the Euphorbia genus of the Euphorbiaceae family. Euphorbia francoisii prefers a warm, dry and sunny environment, is drought-tolerant, not shade-tolerant, and avoids waterlogging. Euphorbia francoisii has a rich variety of horticultural species and hybrids, and is outstanding in ornamental value: the plant is low and bushy, with hypertrophic fleshy rhizomes; the fleshy stems are round and stick-shaped, grayish white, and the epidermis of the side branches is rough; the leaves are whorled, with various leaf shapes, including round, long, lanceolate, diamond-shaped, and ivy, maple, begonia, and other types of leaves. Some varieties have wrinkled leaf edges, even lace edges; the leaf color is bright and colorful, with a variety of colors such as red, pink, white, light green, dark green, and even close to black; some varieties also have obvious leaf veins, such as silver-gray stripes and red midribs; the inflorescence is upright, cup-shaped cymes, with larger bracts, and small flowers are yellow-green or green. Euphorbia fernseri has attracted the attention and love of countless enthusiasts with its excellent ornamental characteristics. It has broad market prospects and the demand for its seedlings has continued to expand in recent years.

[0003] Euphorbia Fisch is usually propagated by sowing and cutting. However, its seed life is not long and cannot be stored for a long time. It should be sown as it is picked, otherwise it will affect the germination rate. Although cutting propagation can maintain the characteristics of the mother plant, the cutting seedling cycle is long, and cutting requires strong and substantial side branches with top leaves, which for large-scale production, the number of maternal plants required also increases. Therefore, the seedlings obtained by sowing or cutting propagation are far from meeting market demand. And tissue culture rapid propagation has the advantages of fast propagation speed, large propagation coefficient, uniform reproduction of offspring, ability to maintain the excellent characteristics of the mother plant and ability to carry out annual factory production. But at present, there is very little research on tissue culture rapid propagation of Euphorbia Fisch. The present invention applies tissue culture rapid propagation technology to the seedling cultivation of Euphorbia Fisch, by screening suitable culture medium and simplifying tissue culture rapid propagation, to achieve the purpose of effectively controlling production costs and quickly producing seedlings, laying the foundation for its large-scale production.

[0004] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0005] The object of the present invention is to provide a rapid seedling raising method for Euphorbia ferox, so as to solve the problems existing in the prior art in the background technology.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] A rapid seedling raising method for Euphorbia freundii comprises the following steps:

[0008] S1. Sterilization of explants

[0009] The young leaves of Euphorbia ferdinandii were used as explants;

[0010] S2. Callus Induction

[0011] The sterilized explants were inoculated into an induction medium and cultured for 30-35 days to obtain callus tissue;

[0012] S3. Callus differentiation

[0013] The callus tissue was inoculated into a differentiation medium and cultured for 40-45 days to obtain tissue culture buds;

[0014] S4. Seedling cultivation

[0015] Cut the tissue culture buds that grow to 3-4 cm in height and place them in a tissue culture bottle filled with seedling medium for 10-15 days.

[0016] S5. Hardening and transplanting

[0017] Keep the tissue culture bottles closed at 25-30℃ and 5000-8000lx for 5-10 days, and loosen the bottle caps 3 days before transplanting.

[0018] S6. Transplanting management.

[0019] Furthermore, the specific operation of step S1 is: take young leaves of Euphorbia freundii, rinse them with tap water for 8-15 minutes; then disinfect them on a clean bench: soak the leaves in 75% alcohol for 15-30 seconds, and rinse them with sterile water 2-3 times; then soak them in 2% sodium hypochlorite solution for 9-13 minutes, and rinse them with sterile water 2-3 times; finally soak them in 2% sodium hypochlorite solution for 5-7 minutes, and rinse them with sterile water 2-3 times.

[0020] Furthermore, in step S2, the formula of the induction culture medium is: MS culture medium + 0.1-1.0 mg / L Lingfasu + 30.0 g / L white sugar + 5.5-6.0 g / L carrageenan + 0.05-0.1 g / L activated carbon; the culture conditions are: daytime temperature 28-30°C, night temperature 25°C, set the lights to turn on at night, lighting time 6-8h / d, light intensity 2000-2500lx.

[0021] Furthermore, in step S3, the formula of the differentiation medium is: modified MS medium + 0.1-0.5 mg / L Lingfasu + 0.1-0.5 mg / L NAA + 55.0 g / L banana + 33.0 g / L potato + 30.0 g / L white sugar + 5.5-6.0 g / L carrageenan; culture conditions: daytime temperature 28-30°C, set the lights to turn on at night, night temperature 25°C, lighting time 6-8h / d, light intensity 2000-2500lx.

[0022] Furthermore, the improved MS medium is obtained by modifying the macroelements and trace elements in the MS medium while keeping the contents of calcium salts, iron salts and organic elements unchanged, as follows:

[0023] Macroelements: add 0.1g / L KCl2 and 0.1g / L (NH4)2SO4, keep the content of NH4NO3 unchanged, and adjust KNO3, MgSO4·7H2O and KH2PO4 to 0.25g / L, 0.25g / L and 0.28g / L respectively;

[0024] Trace elements: Na2MoO4·2H2O, KI, CuSO4·5H2O and CoCl2·6H2O were not added, while MnSO4·4H2O, ZnSO4·7H2O and H3BO4 were adjusted to 0.01g / L, 0.005g / L and 0.005g / L respectively.

[0025] Furthermore, in step S4, the formula of the seedling culture medium is: 1 / 2MS culture medium + 100mL / L coconut milk + 15.0g / L white sugar + 5.5-6.0g / L carrageenan + 0.1-0.2g / L activated carbon; culture conditions: daytime temperature 28-30°C, night temperature 25°C, set the lights to turn on at night, lighting time 6-8h / d, light intensity 2500-3000lx.

[0026] Furthermore, in step S5, when transplanting, the culture medium remaining on the roots is rinsed with clean water, and the roots are soaked in a 0.2-0.5% potassium permanganate solution for 10-15 minutes, taken out, and placed in a ventilated and cool place to dry for 10-15 minutes.

[0027] Furthermore, in step S5, the seedlings are transplanted into a 50-hole tray containing a seedling matrix, and watered thoroughly for rooting after transplanting; wherein the seedling matrix is ​​composed of peat soil with a fiber size of 0.1-10 mm, Akadama soil with a particle size of 1-3 mm, and volcanic rock with a particle size of 3-6 mm, mixed in a volume ratio of 2:1:1 or 3:1:1; and the seedling matrix is ​​sprayed and watered thoroughly with 0.5-1.0% potassium permanganate solution 2-3 days before transplanting.

[0028] Furthermore, the specific operations of step S6 are:

[0029] Move the transplanted seedlings to a place without direct sunlight and manage them according to the following table for 2 consecutive weeks;

[0030]

[0031] After 2 weeks, move the seedlings to a place with sufficient light and manage fertilizer according to the table below.

[0032]

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] The invention selects young and tender leaves of Euphorbia fischeri as explants, and the explants are subjected to the steps of callus induction, callus differentiation, seedling cultivation and seedling hardening to carry out rapid seedling cultivation of Euphorbia fischeri, and the induction culture medium, differentiation culture medium and seedling cultivation culture medium in the seedling cultivation process are optimized and improved, so that the induction effect, growth coefficient and rooting rate of callus can be improved, and the cultivated seedlings can be made strong and have good growth, so as to achieve the purpose of effectively controlling production costs and rapidly producing seedlings, and can lay a foundation for large-scale production of Euphorbia fischeri. DETAILED DESCRIPTION

[0035] The following is a clear and complete description of the technical solution of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by technicians in this field without creative work are within the scope of protection of the present invention.

[0036] 1. Materials and Methods

[0037] A rapid seedling raising method for Euphorbia freundii comprises the following steps:

[0038] S1. Sterilization of explants

[0039] Take the young leaves of Euphorbia freundii as explants; take the young leaves of Euphorbia freundii, rinse them with tap water for 8-15 minutes; then disinfect them on the clean bench: soak the leaves in 75% alcohol for 30 seconds, rinse them with sterile water 3 times; then soak them in 2% sodium hypochlorite solution for 12 minutes, rinse them with sterile water 2 times; finally soak them in 2% sodium hypochlorite solution for 6 minutes, rinse them with sterile water 2 times;

[0040] S2. Callus Induction

[0041] The sterilized explants were inoculated into the induction medium in Table 1 below for culture. The culture conditions were: 28°C during the day and 25°C at night, with the lights turned on at night (it should be noted that the lights were not turned on during the day in the culture room, and there was basically no light in the culture room), the illumination time was 6h / d, and the illumination intensity was 2500lx, and callus tissue was obtained. The results are shown in Table 1.

[0042] Table 1 Effects of different induction media on callus induction

[0043]

[0044]

[0045] As shown in Table 1, the induction medium formula 3 performed better on the callus induced by explants, and the induction medium formula 3 was selected as the induction medium for explant callus to continue the experiment.

[0046] S3. Callus differentiation

[0047] The callus tissue was inoculated into the differentiation medium in Table 2 below and cultured for 45 days. The culture conditions were: 28°C during the day, 25°C at night, the light was set to turn on at night, the illumination time was 6h / d, and the illumination intensity was 2500lx. Tissue culture buds were obtained. The results are shown in Table 2. The modified MS medium in Table 2 was obtained by modifying the macroelements and trace elements in the MS basic medium and keeping the contents of calcium salts, iron salts and organic elements unchanged. The specific contents are as follows:

[0048] Macroelements: add 0.1g / L KCl2 and 0.1g / L (NH4)2SO4, keep the content of NH4NO3 unchanged, and adjust KNO3, MgSO4·7H2O and KH2PO4 to 0.25g / L, 0.25g / L and 0.28g / L respectively;

[0049] Trace elements: Na2MoO4·2H2O, KI, CuSO4·5H2O and CoCl2·6H2O were not added, while MnSO4·4H2O, ZnSO4·7H2O and H3BO4 were adjusted to 0.01g / L, 0.005g / L and 0.005g / L respectively.

[0050] Table 2 Effects of different differentiation media on callus differentiation

[0051]

[0052] As shown in Table 2, the plants cultured with the three different differentiation media all grew well and the seedlings were strong. However, in terms of proliferation coefficient, differentiation medium formula 3 was superior to the other formulas, and differentiation medium formula 3 was selected as the differentiation medium for callus tissue in the subsequent experiments.

[0053] S4. Seedling cultivation

[0054] The tissue culture buds grown to 4 cm in height were cut and inoculated into tissue culture bottles containing the seedling medium in Table 3 below, and cultured for 15 days. The culture conditions were: 30°C during the day and 25°C at night, with the lights set to turn on at night, 8h / d of illumination, and 2500lx of illumination intensity, to obtain tissue culture seedlings;

[0055] Table 3 Effects of different seedling culture media on seedling culture

[0056]

[0057] As shown in Table 3, the seedling medium formula 2 and formula 3 are significantly better than formula 1, and the difference between the two is not large. The seedling medium formula 3 will be used for further testing.

[0058] S5. Hardening and transplanting

[0059] Close the tissue culture bottle at 25℃ and 8000lx light for 5 days, and loosen the bottle cap 3 days before transplanting. When transplanting, rinse the culture medium remaining on the roots with clean water, soak them in 0.5% potassium permanganate solution for 15 minutes, take them out, and place them in a ventilated and cool place to dry for 10 minutes.

[0060] During transplanting, the seedlings are transplanted into a 50-hole tray equipped with a seedling matrix, and watered thoroughly for rooting after transplanting; wherein the seedling matrix is ​​a mixture of peat soil with a fiber size of 0.1-10 mm, red jade soil with a particle size of 1-3 mm, and volcanic rock with a particle size of 3-6 mm in a volume ratio of 3:1:1; the seedling matrix is ​​sprayed and watered thoroughly with 1.0% potassium permanganate solution 2 days before transplanting;

[0061] S6. Transplantation management

[0062] The specific operations are as follows:

[0063] Move the transplanted seedlings to a place without direct sunlight and manage them according to the following table for 2 consecutive weeks;

[0064]

[0065] After 2 weeks, move the seedlings to a place with sufficient light and manage fertilizer according to the table below.

[0066]

[0067] In summary, the present invention selects young and tender leaves of Euphorbia fischeri as explants, and the explants are subjected to the steps of callus induction, callus differentiation, seedling cultivation and seedling hardening to carry out rapid seedling cultivation of Euphorbia fischeri, and the induction culture medium, differentiation culture medium and seedling cultivation culture medium in the seedling cultivation process are optimized and improved, so that the induction effect, growth coefficient and rooting rate of callus can be improved, and the cultivated seedlings can be made strong and have good growth, so as to achieve the purpose of effectively controlling production costs and quickly producing seedlings, and can lay a foundation for large-scale production of Euphorbia fischeri.

[0068] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is obvious that many changes and variations can be made in accordance with the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes.

Claims

1. A method for rapidly raising seedlings of Euphorbia freundii, characterized in that: The following steps are involved: S1. Sterilization of explants The young leaves of Euphorbia ferdinandii were used as explants; S2. Callus Induction The sterilized explants were inoculated into an induction medium and cultured for 30-35 days to obtain callus tissue; S3. Callus differentiation The callus tissue was inoculated into a differentiation medium and cultured for 40-45 days to obtain tissue culture buds; S4. Seedling cultivation Cut the tissue culture buds that grow to 3-4 cm in height and place them in a tissue culture bottle filled with seedling medium for 10-15 days. S5. Hardening and transplanting Keep the tissue culture bottles closed at 25-30℃ and 5000-8000lx for 5-10 days, and loosen the bottle caps 3 days before transplanting. S6. Transplanting management.

2. The rapid seedling raising method of Euphorbia freundii according to claim 1, characterized in that: The specific operation of step S1 is: take young leaves of Euphorbia freundii, rinse them with tap water for 8-15 minutes; then disinfect them on a clean bench: soak the leaves in 75% alcohol for 15-30 seconds, rinse them with sterile water 2-3 times; then soak them in 2% sodium hypochlorite solution for 9-13 minutes, rinse them with sterile water 2-3 times; finally soak them in 2% sodium hypochlorite solution for 5-7 minutes, rinse them with sterile water 2-3 times.

3. The rapid seedling raising method of Euphorbia freundii according to claim 1, characterized in that: In step S2, the formula of the induction medium is: MS medium + 0.1-1.0 mg / L Lingfasu + 30.0 g / L white sugar + 5.5-6.0 g / L carrageenan + 0.05-0.1 g / L activated carbon; The culture conditions are: daytime temperature 28-30℃, nighttime temperature 25℃, set the lights to turn on at night, lighting time 6-8h / d, light intensity 2000-2500lx.

4. The rapid seedling raising method of Euphorbia freundii according to claim 1, characterized in that: In step S3, the formula of the differentiation medium is: modified MS medium + 0.1-0.5 mg / L Lingfasu + 0.1-0.5 mg / L NAA + 55.0 g / L banana + 33.0 g / L potato + 30.0 g / L white sugar + 5.5-6.0 g / L carrageenan; Culture conditions: daytime temperature 28-30℃, nighttime temperature 25℃, set the lights to turn on at night, lighting time 6-8h / d, light intensity 2000-2500lx.

5. The rapid seedling raising method of Euphorbia freundii according to claim 4, characterized in that: The improved MS medium is obtained by modifying the macroelements and trace elements in the MS medium and keeping the contents of calcium salts, iron salts and organic elements unchanged, as follows: Macroelements: add 0.1g / L KCl2 and 0.1g / L (NH4)2SO4, maintain the content of NH4NO3 unchanged, and adjust KNO3, MgSO4·7H2O and KH2PO4 to 0.25g / L, 0.25g / L and 0.28g / L respectively; Trace elements: Na2MoO4·2H2O, KI, CuSO4·5H2O and CoCl2·6H2O were not added, while MnSO4·4H2O, ZnSO4·7H2O and H3BO4 were adjusted to 0.01g / L, 0.005g / L and 0.005g / L respectively.

6. The rapid seedling raising method of Euphorbia freundii according to claim 1, characterized in that: In step S4, the formula of the seedling medium is: 1 / 2MS medium + 100mL / L coconut milk + 15.0g / L white sugar + 5.5-6.0g / L carrageenan + 0.1-0.2g / L activated carbon; Culture conditions: daytime temperature 28-30℃, nighttime temperature 25℃, set the lights to turn on at night, lighting time 6-8h / d, light intensity 2500-3000lx.

7. The rapid seedling raising method of Euphorbia freundii according to claim 1, characterized in that: In step S5, when transplanting, rinse the culture medium remaining on the roots with clean water, soak them in 0.2-0.5% potassium permanganate solution for 10-15 minutes, take them out, and place them in a ventilated and cool place to dry for 10-15 minutes.

8. The rapid seedling raising method of Euphorbia freundii according to claim 1, characterized in that: In step S5, the seedlings are transplanted into a 50-hole hole tray filled with a seedling medium, and watered thoroughly for rooting after transplanting; wherein the seedling medium is formed by mixing peat soil with a fiber size of 0.1-10 mm, Akadama soil with a particle size of 1-3 mm, and volcanic rock with a particle size of 3-6 mm in a volume ratio of 2:1:1 or 3:1:1; and the seedling medium is sprayed and watered thoroughly with 0.5-1.0% potassium permanganate solution 2-3 days before transplanting.

9. The rapid seedling raising method of Euphorbia freundii according to claim 8, characterized in that: The specific operations of step S6 are: Move the transplanted seedlings to a place without direct sunlight and manage them according to the following table for 2 consecutive weeks; After 2 weeks, move the seedlings to a place with sufficient light and manage the fertilizer according to the following table;

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