CN108401905B - Method for rapid propagation culture and annual fresh flower production of gerbera jamesonii - Google Patents
Method for rapid propagation culture and annual fresh flower production of gerbera jamesonii Download PDFInfo
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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
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- 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
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- 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
- A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
- A01G24/25—Dry fruit hulls or husks, e.g. chaff or coir
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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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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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Abstract
The invention belongs to the technical field of gardens, and particularly relates to a method for rapid propagation culture and annual fresh flower production of gerbera jamesonii in the direction of tissue culture and chemical regulation. The method comprises the following steps: primary culture, secondary culture, rooting culture and transplanting and field planting, wherein the primary culture, the secondary culture and the rooting culture all use a basic culture medium and a plant growth regulator as culture mediums. The method sets a new basic culture medium for the gerbera jamesonii, and the basic culture medium is matched with the plant growth hormone, so that the culture efficiency and the tissue culture success rate can be effectively improved, the time required by each step is effectively shortened, the field planting survival rate is high, the time required from field planting to flowering is short, and the single plant flower yield is high.
Description
Technical Field
The invention belongs to the technical field of gardens, and particularly relates to a method for rapid propagation culture and annual fresh flower production of gerbera jamesonii in the direction of tissue culture and chemical regulation.
Background
Gerbera jamesonii is gerbera belonging to the family Compositae. Perennial root herbaceous flowers are called as five fresh cut flowers in the world together with gladiolus, carnation, China rose and chrysanthemum. In the southern part of original Africa, China began to introduce the African chrysanthemum in the 80 th century, but the introduction of the cultivar is completely foreign, which greatly restricts the development of the African chrysanthemum as the fifth cut flower in China.
The traditional propagation method of the gerbera jamesonii is seed propagation and plant division propagation, self-sterility is caused by inconsistent mature periods of male and female stamens in the seed propagation process, the service life of seeds generated by artificial supplementary pollination is short, the germination rate is low, plants are easy to generate characteristic variation, the seed properties are degraded, the flower color becomes light, the flower is small, the yield is low in the short flowering period, and the commodity quality of the flower is reduced. The plant division propagation is limited by seasons, the propagation coefficient is low, and the demands of markets and consumers are difficult to meet. If conventional artificial cross breeding is performed, the use of gerbera resources is limited in foreign countries.
Tissue culture of gerbera jamesonii has been reported, and the basic culture method and operation thereof are not independent of plant tissue culture technology, but almost all of them are cultured by taking MS culture medium as basic culture medium. The prior tissue culture technology has the advantages of low success rate, small quantity of adventitious buds, low efficiency, long culture period and small fresh flower yield.
Disclosure of Invention
In order to solve the problems, the invention provides a method for quickly breeding and producing fresh flowers of gerbera jamesonii, which can effectively improve the success rate of tissue culture, effectively shorten the culture period and improve the yield.
In order to achieve the purpose, the invention adopts the following technical scheme that the method for rapidly propagating and culturing the African daisy and producing fresh flowers annually comprises the following steps: primary culture, subculture, rooting culture and transplanting and field planting, wherein the primary culture and the subculture both use an A culture medium as a culture medium, and the A culture medium comprises the following components: potassium nitrate 100 mg/L, potassium chloride 50 mg/L, four water calcium nitrate 300 mg/L, seven water magnesium sulfate 750 mg/L, sodium sulfate 200 mg/L, sodium dihydrogen phosphate 20 mg/L, four water manganese sulfate 7 mg/L, ferrous sulfate 2.5 mg/L, seven water zinc sulfate 3 mg/L, boric acid 1.5 mg/L, potassium iodide 0.7 mg/L, nicotinic acid 0.5 mg/L, hydrochloric acid pyridoxine 0.3 mg/L, thiamine hydrochloride 0.1 mg/L, glycine 10 mg/L, glucose 50g/L, agar powder 10 g/L; the rooting culture takes a culture medium B as a culture medium, and each component of the culture medium B is consistent with that of the culture medium A, but the concentration of each component is 1/2 of the culture medium A.
Further, the primary culture, the secondary culture and the rooting culture are added with a plant growth regulator in a culture medium.
The plant growth regulator is one or a combination of 6-furfuryl amino purine (6KT), adenine and indolebutyric acid (IBA).
Further, 6-furfuryl amino purine, adenine and indolebutyric acid are added into the culture medium A in the primary culture, and the added 6-furfuryl amino purine is 5 mg/L, adenine is 2 mg/L and indolebutyric acid is 2 mg/L; the subculture is characterized in that 6-furfuryl aminopurine, adenine and indolebutyric acid are added into a culture medium A, and the 6-furfuryl aminopurine is added at a rate of 3-5 mg/L, adenine is added at a rate of 1-2 mg/L and indolebutyric acid is added at a rate of 1.5-2 mg/L; and adding indolebutyric acid into the rooting culture medium B, wherein 5 mg/L of indolebutyric acid is added.
Preferably, 1-2 cm of fine single plant stem tip with pure color and robust growth is selected as an explant, washed by tap water for 5 minutes, soaked by 70% alcohol on a cleaning workbench for 15 minutes, treated by 0.1% mercuric chloride and 0.5% tween 20 for 10 minutes, washed by sterile water, and inoculated in a primary culture medium.
Further, the primary culture is transferred to a new primary culture medium for culture after 1-2 adventitious buds grow on the explant.
Further, the subculture is to cut off and shear the bud from the explant and transfer the bud to a subculture medium for rapid propagation after the adventitious bud grows to 2-3 cm, and the subculture is continued for more than 5 generations.
Further, the rooting culture is that after the number of subcultures is enough, 3-5cm of single plants obtained by the subcultures are cut off and transferred into a rooting culture medium for rooting culture until 2-5 adventitious roots grow on the base of the plantlet.
Further, the transplanting and field planting is that after the residual culture solution is removed from the plantlets obtained by the rooting culture, the plantlets are temporarily planted in a hardening seedling substrate, and new roots and new leaves are grown for field planting.
Further, the seedling hardening matrix is formed by mixing leaf mold, perlite, mushroom bran and vermiculite according to the weight ratio of 5:2:2: 1.
Further, maintenance management is required after field planting, and the maintenance management comprises:
fertilizing: fertilizing is started after planting for one month, biogas liquid fertilizer is mainly applied, direct root irrigation is adopted, and as the seedlings grow, fertilization per plant is gradually increased to 2000mL from 500mL of seedlings to 2-3 times per month;
and (3) promoting flower bud differentiation: after 60 days of planting, foliar fertilization is carried out every 10 days. The leaf fertilizer comprises the following components: an mg/L potassium nitrate, a 150 mg/L potassium dihydrogen phosphate, a 15 mg/L sodium nitrophenolate, a5 mg/L ethephon and a 0.5 mg/L boric acid.
The method of the invention sets a new basic culture medium for the gerbera jamesonii, and the basic culture medium is adopted to be matched with the plant growth hormone, so that the culture efficiency and the tissue culture success rate can be effectively improved, the time required by each step is effectively shortened, the planting survival rate is high, the time required from planting to flowering is short, and the single plant flowering rate is high.
Detailed Description
The present invention is described in further detail below with reference to specific examples.
And at the implementation place, tissue culture of the fast-propagation gerbera jamesonii tissue culture seedlings is finished in a tissue culture room of Yibin institute of occupational technology, and the seedlings are fixedly planted in a plastic greenhouse.
Wherein the components of the culture medium A in the embodiment are as follows: potassium nitrate 100 mg/L, potassium chloride 50 mg/L, four water calcium nitrate 300 mg/L, seven water magnesium sulfate 750 mg/L, sodium sulfate 200 mg/L, sodium dihydrogen phosphate 20 mg/L, four water manganese sulfate 7 mg/L, ferrous sulfate 2.5 mg/L, seven water zinc sulfate 3 mg/L, boric acid 1.5 mg/L, potassium iodide 0.7 mg/L, nicotinic acid 0.5 mg/L, hydrochloric acid pyridoxine 0.3 mg/L, thiamine hydrochloride 0.1 mg/L, glycine 10 mg/L, glucose 50g/L, agar powder 10 g/L; the rooting culture takes a culture medium B as a culture medium. The components of the B culture medium are consistent with those of the A culture medium, but the concentration of each component is 1/2 of the A culture medium.
The comparative example used MS medium.
Example 1: a method for rapid propagation culture and annual fresh flower production of African daisy comprises the following steps:
1. selecting excellent single plants which are pure in red, large in flower and robust in growth from the introduced and cultivated African daisy, shearing 1-2 cm of stem tip as explants, flushing the explants for 5 minutes with tap water, soaking the explants for 15 minutes on a cleaning workbench with 70% alcohol, treating the explants for 10 minutes with 0.1% mercuric chloride and 0.5% Tween 20, flushing the explants for 5 times with sterile water, and inoculating the treated explants in a primary culture medium.
2. 1-2 adventitious buds grow on the explant, and then the explant is transferred to a new primary culture medium for culture.
3. The adventitious bud grows to 2-3 cm, the bud is cut from the explant and cut to be transferred to a subculture medium for rapid propagation, and a certain number of buds can be obtained after 6 successive generations.
4. Cutting a single plant with the seedling height of 5cm, and transferring the single plant into a rooting culture medium for rooting culture. About 10 days, 2-5 adventitious roots grow on 98% of the base parts of the seedlings.
5. And taking out the plantlets, washing the plantlets in clear water to remove residual culture, temporarily planting the plantlets in a seedling hardening matrix, and planting new roots and leaves in a plastic greenhouse after about 30-40 days, wherein the seedling hardening matrix is formed by mixing leaf mold, perlite, mushroom bran and vermiculite according to the weight part ratio of 5:2:2: 1.
And (3) maintaining and managing after field planting:
1. fertilizing: fertilizing is started after planting for one month, biogas liquid fertilizer is mainly applied, direct root irrigation is adopted, and as the seedlings grow, fertilization per plant is gradually increased to 2000mL from 500mL of seedlings, and 2-3 times per month.
2. And (3) promoting flower bud differentiation: after planting for 60 days, carrying out foliar fertilizer application every 10 days, wherein the foliar fertilizer comprises the following components: an mg/L potassium nitrate, a 150 mg/L potassium dihydrogen phosphate, a 15 mg/L sodium nitrophenolate, a5 mg/L ethephon and a 0.5 mg/L boric acid.
Primary culture medium: the A medium +6KT5 mg/L + adenine 2 mg/L + IBA2 mg/L; subculture medium: the A medium +6KT3-5 mg/L + adenine 1-2 mg/L + IBA1.5-2 mg/L; rooting culture medium: b minimal medium + IBA5 mg.
Example 2: a method for rapid propagation culture and annual fresh flower production of African daisy comprises the following steps:
1. selecting excellent single plants which are pure in red, large in flower and robust in growth from the introduced and cultivated African daisy, shearing 1-2 cm of stem tip as explants, flushing the explants for 5 minutes with tap water, soaking the explants for 15 minutes on a cleaning workbench with 70% alcohol, treating the explants for 10 minutes with 0.1% mercuric chloride and 0.5% Tween 20, flushing the explants for 5 times with sterile water, and inoculating the treated explants in a primary culture medium.
2. 1-2 adventitious buds grow on the explant, and then the explant is transferred to a new primary culture medium for culture.
3. The adventitious bud grows to 2-3 cm, the bud is cut from the explant and cut to be transferred to a subculture medium for rapid propagation, and a certain number of buds can be obtained after 6 successive generations.
4. Cutting a single plant with the seedling height of 5cm, and transferring the single plant into a rooting culture medium for rooting culture. About 10 days, 2-5 adventitious roots grow on 98% of the base parts of the seedlings.
5. And taking out the plantlets, washing the plantlets in clear water to remove residual culture, temporarily planting the plantlets in a seedling hardening matrix, and planting new roots and leaves in a plastic greenhouse after about 30-40 days, wherein the seedling hardening matrix is formed by mixing leaf mold, perlite, mushroom bran and vermiculite according to the weight part ratio of 5:2:2: 1.
And (3) maintaining and managing after field planting:
1. fertilizing: fertilizing is started after planting for one month, biogas liquid fertilizer is mainly applied, direct root irrigation is adopted, and as the seedlings grow, fertilization per plant is gradually increased to 2000mL from 500mL of seedlings, and 2-3 times per month.
2. And (3) promoting flower bud differentiation: after 60 days of planting, foliar fertilization is carried out every 10 days.
Primary culture medium: primary culture medium: the A medium +6KT5 mg/L + adenine 2 mg/L + IBA2 mg/L; subculture medium: the A medium +6KT3-5 mg/L + adenine 1-2 mg/L + IBA1.5-2 mg/L; rooting culture medium: b minimal medium + IBA5 mg.
Example 3: a method for rapid propagation culture and annual fresh flower production of African daisy comprises the following steps:
1. selecting excellent individual plants which are pure in red, large in flower and strong in growth from the introduced and cultivated African daisy, and inoculating the individual plants in a primary culture medium after conventional disinfection.
2. 1-2 adventitious buds grow on the explant, and then the explant is transferred to a new primary culture medium for culture.
3. The adventitious bud grows to 2-3 cm, the bud is cut from the explant and cut to be transferred to a subculture medium for rapid propagation, and a certain number of buds can be obtained after 6 successive generations.
4. Cutting a single plant with the seedling height of 5cm, and transferring the single plant into a rooting culture medium for rooting culture. About 10 days, 2-5 adventitious roots grow on 98% of the base parts of the seedlings.
5. Taking out plantlets, washing out residual culture in clear water, temporarily planting in a seedling hardening matrix, and planting in a plastic greenhouse after new roots and leaves grow out in about 30-40 days.
And maintaining and managing after planting by adopting a conventional method.
Primary culture medium: a, a culture medium; subculture medium: a, a culture medium; rooting culture medium: b, basic culture medium.
Comparative example 1: basically the same as example 1, except that: and replacing the culture medium A and the culture medium B with MS culture medium.
Comparative example 2: basically the same as example 2, except that: and replacing the primary culture medium, the secondary culture medium and the rooting culture medium with an MS culture medium.
Comparative example 3: basically the same as example 3, except that: MS culture medium is adopted.
By monitoring the examples and comparative examples, the following table is obtained:
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 present invention, and it will be apparent to those skilled in the art that various modifications and variations can be made in the spirit and principle of the present invention and that equivalent modifications and substitutions and the like are included in the scope of the present invention.
Claims (3)
1. A method for rapid propagation culture and annual fresh flower production of African daisy is characterized by comprising the following steps: primary culture, secondary culture, rooting culture and transplanting and field planting;
the primary culture is to culture 1-2 adventitious buds growing on the explant and transfer the explant to a new primary culture medium;
the subculture is to cut off and cut off the bud from the explant and transfer the bud to a subculture medium for rapid propagation after the adventitious bud grows to 2-3 cm, and the continuous subculture is more than 5 generations;
the rooting culture is that after the number of subcultures is enough, 3-5cm of single plants obtained by the subcultures are cut off and transferred to a rooting culture medium for rooting culture until 2-5 adventitious roots grow on the base of the plantlet;
the transplanting and field planting is that after the residual culture solution is removed from the plantlets obtained by rooting culture, the plantlets are temporarily planted in a seedling hardening matrix, and new roots and leaves are grown for field planting; and (3) maintenance management is required after planting, wherein the maintenance management comprises fertilization and flower bud differentiation promotion:
fertilizing: fertilizing is started after planting for one month, biogas liquid fertilizer is mainly applied, direct root irrigation is adopted, and as the seedlings grow, fertilization per plant is gradually increased to 2000mL from 500mL of seedlings to 2-3 times per month; and (3) promoting flower bud differentiation: after planting for 60 days, carrying out foliar fertilizer application every 10 days, wherein the foliar fertilizer comprises the following components: a potassium nitrate is 500 mg/L, a potassium dihydrogen phosphate is 150 mg/L, a sodium nitrophenolate is 15 mg/L, an ethephon is 5 mg/L, and boric acid is 0.5 mg/L;
the primary medium comprises the components of a medium A, 6-furfuryl aminopurine, adenine and indolebutyric acid, and the added 6-furfuryl aminopurine is 5 mg/L, adenine is 2 mg/L and indolebutyric acid is 2 mg/L;
the components of the subculture medium are A medium, 6-furfuryl aminopurine, adenine and indolebutyric acid, and the 6-furfuryl aminopurine is added for 3-5 mg/L, adenine is added for 1-2 mg/L and indolebutyric acid is added for 1.5-2 mg/L; the rooting medium comprises a culture medium B and indolebutyric acid, and 5 mg/L of indolebutyric acid is added;
the A culture medium comprises the following components: potassium nitrate 100 mg/L, potassium chloride 50 mg/L, four water calcium nitrate 300 mg/L, seven water magnesium sulfate 750 mg/L, sodium sulfate 200 mg/L, sodium dihydrogen phosphate 20 mg/L, four water manganese sulfate 7 mg/L, ferrous sulfate 2.5 mg/L, seven water zinc sulfate 3 mg/L, boric acid 1.5 mg/L, potassium iodide 0.7 mg/L, nicotinic acid 0.5 mg/L, hydrochloric acid pyridoxine 0.3 mg/L, thiamine hydrochloride 0.1 mg/L, glycine 10 mg/L, glucose 50g/L, agar powder 10 g/L;
the components of the B culture medium are consistent with those of the A culture medium, but the concentration of each component is 1/2 of the A culture medium.
2. The method for rapid propagation culture and annual flower production of African daisy according to claim 1, wherein: selecting 1-2 cm of excellent single plant stem tip with pure color and robust growth as an explant, flushing the explant for 5 minutes by tap water, soaking the explant for 15 minutes by 70% alcohol on a cleaning workbench, treating the explant for 10 minutes by 0.1% mercuric chloride and 0.5% tween 20, and flushing the explant by sterile water for primary culture.
3. The method for rapid propagation culture and annual flower production of African daisy according to claim 1, wherein: the hardening-seedling substrate is formed by mixing leaf mold, perlite, mushroom bran and vermiculite according to the weight ratio of 5:2:2: 1.
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