CN115777531A - Ginger tissue culture seedling method - Google Patents
Ginger tissue culture seedling method Download PDFInfo
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- CN115777531A CN115777531A CN202211134268.5A CN202211134268A CN115777531A CN 115777531 A CN115777531 A CN 115777531A CN 202211134268 A CN202211134268 A CN 202211134268A CN 115777531 A CN115777531 A CN 115777531A
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- 235000006886 Zingiber officinale Nutrition 0.000 title claims abstract description 116
- 235000008397 ginger Nutrition 0.000 title claims abstract description 116
- 238000000034 method Methods 0.000 title claims abstract description 21
- 244000273928 Zingiber officinale Species 0.000 title 1
- 241000234314 Zingiber Species 0.000 claims abstract description 115
- 239000001963 growth medium Substances 0.000 claims abstract description 76
- 239000011669 selenium Substances 0.000 claims abstract description 59
- 229910052711 selenium Inorganic materials 0.000 claims abstract description 59
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims abstract description 58
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 42
- 238000012258 culturing Methods 0.000 claims abstract description 26
- 238000004140 cleaning Methods 0.000 claims abstract description 11
- 238000005286 illumination Methods 0.000 claims description 40
- 239000011159 matrix material Substances 0.000 claims description 14
- 238000001784 detoxification Methods 0.000 claims description 13
- 239000002689 soil Substances 0.000 claims description 13
- 230000012010 growth Effects 0.000 claims description 12
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 claims description 11
- 230000035784 germination Effects 0.000 claims description 10
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 7
- 239000000758 substrate Substances 0.000 claims description 7
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims description 6
- 235000019743 Choline chloride Nutrition 0.000 claims description 6
- 229960003178 choline chloride Drugs 0.000 claims description 6
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 6
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- 230000001954 sterilising effect Effects 0.000 claims description 5
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- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 2
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- 230000002354 daily effect Effects 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
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- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
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- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
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- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a ginger tissue culture seedling method, which comprises the following steps: cleaning a ginger explant by using water, and detoxifying the ginger explant by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecules is 10-30 mug/L; in the process of culturing tissue culture seedlings, at least 10-30 mug/L selenium trace water molecule elements are added into an N6 culture medium, and finally domestication and outplanting are carried out. Compared with the prior art, the method has the advantages that the selenium-rich micromolecular water is used for detoxifying the ginger explants, and the selenium trace water molecular element is added into the culture medium, so that the antiviral capability of the ginger in the seedling stage is improved, the harm of ginger blast is fundamentally solved, and the cultivated ginger seedlings are beneficial to producing the selenium-rich ginger in the future; meanwhile, the nutrient components with specific concentration and types, the plant growth regulator and a specific culture environment are combined, so that the cultured ginger seedlings are high in differentiation capacity, high in propagation speed and high in propagation coefficient.
Description
Technical Field
The invention relates to a plant seedling raising technology, in particular to a ginger seedling tissue culture method.
Background
According to the traditional ginger seedling raising method, ginger blocks are buried in soil to bud, and germs such as soft rot and root rot are usually hidden in ginger peel or ginger flesh, so that the aim of sterilization is difficult to achieve by fumigating soil. The bacterial-carrying plants infect the ginger field soil in a long-term and long-term manner, and the bacterial-carrying plants carry a large amount of bacteria, so that the infection rate of ginger bacterial wilt (the sum of soft rot and root rot) is high and is about 10-50%, and the survival rate of ginger seedlings is low and is about 50-70%. And the recurrence rate of the ginger blast is high, the land can not be continuously cultivated, and ginger farmers talk about the blast color change.
In recent years, the development of ginger seedling culture using tissue culture technology has been studied to overcome the above-mentioned drawbacks of the conventional ginger planting method, but further improvement in yield and detoxification has been desired.
Selenium is a strong immunomodulator, almost every kind of immune cells in a human body contains selenium, and selenium supplement can enhance the humoral immune function, the cellular immune function and the non-specific immune function of the human body, so that the disease resistance of the body is integrally enhanced. However, 40 countries around the world are in selenium-deficient areas, 70% of China is in selenium deficiency, and countries with serious selenium deficiency are in countries with 22 provinces and hundreds of millions of people in selenium-deficient or low-selenium areas, and the incidence of tumors, liver diseases, cardiovascular diseases and the like in the population of the areas is high.
The selenium deficiency can be supplemented through two ways of food supplement and medicine supplement. Selenium supplementation with food is a more efficient and superior option for most households. However, selenium is mostly present in animal foods; for a large amount of edible grains and vegetables, only a small amount of selenium-rich products such as selenium-rich rice and potatoes limited to production places are available on the market, the yield is low, the price is high, and the selenium-rich products cannot enter daily dining tables of thousands of households. However, ginger is a daily conventional food material for families, and even ordinary families with a slightly higher price can eat ginger regularly, so that the selenium-rich ginger is a good entry point for introducing selenium into daily dining tables of thousands of households.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a ginger seedling tissue culture method aiming at the defects in the prior art, and apply a biological tissue culture technology to ginger seedling culture to obtain ginger seedlings which are free of virus and can grow selenium-enriched ginger.
The technical scheme adopted by the invention for solving the technical problems is as follows:
the tissue culture seedling method of the ginger is characterized by comprising the following steps:
(1) Accelerating germination, namely selecting strong and strong ginger tubers without rot or diseases, placing the ginger tubers in a non-illumination room, maintaining the temperature at 24-28 ℃ and the humidity at 50-60%, and growing the explants of the ginger buds to 2-3cm;
(2) Detoxification, namely cleaning the ginger explant obtained in the step (1) with water, and performing detoxification treatment on the ginger explant by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecule sterilized water is 10-30 mug/L;
(3) Culturing tissue culture seedling, the first stage, the explant pregermination initial stage, peeling off the stem tip of the ginger explant in the step (2), inoculating the peeled stem tip onto the first culture medium, placing at 24-28 deg.C, irradiating for 6-8 hr per day with illumination intensity of 650-850 μmol/m 2 Culturing for 5-7 days in a culture environment of/s, wherein the first culture medium is N6 culture medium added with at least 2.5mg/L of 6-benzylaminopurine and 10-30 mu g/L of selenium trace water molecular element;
in the second stage, during the primary growth period of leaf and stem, the ginger seedling obtained in the first stage is inoculated to a second culture medium for culture, and the second culture medium is placed at the temperature of 24-28 ℃, the illumination is 8-10 hours per day, and the illumination intensity is 1000 mu mol/m 2 Culturing for 7-10 days in a culture environment of/s; the second culture medium is N6 culture medium added with at least 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid and 10-30 mug/L selenium trace water molecular element;
in the third stage, during the growth period of root hair, the ginger seedlings obtained in the second stage are inoculated into a third culture medium, and are placed at the temperature of 26-29 ℃, the illumination time of 10-12 hours per day and the illumination intensity of 600-800 mu mol/m 2 Culturing for 30-45 days in a culture environment of/s; the third culture medium is N6 culture medium added with at least 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid, 200mg/L choline chloride and 10-30 mug/L selenium trace water molecular element;
(4) Domesticating the tissue culture seedlings.
Further, the method comprises the following steps:
inoculating the ginger seedlings subjected to the second stage in the step (3) into a new second culture medium for subculture.
The step (4) comprises the following steps:
➀ in-bottle acclimation
Placing the ginger seedlings with developed roots obtained in the step 3 in a sterile seedling exercising room, wherein the illumination intensity is 2-2.8 Mo Leke s, unscrewing the bottle caps on the first day, uncovering half of the bottle caps on the second day, uncovering the bottle caps on the third day, taking the ginger seedlings out of the bottles after 5-10 days, and cleaning to remove the culture medium;
➁ seedling domestication in tray
Planting clean ginger seedlings in a seedling tray matrix which is thoroughly watered, then placing the seedling tray in a greenhouse covered with a shading net, arching with bamboo, covering with a film, keeping the temperature in the film at 20-28 ℃, the humidity at 60-80%, gradually uncovering the film after the seedlings grow new leaves, and keeping the illumination intensity in the greenhouse at 2-3 Mo Leke s.
The film is uncovered firstly, two ends of the film are ventilated, then, half of the film is uncovered for ventilation on the next day, and the film is completely uncovered on the third day.
After the film is completely uncovered, water-soluble rooting bacterial manure is applied for one week, and then ammonium sulfate, calcium, magnesium and zinc fertilizers are sprayed every week.
The substrate is prepared from 60% of peat soil 6, 20% of perlite and 20% of thoroughly decomposed dry chicken manure by weight.
Sterilizing the ginger seedlings before planting in a seedling tray matrix, and mixing 20 kilograms of fine soil with 0.5 kilogram of 70% dike pine powder, or uniformly mixing 8-10 grams of 50% carbendazim and a proper amount of soil per square meter of a seedling bed for broadcasting, or spraying 50 kilograms of 25% daphniphora and 50 kilograms of water on 1000 kilograms of matrix while stirring.
The shading rate of the shading net is 60%.
Selecting 50-hole seedling raising trays within 30 days of acclimatization in the seedling tray, and selecting 32-hole seedling raising trays within more than one month of acclimatization in the seedling tray.
Compared with the prior art, the invention has the following beneficial effects:
the explant of the ginger is detoxified by selenium-rich micromolecular water, and meanwhile, a selenium trace water molecular element is added into a culture medium, so that the antiviral capability of the ginger in the seedling stage is improved, the harm of ginger blast is fundamentally solved, and the cultivated ginger seedlings are beneficial to producing the selenium-rich ginger in the future.
The method uses nutrient components and plant growth regulators with specific concentrations and types, and combines a specific culture environment, so that the cultured ginger seedlings have strong differentiation capacity, high propagation speed and large propagation coefficient. Hundreds of millions of ginger seedlings can be produced in a culture factory with 700-800 square meters in one year, and high production benefit can be obtained.
5000 ginger seedlings cultivated by the method are planted in each mu of land, under a proper planting climate, the weight of each root and stem is kept about 1 kg, the yield per mu can reach more than 15000 jin, and the benefit per mu is high.
Detailed Description
The preferred embodiments of the present invention will now be described in detail.
1. Accelerating germination, namely selecting strong and strong ginger tubers without rot or diseases, placing the ginger tubers in a non-illumination room, maintaining the temperature at 24-28 ℃ and the humidity at 50-60%, and growing the explants of the ginger buds to 2-3cm;
2. detoxification, namely cleaning the ginger explant obtained in the step 1 with water, and performing detoxification treatment on the ginger explant by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecules is 10-30 mug/L;
3. culturing tissue culture seedling, the first stage, explant germination accelerating initial stage, peeling stem tip from the ginger explant in step 2, inoculating the peeled stem tip onto the first culture medium, placing at 24-28 deg.C, and illuminating at 650-850 μmol/m for 6-8 hr per day 2 Culturing for 5-7 days in a culture environment of/s to obtain a sterile seedling of 5-8 mm, wherein the first culture medium is N6 culture medium added with 2.5mg/L of 6-benzylaminopurine and 10-30 mu g/L of selenium trace water molecular element;
in the second stage, during the primary growth period of leaves and stems, the aseptic seedling obtained in the first stage is inoculated to a second culture medium for culture, and the second culture medium is placed at the temperature of 24-28 ℃, the illumination is 8-10 hours per day, and the illumination intensity is 1000 mu mol/m 2 Culturing in culture environment for 7-10 days to obtain 30-60mm rhizoma Zingiberis recens seedling; the second culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid and 10-30 mug/L selenium trace water molecular element;
in the third stage, during the growth period of root hair, the ginger seedlings obtained in the second stage are inoculated into a third culture medium, and are placed at the temperature of 26-29 ℃, the illumination time of 10-12 hours per day and the illumination intensity of 600-800 mu mol/m 2 Culturing for 30-45 days in a culture environment of/s; obtaining ginger seedlings with root length of 2-3cm and plant height of 60-80 mm; the third culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine and 0.1mg/L naphthyl ethylAcid, 10-30 mug/L, 200mg/L choline chloride and 10-30 mug/L selenium trace water molecular element;
4. domesticating the tissue culture seedlings.
1) Hardening seedlings in bottles
Firstly, hardening seedlings in a sterile seedling hardening room for 5 to 10 days. The illumination intensity is required to be 2-2.8 Mo Leke s.
The bottle cap is unscrewed and placed in a seedling hardening room on the first day, half of the bottle cap is uncovered on the second day, and the bottle cap is completely uncovered on the third day. After 5-10 days, the bottle seedling is taken out from the bottle and cleaned with the culture medium, and no culture medium is left on the roots and seedlings. The cleaned ginger seedlings are dried indoors for planting. Transplanting is generally carried out the next day, and if the transplanting is carried out for several days, moisture preservation is required.
2) Hardening off seedling with seedling tray substrate
(1) Preparing a matrix: peat soil 6: perlite 2: preparing decomposed dry chicken manure 2; or purchasing a prepared vegetable seedling raising substrate or rice seedling raising substrate.
(2) Preparing a seedling raising tray: 50-hole seedling raising trays are selected within 30 days of planned hardening (32-hole seedling raising trays are selected with the seedlings left in the trays for more than one month).
(3) Film and bamboo chip preparation: agricultural films 2 m wide and bamboo chips 1.8 m long were prepared.
(4) Matrix sterilization: mixing 0.5 kg of 70% dikesong powder with 20 kg of fine soil; or uniformly mixing 8-10 g of 50% carbendazim and a proper amount of soil per square meter of seedbed and spreading; or spraying 50 g of 25% daphniphora urensis with 50 kg of water on 1000 kg of matrix while stirring.
(5) Well laying a seedling raising tray on the ground, applying the prepared substrate, thoroughly watering and sterilizing the substrate one day in advance for later use.
(6) Covering a shading net on the greenhouse for hardening the seedlings by the matrix, wherein the shading net needs to cover the greenhouse with shading of 60 percent, and the illumination intensity in the greenhouse is ensured to be kept between 2 ten thousand and 3 Mo Leke s.
(7) Planting cleaned ginger seedling in water-permeable matrix, watering root-fixing water, and finally arching with bamboo and covering with film. Keeping the temperature in the film at 20-28 deg.C and humidity at 60-80% for about 10 days, and gradually uncovering the film after new leaves grow. The film is uncovered firstly, two ends of the film are ventilated, then, half of the film is uncovered for ventilation on the next day, and the film is completely uncovered on the third day. After the film is removed, the soil is watered properly according to the moisture condition.
(8) Fertilizing: and (3) watering and applying the water-soluble rooting bacterial fertilizer about one week after the film is completely uncovered, and then spraying ammonium sulfate, calcium magnesium zinc fertilizer every week, wherein the application concentration is 0.5-1%.
The formula of the N6 culture medium is as follows (unit mg/L):
potassium nitrate (KNO) 3 】2830，
Ammonium sulfate [ NH ] 4 )2SO 4 】463，
Magnesium sulfate [ MgSO 4 4 ·7H 2 O】185，
Potassium dihydrogen phosphate [ KH ] 2 PO 4 】400，
Calcium chloride [ CaCl ] 2 ·2H 2 O】166，
Manganese sulfate [ MnSO ] 4 ·4H 2 O】4.4，
Zinc sulfate [ ZnSO ] 4 ·7H 2 O】1.5 mg/L，
Potassium iodide [ KI ] 0.8,
boric acid [ H ] 3 BO 3 】1.6，
The content of inositol 100 in the extract is as follows,
2.0 parts of glycine,
nicotinic acid [ VB3 ] 0.5,
pyridoxine hydrochloride [ VB6 ] 0.5,
thiamine hydrochloride [ VB1 ] 0.1,
ferrous sulfate [ FeSO ] 4 ·7H 2 O】27.8，
Sodium edetate 37.3.
Example one
A tissue culture seedling method of ginger comprises the following steps:
1. accelerating germination, namely selecting strong and strong ginger tubers without rot or diseases, placing the ginger tubers in a non-illumination room, maintaining the temperature at 24 ℃ and the humidity at 50%, and allowing the explants of the ginger buds to grow to 2cm;
2. detoxification, namely cleaning the ginger explant obtained in the step 1 with water, and performing detoxification treatment on the ginger explant by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecules is 10 microgram/L;
3. culturing tissue culture seedling, the first stage, accelerating the germination of explantIn the period, the ginger explant obtained in the step 2 is stripped of the stem tip, the stripped stem tip is inoculated to a first culture medium, and the ginger explant is placed at the temperature of 24 ℃, is irradiated for 8 hours every day and has the irradiation intensity of 650 mu mol/m 2 Culturing for 5 days in a culture environment of/s to obtain a sterile seedling of 6 mm, wherein the first culture medium is N6 culture medium added with 2.5mg/L of 6-benzylaminopurine and 10 mu g/L of selenium trace water molecular element;
in the second stage, during the primary growth period of leaves and stems, the aseptic seedling obtained in the first stage is inoculated to a second culture medium for culture, and the second culture medium is placed at the temperature of 24 ℃, the illumination time is 8 hours per day, and the illumination intensity is 1000 mu mol/m 2 Culturing for 7 days in a culture environment of/s to obtain ginger seedlings with the diameter of 35 mm; the second culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid and 10 mug/L selenium trace water molecular element;
in the third stage, during the growth period of root hair, the ginger seedlings obtained in the second stage are inoculated into a third culture medium, and are placed at the temperature of 26 ℃, the illumination is 10 hours per day, and the illumination intensity is 600 mu mol/m 2 Culturing for 30 days in a culture environment of/s; obtaining ginger seedlings with the root length of 2.2 cm and the plant height of 65 mm; the third culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid, 200mg/L choline chloride and 10 mug/L selenium trace water molecular element;
4. domesticating the tissue culture seedlings.
Hardening seedlings in a sterile seedling hardening chamber bottle for 10 days, wherein the illumination intensity is 2 Mo Leke s. Then taking out the bottle seedling from the bottle, cleaning the culture medium, and putting the bottle seedling into a seedling tray matrix for hardening the seedling.
Example two
1. Accelerating germination, namely selecting strong and healthy ginger tubers without rot or diseases, placing the ginger tubers in a non-illumination room, maintaining the temperature at 26 ℃ and the humidity at 55%, and allowing the explants of the ginger buds to grow to 2.5cm;
2. detoxification, namely cleaning the ginger explants obtained in the step 1 by using water, and performing detoxification treatment on the ginger explants by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecules is 20 microgrammes per liter;
3. culturing tissue culture seedling, the first stage, explant germination accelerating initial stage, peeling off stem tip of ginger explant obtained by the step 2, peeling offInoculating the separated stem tip to the first culture medium, and standing at 26 deg.C under illumination intensity of 700 μmol/m for 7 hr per day 2 Culturing for 6 days in a culture environment of/s to obtain a 6.5mm sterile seedling, wherein the first culture medium is N6 culture medium added with 2.5mg/L of 6-benzylaminopurine and 20 mu g/L of selenium trace water molecular element;
in the second stage, during the primary growth period of leaves and stems, the aseptic seedling obtained in the first stage is inoculated to a second culture medium for culture, and the second culture medium is placed at the temperature of 26 ℃, the illumination time of 9 hours per day and the illumination intensity of 1000 mu mol/m 2 Culturing in a culture environment for 8 days to obtain ginger seedlings of 40 mm; the second culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid and 20 mug/L selenium trace water molecular element;
in the third stage, during the growth period of root hair, the ginger seedlings obtained in the second stage are inoculated into a third culture medium, and the third culture medium is placed at the temperature of 27 ℃ and the illumination intensity of 700 mu mol/m for 11 hours per day 2 Culturing for 35 days in a culture environment of/s; obtaining ginger seedlings with root length of 2.5cm and plant height of 70 mm; the third culture medium is N6 culture medium added with 6-benzylaminopurine, 0.1mg/L naphthylacetic acid, 200mg/L choline chloride and 20 microgram of selenium water molecular element;
4. domesticating the tissue culture seedlings.
Hardening seedlings in a sterile seedling hardening chamber bottle for 8 days, wherein the illumination intensity is 2.2 Mo Leke s. Then taking out the bottle seedling from the bottle, cleaning the culture medium, and putting the bottle seedling into a seedling tray matrix for hardening the seedling.
EXAMPLE III
1. Accelerating germination, namely selecting strong and strong ginger tubers without rot or diseases, placing the ginger tubers in a non-illumination room, maintaining the temperature at 28 ℃ and the humidity at 60%, and allowing the explants of the ginger buds to grow to 3cm;
2. detoxification, namely cleaning the ginger explant obtained in the step 1 by using water, and performing detoxification treatment on the ginger explant by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecules is 120 microgram/L;
3. culturing tissue culture seedling, the first stage, explant germination accelerating initial stage, peeling off stem tip of the ginger explant obtained in step 2, inoculating the peeled stem tip onto the first culture medium, placing at 28 deg.C, and culturing at every timeThe light irradiation is carried out for 6 hours in day, and the light intensity is 850 mu mol/m 2 Culturing for 7 days in a culture environment of/s to obtain a 7mm sterile seedling, wherein the first culture medium is N6 culture medium added with 2.5mg/L of 6-benzylaminopurine and 30 mu g/L of selenium trace water molecular elements;
in the second stage, during the primary growth period of leaves and stems, the aseptic seedling obtained in the first stage is inoculated to a second culture medium for culture, and the second culture medium is placed at the temperature of 28 ℃, the illumination time is 10 hours per day, and the illumination intensity is 1000 mu mol/m 2 Culturing in a culture environment for 10 days to obtain 60mm ginger seedlings; the second culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid and 30 mug/L selenium trace water molecular element;
in the third stage, during the growth period of root hair, the ginger seedlings obtained in the second stage are inoculated into a third culture medium, and the third culture medium is placed at the temperature of 29 ℃ and the illumination intensity of 800 mu mol/m for 12 hours per day 2 Culturing for 45 days in a culture environment of/s; obtaining ginger seedlings with the root length of 3cm and the plant height of 80 mm; the third culture medium is N6 culture medium added with 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid, 200mg/L choline chloride and 30 mug/L selenium trace water molecular element;
4. domesticating the tissue culture seedlings.
Hardening seedlings in a sterile seedling hardening chamber bottle for 5 days, wherein the illumination intensity is 2.8 Mo Leke s. Then the bottle seedlings are taken out from the bottle and cleaned, and the culture medium is placed into the seedling tray matrix for seedling hardening.
In the above embodiment, the ginger seedlings passing through the second stage of step 3 do not immediately enter the second stage, i.e. the root hair growth stage, but are inoculated into a new second culture medium for subculture as required.
The biological cell division tissue culture technology has the following advantages:
the plant differentiation capacity is strong, the propagation speed is high, and the propagation coefficient is large;
the complete plant without virus is obtained, the cost is reduced, the variety advantage is kept, most importantly, the antiviral capability of the ginger in the seedling stage is improved, and the harm of ginger blast is fundamentally solved.
It should be understood that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the same, and those skilled in the art can modify the technical solutions described in the above embodiments, or make equivalent substitutions for some technical features; and such modifications and substitutions are intended to be included within the scope of the appended claims.
Claims (9)
1. A tissue culture seedling method of ginger is characterized by comprising the following steps:
(1) Accelerating germination, namely selecting strong ginger tubers without corrosion and diseases, placing the strong ginger tubers in a non-illumination room, maintaining the temperature at 24-28 ℃ and the humidity at 50-60%, and growing the explants of the ginger buds to 2-3cm;
(2) Detoxification, namely cleaning the ginger explant obtained in the step (1) by using water, and performing detoxification treatment on the ginger explant by using selenium-rich micromolecule water, wherein the content of selenium in the selenium-rich micromolecules is 10-30 microgram/L;
(3) Culturing tissue culture seedling, the first stage, the explant pregermination initial stage, peeling off the stem tip of the ginger explant in the step (2), inoculating the peeled stem tip onto the first culture medium, placing at 24-28 deg.C, irradiating for 6-8 hr per day with illumination intensity of 650-850 μmol/m 2 Culturing for 5-7 days in a culture environment of/s, wherein the first culture medium is N6 culture medium added with at least 2.5mg/L of 6-benzylaminopurine and 10-30 mu g/L of selenium trace water molecular element;
in the second stage, during the primary growth period of leaf and stem, the ginger seedling obtained in the first stage is inoculated to a second culture medium for culture, and the second culture medium is placed at the temperature of 24-28 ℃, the illumination is 8-10 hours per day, and the illumination intensity is 1000 mu mol/m 2 Culturing for 7-10 days in a culture environment of/s; the second culture medium is N6 culture medium added with at least 2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid and 10-30 mug/L selenium trace water molecular element;
in the third stage, during the growth period of root hair, the ginger seedlings obtained in the second stage are inoculated into a third culture medium, and are placed at the temperature of 26-29 ℃, the illumination time of 10-12 hours per day and the illumination intensity of 600-800 mu mol/m 2 Culturing for 30-45 days in a culture environment of/s; the third culture medium is N6 culture medium added with at least2.5 mg/L6-benzylaminopurine, 0.1mg/L naphthylacetic acid, 200mg/L choline chloride and 10-30 mu g/L selenium trace water molecular element;
(4) Domesticating the tissue culture seedlings.
2. The tissue culture seedling raising method of ginger as claimed in claim 1, wherein the ginger seedlings after the second stage of step (3) are inoculated into a new second culture medium for subculture.
3. The tissue culture seedling raising method for ginger according to claim 1, wherein the step (4) comprises:
➀ in-bottle acclimation
Placing the ginger seedlings finally obtained in the step (3) in a sterile seedling exercising room, wherein the illumination intensity is 2-2.8 Mo Leke s, unscrewing the bottle caps on the first day, uncovering half of the bottle caps on the second day, uncovering the bottle caps on the third day completely, taking the ginger seedlings out of the bottles after 5-10 days, and cleaning to remove the culture medium;
➁ seedling domestication in tray
Planting clean ginger seedlings in a seedling tray matrix which is thoroughly watered, then placing the seedling tray in a greenhouse covered with a shading net, arching with bamboo, covering with a film, keeping the temperature in the film at 20-28 ℃, the humidity at 60-80%, gradually uncovering the film after the seedlings grow new leaves, and keeping the illumination intensity in the greenhouse at 2-3 Mo Leke s.
4. The tissue culture seedling method of ginger according to claim 3, characterized in that: the film is uncovered firstly, two ends of the film are ventilated, then, half of the film is uncovered for ventilation on the next day, and the film is completely uncovered on the third day.
5. The tissue culture seedling method of ginger according to claim 4, characterized in that: after the film is completely uncovered, water-soluble rooting bacterial manure is applied for one week, and then ammonium sulfate, calcium, magnesium and zinc fertilizers are sprayed every week.
6. The tissue culture seedling method of ginger according to claim 3, characterized in that: the substrate is prepared from 60% of peat soil 6, 20% of perlite and 20% of decomposed dry chicken manure by weight ratio.
7. The tissue culture seedling method of ginger according to claim 6, characterized in that: sterilizing the ginger seedlings before planting in a seedling tray matrix, and mixing 20 kilograms of fine soil with 0.5 kilogram of 70% dike pine powder, or uniformly mixing 8-10 grams of 50% carbendazim and a proper amount of soil per square meter of a seedling bed for broadcasting, or spraying 50 kilograms of 25% daphniphora and 50 kilograms of water on 1000 kilograms of matrix while stirring.
8. The tissue culture seedling method of ginger according to claim 3, characterized in that: the shading rate of the shading net is 60%.
9. The tissue culture seedling raising method for ginger according to claim 3, characterized in that: selecting 50-hole seedling raising trays within 30 days of acclimation in the seedling trays, and selecting 32-hole seedling raising trays within more than one month of acclimation in the seedling trays.
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