CN111109081A - Lycoris radiata rootless tissue culture method and lycoris radiata cultivation method - Google Patents
Lycoris radiata rootless tissue culture method and lycoris radiata cultivation method Download PDFInfo
- Publication number
- CN111109081A CN111109081A CN202010004489.5A CN202010004489A CN111109081A CN 111109081 A CN111109081 A CN 111109081A CN 202010004489 A CN202010004489 A CN 202010004489A CN 111109081 A CN111109081 A CN 111109081A
- Authority
- CN
- China
- Prior art keywords
- rootless
- tissue culture
- culture medium
- culture
- transplanting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
Abstract
The invention belongs to the field of tissue culture, and relates to a lycoris radicless tissue culture method and a lycoris cultivation method, wherein the lycoris radicless tissue culture method comprises the steps of inducing bulb blocks in an induction culture medium to obtain adventitious buds, the induction culture medium is based on MS and further comprises 0.5-3.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 30g/L sucrose and/or white sugar and 6-8 g/L agar, the adventitious buds are cultured in a proliferation culture medium to obtain radicless tissue culture seedlings, the proliferation culture medium is based on MS and further comprises 3.0-5.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 0-3 mg/L forchlorfenuron, 30g/L sucrose and/L white sugar, 0-1 mg/L thidiazuron and 7-8 g/L agar, and the survival rate of the lycoris radicless tissue culture is high for 1-5 times of proliferation culture.
Description
Technical Field
The invention belongs to the technical field of medicinal plant tissue culture, and particularly relates to a lycoris radiata rootless tissue culture method and a lycoris radiata cultivation method.
Background
Lycoris radiata is an important medicinal and ornamental plant in China, has bright color and strong ornamental value, contains lycorine, galanthamine and other alkaloids in bulbs, has important effects of treating myasthenia gravis, senile dementia (AD), tumor resistance, cancer resistance and the like, and has extremely high economic value. Because the lycoris radiata is low in natural propagation coefficient and not easy to fruit normally, only 1-2 seed balls can be generated by bulb-to-bulb propagation every year, and a flowering bulb can be formed in 3-5 years after planting, so that the growth period is long. At present, the propagation period cannot be greatly shortened, so that the method for greatly improving the propagation coefficient becomes an effective way for realizing the rapid propagation of the lycoris radiate, wherein the tissue culture technology is an effective measure for the rapid propagation of the lycoris radiate.
The existing tissue culture seedling hardening method adopts rooted seedlings to harden the seedlings so as to improve the survival rate of the hardened seedlings, namely, the seedlings are cultured in a rooting culture medium of a tissue culture room for a period of time, and then taken to a greenhouse for hardening and transplanting after the seedlings are rooted. For example, when researching the short-tube lycoris tissue culture and rapid propagation technology, the Happy giant Wei firstly carries out rooting culture on test-tube plantlets, and the test-tube plantlets are transplanted into a substrate after the test-tube plantlets grow into a complete root system, so that the short-tube lycoris can be rapidly propagated (the Happy giant Wei, the research on the short-tube lycoris tissue culture and rapid propagation technology, 2010,38(03): 1144-; the method comprises the steps of culturing bulblets on a culture medium containing auxin for 2-4 weeks to root, hardening seedlings for 2-3 days after rooting, taking out the seedlings, washing off agar, transferring the seedlings into a small pot containing vermiculite and sand (1:1), watering the small pot with enough water, giving weak light, and watering the small pot every 4-5 days (research on tissue culture propagation technology of the bulblets and lycoris radiata 2002(04): 46-49).
However, the seedling hardening method needs to take root by using a rooting culture medium, the seedling is taken out of the culture medium after the root is rooted and planted in a seedling hardening matrix, the culture medium attached to the root of the seedling needs to be cleaned before the seedling is planted in the matrix, the seedling is damaged and lost in the cleaning process, and the survival rate of the seedling is reduced.
Therefore, the method for the rootless tissue culture of the lycoris radiate with high survival rate is provided, thereby overcoming the defects existing in the prior art.
Disclosure of Invention
The invention aims to provide a lycoris radiata rootless tissue culture method with high survival rate, and seedlings obtained by the lycoris radiata rootless tissue culture method have the advantages of high survival rate, short rooting time, high rooting rate, good seedling consistency, simple operation and cost and time saving.
In order to achieve the above object, the present invention provides the following technical solutions;
the invention provides a method for rootless tissue culture of lycoris radiata, which comprises the following steps:
(1) inoculating the sterile bulb blocks on an induction culture medium, and performing adventitious bud induction to generate adventitious buds;
the induction culture medium takes an MS culture medium as a basic culture medium, and also comprises 0.5-3.0 mg/L of 6-benzylaminopurine, 0.5-1.5 mg/L of α -naphthylacetic acid, 30g/L of cane sugar and/or white sugar and 6-8 g/L of agar;
(2) transferring the adventitious buds to a multiplication culture medium, performing adventitious bud multiplication culture, and screening to obtain rootless tissue culture seedlings with the diameter of 0.8-1.5 cm;
the enrichment culture medium takes an MS culture medium as a basic culture medium, and also comprises 3.0-5.0 mg/L of 6-benzylaminopurine, 0.5-1.5 mg/L of α -naphthylacetic acid, 0-3 mg/L of forchlorfenuron, 30g/L of cane sugar and/or white sugar, 0-1 mg/L of thidiazuron and 7-8 g/L of agar;
the number of subcultures of the adventitious bud propagation culture is 1-5.
Preferably, the time for inducing the adventitious buds in the step (1) is 45-60 days;
preferably, the subculture period of the adventitious bud propagation culture in the step (2) is 30 d.
The invention also provides a method for cultivating the lycoris, which comprises the following steps: processing the sterile scale tuber blocks by adopting the rootless tissue culture method to obtain rootless tissue culture seedlings of the lycoris radiata; hardening the rootless tissue culture seedling to obtain a rootless test-tube seedling; transplanting the rootless test-tube plantlet to a substrate for culture to obtain the rooted lycoris radiata plantlet.
Preferably, the seedling exercising time is 5-8 days.
Preferably, when the rootless tissue culture process is carried out in the tissue culture container, before seedling exercising, the bottle mouth of the tissue culture container is unscrewed in a greenhouse; the hardening off comprises the steps of injecting 1.5-3.5 mL of water into a bottle on the 3 rd day of hardening off; the tissue culture container comprises a test tube, a triangular flask or a tissue culture bottle.
Preferably, before transplanting, the method further comprises: soaking the rootless test-tube plantlets by adopting a rooting disinfectant; rinsing the rootless test-tube plantlets after soaking and disinfection with clear water;
preferably, the rooting disinfectant comprises the following components in concentration: 0-0.5 mg/L indoleacetic acid and 0.1-0.3 g/L disinfection substance;
preferably, the disinfecting substance comprises one or more of carbendazim, chlorothalonil and potassium permanganate solution;
preferably, the soaking time is 2-5 min.
Preferably, the matrix comprises the following components in parts by volume: 10-12 parts of turf, 8-10 parts of vermiculite and 0.5-1 part of perlite.
Preferably, before transplanting, the method comprises the steps of paving the substrate into a seedbed or a square tray with the thickness of 8-15 cm, or filling a hole tray with 32-72 holes and the height of 6-10 cm, and thoroughly watering the substrate with water.
Preferably, the row spacing of the transplanted plants is 4-6 cm multiplied by 4-6 cm.
Preferably, the bulbs of the rootless test-tube plantlets are completely embedded in the matrix during transplanting, and the overground part has at least 1 leaf.
Preferably, the post-transplanting comprises field management, specifically: the substrate is wet but unsaturated and has no water accumulation; the humidity is 60-70%; the temperature is 15-25 ℃, and after 30 days of transplanting, the Huabao No. 1 fertilizer with the concentration of 1g/L is applied according to the application amount of 3-5L/mu per month.
The invention provides a lycoris radiata rootless tissue culture method which comprises the following steps of (1) inoculating aseptic scaly stem blocks on an induction culture medium, performing adventitious bud induction to generate adventitious buds, wherein the induction culture medium takes an MS culture medium as a basic culture medium, and further comprises 0.5-3.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 30g/L sucrose and/or white sugar and 6-8 g/L agar, (2) transferring the adventitious buds into a propagation culture medium, performing adventitious bud propagation culture and screening to obtain rootless tissue culture seedlings with the diameter of 0.8-1.5 cm, the propagation culture medium takes the MS culture medium as the basic culture medium, and further comprises 3.0-5.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 0-3 mg/L urea, 30g/L sucrose and 0g/L white sugar, and 0 mg/L6-8 g agar, and the root bud propagation coefficient is increased, the root of the rootless seedlings is increased by adopting a high-root propagation medium, the root regeneration culture medium is suitable for a simple rootless tissue culture process, and the transplanting of the rootless seedlings is improved, the root-induced propagation process is improved, the root-induced seedling transplanting cost is reduced, and the root-induced seedling is reduced, the root-induced seedling transplanting process is improved, the root-induced seedling is the rootless seedling is the root-induced seedling-.
Drawings
FIG. 1 shows the growth of adventitious buds of Lycoris radiata of examples 1-5;
FIG. 2 shows the rooting of test-tube plantlets of lycoris radiata transplanted in example 4 and comparative example 7;
FIG. 3 shows the growth of the tube plantlets of lycoris radiata transplanted in example 4 and comparative example 7;
FIG. 4 shows a rootless tissue culture seedling.
Detailed Description
The invention provides a lycoris radiata rootless tissue culture method which comprises the following steps of (1) inoculating aseptic bulb blocks on an induction culture medium, performing adventitious bud induction to generate adventitious buds, wherein the induction culture medium takes an MS culture medium as a basic culture medium, and further comprises 0.5-3.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 30g/L sucrose and/or white sugar and 6-8 g/L agar, (2) transferring the adventitious buds into a propagation culture medium, performing adventitious bud propagation culture and screening to obtain rootless tissue culture seedlings with the diameter of 0.8-1.5 cm, the propagation culture medium takes the MS culture medium as the basic culture medium, and further comprises 3.0-5.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 0-3 mg/L forchlorfenuron, 30g/L sucrose and/L white sugar, 0mg/L thionine and 7-8 g subculture agar.
The invention inoculates the sterile bulb on the induction culture medium, carries on the adventitious bud induction, produces the adventitious bud.
The sterile bulb is not particularly limited in the invention; in the present invention, the sterile bulb is preferably prepared by: based on the underground bulbs of the lycoris radiata growing robustly in the dry leaf stage, the brown outermost scales of the bulbs are peeled off completely, the bulbs are peeled to 1.5-2 cm in size, the bulbs are washed under water for 30-60 minutes, and the roots and the upper half parts of the bulbs are cut off at 1/2-2/3; then, washing for 2-4 times by using an antibacterial agent, wherein the concentration of the antibacterial agent is adjusted according to the type of the antibacterial agent, and particularly when the antibacterial agent is cefadroxil, the concentration is preferably 150-300 mg/L; when the antibacterial agent is gentamicin, the concentration is preferably 50-100 mg/L; when the antibacterial agent is tetracycline, the concentration is preferably 25-50 mg/L; when the antibacterial agent is rifampicin, the concentration is preferably 10-20 mg/L. And then, flushing the bulb disk for 30-60 s by using 75-85 wt.% of alcohol, flushing for 30-45 min by using 0.5-1 wt.% of benzalkonium bromide, cleaning for 4-5 times by using sterile water, and shaking the bulbs continuously during flushing and cleaning to obtain the bulb disk. And cutting the cleaned bulb disk into 2-4 blocks to ensure that each block has 4-6 layers of scales and a basal disk is reserved, and taking the scales as sterile bulb blocks.
In the invention, the induction time of the adventitious bud is preferably 45-60 d, more preferably 50-55 d, the induction culture medium preferably takes an MS culture medium as a basic culture medium, and further comprises 0.5-3.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 30g/L sucrose and/or white sugar and 6-8 g/L agar, further preferably further comprises 1-2.5 mg/L6-benzylaminopurine, 0.7-1.3 mg/L α -naphthylacetic acid, 30g/L sucrose and/or white sugar and 6.5-7.5 g/L agar, and the pH value of the induction culture medium is preferably 5.7-5.9.
After adventitious buds are generated, the adventitious buds are transferred to a multiplication culture medium for multiplication culture of the adventitious buds to obtain a rootless tissue culture seedling, the propagation culture of the adventitious buds is subcultured, the number of times of the subcultured is 1-5, specifically 1, 2, 3, 4 or 5, the subculture period of the propagation culture of the adventitious buds is preferably 30d, the multiplication culture medium preferably takes an MS culture medium as a basic culture medium, the subculture medium further comprises 3.0-5.0 mg/L6-benzylaminopurine, 0.5-1.5 mg/L α -naphthylacetic acid, 0-3 mg/L forchlorfenuron, 30g/L sucrose and/or white sugar, 0-1 mg/L thidiazuron and 7-8 g/L agar, the subculture medium further preferably further comprises 3.5-4.5 mg/L6-aminopurine, 0.7-1.3 mg/L α -naphthylacetic acid, 0.5-2.5 g/L thidiazuron and 7-8 g/L agar, the subculture medium can be cleaned easily, the survival rate of the rootless-root-proliferation culture medium is improved, the subcultured can be cleaned easily, the subcultured can be carried out after the subcultured, the subcultured can be cleaned, the rooting culture medium is preferably cleaned, the rooting culture medium is not easily, the rooting culture medium is cleaned, the rooting culture medium is not easily, the rooting culture medium is cleaned, and the rooting culture medium.
The container for adventitious bud induction and adventitious bud propagation culture is not particularly limited, and preferably comprises a test tube, a triangular flask and a tissue culture flask.
The invention also provides a method for cultivating the lycoris, which comprises the following steps: processing the sterile scaly stem blocks by adopting the rootless tissue culture method in the technical scheme to obtain rootless tissue culture seedlings of the lycoris radiata; hardening the rootless tissue culture seedling to obtain a rootless test-tube seedling; transplanting the rootless test-tube plantlet to a substrate for culture to obtain the rooted lycoris radiata plantlet.
The invention carries out hardening seedling on the rootless tissue culture seedling to obtain the rootless test-tube seedling. In the invention, the seedling exercising time is preferably 5-8 days, and more preferably 6-7 days; the diameter of the obtained rootless tissue culture seedling is preferably 0.8-1.5 cm, and more preferably 1.0-1.3 cm. When the rootless tissue culture process is carried out in a medium tissue culture container, the step of unscrewing the bottle mouth of the tissue culture container in a greenhouse before seedling exercising is preferably carried out, so that the tissue culture seedling adapts to external temperature and humidity conditions and gas exchange, and the tissue culture seedling adapts to external environmental conditions gradually; the hardening off preferably comprises the steps of injecting 1.5-3.5 mL of water into a bottle on the 3 rd day of hardening off; the tissue culture container comprises a test tube, a triangular flask or a tissue culture bottle. The seedling exercising function is to enable the tissue culture seedlings to adapt to external temperature and humidity conditions and gas exchange, and gradually adapt to external environmental conditions.
The container for exercising the seedlings is not particularly limited in the invention, and preferably comprises a test tube, a triangular flask or a tissue culture flask.
After obtaining the rootless test-tube plantlet, transplanting the rootless test-tube plantlet to a substrate for culture to obtain the rooted lycoris radiata plantlet. In the present invention, the matrix preferably comprises the following components in parts by volume: 10-12 parts of turf, 8-10 parts of vermiculite and 0.5-1 part of perlite; before transplanting, the method preferably comprises the steps of paving the substrate into a seedbed or a square tray with the thickness of 8-15 cm, or filling a hole tray with 32-72 holes and the height of 6-10 cm, and thoroughly watering the substrate with water. In the invention, the row spacing of the transplanted plants is preferably 4-6 cm multiplied by 4-6 cm; preferably, the bulbs are completely embedded in the matrix during transplanting, and the overground part preferably has at least 1 leaf. The substrate can ensure the nutrition and the air permeability of the growth of the bulblet, and on the basis that the grass carbon provides nutrients, the perlite and the vermiculite play a role in water retention and air permeability, so that the seedlings can grow healthily and avoid rotting. The appropriate thickness of the substrate can enable the bulbs to be buried under the soil for growth, the appropriate aperture is favorable for providing space for later-stage bulb expansion, the root system can extend downwards and to the periphery for a longer distance and range, and nutrient absorption is facilitated, and bulb growth and expansion are also facilitated.
In the present invention, before the transplanting, it is preferable that: soaking the rootless test-tube plantlets by adopting a rooting disinfectant, and accelerating the rooting speed of the rootless test-tube plantlets on the basis of disinfecting the rootless test-tube plantlets; and preferably rinsing the rootless test-tube plantlets after soaking and disinfection by clear water, so that the rootless test-tube plantlets are prevented from being poisoned by long-time attachment of the disinfectant. In the present invention, the rooting disinfectant preferably comprises the following components in the following concentrations: 0-0.5 mg/L indoleacetic acid and 0.1-0.3 g/L disinfection substance; the disinfecting substance preferably comprises one or more of carbendazim, chlorothalonil and a potassium permanganate solution; the soaking time is preferably 2-5 min, and more preferably 3-4 min.
After the transplanting, the method preferably comprises field management, and specifically comprises the following steps: the substrate is wet but unsaturated and has no water accumulation; the humidity is preferably 60-70%, and more preferably 63-67%; the temperature is preferably 15-25 ℃, and more preferably 18-22 ℃; and after 30 days of transplanting, applying the Huabao No. 1 fertilizer with the concentration of 1g/L according to the application amount of 3-5L/mu per month.
The present invention will be described in detail with reference to examples, but the scope of the present invention is not limited to these examples.
Examples 1 to 1
(1) Selection and sterilization of explants: selecting strong and basically consistent lycoris bulbus as an explant in a dead leaf period for primary culture. Removing the scales on the outermost brown layer, keeping the scales when the scales are peeled to 2cm, washing the scales under running water for 60 minutes, and placing the scales on an ultraclean workbench. Cutting 2/3 the root and upper bulb half, 1/3 the remaining bulb for subsequent disinfection; the bulbs were rinsed 2 times with 200mg/L of cefaloxime water, followed by a 30s wash with 75 wt.% alcohol, a 30min wash with 0.5 wt.% benzalkonium bromide, and a 5-fold wash with sterile water with constant shaking of the bulbs during the wash. Cutting into 4 pieces according to the size of the bulbar disc, and using 6 layers of scales on each piece to reserve a basal disc as an explant.
(2) And (3) inducing adventitious buds, namely inoculating the sterilized explant on an induction culture medium, wherein the induction culture medium takes an MS culture medium as a basic culture medium, and also comprises 3.0mg/L of 6-benzylaminopurine, 1.0mg/L of α -naphthylacetic acid, 30g/L of sucrose and 6g/L of agar, the pH value of the induction culture medium is 5.8, the culture conditions are that the temperature is 25 ℃, the humidity is 70%, the illumination is about 1500lx, and the illumination time is 12h/d, and after induction is carried out for 60d, the adventitious buds are generated.
(3) And (2) adventitious bud proliferation, namely transferring the induced adventitious bud into a proliferation culture medium, wherein the proliferation culture medium takes an MS culture medium as a basic culture medium, and also comprises 5.0mg/L of 6-benzylaminopurine, 0.5mg/L of α -naphthylacetic acid, 1.0mg/L of forchlorfenuron, 0.5mg/L of thidiazuron, 30g/L of cane sugar and 8g/L of agar, the pH value of the proliferation culture medium is 5.8, and the proliferation culture of the adventitious bud is carried out under the conditions of 25 ℃ of temperature, 70% of humidity, 1500lx of illumination, 12h/d of illumination time, every 30d of a cycle, 1 time of the proliferation culture cycle and the selection of proliferation seedlings with the diameter of 0.8-1.5 cm, so as to obtain the rootless tissue culture seedlings.
Examples 1 to 2
The rootless tissue culture seedling obtained in example 1-1 was placed in a greenhouse, the mouth of the tissue culture seedling was unscrewed in the greenhouse, a small amount of water was injected into the bottle after 3 days, and the seedling was acclimatized for 3 days and then taken out. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
Example 2-1
The growth culture period of example 2-1 was 2 times, and the rest of the treatment was the same as that of example 1-1.
Examples 2 to 2
The rootless tissue culture seedling obtained in example 2-1 was placed in a greenhouse, the mouth of the tissue culture seedling was unscrewed in the greenhouse, a small amount of water was injected into the bottle after 3 days, and the seedling was acclimatized for 3 days and then taken out. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
Example 3-1
The growth culture period of example 3-1 was 3 times, and the rest of the treatment was the same as that of example 1-1.
Examples 3 to 2
The rootless tissue culture seedling obtained in example 3-1 was placed in a greenhouse, the mouth of the tissue culture seedling was unscrewed in the greenhouse, a small amount of water was injected into the bottle after 3 days, and the seedling was acclimatized for 3 days and then taken out. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
Example 4-1
The growth culture cycle of example 4-1 was 4 times, and the rest of the treatment was the same as that of example 1-1. And calculating the survival rate of the seedlings.
Example 4 to 2
The rootless tissue culture seedling obtained in example 4-1 was placed in a greenhouse, the mouth of the tissue culture seedling was unscrewed in the greenhouse, a small amount of water was injected into the bottle after 3 days, and the seedling was acclimatized for 3 days and then taken out. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
Example 5-1
The growth culture period of example 5-1 was 5 times, and the rest of the treatment was the same as that of example 1-1.
Examples 5 and 2
The rootless tissue culture seedling obtained in example 5-1 was placed in a greenhouse, the mouth of the tissue culture seedling was unscrewed in the greenhouse, a small amount of water was injected into the bottle after 3 days, and the seedling was acclimatized for 3 days and then taken out. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
Comparative examples 1 to 4
The proliferation culture cycles of comparative examples 1 to 4 were 6 to 9 times, respectively, and the other treatments were the same as in example 1.
The growth of adventitious buds of Lycoris radiata in examples 1-5 and comparative examples 1-4 is shown in Table 1 and FIG. 1, and FIG. 1 shows the growth of adventitious buds of examples 1-5 from left to right. As is clear from Table 1 and FIG. 1, the rootless tissue culture method provided by the present invention is excellent in growth, and the proliferation culture methods of examples 2 to 5 are more suitable.
TABLE 1 subculture frequency of adventitious bud of Lycoris radiata in examples 1-5 and comparative examples 1-4
Note: the normal growth is recorded as +, the vitrification appears, the growth failure begins to appear as- - -, and the growth failure is recorded as.
Comparative example 5
The survival rate of the seedlings was calculated by selecting the proliferated seedlings having a bulb diameter of less than 0.5cm, and the rest of the treatment was the same as in example 4.
Comparative example 6
Selecting rootless test-tube plantlets with bulbs with diameters larger than 0.5cm and smaller than 0.8cm, performing the same treatment as in example 4, and calculating the survival rate of the plantlets.
And (3) test results: the results of the determination of the survival rate of transplanting of example 4 and comparative examples 5 and 6 are shown in table 4, and it can be seen from table 4 that the survival rate of example 4 is as high as 95.6% which is higher than that of comparative examples 5 and 6.
TABLE 2 Effect of different bulb diameter sizes on the survival rate of transplantation
Comparative example 7
According to the method recorded in the research on tissue culture propagation technology of the Zhujin and the lycoris radiate (J) Zhejiang forestry technology 2002(04) 46-49, transplanting is carried out after roots grow in a bottle, the cultivation management after transplanting is the same as that in the example 4, and 1 square meter is taken for data statistics in each treatment.
The rooting conditions of the test-tube plantlets of the lycoris radiata transplanted in the example 4 and the comparative example 7 are shown in fig. 2, the growth conditions are shown in fig. 3 and table 3, and as can be seen from fig. 2 and fig. 3, the method in the example 4 of the invention can realize the ex vitro rooting of the lycoris radiata and can survive. As can be seen from Table 3, in example 4 of the present invention, the yield and quality of seedlings can be improved, and the obtained lycoris radiata seed balls have uniform sizes, and the yield is improved by 1.22 times compared with comparative example 7. In the transplanting link, the invention can save the cleaning time, is not easy to damage the test-tube plantlet and reduces the production labor cost. The seed balls of Lycoris radiata in the control group have different sizes.
TABLE 3 results of two different methods for transplanting Lycoris radiata test-tube plantlets
Example 6
(1) Selection and sterilization of explants: selecting strong and basically consistent lycoris bulbus as an explant in a dead leaf period for primary culture. Removing the scales on the outermost brown layer, keeping the scales when the scales are peeled to 2cm, washing the scales under running water for 60 minutes, and placing the scales on an ultraclean workbench. Cutting 2/3 the root and upper bulb half, 1/3 the remaining bulb for subsequent disinfection; the bulbs were rinsed 2 times with 200mg/L of cefaloxime water, followed by a 30s wash with 75 wt.% alcohol, a 30min wash with 0.5 wt.% benzalkonium bromide, and a 5-fold wash with sterile water with constant shaking of the bulbs during the wash. Cutting into 4 pieces according to the size of the bulb dish, reserving the basal disc with 6 layers of scales for each piece, inoculating on a sterilized culture medium, and counting the pollution rate and the death rate after 2 weeks.
(2) And (3) inducing adventitious buds, namely inoculating the sterilized explant on an induction culture medium, wherein the induction culture medium takes an MS culture medium as a basic culture medium, and also comprises 2.0mg/L of 6-benzylaminopurine, 1.0mg/L of α -naphthylacetic acid, 30g/L of sucrose and 6g/L of agar, the pH value of the induction culture medium is 5.8, the culture conditions are that the temperature is 25 ℃, the humidity is 70%, the illumination is about 1500lx, and the illumination time is 12h/d, and after induction is carried out for 60d, the adventitious buds are generated.
(3) And (2) adventitious bud proliferation, namely transferring the induced adventitious bud into a proliferation culture medium, wherein the proliferation culture medium takes an MS culture medium as a basic culture medium, and also comprises 4.0mg/L of 6-benzylaminopurine, 1.0mg/L of α -naphthylacetic acid, 0.4mg/L of forchlorfenuron, 0mg/L of thidiazuron, 30g/L of cane sugar and 7.5g/L of agar, the pH value of the proliferation culture medium is 5.8, performing proliferation culture on the adventitious bud under the conditions of 25 ℃ of temperature, 70% of humidity and 1500lx of illumination, 12h/d of illumination time, every 30d of a period, the proliferation culture period is 1 time, and screening proliferation seedlings with the diameter of 0.8-1.5 cm to obtain the rootless tissue culture seedlings.
(4) Unscrewing the mouth of the tissue culture seedling in a greenhouse, injecting a small amount of water into the bottle after 3 days, and taking out the tissue culture seedling after hardening the seedling for 4 days. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
Comparative example 8
The 0.5 wt.% benzalkonium bromide wash of example 6 was replaced by a 30min wash with 0.1% (w/v) mercuric chloride solution for 20min, the other treatments were the same as example 6, and the explant contamination and mortality were counted after 2 weeks.
The contamination rate is the number of contaminated explants/inoculated explants x 100%;
mortality rate-number of dead explants/number of inoculated explants × 100%
And (3) test results: the explant culture conditions of example 6 and comparative example 8 are shown in Table 4, and from Table 4, it can be seen that the disinfection method of example 6 of the present application has a lower contamination rate and a lower death rate than the mercury-liter disinfection treatment of comparative example 8.
TABLE 4 Effect of different sterilization modes on explant culture
Example 7
(1) Selection and sterilization of explants: selecting strong and basically consistent lycoris bulbus as an explant in a dead leaf period for primary culture. Removing the scales on the outermost brown layer, keeping the scales when the scales are peeled to 2cm, washing the scales under running water for 60 minutes, and placing the scales on an ultraclean workbench. Cutting 2/3 the root and upper bulb half, 1/3 the remaining bulb for subsequent disinfection; the bulbs were rinsed 2 times with 200mg/L of cefaloxime water, followed by a 30s wash with 75 wt.% alcohol, a 30min wash with 0.5 wt.% benzalkonium bromide, and a 5-fold wash with sterile water with constant shaking of the bulbs during the wash. Cutting into 4 pieces according to the size of the bulbar disc, and using 6 layers of scales on each piece to reserve a basal disc as an explant.
(2) Induction of adventitious buds: inoculating the sterilized explants on different induction culture media, wherein the pH values are 5.8, the culture conditions are that the temperature is 25 ℃, the humidity is 70%, the illumination is about 1500lx, and the illumination time is 12 h/d. After induction for 60 days, adventitious buds were generated.
(3) And (2) adventitious bud proliferation, namely transferring the induced adventitious bud into a proliferation culture medium, wherein the proliferation culture medium takes an MS culture medium as a basic culture medium, and also comprises 5.0mg/L of 6-benzylaminopurine, 0.5mg/L of α -naphthylacetic acid, 1.0mg/L of forchlorfenuron, 0.5mg/L of thidiazuron, 30g/L of cane sugar and 8g/L of agar, the pH value of the proliferation culture medium is 5.8, and the proliferation culture of the adventitious bud is carried out under the conditions of 25 ℃ of temperature, 70% of humidity, 1500lx of illumination, 12h/d of illumination time, every 30d of a cycle, 1 time of the proliferation culture cycle and the selection of proliferation seedlings with the diameter of 0.8-1.5 cm, so as to obtain the rootless tissue culture seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
And (3) test results: the induction rate of different induction culture media on adventitious buds is shown in table 5, and the induction rate of the induction culture medium of the invention on the adventitious buds of lycoris radiata can reach 82.5 percent at most.
TABLE 5 Effect of different induction media on Amaryllidaceae adventitious bud induction
Example 8
(1) Selection and sterilization of explants: selecting strong and basically consistent lycoris bulbus as an explant in a dead leaf period for primary culture. Removing the scales on the outermost brown layer, keeping the scales when the scales are peeled to 2cm, washing the scales under running water for 60 minutes, and placing the scales on an ultraclean workbench. Cutting 2/3 the root and upper bulb half, 1/3 the remaining bulb for subsequent disinfection; the bulbs were rinsed 2 times with 200mg/L of cefaloxime water, followed by a 30s wash with 75 wt.% alcohol, a 30min wash with 0.5 wt.% benzalkonium bromide, and a 5-fold wash with sterile water with constant shaking of the bulbs during the wash. Cutting into 4 pieces according to the size of the bulb dish, reserving the basal disc with 6 layers of scales for each piece, inoculating on a sterilized culture medium, and counting the pollution rate and the death rate after 2 weeks.
(2) And (3) inducing adventitious buds, namely inoculating the sterilized explant on an induction culture medium, wherein the induction culture medium takes an MS culture medium as a basic culture medium, and also comprises 3.0mg/L of 6-benzylaminopurine, 1.0mg/L of α -naphthylacetic acid, 30g/L of sucrose and 6g/L of agar, the pH value of the induction culture medium is 5.8, the culture conditions are that the temperature is 25 ℃, the humidity is 70%, the illumination is about 1500lx, and the illumination time is 12h/d, and after induction is carried out for 60d, the adventitious buds are generated.
(3) Proliferation of adventitious buds: transferring the induced adventitious buds to different proliferation culture media, wherein the pH values of the proliferation culture media are 5.8, and performing proliferation culture on the adventitious buds under the conditions of 25 ℃ of temperature, 70% of humidity, about 1500lx of illumination and 12h/d of illumination time. And (4) selecting the propagation seedlings with the diameter of 0.8-1.5 cm from every 30 days as a period, wherein the propagation culture period is 1 time, and obtaining the rootless tissue culture seedlings.
(4) Unscrewing the mouth of the tissue culture seedling in a greenhouse, injecting a small amount of water into the bottle after 3 days, and taking out the tissue culture seedling after 3 days of hardening. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. And (3) soaking the rootless seedlings in a potassium permanganate solution containing 0.2mg/L IBA and one ten thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
And (3) test results: the influence of different proliferation culture media on the proliferation culture of lycoris radiata is shown in table 6, and the proliferation fold of the proliferation culture medium of the invention on lycoris radiata can reach 8.42 times at most.
TABLE 6 Effect of different propagation media on the propagation of Lycoris radiata
Example 9
(1) Selection and sterilization of explants: selecting strong and basically consistent lycoris bulbus as an explant in a dead leaf period for primary culture. Removing the scales on the outermost brown layer, keeping the scales when the scales are peeled to 2cm, washing the scales under running water for 60 minutes, and placing the scales on an ultraclean workbench. Cutting 2/3 the root and upper bulb half, 1/3 the remaining bulb for subsequent disinfection; the bulbs were rinsed 2 times with 200mg/L of cefaloxime water, followed by a 30s wash with 75 wt.% alcohol, a 30min wash with 0.5 wt.% benzalkonium bromide, and a 5-fold wash with sterile water with constant shaking of the bulbs during the wash. Cutting into 4 pieces according to the size of the bulb dish, reserving the basal disc with 6 layers of scales for each piece, inoculating on a sterilized culture medium, and counting the pollution rate and the death rate after 2 weeks.
(2) And (3) inducing adventitious buds, namely inoculating the sterilized explant on an induction culture medium, wherein the induction culture medium takes an MS culture medium as a basic culture medium, and also comprises 3.0mg/L of 6-benzylaminopurine, 1.0mg/L of α -naphthylacetic acid, 30g/L of sucrose and 6g/L of agar, the pH value of the induction culture medium is 5.8, the culture conditions are that the temperature is 25 ℃, the humidity is 70%, the illumination is about 1500lx, and the illumination time is 12h/d, and after induction is carried out for 60d, the adventitious buds are generated.
(3) And (2) adventitious bud proliferation, namely transferring the induced adventitious bud into a proliferation culture medium, wherein the proliferation culture medium takes an MS culture medium as a basic culture medium, and also comprises 5.0mg/L of 6-benzylaminopurine, 0.5mg/L of α -naphthylacetic acid, 1.0mg/L of forchlorfenuron, 0.5mg/L of thidiazuron, 30g/L of cane sugar and 8g/L of agar, the pH value of the proliferation culture medium is 5.8, and the proliferation culture of the adventitious bud is carried out under the conditions of 25 ℃ of temperature, 70% of humidity, 1500lx of illumination, 12h/d of illumination time, every 30d of a cycle, 1 time of the proliferation culture cycle and the selection of proliferation seedlings with the diameter of 0.8-1.5 cm, so as to obtain the rootless tissue culture seedlings.
(4) Unscrewing the mouth of the tissue culture seedling in a greenhouse, injecting a small amount of water into the bottle after 3 days, and taking out the tissue culture seedling after 3 days of hardening. And (4) lightly taking out the domesticated lycoris rootless test-tube plantlet by using forceps. Soaking rootless seedlings in a potassium permanganate solution containing IBA with different concentrations and one ten-thousandth of IBA for 5min, rinsing with clear water, transplanting the seedlings onto a substrate, and culturing to obtain rooted lycoris radiata seedlings.
The transplanting matrix comprises grass peat: vermiculite: perlite, which is mixed and packed in a square disc with the height of 10cm according to the volume ratio of 12:10: 1; after watering the square plate matrix, planting the test-tube plantlets into a seedbed according to the plant row spacing of 5 multiplied by 6cm, and when the test-tube plantlets are transplanted, burying all bulbils into the matrix, wherein the overground part has 3 leaves; when watering, the substrate is moistened but not saturated and does not accumulate water, the humidity after transplanting is controlled at 60 percent, the temperature is controlled at 20 ℃, and conventional field management of water, fertilizer, plant diseases and insect pests and the like is carried out.
And (3) test results: the influence of different hormone ratios on the transplanting and rooting of the lycoris radiata rootless seedlings is shown in table 7 and figure 4, and the survival rate of the transplanting and rooting of the lycoris radiata rootless seedlings by utilizing the hormone ratio of the invention can reach 92 percent.
TABLE 7 influence of different hormone ratios on root-taking of Lycoris radiata seedlings in transplanting
The invention provides a lycoris radiata rootless tissue culture method and a lycoris radiata cultivation method with high survival rate.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. A lycoris radiata rootless tissue culture method is characterized by comprising the following steps:
(1) inoculating the sterile bulb blocks on an induction culture medium, and performing adventitious bud induction to generate adventitious buds;
the induction culture medium takes an MS culture medium as a basic culture medium, and also comprises 0.5-3.0 mg/L of 6-benzylaminopurine, 0.5-1.5 mg/L of α -naphthylacetic acid, 30g/L of cane sugar and/or white sugar and 6-8 g/L of agar;
(2) transferring the adventitious buds to a multiplication culture medium, performing adventitious bud multiplication culture, and screening to obtain rootless tissue culture seedlings with the diameter of 0.8-1.5 cm;
the enrichment culture medium takes an MS culture medium as a basic culture medium, and also comprises 3.0-5.0 mg/L of 6-benzylaminopurine, 0.5-1.5 mg/L of α -naphthylacetic acid, 0-3 mg/L of forchlorfenuron, 30g/L of cane sugar and/or white sugar, 0-1 mg/L of thidiazuron and 7-8 g/L of agar;
the number of subcultures of the adventitious bud propagation culture is 1-5.
2. The method for the rootless tissue culture of the lycoris radiata according to claim 1, wherein the induction time of the adventitious bud in the step (1) is 45-60 days;
the subculture period of the adventitious bud multiplication culture in the step (2) is 30 d.
3. A method for cultivating lycoris radiata is characterized by comprising the following steps: processing the sterile bulb blocks by adopting the rootless tissue culture method of claim 1 or 2 to obtain rootless tissue culture seedlings of the lycoris radiata; hardening the rootless tissue culture seedling to obtain a rootless test-tube seedling; transplanting the rootless test-tube plantlet to a substrate for culture to obtain the rooted lycoris radiata plantlet.
4. The cultivation method according to claim 3, wherein the hardening time is 5-8 days.
5. The cultivation method according to claim 3, wherein when the rootless tissue culture process is carried out in a tissue culture container, the acclimatization comprises unscrewing a mouth of the tissue culture container in a greenhouse;
the hardening off comprises the steps of injecting 1.5-3.5 mL of water into a bottle on the 3 rd day of hardening off;
the tissue culture container comprises a test tube, a triangular flask or a tissue culture bottle.
6. The method of claim 3, further comprising, prior to said transplanting: soaking the rootless test-tube plantlets by adopting a rooting disinfectant; rinsing the rootless test-tube plantlets after soaking and disinfection with clear water;
the rooting disinfectant comprises the following components in concentration: 0-0.5 mg/L indoleacetic acid and 0.1-0.3 g/L disinfection substance;
the disinfectant substance comprises one or more of carbendazim, chlorothalonil and potassium permanganate solution;
the soaking time is 2-5 min.
7. The cultivation method as claimed in claim 3, wherein the substrate comprises the following components in parts by volume: 10-12 parts of turf, 8-10 parts of vermiculite and 0.5-1 part of perlite.
8. The cultivation method according to claim 3, wherein before transplanting, the substrate is paved into a seedbed or a square tray with the thickness of 8-15 cm, or a plug tray with 32-72 holes and the height of 6-10 cm is filled, and the substrate is drenched by water;
the row spacing of the transplanted plants is 4-6 cm multiplied by 4-6 cm.
9. The method according to claim 3, wherein the bulbs of the rootless test-tube plantlets are completely embedded in the substrate during transplanting, and the aerial part has at least 1 leaf.
10. The cultivation method according to claim 3, wherein the post-transplanting comprises field management, specifically: the substrate is wet but unsaturated and has no water accumulation; the humidity is 60-70%; the temperature is 15-25 ℃, and after 30 days of transplanting, the Huabao No. 1 fertilizer with the concentration of 1g/L is applied according to the application amount of 3-5L/mu per month.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010004489.5A CN111109081B (en) | 2020-01-03 | 2020-01-03 | Lycoris radiata rootless tissue culture method and lycoris radiata cultivation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010004489.5A CN111109081B (en) | 2020-01-03 | 2020-01-03 | Lycoris radiata rootless tissue culture method and lycoris radiata cultivation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111109081A true CN111109081A (en) | 2020-05-08 |
| CN111109081B CN111109081B (en) | 2022-03-22 |
Family
ID=70507707
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010004489.5A Active CN111109081B (en) | 2020-01-03 | 2020-01-03 | Lycoris radiata rootless tissue culture method and lycoris radiata cultivation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111109081B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112243729A (en) * | 2020-10-19 | 2021-01-22 | 上海市农业科学院 | Method for regulating and controlling flowering phase of lycoris plants |
| CN112273210A (en) * | 2020-11-13 | 2021-01-29 | 上海上房园艺有限公司 | Seedling hardening method for brocade-changing tissue culture seedlings |
| CN112753410A (en) * | 2021-01-13 | 2021-05-07 | 上海科立特农科（集团）有限公司 | Method for improving alkaloid content in lycoris by using exogenous substances |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100242134A1 (en) * | 2007-07-26 | 2010-09-23 | Sathish Puthigae | Methods and polynucleotides for improving plants |
| CN102144554A (en) * | 2011-01-25 | 2011-08-10 | 江苏九久环境科技有限公司 | Method for producing lycoris by plant tissue culture |
| CN102696479A (en) * | 2012-04-24 | 2012-10-03 | 江苏省中国科学院植物研究所 | Method for propagating stonegarlic quickly and efficiently |
| CN104663451A (en) * | 2015-03-10 | 2015-06-03 | 朱海燕 | Tissue culture and rapid propagation method for lycoris albiflora |
| CN105379618A (en) * | 2015-10-20 | 2016-03-09 | 韦丽 | In-vitro rapid propagation method for lycoris radiata |
| CN106561450A (en) * | 2016-10-20 | 2017-04-19 | 江苏省中国科学院植物研究所 | Method for inducing adventitious buds by adopting Lycoris radiate rachis as explant |
| CN107047259A (en) * | 2017-04-21 | 2017-08-18 | 南京林业大学 | A kind of method of bulb block air culture cutting propagation Lycoris radiata |
| CN107864853A (en) * | 2016-09-27 | 2018-04-03 | 上海上房园艺有限公司 | Brocade flower fast breeding method is changed based on tissue cultures |
| CN110073835A (en) * | 2019-05-29 | 2019-08-02 | 上海市农业科学院 | A kind of cultural method of lycoris plants |
| CN110583488A (en) * | 2019-10-24 | 2019-12-20 | 杭州植物园（杭州市园林科学研究院） | Method for establishing tissue culture rapid propagation technical system of new lycoris variety' pink |
-
2020
- 2020-01-03 CN CN202010004489.5A patent/CN111109081B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100242134A1 (en) * | 2007-07-26 | 2010-09-23 | Sathish Puthigae | Methods and polynucleotides for improving plants |
| CN102144554A (en) * | 2011-01-25 | 2011-08-10 | 江苏九久环境科技有限公司 | Method for producing lycoris by plant tissue culture |
| CN102696479A (en) * | 2012-04-24 | 2012-10-03 | 江苏省中国科学院植物研究所 | Method for propagating stonegarlic quickly and efficiently |
| CN104663451A (en) * | 2015-03-10 | 2015-06-03 | 朱海燕 | Tissue culture and rapid propagation method for lycoris albiflora |
| CN105379618A (en) * | 2015-10-20 | 2016-03-09 | 韦丽 | In-vitro rapid propagation method for lycoris radiata |
| CN107864853A (en) * | 2016-09-27 | 2018-04-03 | 上海上房园艺有限公司 | Brocade flower fast breeding method is changed based on tissue cultures |
| CN106561450A (en) * | 2016-10-20 | 2017-04-19 | 江苏省中国科学院植物研究所 | Method for inducing adventitious buds by adopting Lycoris radiate rachis as explant |
| CN107047259A (en) * | 2017-04-21 | 2017-08-18 | 南京林业大学 | A kind of method of bulb block air culture cutting propagation Lycoris radiata |
| CN110073835A (en) * | 2019-05-29 | 2019-08-02 | 上海市农业科学院 | A kind of cultural method of lycoris plants |
| CN110583488A (en) * | 2019-10-24 | 2019-12-20 | 杭州植物园（杭州市园林科学研究院） | Method for establishing tissue culture rapid propagation technical system of new lycoris variety' pink |
Non-Patent Citations (2)
| Title |
|---|
| 常乐等: ""石蒜属植物鳞片扦插繁殖研究"", 《中国球根花卉研究进展》 * |
| 朱锦等: ""石蒜组培繁殖技术的研究"", 《浙江林业科技》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112243729A (en) * | 2020-10-19 | 2021-01-22 | 上海市农业科学院 | Method for regulating and controlling flowering phase of lycoris plants |
| CN112243729B (en) * | 2020-10-19 | 2022-04-22 | 上海市农业科学院 | Method for regulating and controlling flowering phase of lycoris plants |
| CN112273210A (en) * | 2020-11-13 | 2021-01-29 | 上海上房园艺有限公司 | Seedling hardening method for brocade-changing tissue culture seedlings |
| CN112753410A (en) * | 2021-01-13 | 2021-05-07 | 上海科立特农科（集团）有限公司 | Method for improving alkaloid content in lycoris by using exogenous substances |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111109081B (en) | 2022-03-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107980635B (en) | Two-step transplanting method for high-survival-rate apple tissue culture seedlings | |
| CN111109081B (en) | Lycoris radiata rootless tissue culture method and lycoris radiata cultivation method | |
| CN103125386B (en) | Industrial horseradish planting method | |
| CN109601213A (en) | A kind of open country crow Chinese toon container seedling culture method | |
| CN113331055A (en) | Cutting method of stingless pepper tissue culture seedlings | |
| CN111990200A (en) | Method for planting epiphytic orchid on tree | |
| JP2700741B2 (en) | Cultivation method of moss using cultured species | |
| CN103430845A (en) | Strawberry tissue culturing method | |
| CN106359087A (en) | Tissue culture quick-breeding seedling raising method for radix asparagi | |
| CN103039366A (en) | Industrial production method of mycorrhizal seedlings of Changbai Mountain rhododendron plants | |
| CN110810250A (en) | Method for rapid propagation in tissue culture of date palm | |
| CN111480578B (en) | Tissue culture and rapid propagation method for seed embryo of Epimedium sagittatum | |
| CN109618800B (en) | Light medium seedling culture method for two-year-old camellia oleifera | |
| CN101564010B (en) | Method for rapidly propagating tupelos | |
| CN116636350A (en) | Garlic fruit mixed seedling raising method | |
| CN107155843B (en) | Improved method for industrially transplanting tissue culture seedlings of Chinese yams and cultivating potato seeds | |
| CN114532225B (en) | Tissue culture rapid propagation and cultivation method for paphiopedilum delbrueckii | |
| CN115589948A (en) | Bletilla striata non-symbiotic germination culture medium and propagation method | |
| CN114027182A (en) | Tissue culture propagation method for dolichos succulent plants in crassulaceae echeveria | |
| CN101032224A (en) | Tissue cultivating and seedling method of fast growing anthocephalus | |
| CN113678736A (en) | Tissue culture rapid propagation method of amorphophallus rivieri | |
| CN109329047B (en) | Method for improving seedling efficiency of tissue culture container of Helianthus tuberosus | |
| CN113728845A (en) | Method for rooting of Ardisia mamillata Hance tissue culture seedlings outside bottles | |
| CN107306740B (en) | Method for breeding false magpie kidney tree seedlings | |
| CN104221851A (en) | Method for in-vitro culture and rapid propagation of gunnera manlcata l |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |