CN114303952A - Water lily tuber induction tissue culture rapid propagation method - Google Patents
Water lily tuber induction tissue culture rapid propagation method Download PDFInfo
- Publication number
- CN114303952A CN114303952A CN202111675282.1A CN202111675282A CN114303952A CN 114303952 A CN114303952 A CN 114303952A CN 202111675282 A CN202111675282 A CN 202111675282A CN 114303952 A CN114303952 A CN 114303952A
- Authority
- CN
- China
- Prior art keywords
- culture
- culture medium
- water lily
- induction
- explant
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/40—Afforestation or reforestation
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a method for inducing tissue culture and rapid propagation of water lily tubers, which comprises the following steps: (1) explant disinfection: sterilizing the water lily tuber as an explant; (2) induction: inoculating the sterilized explant into an induction culture medium, and then carrying out induction culture; the induction culture medium contains 6-BA, IBA and S106 bactericides; (3) and (3) proliferation culture: inoculating the bud strains obtained by induction into a proliferation culture medium, and then performing proliferation culture; the proliferation culture medium contains 6-BA, IBA and S106 bactericides; (4) rooting culture: inoculating the obtained bud seedling into a rooting culture medium for rooting culture; the rooting culture medium contains IBA; (5) and (5) hardening seedlings. The invention successfully establishes the water lily tissue culture system with tubers as explants, the induction rate and the rooting rate are both as high as 80 percent, and the time consumption in the process is short.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to a method for inducing tissue culture and rapid propagation of water lily tubers.
Background
Nymphaea tetragona (Nymphaea tetragona) is a perennial leafy aquatic herbaceous plant, and is mainly propagated by plants in a separated mode, so that the price of a water lily seedling is high due to the limited propagation quantity and the like, and the industrialization and large-scale development of the water lily are restricted. Tissue culture of water lily is an effective means for obtaining a large number of seedlings in a short period, however, tissue culture research of water lily is only reported so far, and the problems of serious pollution, low induction rate and rooting rate, low proliferation and rooting speed and the like in the water lily tissue culture process cannot be effectively solved.
In the aspect of explant disinfection, because different plants are different in pollution microorganisms involved in the tissue culture process, the disinfection method adopted in the tissue culture process of other plants is not very suitable for the disinfection of water lily explants. The whole growth cycle of the water lily grows in the water body environment, and roots and stems grow in sludge, so that the water lily explant is associated with a large amount of fungi and bacteria and a large amount of endophytes, and the microorganisms are not easy to remove, thereby bringing great difficulty to the tissue culture work of the water lily. Chiffon and the like use rhizomes, roots, leaves, petioles and seeds of blue drug water lily as explants to carry out disinfection test research. The results show that: the adopted rhizome and seed have good disinfection effect. However, when the rhizome is used as the explant, the pollution rate is still as high as 70%, and the browning rate is as high as 45%.
Patent document CN104488716A discloses a method for tissue culture of water lily, which comprises the following specific steps: 1) preparing a culture medium, including a basic culture medium and components of the culture medium at each stage of tissue culture; 2) selecting and disinfecting explants; 3) induction and proliferation of adventitious buds; 4) rooting culture; 5) and (5) transplanting. The method utilizes plant tissue culture technology to perform adventitious bud induction and multiplication culture on the water lily, can obtain a large number of high-quality strong seedlings with consistent genetic characters in a short time, and overcomes the defect of slow conventional propagation method. However, the bud is generated after the induction culture for 1-2 months, and the rooting effect is still to be further improved.
Disclosure of Invention
In view of the defects of the prior art, the invention provides a method for inducing tissue culture and rapid propagation of water lily tubers.
The technical scheme of the invention mainly comprises the following contents:
a method for inducing tissue culture and rapid propagation of water lily tubers comprises the following steps:
(1) explant disinfection: sterilizing the water lily tuber as an explant;
(2) induction: inoculating the explant sterilized in the step (1) into an induction culture medium, and then carrying out induction culture;
the induction culture medium is prepared by adding 2.0-3.0 mg/L6-BA, 0.5-1.0 mg/L IBA and 0.5-1.0 mL/L S106 bactericide into a basic culture medium;
(3) and (3) proliferation culture: inoculating the bud strains obtained by the induction in the step (2) into a proliferation culture medium, and then performing proliferation culture;
the proliferation culture medium is prepared by adding 2.0-3.0 mg/L6-BA, 0.5-1.0 mg/L IBA and 0.5-1.0 mL/L S106 bactericide into a basic culture medium;
(4) rooting culture: inoculating the bud seedling obtained after the culture in the step (3) into a rooting culture medium for rooting culture;
the rooting culture medium is obtained by adding 0.5-1.0 mg/L IBA into a basic culture medium;
(5) and (5) hardening seedlings.
Preferably, the basal medium is MS medium or 1/2MS medium.
One of the difficult problems in the water lily tissue culture process is fungus pollution, and the reduction of the pollution rate is the premise of establishing a water lily tissue culture rapid propagation system. The invention has been tested to determine the optimal disinfection method. The disinfection method comprises the following steps: soaking the cleaned explant in 75% ethanol solution for 1min, and washing with sterilized water for 3-5 times; adding the explant into 0.1% mercuric chloride solution by mass concentration, placing the explant in a shaking table with 100rpm for shaking for 15min, pouring the mercuric chloride solution, cleaning the explant with sterile water for 3-5 min, soaking the explant with sterile water for 5min, and cleaning the explant with sterile water for 3-5 times.
Preferably, in step (2), the conditions for inducing culture are: sterile water is added into the induction culture medium to simulate the growth environment of the water lily; placing the induction culture medium in a 28 ℃ culture room for dark culture for 5-7 d; and then continuously culturing at the temperature of 28 ℃, under the conditions of illumination intensity of 4000lx and light cycle of 10-14 h/d.
Preferably, in step (3), the conditions for propagation culture are: sterile water is added into the proliferation culture medium to simulate the growth environment of the water lily; placing the proliferation culture medium in a 28 ℃ culture room for dark culture for 2-3 d; and then placing the mixture at the temperature of 28 ℃, the illumination intensity of 4000lx and the photoperiod of 10-14 h/d for continuous culture.
Preferably, in the step (4), the rooting culture conditions are as follows: placing the rooting culture medium in a 28 ℃ culture room for dark culture for 2-3 d; and then placing the mixture at the temperature of 28 ℃, the illumination intensity of 4000lx and the photoperiod of 10-14 h/d for continuous culture.
Preferably, in the step (5), the seedling exercising step is as follows: and (4) culturing the rooted seedlings obtained after the culture in the step (4) in a shading place for 1 week, then transferring to the light for 1 week, removing the culture medium at the root parts of the seedlings, planting the seedlings in a non-porous flowerpot filled with clay or sludge for culture, and transferring to the sun for culture after shading culture until new leaves grow out.
Preferably, the induction culture medium is a basal culture medium added with 2.0 mg/L6-BA, 0.5mg/L IBA and 0.5mL/LS106 bactericide.
Preferably, the proliferation culture medium is a basal culture medium added with 3.0 mg/L6-BA, 0.5mg/L IBA and 1.0mL/L S106 bactericide.
Preferably, the rooting medium is a basal medium added with 0.5mg/L IBA.
The invention has the following beneficial effects:
the selection of explants is an important factor influencing the success of water lily tissue culture, and the tissue culture work of different explants of different plants is difficult to work to different degrees. The invention successfully establishes the water lily tissue culture system with tubers as explants.
The invention takes the water lily tubers as explants, directly induces and generates cluster buds, and then obtains tissue culture seedlings through cluster bud induction and rooting culture, no callus is generated in the process, the whole tissue culture process consumes short time and is simple and convenient to operate.
In the known technology, the sterilizing effect on the water lily stems is not ideal, and the pollution rate and the browning rate are high. By adopting the disinfection method provided by the invention (2 min of 75% alcohol and 15min of 0.1% mercury bichloride and adding S106 bactericide in the culture medium) to disinfect the water lily tubers, the average pollution rate is as low as 30%, the browning rate is as low as 25%, and the disinfection effect is good.
The S106 bactericide is used in the induction and proliferation stages of the water lily tissue culture for the first time, so that the pollution rate is reduced, and the improvement of the induction rate is promoted.
Drawings
FIG. 1: the water lily tissue culture process effect picture is provided.
Detailed Description
In order to better understand the technical content of the invention, specific examples are provided below to further illustrate the invention.
Test 1: influence of different disinfectants and disinfection time on water lily explant disinfection
Collecting materials on sunny days, taking healthy delay-drug water lily tubers without diseases and insect pests, filling the tubers into a bucket with water, and taking the tubers back to a laboratory. Washing away mud on the surface of the water lily tubers by using tap water, cleaning the mud on the surface of the water lily tubers, clamping root hairs on the surface of the root tubers by using sharp tweezers, cleaning by using washing powder, washing by using the washing powder, washing for 30min by using the tap water below a tap, and then placing on a clean bench for a disinfection test.
Each disinfectant solution is treated and then thoroughly washed with sterile water to remove the disinfectant solution. Explants sterilized with various sterilization methods were inoculated into MS + S106 medium in 3 replicates per treatment, and 20 flasks with 1 explant per flask were inoculated per replicate. The results were recorded after 10 d. Contamination rate (%) — number of contaminated explants/number of inoculated explants × 100%; browning rate (%) -, number of browned explants/number of inoculated explants × 100%.
The test results show (table 1): the disinfection method A6 is most suitable for the disinfection method of the water lily tubers, the pollution rate is 30 percent, and the browning rate is 25 percent; although the contamination rate of the disinfection rate of A7 is 26.7% at the lowest, the browning rate is severe and reaches 58.3%. Therefore, A6 is most suitable as a sterilization method for the water lily tubers.
TABLE 1 Effect of different disinfectants and disinfection time on the disinfection of Water lily explants
Test 2: effect of different plant growth regulators on Water lily explant Induction
The non-polluted and browned explants sterilized by the A6 sterilization method are inoculated into a B1-B4 culture medium to simulate the water lily growth environment, each bottle is just submerged in water, the water lily growth environment is simulated, each treatment is inoculated with 60 bottles, and each bottle contains 1 explant. Placing in a 28 ℃ culture room for dark culture for 7 days, and then placing in a 28 ℃ culture room for culture under the culture conditions of illumination intensity of 4000lx and light cycle of 14 h/d. Observed daily after inoculation and the results were recorded after 20 days. Induction rate (%) -. number of explants in which adventitious shoots were induced/number of explants inoculated × 100%.
The test results show (table 2): the plant growth regulator 6-BA + IBA is most suitable for being a hormone induced by the water lily tubers, the induction rate reaches 76.7 percent, and more than 2 cluster buds exist; while the control, 6-BA + IAA and 6-BA + NAA also showed some buds, but they all showed more single buds, and the buds showed good growth without the addition of 6-BA + IBA. Therefore, 6-BA + IBA is most suitable as a plant growth regulator for inducing the water lily tubers.
TABLE 2 Effect of different plant growth regulators on the induction of Water lily explants
Test 3: influence of different S106 bactericide dosage on induction of water lily explant
The non-polluted and browned explants sterilized by the A6 sterilization method are inoculated into a B5-B8 culture medium to simulate the water lily growth environment, each bottle is just submerged in water, the water lily growth environment is simulated, each treatment is inoculated with 60 bottles, and each bottle contains 1 explant. Placing in a 28 ℃ culture room for dark culture for 7 days, and then placing in a 28 ℃ culture room for culture under the culture conditions of illumination intensity of 4000lx and light cycle of 14 h/d. Observed daily after inoculation and the results were recorded after 20 days.
The test results show (table 3): when the dosage of the S106 bactericide is 0.5-1.0 mL/L, the inductivity is as high as 83.3%, and the germination is fast and the bud growth vigor is good.
Further experiments confirm that the average induction rate reaches more than 70 percent when the induction culture medium is MS +6-BA 2.0-3.0 mg/L + IBA 0.5-1.0 mg/L + S1060.5-1 mL/L.
TABLE 3 Effect of different amounts of S106 fungicide on the induction of Water lily explants
Test 4: effect of different plant growth regulator concentrations on the multiplication of Water lily clumped buds
Dividing the cluster buds obtained by the B2 induction method into two or four or six (according to the size of the cluster buds), continuously inoculating into a proliferation culture medium C1-C5, and adding water into the culture medium to simulate the growth environment of the water lily. 60 flasks were inoculated per treatment, 1 explant per flask. Placing in a 28 ℃ culture room for dark culture for 7 days, and then placing in a 28 ℃ culture room for culture under the culture conditions of illumination intensity of 4000lx and light cycle of 14 h/d. The results were recorded by observation after 20d incubation.
The test results show (table 4): the C2 and C3 are suitable for being used as the multiplication culture medium of the water lily induction bud, the induction rate reaches 80 percent, and 2-8 bud seedlings are generated. The induction rate is increased along with the increase of the concentration of the plant growth regulator 6-BA, but the induction rate is reduced when the concentration of the 6-BA reaches a certain level, and the higher the concentration of the 6-BA is, the callus can be generated to influence the induction of buds and even die.
TABLE 4 Effect of different plant growth regulator concentrations on the multiplication of Water lily clumped buds
Test 5: effect of different amounts of S106 Fungicide on multiplication of Water lily clumped buds
Dividing the cluster buds obtained by the B2 induction method into two or four or six (according to the sizes of the cluster buds), continuously inoculating the cluster buds into a multiplication medium C6-C9, simulating the growth environment of the water lily, and adding water into each bottle to just submerge tubers so as to simulate the growth environment of the water lily. 60 flasks were inoculated per treatment, 1 explant per flask. Placing in a 28 ℃ culture room, and performing dark culture for 7 d; then culturing under the culture conditions of 28 ℃, the illumination intensity of 4000lx and the light cycle of 14 h/d. The recorded results were observed after 20 d.
The test results show (table 5): with the increase of the addition amount of S106, the inductivity shows a trend of increasing firstly and then decreasing, and when the addition amount of S106 is 0.5-1.0 mL/L, the inductivity reaches more than 80%, the number of buds is large, and the growth vigor is good.
Further experiments confirm that when the induction culture medium is MS +6-BA 2.0-3.0 mg/L + IBA 0.5-1.0 mg/L + S1060.5-1 mL/L, the average induction rate reaches more than 76%.
TABLE 5 Effect of different amounts of S106 fungicide on the proliferation of Water lily clumped buds
Test 6: influence of different plant growth regulators and concentrations on rooting of water lily tissue culture seedlings
Selecting a bud seedling which is processed by C8 and is 3-5 cm high, grows consistently and robustly, cutting a single plant, inoculating the cut single plant into a rooting culture medium D1-D7, inoculating 30 bottles of tissue culture seedlings in each bottle, placing 1 plant of tissue culture seedlings in each bottle, and culturing under the conditions of 16 hours/day illumination of an LED illuminating lamp, 4000lx illumination intensity and 28 ℃. And 20d, recording the result and calculating the rooting rate. The rooting rate (%) is the number of rooted seedlings/inoculation number × 100%.
The test results show (table 6): IBA is more suitable to be used as a water lily rooting hormone, and IBA is used as the rooting hormone, so that the water lily tissue culture seedlings have more roots, are long and thick, have the root length of more than 2cm and are robust; NAA as the rooting hormone of the water lily tissue culture seedling also has a small amount of roots, but is thin and short and has slow rooting speed. Therefore, IBA is more suitable as a rooting hormone of the water lily tissue culture seedlings.
TABLE 6 influence of different plant growth regulators and concentrations on rooting of water lily tissue culture seedlings
Test 7: influence of different proliferation treatments on rooting of subsequent water lily tissue culture seedlings
Selecting C6 and C7, processing the seedlings to be 3-5 cm high, cutting single plants of the strong and consistent growing bud seedlings, inoculating the single plants into a rooting culture medium D3, inoculating 30 bottles of tissue culture seedlings in each bottle, placing 1 plant of tissue culture seedlings in each bottle, and culturing the tissue culture seedlings under the conditions of 16 hours/day illumination of an LED illuminating lamp, 4000lx illumination intensity and 28 ℃. And 20d, recording the result and calculating the rooting rate. The rooting rate (%) is the number of rooted seedlings/inoculation number × 100%.
The test results show (table 7): when well-growing and non-callus seedlings obtained by the treatment of C6 and C7 in the test 5 are inoculated to a D3 culture medium for rooting culture, the rooting conditions are obviously different. When the bud seedlings obtained after the C6 treatment are subjected to rooting culture, the rooting time is relatively late, and the rooting rate is relatively low. Therefore, D3 is the most suitable culture medium for the water lily tissue culture seedlings.
TABLE 7 Effect of different proliferation treatments on rooting of subsequent Water lily tissue culture seedlings
According to the experimental result of the invention, the method for inducing tissue culture and rapid propagation of the water lily tubers is provided, and the detailed operation steps are summarized as follows:
(1) explant sterilization
The water lily root tuber is used as an explant, the explant with the clean surface is soaked in 75% ethanol for 1min in an operation table, and then is washed with sterilized water for 3-5 times for later use. Adding 0.1% mercuric chloride solution into the explant, placing the explant in a shaking table with the speed of 100r/min, shaking for 15min, pouring the mercuric chloride solution, washing for 3-5 times by using sterile water, soaking for 5min by using the sterile water, removing the mercuric chloride solution in the explant so as to avoid poisoning the explant, increasing the browning rate, washing for 3-5 times by using the sterile water, then sucking water by using sterile filter paper, and removing a layer of the surface of the root tuber contacting the mercuric chloride by using a surgical knife.
(2) Induction of the root tuber of Helianthus tuberosus
Inoculating the explant treated in the step (1) into an induction culture medium (MS + 2.0-3.0 mg/L6-BA + 0.5-1.0 mg/LIBA + 0.5-1.0 mL/L S106), adding sterile water (just submerging the surface of a tuber) into the culture medium to simulate the growth environment of water lily, and placing the culture medium in a 28 ℃ culture chamber for dark culture for 5-7 days; and then culturing under the culture conditions of 28 ℃, illumination intensity of 4000lx and light cycle of 10-14 h/d.
(3) Proliferation culture
Dividing the bud obtained in the step (2) into two parts, one part and three parts or one part and four parts (according to the size of the bud), inoculating the bud into a multiplication culture medium (MS + 2.0-3.0 mg/L6-BA + 0.5-1.0 mg/L IBA + 0.5-1.0 mL/L S106), adding sterile water into the culture medium to simulate the growth environment of water lily, and placing the culture medium in a 28 ℃ culture room for dark culture for 5-7 days; and then culturing under the culture conditions of 28 ℃, illumination intensity of 4000lx and light cycle of 10-14 h/d.
(4) Rooting culture
Dividing the induced tissue culture seedlings with the height of 3-5 cm into single plants, inoculating the single plants into a rooting culture medium (1/2MS + 0.5-1.0 mg/LIBA) for strong seedling rooting culture, wherein the rooting culture medium is a semi-solid culture medium, and placing the semi-solid culture medium in a 28 ℃ culture room for dark culture for 2-3 days; and then culturing under the culture conditions of 28 ℃, illumination intensity of 4000lx and light cycle of 10-14 h/d.
(5) Hardening seedlings of water lily
The bottle mouth of the water lily rooted seedling is opened, the water lily rooted seedling is placed and moved to a shading position for about one week, the water lily rooted seedling is slowly moved to a position with good light to be hardened for about one week, the bottle body or the bag surface is frequently rotated to enable the tissue culture seedling to be uniformly lighted, and the hardened tissue culture seedling is uniformly lighted and healthy. Taking out refined water lily seedlings, cleaning root culture medium, planting in soil with large viscosity or sludge in a pond, culturing in a non-porous flowerpot, culturing in a shade shed until new leaves grow out, and transferring to sunlight for normal culture (20-30d), wherein the shade degree of the shade shed is about 50%, and the seedling culture medium is clay or sludge. The survival rate of the acclimated seedlings reaches 99 percent after 20 days of culture.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A method for inducing tissue culture and rapid propagation of water lily tubers is characterized by comprising the following steps:
(1) explant disinfection: sterilizing the water lily tuber as an explant;
(2) induction: inoculating the explant sterilized in the step (1) into an induction culture medium for induction culture;
the induction culture medium is prepared by adding 2.0-3.0 mg/L6-BA, 0.5-1.0 mg/L IBA and 0.5-1.0 mL/L S106 bactericide into a basic culture medium;
(3) and (3) proliferation culture: carrying out enrichment culture on the bud-grafted seeds obtained by inducing in the step (2) in an enrichment culture medium;
the proliferation culture medium is prepared by adding 2.0-3.0 mg/L6-BA, 0.5-1.0 mg/L IBA and 0.5-1.0 mL/L S106 bactericide into a basic culture medium;
(4) rooting culture: inoculating the bud seedling obtained after the culture in the step (3) into a rooting culture medium for rooting culture;
the rooting culture medium is obtained by adding 0.5-1.0 mg/L IBA into a basic culture medium;
(5) and (5) hardening seedlings.
2. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein the basic culture medium is an MS culture medium or an 1/2MS culture medium.
3. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein the disinfection in the step (1) is as follows: soaking the cleaned explant in 75% ethanol solution for 1min, and washing with sterilized water for 3-5 times; adding the explant into 0.1% mercuric chloride solution by mass concentration, placing the explant in a shaking table with 100rpm for shaking for 15min, pouring the mercuric chloride solution, cleaning the explant with sterile water for 3-5 min, soaking the explant with sterile water for 5min, and cleaning the explant with sterile water for 3-5 times.
4. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein in the step (2), the conditions of the induction culture are as follows: sterile water is added into the induction culture medium to simulate the growth environment of the water lily; placing the induction culture medium in a 28 ℃ culture room for dark culture for 5-7 d; and then continuously culturing at the temperature of 28 ℃, under the conditions of illumination intensity of 4000lx and light cycle of 10-14 h/d.
5. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein in the step (3), the conditions of propagation culture are as follows: sterile water is added into the proliferation culture medium to simulate the growth environment of the water lily; placing the proliferation culture medium in a 28 ℃ culture room for dark culture for 2-3 d; and then placing the mixture at the temperature of 28 ℃, the illumination intensity of 4000lx and the photoperiod of 10-14 h/d for continuous culture.
6. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein in the step (4), the rooting culture conditions are as follows: placing the rooting culture medium in a 28 ℃ culture room for dark culture for 2-3 d; and then placing the mixture at the temperature of 28 ℃, the illumination intensity of 4000lx and the photoperiod of 10-14 h/d for continuous culture.
7. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein in the step (5), the seedling hardening step is as follows: and (4) culturing the rooted seedlings obtained after the culture in the step (4) in a shading place for 1 week, then transferring to the light for 1 week, removing the culture medium at the root parts of the seedlings, planting the seedlings in a non-porous flowerpot filled with clay or sludge for culture, and transferring to the sun for culture after shading culture until new leaves grow out.
8. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein the induction culture medium is a basal culture medium added with 2.0 mg/L6-BA, 0.5mg/L IBA and 0.5mL/L S106 bactericide.
9. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein the propagation medium is a basal medium supplemented with 3.0 mg/L6-BA, 0.5mg/L IBA and 1.0mL/L S106 bactericide.
10. The method for inducing tissue culture and rapid propagation of the water lily tubers according to claim 1, wherein the rooting medium is a basal medium supplemented with 0.5mg/L IBA.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111675282.1A CN114303952B (en) | 2021-12-31 | 2021-12-31 | Water lily tuber induction tissue culture rapid propagation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111675282.1A CN114303952B (en) | 2021-12-31 | 2021-12-31 | Water lily tuber induction tissue culture rapid propagation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114303952A true CN114303952A (en) | 2022-04-12 |
| CN114303952B CN114303952B (en) | 2023-03-31 |
Family
ID=81020472
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111675282.1A Active CN114303952B (en) | 2021-12-31 | 2021-12-31 | Water lily tuber induction tissue culture rapid propagation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114303952B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104488716A (en) * | 2014-12-19 | 2015-04-08 | 浙江省农业科学院 | Method for tissue culture of water nymph |
| CN104996298A (en) * | 2015-07-06 | 2015-10-28 | 三明学院 | Method for cultivating polygala fallax hemsl tissue culture seedlings based on multiple internodal stem segments integration |
| CN106577272A (en) * | 2015-10-14 | 2017-04-26 | 西南大学 | Method for producing brasenia schreberi j.f.gmel seedlings by using plant tissue culture technology |
| CN109122311A (en) * | 2018-07-31 | 2019-01-04 | 海南大学 | A kind of culture medium and its cultural method of water lily viviparity stem tuber induction sterile bud |
-
2021
- 2021-12-31 CN CN202111675282.1A patent/CN114303952B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104488716A (en) * | 2014-12-19 | 2015-04-08 | 浙江省农业科学院 | Method for tissue culture of water nymph |
| CN104996298A (en) * | 2015-07-06 | 2015-10-28 | 三明学院 | Method for cultivating polygala fallax hemsl tissue culture seedlings based on multiple internodal stem segments integration |
| CN106577272A (en) * | 2015-10-14 | 2017-04-26 | 西南大学 | Method for producing brasenia schreberi j.f.gmel seedlings by using plant tissue culture technology |
| CN109122311A (en) * | 2018-07-31 | 2019-01-04 | 海南大学 | A kind of culture medium and its cultural method of water lily viviparity stem tuber induction sterile bud |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114303952B (en) | 2023-03-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106106163A (en) | A kind of iris cultivates propagation method | |
| CN113331059B (en) | Method for establishing efficient regeneration system by taking bird king tea tree hypocotyls as explants | |
| CN111657151A (en) | Rapid seedling method for acer truncatum | |
| CN112868527A (en) | Method for rapidly propagating flamingo pepper grass | |
| CN1771796A (en) | In vitro broad bean culturing method | |
| CN114303952B (en) | Water lily tuber induction tissue culture rapid propagation method | |
| CN104357374B (en) | A kind of pepper anther method for tissue culture | |
| CN114431149B (en) | Non-symbiotic germination method for seeds of rare or endangered plant large yellow croaker calanthe | |
| CN110663554A (en) | Commercial tissue culture seedling raising method for Hosta chinensis Redlar | |
| CN113317206B (en) | Culture medium combination for tissue culture and rapid propagation of lilium martagon, tissue culture and rapid propagation method and application | |
| CN110122331A (en) | It is a kind of using leaf as the quick-breeding method of the U.S. Liriodendron chinense in Asia of explant | |
| CN1149921C (en) | Morinda tissue culture method | |
| CN101543184A (en) | Method for culturing open type tissue of konjak | |
| CN111165356B (en) | Tissue culture propagation method of peony | |
| CN108450333B (en) | Induction method of lilium tigrinum callus | |
| CN109258463B (en) | Vegetative propagation method of paphiopedilum armeniacum | |
| CN109526748B (en) | Tissue culture method for anthurium andraeanum inflorescence | |
| CN112167064A (en) | Tissue culture and rapid propagation method for salvia miltiorrhiza | |
| CN114568305B (en) | Treatment method for improving regeneration efficiency by tissue culture of quercus acutissima | |
| CN111248087B (en) | Tissue culture rapid propagation method for primary seedling formation of radix glehniae | |
| NL2032821B1 (en) | Efficient method for micropropagation of sesuvium portulacastrum through axillary buds | |
| CN116806694B (en) | Eggplant doubled haploid anther culture method | |
| CN114208673B (en) | Tissue culture method of north China blue potted flowers | |
| CN110199872B (en) | Tissue culture and rapid propagation method for stem tips of anthurium andraeanum with short disinfection time | |
| CN111887152B (en) | Method for establishing plant regeneration system of swallow flower bud growing way |
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 |