CN110537489A - Test tube micro-cuttage aseptic propagation method for eggplant rootstocks - Google Patents
Test tube micro-cuttage aseptic propagation method for eggplant rootstocks Download PDFInfo
- Publication number
- CN110537489A CN110537489A CN201910863985.3A CN201910863985A CN110537489A CN 110537489 A CN110537489 A CN 110537489A CN 201910863985 A CN201910863985 A CN 201910863985A CN 110537489 A CN110537489 A CN 110537489A
- Authority
- CN
- China
- Prior art keywords
- culture
- culture medium
- aseptic
- rooting
- eggplant
- 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.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
-
- 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
-
- 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
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Soil Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a test tube micro-cuttage aseptic propagation method for eggplant stocks, which comprises the following steps: obtaining aseptic seedlings, bud induction culture, subculture proliferation culture, rooting culture and hardening seedling transplantation. The bud induction culture medium is MS + KT 0.4-0.6 mg/L + IBA 0.05-0.15 mg/L, the subculture multiplication culture medium is MS + IBA 0.35-0.45 mg/L, and the rooting culture medium is 1/2MS + IBA 0.15-0.25 mg/L. The stem section is used as the explant, and the used explant is simple and easy to take; the plant growth regulator has simple proportioning, low price of the used reagent and low cost; the bud inductivity reaches 90 percent, and lateral buds about 1cm can grow after 15 days; subculture multiplication is carried out for 30 days, the multiplication coefficient reaches 5-6 times, the rooting effect is obvious, and the survival rate of transplanting is high. The invention establishes the whole tissue culture and rapid propagation technical system of the solanum torvum, has short period of the whole production process, simple and convenient operation and high multiplication factor, can completely meet the requirement of large-scale seedling propagation and has good application prospect.
Description
Technical Field
the invention relates to the technical field of plant tissue culture and rapid propagation, in particular to a test tube micro-cuttage aseptic propagation method for eggplant stocks.
background
solanum torvum (Solanum torvum), also called "Toolubam", is a closely related wild species of the cultivated species eggplant (Solanum melogena) and is native to America. The plant has high resistance to bacterial wilt, verticillium wilt, blight, root-knot nematode and other main soil-borne diseases and pests, and is a commonly used grafting stock in eggplant and tomato production.
although the growth potential of the plant is extremely strong, the solanum torvum seeds are extremely small and have low endogenous hormone content, so that the solanum torvum seeds have extremely strong dormancy after being mature, and the normally sown solanum torvum seeds bud for about one month. After the germination and the soil breaking, the seedlings grow very slowly in the initial stage, and particularly the growth speed is nearly stopped under the low-temperature condition.
Due to the problems of long seedling age, low germination rate, germination vigor and germination index, slow seedling growth speed and the like of the solanum torvum bunge, the large-scale application of the solanum torvum bunge on industrial seedling culture is limited, so that the development of other methods for improving the seedling culture efficiency of the solanum torvum bunge and reducing the seedling culture cost is urgently needed.
disclosure of Invention
The invention aims to: aiming at the problems, the method for the test tube micro-cuttage sterile propagation of the eggplant stocks is provided, the method is simple to operate, low in production cost, easy to obtain explants, high in multiplication coefficient and short in growth period, and is an effective way for obtaining a large number of solanum torvum in a short time.
In order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows:
a method for the test tube micro-cuttage aseptic propagation of eggplant stocks comprises the following steps:
(1) obtaining aseptic seedlings: pretreating solanum torvum seeds, sowing the solanum torvum seeds on the surface of an MS culture medium, and obtaining primary sterile seedlings when the seeds germinate and grow to 3-4 true leaves;
(2) Inducing lateral buds: taking the stem segment of the aseptic seedling obtained in the step (1) as an explant, cutting the stem segment of the side bud of the aseptic seedling into 0.5cm in a superclean bench by using a sterilization blade, inoculating the stem segment of the side bud of the aseptic seedling into a bud induction culture medium in a micro-cuttage mode, and culturing until the side bud grows into 5-6 true leaves;
(3) Subculture multiplication culture: micro-cuttage is carried out on the stem segments growing out the buds in the step (2) into a subculture multiplication culture medium, the buds can be continuously multiplied in the culture medium, subculture is carried out every 25-30 days, and the step is repeated to realize mass multiplication;
(4) rooting culture: cutting the buds propagated in the step (3) when the buds grow to 1-2 cm long into a rooting culture medium for rooting culture, and obtaining tissue culture seedlings for hardening seedling and transplanting after 25-30 d of rooting;
(5) hardening and transplanting seedlings: transferring the tissue culture seedlings obtained in the step (4) from culture to natural light for hardening for 3 days, opening a bottle cover, continuing hardening for 3 days, taking out the plants, washing a root culture medium with clear water, punching a hole in the substrate, implanting the tissue culture seedlings, covering the substrate, slightly compacting to enable the plants to be upright, and keeping the substrate moist.
preferably, in step (1), the pretreatment is: soaking solanum torvum seeds in 1g/L gibberellin for 3 hours, washing with sterile water on a super clean workbench for 3 times, soaking with 70% ethanol for 15 seconds, sterilizing with 0.1% HgCl2 for 10min, and washing with sterile water for 5-6 times.
Preferably, in step (2), the bud induction medium is MS medium added with 0.4-0.6 mg/L, IBA 0.05.05-0.15 mg/L of KT, 25-32 g/L of sucrose, 4.5-5.0 g/L of agar, and pH 5.8-6.0.
Preferably, in the step (3), the subculture multiplication medium is MS medium added with 0.35-0.45 mg/L IBA, 25-32 g/L sucrose, 4.5-5.0 g/L agar, and pH 5.8-6.0.
Preferably, in the step (4), 0.15-0.25 mg/L of IBA, 25-32 g/L of sucrose, 4.5-5.0 g/L of agar and pH 5.8-6.0 are added into the 1/2MS culture medium.
Preferably, in step (5), the substrate is peat: coconut husk: perlite is 3: 2: 1.
preferably, in the culture process, the temperature of the culture chamber is set to be 25 +/-2 ℃, the relative humidity is set to be 50-60%, the illumination time is 16h/d, and the illumination intensity is set to be 2000-3000 lx.
the pH value of the culture medium is adjusted by 1mol/L HCl or NaOH.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
(1) the stem section is used as the explant, and the used explant is simple and easy to take; the plant growth regulator has simple proportioning, low price of the used reagent and low cost; the bud inductivity reaches more than 90 percent, and side buds about 1cm can grow after 15 days; subculture multiplication is carried out for 30 days, the multiplication coefficient reaches 5-6 times, the rooting effect is obvious, and the survival rate of transplanting is high.
(2) the invention establishes the whole tissue culture and rapid propagation technical system of the solanum torvum, has short period of the whole production process, simple and convenient operation and high multiplication factor, can completely meet the requirement of large-scale seedling propagation and has good application prospect.
drawings
FIG. 1 is a schematic diagram of the induction of Solanum torvum buds;
FIG. 2 is a schematic diagram of the subculture proliferation of Solanum torvum;
FIG. 3 is another schematic diagram of the subculture proliferation of Solanum torvum;
FIG. 4 is a schematic view of the rooting effect of Solanum torvum.
Detailed Description
in order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to preferred embodiments. It should be noted, however, that the numerous details set forth in the description are merely for the purpose of providing the reader with a thorough understanding of one or more aspects of the present invention, which may be practiced without these specific details.
example 1
a method for the test tube micro-cuttage aseptic propagation of eggplant stocks comprises the following steps:
The method comprises the following steps: preparing a culture medium, wherein a bud differentiation culture medium is prepared by adding KT0.5mg/L, IBA 0.10.10 mg/L, sucrose 30g/L and agar 4.8g/L to an MS culture medium; adding IBA0.40mg/L, sucrose 30g/L and agar 4.5-5.0 g/L into the subculture multiplication medium by using an MS culture medium; the rooting medium is 1/2MS medium with additional IBA0.20mg/L, cane sugar 30g/L and agar 4.5-5.0 g/L. Adjusting the pH of the culture medium to 5.8, subpackaging 50mL of each bottle into a tissue culture bottle, sealing, and sterilizing in an autoclave at 121 ℃ for 20 min;
Step two: obtaining aseptic seedlings, namely soaking solanum torvum seeds in 1g/L gibberellin for 3h, transferring the solanum torvum seeds into a super clean workbench for disinfection after the solanum torvum seeds are completely imbibed, firstly washing the solanum torvum seeds for 15s with 70% ethanol, washing the solanum torvum seeds for 3 times with sterile water, then disinfecting the solanum torvum seeds for 10min with 0.1% HgCl2, washing the solanum torvum seeds for 5 times with the sterile water, then sucking surface moisture with the sterilized filter paper, carefully dibbling the seeds onto the surface of an MS basic culture medium by using sterilized tweezers, and obtaining primary aseptic seedlings when the seeds germinate and grow;
Step three: inducing lateral buds, carefully taking out aseptic seedlings in an ultra-clean workbench, cutting stem sections containing growing points of the aseptic seedlings into about 0.5cm, simultaneously cutting off leaves, sterilizing the stem sections with 0.05% mercuric chloride for 10min, clamping the upper ends of the stem sections with sterilization forceps, directly inserting the stem sections into a bud induction culture medium, inserting 5 plants in each bottle at a depth of about 0.3cm, covering a cover on the bottle, placing the bottle in a culture room for culture, and carrying out subculture proliferation when 5-6 true leaves grow out of new buds;
Step four: carrying out subculture proliferation, namely taking out the buds in the third step, clamping the buds by using a sterilization forceps to carry out micro cuttage into a subculture proliferation culture medium, inserting the buds into the subculture proliferation culture medium to a depth of about 0.3cm, carrying out cuttage on 5 plants in each bottle, covering the bottles with a cover, putting the bottles into a culture chamber for culture, continuously growing the buds from the plants in the culture process, carrying out subculture for 30d, wherein the proliferation coefficient can reach 6.11, and repeating the step every 30d to realize proliferation;
step five: rooting culture, namely cutting the buds subjected to the subculture propagation in the fourth step into a rooting culture medium when the buds grow to 1-2 cm long for rooting culture, inserting the buds into a rooting culture medium with the depth of about 0.3cm, and cutting 5 plants in each bottle; covering the cover, putting the cover into a culture chamber for culturing, starting rooting after one week, wherein the rooting rate is 100%, the number of first-stage roots reaches 4.000 after 30d of rooting process, the root length is 145.09mm, the root thickness is 0.71mm, and the fibrous root is dense;
Step six: hardening and transplanting the seedlings, transferring the tissue culture seedlings obtained in the fifth step from culture to natural light for hardening for 3 days to enable the plants to adapt to external illumination conditions, then opening a bottle cap to continue hardening for 3 days, taking out the plants, washing a root culture medium with clear water, punching holes in a matrix to insert the tissue culture seedlings, covering the matrix and slightly compacting to enable the plants to be upright; the mixture ratio of the substrate is peat: coconut husk: perlite is 3: 2: 1, covering the three days before, placing the substrate in a culture room at 25 ℃ by using a plastic film to keep the substrate moist, wherein the relative humidity is 55%, the illumination time is 16h/d, the illumination intensity is 2500lx, and the stress resistance of the plants is enhanced and then transferring the plants into a greenhouse.
The method for establishing the solanum torvum fast propagation system comprises the processes of obtaining aseptic seedlings outside the solanum torvum, bud induction, subculture multiplication, rooting culture and hardening seedling transplantation. Wherein the bud induction culture medium is MS + KT0.5mg/L + IBA0.10 mg/L, the average induction rate reaches 92.5%, the successive transfer multiplication culture medium is MS + IBA0.40mg/L, the multiplication coefficient reaches 6.11 times, the rooting culture is 1/2MS + IBA0.20mg/L, and the rooting rate is 100%. The culture medium can improve the multiplication coefficient, has obvious rooting effect, greatly shortens the conventional breeding time, is simple to operate and low in cost, can meet the requirement of large-scale production, and provides a basis for the industrial production of the solanum torvum tissue culture seedlings.
Fig. 1 is a schematic diagram illustrating induction of solanum torvum buds in the present embodiment, and fig. 1 shows that solanum torvum buds have good induction effect; fig. 2 is a schematic diagram of the secondary proliferation of solanum torvum, fig. 3 is another schematic diagram of the secondary proliferation of solanum torvum, and as can be seen from fig. 2 and fig. 3, the secondary proliferation coefficient of solanum torvum is high; fig. 4 is a schematic view of the rooting effect of solanum torvum, and as can be seen from fig. 4, the solanum torvum has flourishing root system and remarkable rooting effect.
example 2
The method comprises the following steps: preparing a culture medium, wherein a bud induction culture medium is added with KT0.4mg/L, IBA 0.05.05 mg/L, sucrose 25g/L and agar 4.5g/L by an MS culture medium; in the subculture multiplication medium, IBA0.15mg/L, sucrose 30g/L and agar 4.5g/L are added to an MS culture medium; the rooting culture medium is 1/2MS culture medium with IBA0.15mg/L, sucrose 30g/L and agar 4.8 g/L. Adjusting the pH of the culture medium to 5.8, subpackaging 50mL of each bottle into a tissue culture bottle, sealing, and sterilizing in an autoclave at 121 ℃ for 20 min;
Step two: obtaining aseptic seedlings, namely soaking solanum torvum seeds in 1g/L gibberellin for 3h, transferring the solanum torvum seeds into a super clean workbench for disinfection after the solanum torvum seeds are completely imbibed, firstly washing the solanum torvum seeds for 15s with 70% ethanol, washing the solanum torvum seeds for 3 times with sterile water, then disinfecting the solanum torvum seeds for 10min with 0.1% HgCl2, washing the solanum torvum seeds for 5 times with the sterile water, then sucking surface moisture with the sterilized filter paper, carefully dibbling the seeds onto the surface of an MS basic culture medium by using sterilized tweezers, and obtaining primary aseptic seedlings when the seeds germinate and grow;
Step three: inducing lateral buds, carefully taking out aseptic seedlings in an ultra-clean workbench, cutting stem sections containing growing points of the aseptic seedlings into about 0.5cm, simultaneously cutting off leaves, sterilizing the stem sections with 0.05% mercuric chloride for 10min, clamping the upper ends of the stem sections with sterilization forceps, directly inserting the stem sections into a bud induction culture medium, inserting 5 plants in each bottle at a depth of about 0.3cm, covering a cover on the bottle, placing the bottle in a culture room for culture, and carrying out subculture proliferation when 5-6 true leaves grow out of new buds;
Step four: carrying out subculture proliferation, namely taking out the buds in the third step, clamping the buds by using a sterilization forceps to carry out micro cuttage into a subculture proliferation culture medium, inserting the buds into the subculture proliferation culture medium to a depth of about 0.3cm, carrying out cuttage on 5 plants in each bottle, covering the bottles with a cover, putting the bottles into a culture chamber for culture, continuously growing the buds from the plants in the culture process, carrying out subculture for 30d, wherein the proliferation coefficient can reach 6.11, and repeating the step every 30d to realize proliferation;
Step five: rooting culture, namely cutting the buds subjected to the subculture propagation in the fourth step into a rooting culture medium when the buds grow to 1-2 cm long for rooting culture, inserting the buds into a rooting culture medium with the depth of about 0.3cm, and cutting 5 plants in each bottle; covering the cover, putting the cover into a culture chamber for culturing, starting rooting after one week, wherein the rooting rate is 100%, the number of first-stage roots reaches 5.00 after 30d of rooting process, the root length is 119.68mm, the root thickness is 0.43mm, and the fibrous root is dense;
Step six: hardening and transplanting the seedlings, transferring the tissue culture seedlings obtained in the fifth step from culture to natural light for hardening for 3 days to enable the plants to adapt to external illumination conditions, then opening a bottle cap to continue hardening for 3 days, taking out the plants, washing a root culture medium with clear water, punching holes in a matrix to insert the tissue culture seedlings, covering the matrix and slightly compacting to enable the plants to be upright; the mixture ratio of the substrate is peat: coconut husk: perlite is 3: 2: 1, covering the three days before, placing the substrate in a culture room at 25 ℃ by using a plastic film to keep the substrate moist, wherein the relative humidity is 55%, the illumination time is 16h/d, the illumination intensity is 2500lx, and the stress resistance of the plants is enhanced and then transferring the plants into a greenhouse.
the method for establishing the solanum torvum fast propagation system comprises the processes of obtaining aseptic seedlings outside the solanum torvum, bud differentiation induction, subculture proliferation, rooting culture and hardening seedling transplantation. Wherein the bud induction culture medium is MS + KT0.4mg/L + IBA0.05mg/L, the average induction rate reaches 91.2%, the subculture multiplication culture medium is MS + IBA0.15mg/L, the multiplication coefficient reaches 6.03 times, the rooting culture is 1/2MS + IBA0.15mg/L, and the rooting rate is 100%. The culture medium can improve the multiplication coefficient, has obvious rooting effect, greatly shortens the conventional breeding time, is simple to operate and low in cost, can meet the requirement of large-scale production, and provides a basis for the industrial production of the solanum torvum tissue culture seedlings.
example 3
The method comprises the following steps: preparing a culture medium, wherein a bud induction culture medium is prepared by adding KT0.45mg/L, IBA 0.15.15 mg/L, sucrose 32g/L and agar 5.0g/L to an MS culture medium; the subculture multiplication medium is characterized in that IBA0.25mg/L, sucrose 32g/L and agar 5.0g/L are added to an MS culture medium; the rooting medium is 1/2MS medium with IBA0.25mg/L, sucrose 32g/L and agar 5.0 g/L. Adjusting pH of the culture medium to 6.0, subpackaging 50mL per bottle into tissue culture bottles, sealing, and sterilizing in autoclave at 121 deg.C for 20 min;
Step two: obtaining aseptic seedlings, namely soaking solanum torvum seeds in 1g/L gibberellin for 3h, transferring the solanum torvum seeds into a super clean workbench for disinfection after the solanum torvum seeds are completely imbibed, firstly washing the solanum torvum seeds for 15s with 70% ethanol, washing the solanum torvum seeds for 3 times with sterile water, then disinfecting the solanum torvum seeds for 10min with 0.1% HgCl2, washing the solanum torvum seeds for 5 times with the sterile water, then sucking surface moisture with the sterilized filter paper, carefully dibbling the seeds onto the surface of an MS basic culture medium by using sterilized tweezers, and obtaining primary aseptic seedlings when the seeds germinate and grow;
Step three: inducing lateral buds, carefully taking out aseptic seedlings in an ultra-clean workbench, cutting stem sections containing growing points of the aseptic seedlings into about 0.5cm, simultaneously cutting off leaves, sterilizing the stem sections with 0.05% mercuric chloride for 10min, clamping the upper ends of the stem sections with sterilization forceps, directly inserting the stem sections into a bud induction culture medium, inserting 5 plants in each bottle at a depth of about 0.3cm, covering a cover on the bottle, placing the bottle in a culture room for culture, and carrying out subculture proliferation when 5-6 true leaves grow out of new buds;
Step four: carrying out subculture proliferation, namely taking out the buds in the third step, clamping the buds by using a sterilization forceps to carry out micro cuttage into a subculture proliferation culture medium, inserting the buds into the subculture proliferation culture medium to a depth of about 0.3cm, carrying out cuttage on 5 plants in each bottle, covering the bottles with a cover, putting the bottles into a culture chamber for culture, continuously growing the buds from the plants in the culture process, carrying out subculture for 30d, wherein the proliferation coefficient can reach 6.11, and repeating the step every 30d to realize proliferation;
step five: rooting culture, namely cutting the buds subjected to the subculture propagation in the fourth step into a rooting culture medium when the buds grow to 1-2 cm long for rooting culture, inserting the buds into a rooting culture medium with the depth of about 0.3cm, and cutting 5 plants in each bottle; covering the cover, putting the cover into a culture chamber for culturing, starting rooting after one week, wherein the rooting rate is 100%, the number of first-stage roots reaches 4.67 after 30d of rooting process, the root length is 112.64mm, the root thickness is 0.36mm, and the fibrous root is dense;
step six: hardening and transplanting the seedlings, transferring the tissue culture seedlings obtained in the fifth step from culture to natural light for hardening for 3 days to enable the plants to adapt to external illumination conditions, then opening a bottle cap to continue hardening for 3 days, taking out the plants, washing a root culture medium with clear water, punching holes in a matrix to insert the tissue culture seedlings, covering the matrix and slightly compacting to enable the plants to be upright; the mixture ratio of the substrate is peat: coconut husk: perlite is 3: 2: 1, covering the three days before, placing the substrate in a culture room at 25 ℃ by using a plastic film to keep the substrate moist, wherein the relative humidity is 55%, the illumination time is 16h/d, the illumination intensity is 2500lx, and the stress resistance of the plants is enhanced and then transferring the plants into a greenhouse.
the method for establishing the solanum torvum fast propagation system comprises the processes of obtaining aseptic seedlings outside the solanum torvum, bud induction, subculture multiplication, rooting culture and hardening seedling transplantation. Wherein the bud induction culture medium is MS + KT0.45mg/L + IBA0.15mg/L, the average induction rate reaches 90.4%, the subculture multiplication culture medium is MS + IBA0.25mg/L, the multiplication coefficient reaches 5.23 times, the rooting culture is 1/2MS + IBA0.25mg/L, and the rooting rate is 100%. The culture medium can improve the multiplication coefficient, has obvious rooting effect, greatly shortens the conventional breeding time, is simple to operate and low in cost, can meet the requirement of large-scale production, and provides a basis for the industrial production of the solanum torvum tissue culture seedlings.
the foregoing is only a preferred embodiment of the present invention, and it should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (7)
1. A method for the test tube micro-cuttage aseptic propagation of eggplant stocks is characterized by comprising the following steps: the method comprises the following steps:
(1) Obtaining aseptic seedlings: pretreating solanum torvum seeds, sowing the solanum torvum seeds on the surface of an MS culture medium, and obtaining primary sterile seedlings when the seeds germinate and grow to 3-4 true leaves;
(2) Inducing lateral buds: taking the stem segment of the aseptic seedling obtained in the step (1) as an explant, cutting the stem segment of the side bud of the aseptic seedling into 0.5cm in a superclean bench by using a sterilization blade, inoculating the stem segment of the side bud of the aseptic seedling into a bud induction culture medium in a micro-cuttage mode, and culturing until the side bud grows into 5-6 true leaves;
(3) subculture multiplication culture: micro-cuttage is carried out on the stem segments growing out the buds in the step (2) into a subculture multiplication culture medium, the buds can be continuously multiplied in the culture medium, subculture is carried out every 25-30 days, and the step is repeated to realize mass multiplication;
(4) Rooting culture: cutting the buds propagated in the step (3) when the buds grow to 1-2 cm long into a rooting culture medium for rooting culture, and obtaining tissue culture seedlings for hardening seedling and transplanting after 25-30 d of rooting;
(5) hardening and transplanting seedlings: transferring the tissue culture seedlings obtained in the step (4) from culture to natural light for hardening for 3 days, opening a bottle cover, continuing hardening for 3 days, taking out the plants, washing a root culture medium with clear water, punching a hole in the substrate, implanting the tissue culture seedlings, covering the substrate, slightly compacting to enable the plants to be upright, and keeping the substrate moist.
2. The method for test-tube micro-cutting aseptic propagation of eggplant stocks according to claim 1, characterized in that: in step (1), the pretreatment is: soaking solanum torvum seeds in 1g/L gibberellin for 3 hours, washing with sterile water on a super clean workbench for 3 times, soaking with 70% ethanol for 15 seconds, sterilizing with 0.1% HgCl2 for 10min, and washing with sterile water for 5-6 times.
3. the method for test-tube micro-cutting aseptic propagation of eggplant stocks according to claim 1, characterized in that: in the step (2), the bud induction culture medium is MS culture medium added with 0.4-0.6 mg/L, IBA 0.05.05-0.15 mg/L of KT, 25-32 g/L of sucrose, 4.5-5.0 g/L of agar, and pH is 5.8-6.0.
4. The method for test-tube micro-cutting aseptic propagation of eggplant stocks according to claim 1, characterized in that: in the step (3), the subculture multiplication medium is MS medium added with 0.35-0.45 mg/L of IBA, 25-32 g/L of sucrose, 4.5-5.0 g/L of agar and with the pH value of 5.8-6.0.
5. The method for test-tube micro-cutting aseptic propagation of eggplant stocks according to claim 1, characterized in that: in the step (4), 0.15-0.25 mg/L of IBA, 25-32 g/L of cane sugar, 4.5-5.0 g/L of agar and pH 5.8-6.0 are added into 1/2MS culture medium.
6. The method for test-tube micro-cutting aseptic propagation of eggplant stocks according to claim 1, characterized in that: in step (5), the substrate is peat: coconut husk: perlite is 3: 2: 1.
7. The method for test-tube micro-cutting aseptic propagation of eggplant stocks according to claim 1, characterized in that: in the culture process, the temperature of the culture room is set to be 25 +/-2 ℃, the relative humidity is set to be 50-60%, the illumination time is 16h/d, and the illumination intensity is 2000-3000 lx.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910863985.3A CN110537489A (en) | 2019-09-12 | 2019-09-12 | Test tube micro-cuttage aseptic propagation method for eggplant rootstocks |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910863985.3A CN110537489A (en) | 2019-09-12 | 2019-09-12 | Test tube micro-cuttage aseptic propagation method for eggplant rootstocks |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110537489A true CN110537489A (en) | 2019-12-06 |
Family
ID=68713495
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910863985.3A Pending CN110537489A (en) | 2019-09-12 | 2019-09-12 | Test tube micro-cuttage aseptic propagation method for eggplant rootstocks |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110537489A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116369207A (en) * | 2023-06-05 | 2023-07-04 | 西南林业大学 | Solanum torvum tissue culture rapid propagation method and application |
-
2019
- 2019-09-12 CN CN201910863985.3A patent/CN110537489A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 张红: "水茄的组织培养与快速繁殖", 《植物生理学通讯》 * |
| 张红: "茄子砧木"托鲁巴姆"组培快繁简化技术", 《北方园艺》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116369207A (en) * | 2023-06-05 | 2023-07-04 | 西南林业大学 | Solanum torvum tissue culture rapid propagation method and application |
| CN116369207B (en) * | 2023-06-05 | 2023-08-04 | 西南林业大学 | Solanum torvum tissue culture rapid propagation method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107980635B (en) | Two-step transplanting method for high-survival-rate apple tissue culture seedlings | |
| CN110352853B (en) | Method for improving quality and transplanting survival rate of atractylis lancea test-tube plantlet | |
| CN111280056A (en) | Subculture breeding method of stingless pepper tissue culture seedlings | |
| CN111616052A (en) | Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei | |
| CN103430845A (en) | Strawberry tissue culturing method | |
| CN108029559B (en) | Method for quickly cultivating ilex latifolia tissue culture seedlings | |
| CN112189566A (en) | Rapid breeding method of cherry seedlings for rootstocks | |
| CN105532467B (en) | Endangered rhododendron molle in-vitro tissue culture propagation and preservation method | |
| CN103155868B (en) | Rapid seeding raising method of cherry rootstock ZY-1 tissue culture | |
| CN103155869A (en) | Sweet cherry rootstock Colt tissue culture method | |
| CN111034617B (en) | Method for breeding tea seedlings by culturing young embryo tissues of Yunnan large-leaf tea trees | |
| CN109247235B (en) | Rapid breeding and seedling raising method for cymbidium faberi Rolfe | |
| CN116602213A (en) | Tissue culture method for one-step seedling formation of bulbil konjak leaf surface corm | |
| CN110537489A (en) | Test tube micro-cuttage aseptic propagation method for eggplant rootstocks | |
| WO2022171212A2 (en) | Method for ex vivo culturing of thornless green prickly ash zanthoxylum armatum dc | |
| CN103155866B (en) | Malus zumi tissue culture rapid propagation seedling raising method | |
| CN113383706B (en) | Efficient eucommia bark regeneration method based on LED light quality regulation | |
| CN112931226B (en) | Tissue culture rapid propagation method for alnus ferox | |
| CN111557242B (en) | Method for culturing and rapidly propagating lotus tissue culture seedlings | |
| CN112106664B (en) | Sterile germination and rapid propagation method for michelia spectabilis seeds | |
| CN101543184A (en) | Method for culturing open type tissue of konjak | |
| CN1768576A (en) | Peony standardized quick-breeding method | |
| CN113678736A (en) | Tissue culture rapid propagation method of amorphophallus rivieri | |
| CN106900552B (en) | Culture medium kit and method for promoting rapid in-vitro propagation of fish | |
| CN110999786A (en) | Pear tissue culture seedling rooting and domestication method |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191206 |