CN108077067B - Tissue culture and rapid propagation method of cotton rose - Google Patents
Tissue culture and rapid propagation method of cotton rose Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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Abstract
The invention discloses a tissue culture and rapid propagation method of cotton rose, which comprises the following steps: (1) and (3) bud induction culture: cutting a stem section with bud points of cotton rose hibiscus as an explant, sterilizing, inoculating the stem section into an induction culture medium, and culturing until axillary buds germinate and grow to a certain length; (2) rooting culture: cutting the axillary buds obtained in the step (1), inoculating the axillary buds into a rooting culture medium, and culturing to form rooted seedlings; (3) and (3) proliferation culture: cutting stem sections or stem tips with single buds of the rooted seedlings in the step (2), inoculating the stem sections or stem tips into a multiplication culture medium, and culturing to form cotton rose hibiscus seedlings; (4) hardening and transplanting seedlings: and (4) moving the cotton rose hibiscus seedlings in the step (3) outdoors, closing bottles, hardening seedlings for 2-3 days, transplanting the cotton rose hibiscus seedlings to a seedbed, covering films, covering a sunshade net for culturing for 8-12 days until the cotton rose hibiscus seedlings grow new roots, and then removing the films and the sunshade net for culturing. The method can promote growth, the survival rate of the cultured seedlings is high, the rooting rate is up to 100%, the culture time is short, and the rapid propagation of the cotton rose hibiscus can be realized.
Description
Technical Field
The invention relates to the technical field of plant tissue culture and rapid propagation, in particular to a tissue culture and rapid propagation method of cotton rose.
Background
Hibiscus mutabilis is native to yellow river basin to south China and is distributed in Hunan, Sichuan, Yunnan, Guangdong provinces and the like. The lotus has a cultivation history of nearly three thousand years in China, is mainly planted in Hunan, Sichuan and other places at present, is also a city flower in metropolis, and is mostly planted in ponds by people. Besides being used as an ornament, the flowers and leaves of the cotton rose can be used as a medicine, and have the effects of clearing away heat and toxic materials, relieving swelling and the like. The existing hibiscus manihot propagation and cultivation technology is generally a cutting or grafting mode, has long propagation time and low survival rate and cannot be popularized.
Disclosure of Invention
In view of the above, the tissue culture and rapid propagation method of cotton rose provided by the invention has the advantages that the obtained seedlings are short in culture time, high in survival rate, high in rooting rate up to 100%, neat in seedlings, convenient for later-stage uniform transplanting and planting management, and capable of realizing rapid propagation of cotton rose.
In order to solve the technical problems, the invention provides a tissue culture rapid propagation method of cotton rose hibiscus, which comprises the following steps:
(1) and (3) bud induction culture: cutting a stem section with a bud point of cotton rose hibiscus to be used as an explant, sterilizing, inoculating the stem section into an induction culture medium, and culturing for 2-3 weeks until axillary buds on the explant germinate and grow to a certain length;
(2) rooting culture: cutting the axillary buds obtained in the step (1), inoculating the axillary buds into a rooting culture medium, culturing for 3-6 weeks until the base parts of the axillary buds root, and the roots grow to a certain length to form rooted seedlings;
(3) and (3) proliferation culture: cutting stem segments or stem tips with single buds of the rooted seedlings in the step (2), inoculating the stem segments or stem tips into a multiplication culture medium, culturing for 2-3 weeks until the stem segments or stem tips root, and growing the single buds to a certain length to form cotton rose hibiscus seedlings;
(4) hardening and transplanting seedlings: and (4) moving the cotton rose hibiscus seedlings in the step (3) outdoors, closing bottles, hardening seedlings for 2-3 days, transplanting the cotton rose hibiscus seedlings to a seedbed, covering films, covering a sunshade net for culturing for 8-12 days until the cotton rose hibiscus seedlings grow new roots, and then removing the films and the sunshade net for culturing.
Wherein the length of the axillary bud growth in the step (1) is 2-3 cm; the length of the root growth in the step (2) is 5-7 cm; the length of the single bud growth in the step (3) is 4-6 cm.
Preferably, the composition of the induction medium in step (1) is: MS minimal medium, 0.1-0.5mg/L6-BA, 0.05-0.15mg/L NAA, 10-30g/L sucrose, 4-6g/L agar, the concentration ratio of the 6-BA and the NAA is more than 1.
More preferably, the composition of the induction medium in step (1) is: MS minimal medium, 0.2mg/L6-BA, 0.1mg/L NAA, 20g/L sucrose, 6g/L agar.
Preferably, the sterilization treatment of the explant in the step (1) is as follows: and (3) cleaning the explant, treating the explant with 75% alcohol for 25-35s, cleaning with sterile water for 1-2 times, treating with 0.1% mercuric chloride for 8-10min, cleaning with sterile water for 3-5 times, and finally absorbing water with sterile filter paper.
Preferably, the rooting medium in step (2) and the proliferation medium in step (3) both have the following composition: 1/2MS minimal medium, which is supplemented with 0.1-0.5mg/L NAA, 0.05-0.15mg/L6-BA, 10-30g/L sucrose, 4-6g/L agar, and the concentration ratio of NAA to 6-BA is larger than 1.
More preferably, the rooting medium in step (2) and the proliferation medium in step (3) both have the following composition: 1/2MS minimal medium, supplemented with 0.2mg/L NAA, 0.1mg/L6-BA, 20g/L sucrose, 6g/L agar.
Preferably, the temperature of the bud induction culture, the rooting culture and the proliferation culture is 23-27 ℃, the illumination intensity is 1500-2000Lx, and the light cycle is 14/10 h.
Preferably, the matrix of the seedbed in the step (4) is prepared by mixing the components in a volume ratio of 1: (0.5-1) perlite and vermiculite.
More preferably, the matrix of the seedbed in the step (4) is prepared from a mixture of the following components in a volume ratio of 1: 1 perlite and vermiculite.
Preferably, the substrate is sterilized by chlorothalonil before transplanting the cotton rose seedlings.
Preferably, the culture temperature of the membrane covering and sun shading net covering in the step (4) is 23-27 ℃, the relative humidity is 90-99%, and the culture temperature of the membrane removing and sun shading net removing is 23-27 ℃, and the relative humidity is 70-80%.
More preferably, the culture temperature of the membrane covering and sun shading net covering in the step (4) is 25 ℃, the relative humidity is 95%, and the culture temperature of the membrane removing and sun shading net removing is 25 ℃, and the relative humidity is 75%.
Preferably, the shading rate of the shading net is 75%.
Preferably, after transplanting the cotton rose hibiscus seedlings in the step (4), spraying and watering the cotton rose hibiscus seedlings by using 1000-time dilution solution of carbendazim.
The MS minimal medium has high inorganic salt concentration, can ensure mineral nutrition required by tissue growth, is a stable ion balance solution, has high nitrate content and proper nutrient quantity and proportion, and can meet the nutritional and physiological needs of plant cells, so the application range is wide, and most of plant tissues are cultured and rapidly propagated to use the MS minimal medium as the minimal medium of the medium.
The 1/2MS minimal medium is a medium formed by halving macroelements and keeping the rest unchanged in the MS minimal medium.
The 6-BA is 6-benzylamino adenine, is an artificially synthesized plant cytokinin, has the main functions of promoting the formation of buds, inducing callus to generate, promoting cell division, promoting differentiation of non-differentiated tissues, promoting the accumulation of substances in organisms, promoting the generation of lateral buds, preventing aging, and is commonly used for plant tissues and cell culture.
NAA is naphthylacetic acid, is an auxin analogue in a plant growth regulator, is used when plants are propagated by a cutting method, can also be used for plant tissue culture, can promote cell division and expansion, induce to form adventitious roots to increase fruit setting, prevent fruit dropping, change the ratio of female flowers and male flowers and the like, can enter the plants through the tender epidermis and seeds of leaves and branches, and is guided to the whole plants along with nutrient flow.
The technical scheme mainly utilizes the principle of micro-cuttage to carry out tissue culture and propagation on the cotton rose hibiscus, the 6-BA is mainly used for promoting the growth of buds, and the NAA is mainly used for the growth of adventitious roots. In the bud induction culture, the concentration ratio of 6-BA/NAA is more than 1, and the germination of axillary buds is induced; in rooting and proliferation culture, NAA and 6-BA are added, and the concentration ratio of NAA/6-BA is greater than 1, so that the generation of root is induced, and the development of bud is promoted.
Compared with the prior rapid propagation technical research, the technical scheme has the advantages that the propagation period can be obviously shortened, the propagation and the rooting in the scheme are the same culture medium, the culture medium can meet the rooting requirement while promoting the bud development, and the step of rooting before seedling hardening is omitted.
In addition, the substrate treatment, the water management, the disinfection measures and the humidity control method before and during the transplanting in the technical scheme can ensure that the transplanting survival rate reaches 100 percent and is higher than the prior art level.
According to the technical scheme, the stem section with the bud point of cotton rose hibiscus is used as an explant to be subjected to bud induction culture, axillary buds germinate, then the axillary buds are subjected to rooting culture to form rooted seedlings, the rooted seedlings are cut into the stem section with the single bud or the stem tip to be subjected to proliferation culture to form cotton rose hibiscus seedlings, then seedling hardening and transplanting are carried out, the rapid propagation of the cotton rose hibiscus is realized through the method, and the growth period is shortened. The optimal culture medium components and proportion are obtained by screening the induction culture medium, the rooting culture medium and the proliferation culture medium, and the culture medium prepared by adopting the plant growth regulator has the advantages of short culture time of obtained seedlings, capability of promoting growth, high survival rate, rooting rate up to 100%, neat seedlings, convenience for later-stage uniform transplanting and planting management and realization of rapid propagation of cotton rose.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following detailed description of the present invention is provided with reference to specific embodiments.
Example 1
The tissue culture and rapid propagation method of cotton rose in the embodiment comprises the following steps:
(1) and (3) bud induction culture: the stem segment of the cotton rose bud is cut as an explant, after the stem segment is cleaned, the stem segment is firstly treated by 75% of alcohol for 30s, is cleaned by sterile water for 2 times, is treated by 0.1% of mercury bichloride for 9min, is cleaned by the sterile water for 4 times, and finally absorbs water by sterile filter paper, is inoculated to an induction culture medium which is adjusted to pH to 6.0 and is subjected to sterilization treatment for culture until axillary buds germinate and grow to a certain length on the explant, wherein the induction culture medium comprises the following components: MS minimal medium, supplement 0.2mg/L6-BA, 0.1mg/L NAA, 20g/L cane sugar, 6g/L agar;
(2) rooting culture: cutting the axillary buds obtained in the step (1), inoculating the axillary buds into a rooting culture medium for culturing until the base parts of the axillary buds root, and the roots grow to a certain length to form rooted seedlings, wherein the rooting culture medium comprises the following components: 1/2MS minimal medium, supplemented with 0.2mg/L NAA, 0.1mg/L6-BA, 20g/L sucrose, 6g/L agar;
(3) and (3) proliferation culture: cutting stem segments or stem tips with single buds of the rooted seedlings in the step (2), inoculating the stem segments or stem tips into a multiplication culture medium for culturing until the stem segments or stem tips root, and growing the single buds to a certain length to form the hibiscus mutabilis seedlings, wherein the multiplication culture medium comprises the following components: 1/2MS minimal medium, supplemented with 0.2mg/L NAA, 0.1mg/L6-BA, 20g/L sucrose, 6g/L agar;
(4) hardening and transplanting seedlings: and (4) transferring the cotton rose hibiscus seedlings in the step (3) outdoors, closing bottles, hardening seedlings for 2-3 days, and then transplanting the cotton rose hibiscus seedlings to a seedbed, wherein the substrate of the seedbed is prepared by mixing the following components in a volume ratio of 1: 1, sterilizing by using chlorothalonil, spraying and watering by using 1000 times of carbendazim solution after transplanting is finished, covering and moisturizing by using a plastic film, shading by using a shading net with the shading rate of 75%, controlling the temperature to be 25 ℃ and the relative humidity to be 95%, culturing for 8-12 days until new roots grow out of the cotton rose hibiscus seedlings, then removing the film, removing the shading net, and culturing at the temperature of 25 ℃ and the relative humidity of 75%.
Wherein the temperature of bud induction culture, rooting culture and proliferation culture is 25 ℃, the illumination intensity is 2000Lx, and the light cycle is 14/10 h.
In this example, after the bud induction culture of step (1) for 2 weeks, the axillary buds sprouting from the explants grow to 2-3 cm; after the rooting culture of the step (2) is carried out for 1 week, the base part of the axillary bud takes root, the root length is 2-4 cm after 3 weeks, the rooting rate is 100%, the upper axillary bud can be stretched to 5cm, and the axillary bud can be stretched to 10cm after 6 weeks; the stem segments or stem tips root after the multiplication culture for 1 week in the step (3), and the single buds grow to 5cm after 3 weeks, at the moment, the hibiscus mutabilis seedlings can be acclimatized and transplanted; after the cottonrose hibiscus seedlings are continuously cultured for 6 weeks, subculture can be carried out again, and the multiplication coefficient is 5 on average; and (4) growing new roots of the hibiscus seedlings 10 days after transplanting, wherein the transplanting survival rate reaches 100%.
Example 2
The tissue culture and rapid propagation method of cotton rose in this embodiment is different from that in embodiment 1 in that:
the composition of the induction culture medium in the step (1) is as follows: MS minimal medium, supplement 0.1mg/L6-BA, 0.05mg/LNAA, 10g/L sucrose, 4g/L agar;
the rooting culture medium in the step (2) comprises the following components: 1/2MS minimal medium, supplemented with 0.1mg/L NAA, 0.05mg/L6-BA, 10g/L sucrose, 4g/L agar;
the proliferation culture medium in the step (3) comprises the following components: 1/2MS minimal medium, supplemented with 0.1mg/L NAA, 0.05mg/L6-BA, 10g/L sucrose, 4g/L agar.
In this example, after 2 weeks of the bud induction culture in step (1), axillary buds sprouting from the explants grow to 1-2 cm; after 10 days of rooting culture in the step (2), the base parts of the axillary buds take roots, the root length is 2-3 cm after 3 weeks, the rooting rate is 100%, the upper axillary buds can be stretched to 4cm, and the axillary buds can be stretched to 8cm after 6 weeks; after 10 days of enrichment culture, the stem segments or stem tips take roots, after 4 weeks, single buds grow to 5-6 cm, and then the cottonrose hibiscus seedlings can be acclimatized and transplanted; after the cottonrose hibiscus seedlings are continuously cultured for 7 weeks, subculture can be carried out again, and the multiplication coefficient is 5 on average; and (4) growing new roots of the hibiscus seedlings 12 days after transplantation, wherein the transplanting survival rate reaches 95%.
Example 3
The tissue culture and rapid propagation method of cotton rose in this embodiment is different from that in embodiment 1 in that:
the composition of the induction culture medium in the step (1) is as follows: MS minimal medium, supplement 0.5mg/L6-BA, 0.15mg/LNAA, 30g/L sucrose, 6g/L agar;
the rooting culture medium in the step (2) comprises the following components: 1/2MS minimal medium, supplemented with 0.5mg/L NAA, 0.15mg/L6-BA, 30g/L sucrose, 6g/L agar;
the proliferation culture medium in the step (3) comprises the following components: 1/2MS minimal medium, supplemented with 0.5mg/L NAA, 0.15mg/L6-BA, 30g/L sucrose, 6g/L agar.
In this example, after the bud induction culture of step (1) for 2 weeks, the axillary buds sprouting from the explants grow to 2 cm; after the rooting culture of the step (2) is carried out for 1 week, the base part of the axillary bud takes root, the root length is 2-4 cm after 3 weeks, the rooting rate is 100%, the upper axillary bud can be stretched to 4cm, and the axillary bud can be stretched to 9cm after 6 weeks; the stem segments or stem tips root after the multiplication culture for 1 week in the step (3), and the single buds grow to 4cm after 3 weeks, at the moment, the hibiscus mutabilis seedlings can be acclimatized and transplanted; after continuously culturing for 6-7 weeks, the cottonrose hibiscus seedlings can be subjected to subculture multiplication again, and the multiplication coefficient is 5 on average; and (4) growing new roots of the hibiscus seedlings 11 days after transplanting, wherein the transplanting survival rate reaches 98%.
Example 4
The tissue culture and rapid propagation method of cotton rose in this embodiment is different from that in embodiment 1 in that:
in the hardening-seedling transplanting in the step (4), the matrix of the seedbed is prepared by mixing the following components in a volume ratio of 1: 0.5 perlite and vermiculite.
In the embodiment, new roots grow out from the cottonrose hibiscus seedlings 15 days after the transplanting, and the transplanting survival rate reaches 90%.
Example 5
Effect of plant growth regulator Components and concentrations in Induction Medium on shoot Induction culture
The stem segments with bud points of the cotton rose hibiscus are cut and taken as explants, after the stem segments are cleaned, 75% of alcohol is firstly used for treating for 30s, the stem segments are cleaned with sterile water for 2 times, then 0.1% of mercury bichloride is used for treating for 9min, the stem segments are cleaned with the sterile water for 4 times, finally, the stem segments are absorbed by sterile filter paper, and the stem segments are inoculated into an induction culture medium for culture until axillary buds germinate on the explants and grow to a certain length, wherein the induction culture medium comprises the following components: MS basic culture medium, supplementary plant growth regulator, sucrose, agar, under MS basic culture medium, sucrose, agar all the same condition, according to the composition and concentration of plant growth regulator grouping, observe and record the culture condition of the explant in the culture medium, specific grouping and culture result see Table 1.
TABLE 1 Effect of plant growth regulator Components and concentrations in Induction Medium on shoot Induction culture results
| Grouping | 6-BA(mg/L) | NAA(mg/L) | Number of explants | Number of sprouts (pieces) |
| 1 | 0.1 | -- | 20 | 14 |
| 2 | 0.2 | -- | 20 | 16 |
| 3 | 0.5 | -- | 20 | 16 |
| 4 | -- | 0.05 | 20 | 13 |
| 5 | -- | 0.10 | 20 | 13 |
| 6 | -- | 0.15 | 20 | 12 |
| 7 | 0.2 | 0.05 | 20 | 18 |
| 8 | 0.2 | 0.10 | 20 | 20 |
| 9 | 0.2 | 0.15 | 20 | 17 |
As can be seen from the above table, the number of sprouts when 6-BA and NAA are used simultaneously is higher than that when either one is used alone, and from 7-9 groups, when both are used simultaneously, the 6-BA is 0.2mg/L, and when NAA is 0.1mg/L, the sprouting rate is the highest, and the growth regulator is the most suitable composition condition for the plant growth regulator of the induction culture medium.
Example 6
Influence of plant growth regulator components and concentration in rooting medium on axillary bud rooting culture and bud number
A plurality of axillary buds formed after induction are inoculated into a rooting culture medium for culture until the base part of the axillary buds roots and the roots grow to a certain length to form a rooted seedling, wherein the rooting culture medium comprises the following components: 1/2MS minimal medium, supplementing plant growth regulator, sucrose and agar, grouping according to the components and concentration of the plant growth regulator under the condition that 1/2MS minimal medium, sucrose and agar are the same, observing and recording the culture condition of axillary buds in the medium, wherein the specific grouping and culture results are shown in Table 2.
TABLE 2 Effect of plant growth regulator composition and concentration in rooting Medium on axillary bud rooting culture and bud number
As can be seen from the table above, when the NAA is used alone, the same effect as that of a control group IBA (the existing commonly used plant growth regulator, but the IBA price is far higher than that of the NAA) can be achieved, the rooting rate is basically consistent, the rooting requirement can be met, but the average number of the bud points is small, and the next step proliferation requirement cannot be met. The number of buds of NAA and 6-BA used at the same time is higher than that of the buds of NAA used alone, and from 7-9 groups, the NAA is 0.2mg/L, and the 6-BA is 0.1mg/L, so that the number of roots, roots and buds is the largest, and the NAA and the 6-BA are the most suitable composition conditions of the plant growth regulator for the rooting and proliferation culture medium.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (8)
1. A tissue culture and rapid propagation method of cotton rose hibiscus is characterized in that: the tissue culture rapid propagation method comprises the following steps:
(1) and (3) bud induction culture: cutting a stem section with a bud point of cotton rose hibiscus to be used as an explant, sterilizing, inoculating the stem section into an induction culture medium, and culturing for 2-3 weeks until axillary buds on the explant germinate and grow to a certain length;
(2) rooting culture: cutting the axillary buds obtained in the step (1), inoculating the axillary buds into a rooting culture medium, culturing for 3-6 weeks until the base parts of the axillary buds root, and the roots grow to a certain length to form rooted seedlings;
(3) and (3) proliferation culture: cutting stem segments or stem tips with single buds of the rooted seedlings in the step (2), inoculating the stem segments or stem tips into a multiplication culture medium, culturing for 2-3 weeks until the stem segments or stem tips root, and growing the single buds to a certain length to form cotton rose hibiscus seedlings;
(4) hardening and transplanting seedlings: transferring the cotton rose hibiscus seedlings in the step (3) outdoors, closing bottles, hardening seedlings for 2-3 days, transplanting the cotton rose hibiscus seedlings to a seedbed, covering films on the cotton rose hibiscus seedlings, covering sun-shading nets on the cotton rose hibiscus seedlings, culturing the cotton rose hibiscus seedlings for 8-12 days until new roots grow on the cotton rose hibiscus seedlings, and removing the films and the sun-shading nets;
the composition of the induction medium is as follows: MS minimal medium, which is supplemented with 0.1-0.5mg/L6-BA, 0.05-0.15mg/L LNAA, 10-30g/L sucrose and 4-6g/L agar, wherein the concentration ratio of the 6-BA to the NAA is more than 1; the rooting medium and the proliferation medium both consist of: 1/2MS minimal medium, which is supplemented with 0.1-0.5mg/LNAA, 0.05-0.15mg/L6-BA, 10-30g/L sucrose, 4-6g/L agar, the concentration ratio of NAA to 6-BA is more than 1; the 1/2MS minimal medium is a medium formed by halving macroelements and keeping the rest unchanged in the MS minimal medium.
2. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: the induction culture medium in the step (1) comprises the following components: MS minimal medium, supplemented with 0.2mg/L6-BA, 0.1mg/L LNAA, 20g/L sucrose, 6g/L agar.
3. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: the explant sterilization treatment in the step (1) comprises the following steps: cleaning the explant, treating with 75% alcohol for 25-35s, washing with sterile water for 1-2 times, treating with 0.1% mercuric chloride for 8-10min, washing with sterile water for 3-5 times, and removing water with sterile filter paper.
4. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: the rooting culture medium in the step (2) and the proliferation culture medium in the step (3) both comprise the following components: 1/2MS minimal medium, supplemented with 0.2mg/LNAA, 0.1mg/L6-BA, 20g/L sucrose, 6g/L agar; the 1/2MS minimal medium is a medium formed by halving macroelements and keeping the rest unchanged in the MS minimal medium.
5. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: the temperature of the bud induction culture, the rooting culture and the proliferation culture is 23-27 ℃, the illumination intensity is 1500-.
6. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: and (4) the matrix of the seedbed in the step (4) is prepared from the following components in a volume ratio of 1: (0.5-1) perlite and vermiculite.
7. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: and (4) culturing the membrane covering and sun shading net at the temperature of 23-27 ℃ and the relative humidity of 90-99%, and culturing the membrane removing and sun shading net at the temperature of 23-27 ℃ and the relative humidity of 70-80%.
8. The tissue culture and rapid propagation method of cotton rose hibiscus as claimed in claim 1, wherein: and (4) after transplanting the cotton rose hibiscus seedlings, spraying and watering the cotton rose hibiscus seedlings by using a 1000-time carbendazim diluted solution.
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