CN113661925A - Method for promoting rapid proliferation of dendrobium officinale protocorm - Google Patents
Method for promoting rapid proliferation of dendrobium officinale protocorm Download PDFInfo
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- CN113661925A CN113661925A CN202111170125.5A CN202111170125A CN113661925A CN 113661925 A CN113661925 A CN 113661925A CN 202111170125 A CN202111170125 A CN 202111170125A CN 113661925 A CN113661925 A CN 113661925A
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- dendrobium officinale
- culture medium
- protocorm
- dendrobium
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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
- A01H4/002—Culture media for tissue culture
Abstract
The invention discloses a method for promoting the rapid propagation of dendrobium officinale protocorm, which comprises the following steps: s1: collecting mature dendrobium officinale Kimura et Migo without plant diseases and insect pests, wherein the shell of the fruit is yellow green, and the uncracked capsule with the seed age of 6 months is taken as an explant, and carrying out pretreatment; s2: transversely cutting the pretreated capsule S1 with a fully-burned scalpel, clamping the capsule with forceps, uniformly scattering sterile seeds in an induction culture medium, and culturing for 30-60 days to obtain the dendrobium officinale protocorm; s3: the dendrobium officinale protocorm induced by the S2 is transferred to a multiplication culture medium to be rapidly propagated and cultured, and the dendrobium officinale protocorm which is green, large in individual, full and has conical buds is obtained after being cultured for 30-40 days.
Description
Technical Field
The invention relates to the technical field of plant culture, in particular to a method for promoting the rapid proliferation of dendrobium officinale protocorm.
Background
Dendrobium Yunnanense (Flickingenia albopurpurea Seidenf.) is a plant of the genus Dendrobium of Orchidaceae. The stem is yellow brown, drooping and multi-branch, the leaf leathers, the flower stalks and the ovaries are light yellow, the sepals and the petals are white, the flower texture is thin, the flower fades only in half a day when the flower is opened, the rhizome of the primary plant is bent and crawled on the trunk or on the understory rocks in a mountain sparse forest with the altitude of 800 plus 1200 meters, the primary plant is mainly produced in the south of Yunnan province, Thailand, Vietnam and Laos in China, and the primary plant can be used as pot flower and bonsai and has higher gardening value and medicinal value. The Dendrobium Yunnanense is listed in the International Trade Convention of Wild animals and plants (Convention on International Trade in enhanced specialties of Wild Fauna and Flora) Endangered by dead Species, belongs to the national secondary protection Wild plant, and is a Dendrobium plant belonging to the genus of the kindred genus of the Dendrobium plant. However, the problems of small propagation coefficient, long time consumption, low propagation rate and low yield of the dendrobium officinale are serious, and the increasing market demand cannot be met. Therefore, there is an urgent need to develop a method for promoting the rapid proliferation of dendrobium officinale protocorm to solve the above technical problems.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a method for promoting the rapid propagation of the dendrobium yunnanense protocorm, which ensures that the dendrobium yunnanense is rapidly propagated to plants with the same material metabolism and morphological development potential in tissue culture by promoting the rapid propagation of the dendrobium yunnanense protocorm, has high propagation multiple and rapid growth, does not basically cause variety variation, can maintain the stability of the variety, can greatly improve the propagation rate of the dendrobium yunnanense, solves the problem of low yield of the dendrobium yunnanense, and is a feasible measure for protecting endangered plants and relieving resource scarcity.
In order to realize the aim, the method for promoting the rapid propagation of the dendrobium officinale protocorm provided by the invention comprises the following steps:
s1: collecting mature dendrobium officinale Kimura et Migo without plant diseases and insect pests, wherein the shell of the fruit is yellow green, and the uncracked capsule with the seed age of 6 months is taken as an explant, and carrying out pretreatment;
s2: transversely cutting the pretreated capsule S1 with a fully-burned scalpel, clamping the capsule with forceps, uniformly scattering sterile seeds in an induction culture medium, and culturing for 30-60 days to obtain the dendrobium officinale protocorm;
s3: transferring the dendrobium officinale protocorm induced by the S2 to a proliferation culture medium to enable the dendrobium officinale protocorm to be rapidly propagated and cultured, and obtaining the dendrobium officinale protocorm which is green, large in individual, full and provided with conical buds after culturing for 30-40 days.
Preferably, in S1, the preprocessing method includes: the capsule is washed by tap water repeatedly, soaked for 10min by using detergent, washed by tap water for 6 times, drained, placed on a super clean workbench for later use, sterilized by using 75% alcohol solution for 30s on the super clean workbench, washed by using sterile water for 3 times, treated by using 3.5% sodium hypochlorite solution for 8min, washed by using sterile water for 6 times, placed on sterile paper for sucking water, and placed into a bottle.
Preferably, in S2, the method for preparing the induction medium includes: taking MS as a basic culture medium, adding 1.0mg/L of 6-benzylaminopurine, 0.1mg/L of naphthylacetic acid, 3% of sucrose and 8g/L of agar powder, subpackaging an induction culture medium, sterilizing by an autoclave and cooling for use, wherein the pH value of the induction culture medium is 5.8.
Preferably, in S3, the method for preparing the propagation medium includes: taking MS as a basic culture medium, adding 0.5 mg/L6-benzylaminopurine, 0.5mg/L naphthylacetic acid, 3% sucrose and 8g/L agar powder, subpackaging a proliferation culture medium, sterilizing by an autoclave and cooling for use, wherein the pH value of the proliferation culture medium is 5.8.
Preferably, in S2 and S3, both the induction culture and the proliferation culture are performed in a culture chamber, wherein the temperature of the culture chamber is (24 +/-2) DEG C, the illumination is 12h/d, and the illumination intensity is 1500-2000 Lx.
The method for promoting the rapid proliferation of the dendrobium officinale protocorm has the following beneficial effects.
1. The invention promotes the rapid propagation of the protocorm of the dendrobium candidum, so that the dendrobium candidum can be rapidly propagated into plants with the same material metabolism and morphological development potential in tissue culture, the propagation multiple is high, the growth is rapid, the variety variation is basically not caused, the stability of the variety can be maintained, the propagation rate of the dendrobium candidum can be greatly improved, the problem of low yield of the dendrobium candidum is solved, and the invention is a feasible measure for protecting endangered plants and relieving the resource scarcity.
2. The method can ensure that the dendrobium officinale can be rapidly propagated and grown in multiples within 3-4 months, improves the propagation rate and the yield of the dendrobium officinale, does not cause variety variation by the technology, can keep the stability of the dendrobium officinale variety, is suitable for industrial production, and is beneficial to popularization and application.
Drawings
FIG. 1 is a schematic diagram of mature and uncracked capsule of Dendrobium Yunnanense;
FIG. 2 is a schematic diagram of the sterile seeds of Dendrobium Yunnanense after being broadcast to an induction medium;
FIG. 3 is a schematic view showing the growth of the sterile seeds of Dendrobium Yunnanense after being broadcast to the induction medium for 30 days;
FIG. 4 is a schematic view showing the growth of the sterile seeds of Dendrobium Yunnanense after being broadcast to the induction medium for 60 days;
FIG. 5 is a schematic diagram of the Dendrobium monitum protocorm after being inoculated into a proliferation medium;
FIG. 6 is a schematic view showing the growth of Dendrobium Yunnanense protocorm after being inoculated into a proliferation culture medium for 30 days.
Detailed Description
The present invention will be further described with reference to the following specific embodiments and accompanying drawings to assist in understanding the contents of the invention.
The invention provides a method for promoting the rapid propagation of dendrobium officinale protocorm, which comprises the following steps:
s1: collecting mature dendrobium officinale Kimura et Migo without plant diseases and insect pests, wherein the shell of the fruit is yellow green, and the uncracked capsule with the seed age of 6 months is taken as an explant, and carrying out pretreatment; as shown in figure 1, is a schematic diagram of mature uncased capsule of dendrobium officinale. The pretreatment method comprises the following steps: the capsule is washed by tap water repeatedly, soaked for 10min by using detergent, washed by tap water for 6 times, drained, placed on a super clean workbench for later use, sterilized by using 75% alcohol solution for 30s on the super clean workbench, washed by using sterile water for 3 times, treated by using 3.5% sodium hypochlorite solution for 8min, washed by using sterile water for 6 times, placed on sterile paper for sucking water, and placed into a bottle.
S2: the S1 pretreated capsule was transversely cut using a fully burned scalpel, the capsule was grasped with forceps and the sterile seeds were evenly broadcast in induction medium. As shown in fig. 2, the schematic diagram is shown after the dendrobium yunnanense sterile seeds are sowed in the induction culture medium. Culturing for 30-60 days to obtain protocorm of herba Dendrobii Yunnanensis; the induction culture is carried out in a culture chamber, the temperature of the culture chamber is (24 +/-2) DEG C, the illumination is 12h/d, and the illumination intensity is 1500-2000 Lx. As shown in FIGS. 3-4, the growth of the sterile seeds of Dendrobium Yunnanense is shown in the schematic diagram 30 days and 60 days after the seeds are sowed in the induction medium.
The preparation method of the induction culture medium comprises the following steps: taking MS as a basic culture medium, adding 1.0mg/L of 6-benzylaminopurine, 0.1mg/L of naphthylacetic acid, 3% of sucrose and 8g/L of agar powder, subpackaging an induction culture medium, sterilizing by an autoclave and cooling for use, wherein the pH value of the induction culture medium is 5.8.
S3: transferring the dendrobium officinale protocorm induced by the S2 to a proliferation culture medium for rapid propagation culture, as shown in FIG. 5, which is a schematic diagram of the dendrobium officinale protocorm inoculated to the proliferation culture medium. Culturing for 30-40 days to obtain green Dendrobium Yunnanense protocorm with large individual, plump and conical bud; the proliferation culture is carried out in a culture chamber, the temperature of the culture chamber is (24 +/-2) DEG C, the illumination is 12h/d, and the illumination intensity is 1500-2000 Lx. As shown in FIG. 6, the growth of the Dendrobium loddigesii protocorm after being inoculated into the proliferation medium for 30 days is shown.
The preparation method of the proliferation culture medium comprises the following steps: taking MS as a basic culture medium, adding 0.5 mg/L6-benzylaminopurine, 0.5mg/L naphthylacetic acid, 3% sucrose and 8g/L agar powder, subpackaging a proliferation culture medium, sterilizing by an autoclave and cooling for use, wherein the pH value of the proliferation culture medium is 5.8.
In the process of increment culture, the influence of different hormone combinations on the rapid propagation of the dendrobium officinale protocorm is tested: MS is taken as a basic culture medium, 6-benzylaminopurine (6-BA) and naphthylacetic acid (NAA) with different concentrations are added, the influence of different hormone combinations on the increment rate and the growth condition of the dendrobium officinale protocorm is researched, and the test results are shown in table 1:
TABLE 1 Effect of different hormone combinations on the Rapid propagation of Dendrobium Dianthum protocorm
As is clear from Table 1, in the experimental treatments 1 to 6, the hormone ratios of the growth media selected in the present invention were optimal, and the growth rate was the highest and the growth was the best when 0.5 mg/L6-benzylaminopurine and 0.5mg/L naphthylacetic acid were selected.
The invention promotes the rapid propagation of the protocorm of the dendrobium candidum, so that the dendrobium candidum can be rapidly propagated into plants with the same material metabolism and morphological development potential in tissue culture, the propagation multiple is high, the growth is rapid, the variety variation is basically not caused, the stability of the variety can be maintained, the propagation rate of the dendrobium candidum can be greatly improved, the problem of low yield of the dendrobium candidum is solved, and the invention is a feasible measure for protecting endangered plants and relieving the resource scarcity. The method can ensure that the dendrobium officinale can be rapidly propagated and grown in multiples within 3-4 months, improves the propagation rate and the yield of the dendrobium officinale, does not cause variety variation by the technology, can keep the stability of the dendrobium officinale variety, is suitable for industrial production, and is beneficial to popularization and application.
The inventive concept is explained in detail herein using specific examples, which are given only to aid in understanding the core concepts of the invention. It should be understood that any obvious modifications, equivalents and other improvements made by those skilled in the art without departing from the spirit of the present invention are included in the scope of the present invention.
Claims (5)
1. A method for promoting the rapid propagation of dendrobium officinale protocorm is characterized by comprising the following steps:
s1: collecting mature dendrobium officinale Kimura et Migo without plant diseases and insect pests, wherein the shell of the fruit is yellow green, and the uncracked capsule with the seed age of 6 months is taken as an explant, and carrying out pretreatment;
s2: transversely cutting the pretreated capsule S1 with a fully-burned scalpel, clamping the capsule with forceps, uniformly scattering sterile seeds in an induction culture medium, and culturing for 30-60 days to obtain the dendrobium officinale protocorm;
s3: transferring the dendrobium officinale protocorm induced by the S2 to a proliferation culture medium to enable the dendrobium officinale protocorm to be rapidly propagated and cultured, and obtaining the dendrobium officinale protocorm which is green, large in individual, full and provided with conical buds after culturing for 30-40 days.
2. The method for promoting the rapid propagation of dendrobium officinale protocorm according to claim 1, wherein in S1, the pretreatment method comprises the following steps: the capsule is washed by tap water repeatedly, soaked for 10min by using detergent, washed by tap water for 6 times, drained, placed on a super clean workbench for later use, sterilized by using 75% alcohol solution for 30s on the super clean workbench, washed by using sterile water for 3 times, treated by using 3.5% sodium hypochlorite solution for 8min, washed by using sterile water for 6 times, placed on sterile paper for sucking water, and placed into a bottle.
3. The method for promoting the rapid propagation of dendrobium yunnanense protocorm according to claim 1, wherein the preparation method of the induction medium in S2 comprises the following steps: taking MS as a basic culture medium, adding 1.0mg/L of 6-benzylaminopurine, 0.1mg/L of naphthylacetic acid, 3% of sucrose and 8g/L of agar powder, subpackaging an induction culture medium, sterilizing by an autoclave and cooling for use, wherein the pH value of the induction culture medium is 5.8.
4. The method for promoting the rapid propagation of the dendrobium officinale protocorm according to claim 1, wherein the proliferation medium in the S3 is prepared by the following steps: taking MS as a basic culture medium, adding 0.5 mg/L6-benzylaminopurine, 0.5mg/L naphthylacetic acid, 3% sucrose and 8g/L agar powder, subpackaging a proliferation culture medium, sterilizing by an autoclave and cooling for use, wherein the pH value of the proliferation culture medium is 5.8.
5. The method according to claim 1, wherein the inducing culture and the proliferation culture are both performed in the culture chamber in S2 and S3, the temperature of the culture chamber is (24 + -2) deg.C, the illumination is 12h/d, and the illumination intensity is 1500-2000 Lx.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114303950A (en) * | 2021-12-31 | 2022-04-12 | 上海应用技术大学 | Tissue culture and rapid propagation method for dendrobium officinale seeds |
Citations (2)
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| CN101213940A (en) * | 2008-01-18 | 2008-07-09 | 中国科学院昆明植物研究所 | Fast replication method for dendrobium |
| CN109122313A (en) * | 2018-08-02 | 2019-01-04 | 广西壮族自治区中国科学院广西植物研究所 | A kind of culture medium and method for making wing calyx dendrobium nobile seed quickly breed seedling |
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2021
- 2021-10-08 CN CN202111170125.5A patent/CN113661925A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101213940A (en) * | 2008-01-18 | 2008-07-09 | 中国科学院昆明植物研究所 | Fast replication method for dendrobium |
| CN109122313A (en) * | 2018-08-02 | 2019-01-04 | 广西壮族自治区中国科学院广西植物研究所 | A kind of culture medium and method for making wing calyx dendrobium nobile seed quickly breed seedling |
Non-Patent Citations (2)
| Title |
|---|
| G. S. NAGANANDA等: ""Brick pieces soaked in liquid culture medium – a new matrix for seed germination and plantlet development for orchid Flickingeria nodosa (Dalz.) Seidenf."", 《CURRENT SCIENCE》 * |
| P. SERVIN WESLEY等: ""In vitro propagation of Coelogyne breviscapa Lindl., Dendrobium aqueum Lindl., and Flickingeria nodosa (Dalz.) Seidenf. via asymbiotic seed germination"", 《ASPAC J. MOL. BIOL. BIOTECHNOL.》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114303950A (en) * | 2021-12-31 | 2022-04-12 | 上海应用技术大学 | Tissue culture and rapid propagation method for dendrobium officinale seeds |
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