CN113412787A - Method for improving in-vitro flowering of dendrobium officinale - Google Patents
Method for improving in-vitro flowering of dendrobium officinale Download PDFInfo
- Publication number
- CN113412787A CN113412787A CN202110776148.4A CN202110776148A CN113412787A CN 113412787 A CN113412787 A CN 113412787A CN 202110776148 A CN202110776148 A CN 202110776148A CN 113412787 A CN113412787 A CN 113412787A
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- China
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- dendrobium officinale
- bud
- flower bud
- differentiation
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- 241001076416 Dendrobium tosaense Species 0.000 title claims abstract description 32
- 238000000338 in vitro Methods 0.000 title claims abstract description 18
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title claims abstract description 16
- 230000004069 differentiation Effects 0.000 claims abstract description 34
- 239000001963 growth medium Substances 0.000 claims abstract description 33
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 23
- 239000002609 medium Substances 0.000 claims abstract description 19
- 230000001737 promoting effect Effects 0.000 claims abstract description 15
- 230000001954 sterilising effect Effects 0.000 claims abstract description 11
- 230000006698 induction Effects 0.000 claims abstract description 10
- 238000005406 washing Methods 0.000 claims abstract description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229960002523 mercuric chloride Drugs 0.000 claims abstract description 4
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims abstract description 4
- 239000008399 tap water Substances 0.000 claims abstract description 4
- 235000020679 tap water Nutrition 0.000 claims abstract description 4
- 238000002791 soaking Methods 0.000 claims abstract description 3
- 230000000249 desinfective effect Effects 0.000 claims description 3
- 239000006870 ms-medium Substances 0.000 claims 2
- 241000196324 Embryophyta Species 0.000 description 7
- 239000000463 material Substances 0.000 description 3
- UXDFUVFNIAJEGM-UHFFFAOYSA-N 2-methoxy-5-[2-(3,4,5-trimethoxyphenyl)ethyl]phenol Chemical compound C1=C(O)C(OC)=CC=C1CCC1=CC(OC)=C(OC)C(OC)=C1 UXDFUVFNIAJEGM-UHFFFAOYSA-N 0.000 description 2
- 241001523681 Dendrobium Species 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241001122767 Theaceae Species 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 235000015961 tonic Nutrition 0.000 description 2
- 230000001256 tonic effect Effects 0.000 description 2
- 229960000716 tonics Drugs 0.000 description 2
- 229920001917 Ficoll Polymers 0.000 description 1
- 241000233855 Orchidaceae Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 239000003864 humus Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—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
Abstract
The invention discloses a method for improving in-vitro flowering of dendrobium officinale, which comprises the following steps of 1: in the blooming period of the dendrobium officinale, taking the bud of the unopened flower bud of the dendrobium officinale; washing with washing powder under tap water for half an hour; step 2: cutting the flower bud buds obtained in the step 1 into bud bodies with the length of 5cm, quickly soaking the bud bodies in 70% alcohol for 30 seconds, and then placing the bud bodies in 0.1% mercuric chloride solution for sterilization for 5 minutes; and step 3: cutting the buds in the step 2 into explants with the length of 1cm, and inoculating the explants into a callus induction culture medium; and 4, step 4: after the callus in the step 3 is induced, taking out the callus, and cutting the callus into explants with the length of 1cm and inoculating the explants into a differentiation culture medium; and 5: in the differentiation medium in the step 4, when the bud body differentiates, the flower bud is continuously inoculated into the flower bud differentiation promoting medium, and the flower bud differentiation in the flower bud differentiation promoting medium is obviously increased; steps 1, 2, 3, 4 and 5 were all performed on a sterile worktop.
Description
Technical Field
The invention belongs to the technical field of plant biology, and particularly relates to a method for promoting and improving in-vitro flowering of dendrobium officinale by a plant in-vitro culture technology.
Background
Dendrobium officinale is a plant of the genus Dendrobium of the family Orchidaceae. The famous Dendrobium of ficoll Kimura et Migo Green is named because the original environment is in a mountain land with the altitude of more than 1600 meters, the mountain land usually grows on semi-yin wet rock and the epidermis is iron Green. Dendrobium officinale is naturally distributed in Anhui, Zhejiang, Fujian, etc. places in China. The organs of the stems, flowers and the like contain polysaccharide, erianin and the like which can be used as medicines to play the roles of benefiting the stomach and promoting the production of body fluid, nourishing yin and clearing heat, and belong to yin tonics in tonics.
The dendrobium officinale is a herbaceous plant, and the stem is cylindrical and upright, is usually not branched and has multiple sections; the inflorescence is a raceme which usually comes from the upper part of an old stem and 2-3 flowers of a small flower. The dendrobium officinale is an epiphytic plant, and can simulate natural environment artificial breeding and cultivation by domestication. The artificial attached main planting of the dendrobium officinale can be divided into tree planting, stone planting, humus soil planting and other forms according to different characteristics of attached main plants.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a method for improving in-vitro flowering of dendrobium officinale, which promotes and improves in-vitro flowering of the dendrobium officinale by a plant in-vitro culture technology.
In order to achieve the purpose, the method for improving in-vitro flowering of the dendrobium officinale provided by the invention comprises the following steps:
step 1: in the blooming period of the dendrobium officinale, taking the bud of the unopened flower bud of the dendrobium officinale; washing with washing powder under tap water for half an hour;
step 2: cutting the flower bud buds obtained in the step 1 into bud bodies with the length of 5cm, and sterilizing and disinfecting;
and step 3: cutting the buds in the step 2 into explants with the length of 1cm, and inoculating the explants into a callus induction culture medium;
and 4, step 4: after the callus in the step 3 is induced, taking out the callus, and cutting the callus into explants with the length of 1cm and inoculating the explants into a differentiation culture medium;
and 5: in the differentiation medium of the step 4, when the bud body differentiates, the flower bud is continuously inoculated into the flower bud differentiation promoting medium, and the flower bud differentiation in the flower bud differentiation promoting medium is obviously increased.
Preferably, said steps 1, 2, 3, 4 and 5 are all performed on a sterile worktop;
preferably, the sterilization in the step 2 is performed by rapidly soaking the substrate in 70% alcohol for 30 seconds, and then placing the substrate in 0.1% mercuric chloride solution for sterilization for 5 minutes;
preferably, the callus induction culture medium takes an MS culture medium as a basic culture medium, and further comprises the following components in percentage by weight: 0.5 mg/L6-BA, 0.5 mg/L2.4-D and 0.1mg/L ZT;
preferably, the differentiation medium takes an MS culture medium as a basic culture medium, and further comprises the following components in percentage by weight: 1mg/l 6-BA and 0.1mg/l NAA;
preferably, the flower bud differentiation promoting medium takes an MS culture medium as a basic culture medium, and further comprises the following components in percentage by weight: 1mg/l 6-BA, 0.1mg/l NAA and 0.1mg/l AD.
The method for improving in-vitro flowering of dendrobium officinale provided by the invention has the following beneficial effects:
the method promotes and improves the in vitro flowering of the dendrobium officinale by sterile tube-entering treatment of buds, flower bud induction and bud differentiation promotion, and through a dendrobium officinale plant tissue culture technology, material selection time selection, a nutritional formula and hormone combination.
Detailed Description
The present invention will be further described with reference to specific examples to assist understanding of the invention.
The invention provides a method for improving in-vitro flowering of dendrobium officinale, which comprises the following steps:
1. in the flowering period of the dendrobium officinale, adopting germinated buds to perform sterile treatment on the bud explants to obtain sterile bud explants;
2. preparing callus induction culture medium, bud differentiation culture medium, flower bud differentiation promoting culture medium, etc., and sterilizing at high temperature and high pressure for use;
3. inoculating the sterile bud explants into an induction culture medium for inducing callus, and inducing the callus of the dendrobium officinale;
4. after the callus is induced, cutting on a sterile table, and inoculating a bud differentiation culture medium to promote bud differentiation;
5. on the bud differentiation culture medium, a material with a nutrient bud and a flower bud appears simultaneously, and the material is inoculated into the flower bud differentiation promoting culture medium, so that the flower bud differentiation proportion is further improved.
The method specifically comprises the following steps:
step 1: in the blooming period of the dendrobium officinale, taking the bud of the unopened flower bud of the dendrobium officinale; washing with washing powder under tap water for half an hour;
step 2: cutting the flower bud buds obtained in the step 1 into bud bodies with the length of 5cm, and sterilizing and disinfecting;
and step 3: cutting the buds in the step 2 into explants with the length of 1cm, and inoculating the explants into a callus induction culture medium;
and 4, step 4: after the callus in the step 3 is induced, taking out the callus, and cutting the callus into explants with the length of 1cm and inoculating the explants into a differentiation culture medium;
and 5: in the differentiation medium of the step 4, when the bud body differentiates, the flower bud is continuously inoculated into the flower bud differentiation promoting medium, and the flower bud differentiation in the flower bud differentiation promoting medium is obviously increased.
Steps 1, 2, 3, 4 and 5 are all operated on a sterile operating platform;
the sterilization operation in the step 2 is to rapidly soak the tea leaves in 70 percent alcohol for 30 seconds and then put the tea leaves in 0.1 percent mercuric chloride solution for sterilization for 5 minutes;
the callus induction culture medium takes an MS culture medium as a basic culture medium, and also comprises the following components by weight percent: 0.5 mg/L6-BA, 0.5 mg/L2.4-D and 0.1mg/L ZT;
the differentiation culture medium takes an MS culture medium as a basic culture medium and also comprises the following components in percentage by weight: 1mg/l 6-BA and 0.1mg/l NAA;
the flower bud differentiation promoting culture medium takes an MS culture medium as a basic culture medium, and also comprises the following components in percentage by weight: 1mg/l 6-BA, 0.1mg/l NAA and 0.1mg/l AD.
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 (6)
1. A method for improving in-vitro flowering of dendrobium officinale is characterized by comprising the following steps:
step 1: in the blooming period of the dendrobium officinale, taking the bud of the unopened flower bud of the dendrobium officinale; washing with washing powder under tap water for half an hour;
step 2: cutting the flower bud buds obtained in the step 1 into bud bodies with the length of 5cm, and sterilizing and disinfecting;
and step 3: cutting the buds obtained in the step 2 into explants with the length of 1cm, and inoculating the explants into a callus induction culture medium;
and 4, step 4: after the callus in the step 3 is induced, taking out the callus, and cutting the callus into explants with the length of 1cm and inoculating the explants into a differentiation culture medium;
and 5: in the differentiation medium of the step 4, when the bud body differentiates, the flower bud is continuously inoculated into the flower bud differentiation promoting medium, and the flower bud differentiation in the flower bud differentiation promoting medium is obviously increased.
2. The method for improving in-vitro flowering of dendrobium officinale according to claim 1, wherein the steps 1, 2, 3, 4 and 5 are all operated on a sterile operating platform.
3. The method for improving in-vitro flowering of dendrobium officinale according to claim 1, wherein the sterilization in the step 2 is performed by rapidly soaking in 70% alcohol for 30 seconds, and then placing in 0.1% mercuric chloride solution for sterilization for 5 minutes.
4. The method for improving in-vitro flowering of dendrobium officinale according to claim 1, wherein the callus induction medium is an MS medium as a basic medium, and further comprises the following components in percentage by weight: 0.5 mg/L6-BA, 0.5 mg/L2.4-D and 0.1mg/L ZT.
5. The method for improving in-vitro flowering of dendrobium officinale according to claim 1, wherein the differentiation medium takes an MS culture medium as a basic culture medium, and further comprises the following components in percentage by weight: 1mg/l 6-BA and 0.1mg/l NAA.
6. The method for improving in-vitro flowering of dendrobium officinale according to claim 1, wherein the flower bud differentiation promoting medium takes an MS medium as a basic medium, and further comprises the following components in percentage by weight: 1mg/l 6-BA, 0.1mg/l NAA and 0.1mg/l AD.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110776148.4A CN113412787B (en) | 2021-07-08 | 2021-07-08 | Method for improving in-vitro flower development of dendrobium candidum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110776148.4A CN113412787B (en) | 2021-07-08 | 2021-07-08 | Method for improving in-vitro flower development of dendrobium candidum |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113412787A true CN113412787A (en) | 2021-09-21 |
| CN113412787B CN113412787B (en) | 2024-01-30 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110776148.4A Active CN113412787B (en) | 2021-07-08 | 2021-07-08 | Method for improving in-vitro flower development of dendrobium candidum |
Country Status (1)
| Country | Link |
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| CN (1) | CN113412787B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104938331A (en) * | 2015-05-27 | 2015-09-30 | 广西健宝石斛有限责任公司 | Tissue culture propagation method of dendrobium officinale |
| CN105613292A (en) * | 2016-01-06 | 2016-06-01 | 江苏远鹏基因组研究院有限公司 | Special dendrobium officinale leaf sheath tissue culture seedling forming method |
| CN106718942A (en) * | 2017-02-15 | 2017-05-31 | 商丘人源农业科技有限公司 | The tissue-culturing rapid propagation and domestication culture techniques of North China's dendrobium candidum |
-
2021
- 2021-07-08 CN CN202110776148.4A patent/CN113412787B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104938331A (en) * | 2015-05-27 | 2015-09-30 | 广西健宝石斛有限责任公司 | Tissue culture propagation method of dendrobium officinale |
| CN105613292A (en) * | 2016-01-06 | 2016-06-01 | 江苏远鹏基因组研究院有限公司 | Special dendrobium officinale leaf sheath tissue culture seedling forming method |
| CN106718942A (en) * | 2017-02-15 | 2017-05-31 | 商丘人源农业科技有限公司 | The tissue-culturing rapid propagation and domestication culture techniques of North China's dendrobium candidum |
Non-Patent Citations (4)
| Title |
|---|
| 何静茹;李振坚;鲁琳;: "春石斛离体快繁与瓶内开花", 广东农业科学 * |
| 王光远等: "石斛离体培养中ABA对诱导花芽形成的影响", 植物学报 * |
| 王光远等: "铁皮石斛的离体开花", 中国科学 * |
| 董璐;李青;: "春石斛试管花芽分化与开花的影响因素", 东北林业大学学报 * |
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| Publication number | Publication date |
|---|---|
| CN113412787B (en) | 2024-01-30 |
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