CN116326483A - Application of nerolidol in rooting culture of Mo Langen-shaped stems - Google Patents
Application of nerolidol in rooting culture of Mo Langen-shaped stems Download PDFInfo
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- CN116326483A CN116326483A CN202310448738.3A CN202310448738A CN116326483A CN 116326483 A CN116326483 A CN 116326483A CN 202310448738 A CN202310448738 A CN 202310448738A CN 116326483 A CN116326483 A CN 116326483A
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- CN
- China
- Prior art keywords
- nerolidol
- rooting culture
- langen
- rooting
- culture medium
- Prior art date
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- FQTLCLSUCSAZDY-UHFFFAOYSA-N (+) E(S) nerolidol Natural products CC(C)=CCCC(C)=CCCC(C)(O)C=C FQTLCLSUCSAZDY-UHFFFAOYSA-N 0.000 title claims abstract description 29
- FQTLCLSUCSAZDY-ATGUSINASA-N Nerolidol Chemical compound CC(C)=CCC\C(C)=C\CC[C@](C)(O)C=C FQTLCLSUCSAZDY-ATGUSINASA-N 0.000 title claims abstract description 29
- WASNIKZYIWZQIP-AWEZNQCLSA-N nerolidol Natural products CC(=CCCC(=CCC[C@@H](O)C=C)C)C WASNIKZYIWZQIP-AWEZNQCLSA-N 0.000 title claims abstract description 29
- 239000012883 rooting culture medium Substances 0.000 claims abstract description 24
- 241000173252 Cymbidium sinense Species 0.000 claims abstract description 21
- 238000012258 culturing Methods 0.000 claims abstract description 3
- 239000001963 growth medium Substances 0.000 claims description 11
- 229920001817 Agar Polymers 0.000 claims description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- 238000005286 illumination Methods 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 241000272194 Ciconiiformes Species 0.000 claims description 7
- 239000012882 rooting medium Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 2
- 206010034878 phimosis Diseases 0.000 claims 3
- 230000006378 damage Effects 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 5
- 241000196324 Embryophyta Species 0.000 abstract description 3
- 208000027418 Wounds and injury Diseases 0.000 abstract description 3
- 230000007123 defense Effects 0.000 abstract description 3
- 208000014674 injury Diseases 0.000 abstract description 3
- 229930000044 secondary metabolite Natural products 0.000 abstract description 3
- 241000238631 Hexapoda Species 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 230000012010 growth Effects 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract description 2
- 238000001727 in vivo Methods 0.000 abstract description 2
- 238000009776 industrial production Methods 0.000 abstract description 2
- 210000000056 organ Anatomy 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 150000001875 compounds Chemical class 0.000 abstract 1
- 229930003658 monoterpene Natural products 0.000 abstract 1
- 150000002773 monoterpene derivatives Chemical class 0.000 abstract 1
- 235000002577 monoterpenes Nutrition 0.000 abstract 1
- 239000000306 component Substances 0.000 description 9
- 241000233855 Orchidaceae Species 0.000 description 5
- 230000008901 benefit Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 2
- 238000004161 plant tissue culture Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 241000732800 Cymbidium Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000003505 terpenes Chemical class 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
- A01H4/002—Culture media 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)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses application of nerolidol in rooting culture of Mo Langen corm. The invention cultures the cymbidium sinense rootstock in rooting culture medium for rooting culture, wherein the rooting culture medium contains monoterpene compound nerolidol. Plant organs respond after being subjected to factors such as mechanical injury or insect damage, and secondary metabolites are released for indirect defense through a series of complex regulation and control in vivo. In the process of culturing Mo Langen corms in vitro, physical damage is unavoidable, and a series of important endogenous signal molecules for defending reaction are activated, so that the growth and development of the corms are affected. The invention cultures the cymbidium sinense in the rooting culture medium containing nerolidol component, which can promote the cymbidium sinense to root, thereby overcoming the problem that the cymbidium sinense root is not differentiated or is difficult to differentiate, and having important practical significance for realizing the tissue culture industrial production of cymbidium sinense seedlings.
Description
Technical Field
The invention relates to a plant tissue culture method in agricultural biotechnology, in particular to application of nerolidol in Mo Langen-like stem rooting culture.
Background
The national orchid is a flower with the characteristic advantages of Guangxi provinces, has long history and deep culture foundation, and has important ornamental, economic and cultural values. At present, the propagation of the cymbidium is mainly carried out by dividing plant propagation, and the problems of low propagation coefficient, long period, high production cost and the like exist, so that the market demand can not be met. The seedling industrialized propagation based on the tissue culture technology has the advantages of high propagation coefficient, high speed, high benefit and the like. However, the orchid with the root-shaped stem as an intermediate propagule is one of the orchid types which are most difficult to be industrially propagated at present, so that the problem of difficult industrial propagation of orchid seedlings is not solved fundamentally so far, and the industrial development of the orchid is severely restricted.
Disclosure of Invention
In order to solve the problem that the root stem root is difficult to differentiate or does not differentiate in the tissue culture of the cymbidium sinense, the nerolidol is added into a rooting culture medium so as to promote the rooting of the cymbidium sinense.
The invention is realized by the following technical scheme: application of nerolidol in rooting culture of Mo Langen corm, and rooting culture of Mo Langen corm in rooting culture medium containing nerolidol (CAS number 7212-44-4).
In one embodiment of the invention, the rooting medium contains 2-6 micromoles of nerolidol, 1.0-2.0 milligrams of NAA, 0.1-0.5 milligrams of 6-BA, 20-30 grams of sucrose, 5-7 grams of agar powder, and the balance of 1/2MS medium, and the pH value is 5.4-6.0.
The application method of the nerolidol comprises the following steps: and sterilizing the components of the rooting culture medium except the nerolidol by high-temperature steam, and adding the filtered and sterilized nerolidol into a sterile operation table after the rooting culture medium is cooled to below 50 ℃. The high temperature steam sterilization condition is 121 ℃ for 20 minutes.
The rooting culture conditions of the invention are as follows: inoculating Mo Langen stems into a root stem root differentiation medium, inoculating Mo Langen stems into a root growth medium, performing dark culture at 25-22 ℃ for 7-14 days, transferring to the culture temperature of 25-22 ℃ and the illumination intensity of 2000-3000lx, and culturing for 30-40 days at the illumination length of 12 hours/day.
The Mo Langen corms refer to the root corms of the cymbidium sinense variety 'penguin Bai Momo blue' bud differentiated.
Plant organs respond after being subjected to factors such as mechanical injury or insect damage, and secondary metabolites are released for indirect defense through a series of complex regulation and control in vivo. Terpenoids such as nerolidol are one of the indirect defenses against secondary metabolites. In the process of in vitro culture of Mo Langen corms, physical injury is unavoidable, and a series of important endogenous signal molecules for defending reaction are activated, so that biosynthesis of endogenous hormones is blocked, and growth and development of the corms are affected. The invention cultures the cymbidium sinense in the rooting culture medium containing nerolidol component, which can promote the cymbidium sinense to root, thereby overcoming the problem that the cymbidium sinense root is not differentiated or is difficult to differentiate, and having important practical significance for realizing the tissue culture industrial production of cymbidium sinense seedlings.
Detailed Description
The technical scheme of the present invention will be described in further detail below with reference to the specific embodiments, but the present invention is not limited thereto.
MS is an international culture medium, and the components and preparation method can refer to literature (Tan Wencheng, dai Cegang, main editions, ornamental plant tissue culture technique, beijing: china forestry Press, 1991); the 1/2MS culture medium is formed by halving the concentration of major elements in an MS basic culture medium, and the concentration of other components is unchanged.
Example 1
The bud-differentiated cymbidium sinense variety 'penguin Bai Momo cymbidium sinense rootstock is respectively cultivated in the 1-3 groups of rooting culture mediums, firstly, dark cultivation is carried out for 7 days at the cultivation temperature of 25 ℃, then, the culture is carried out for 30 days under the conditions that the cultivation temperature is 25 ℃, the illumination intensity is 2000lx and the illumination length is 12 hours/day, and the rooting rate of the' penguin Bai Momo cymbidium sinense rootstock cultivated in the 1-3 groups of rooting culture mediums is 31.5%, 2.47% and 23.9%, respectively. Wherein the culture medium component of the rooting culture medium 1 is 1/2MS+2 mu mol/L nerolidol+
0.1 mg/L6-BA+1.0 mg/L NAA+20g/L sucrose+5 g/L agar, pH value is 5.4; the rooting culture medium 2 has the culture medium components of 1/2MS+0.1 mg/L6-BA+1.0 mg/L NAA+20g/L sucrose+5 g/L agar and the pH value of 5.4; the rooting medium 3 comprises 1/2MS+1. Mu. Mol/L nerolidol+0.1 mg/L6-BA+1.0 mg/L NAA+20g/L sucrose+5 g/L agar, and has pH value of 5.4
Example 2
The bud-differentiated cymbidium sinense variety' penguin Bai Momo cymbidium sinense rootstock is respectively cultivated in a rooting culture medium 4-5 group, firstly, dark cultivation is carried out for 10 days at the cultivation temperature of 27 ℃, then, the culture temperature is 25 ℃, the illumination intensity is 2500lx, the illumination length is 12 hours/day, and after the culture is carried out for 35 days, the rooting rate of the cymbidium sinense Bai Momo rootstock cultivated in the rooting culture medium 4-5 group is 36.9 percent and 3.52 percent respectively. Wherein the culture medium component of the rooting culture medium 4 is 1/2MS+4 mu mol/L nerolidol+0.3 mg/L6-BA+1.5 mg/L NAA+25g/L sucrose+6 g/L agar, and the pH value is 5.2; the rooting culture medium 5 has culture medium components of 1/2MS+0.3mg/L6-BA+1.5 mg/L NAA+25g/L sucrose+6 g/L agar and pH value of 5.2.
Example 3
The bud-differentiated cymbidium sinense variety 'penguin Bai Momo' root-like stems are respectively cultivated in a rooting culture medium 6-2 group, firstly, dark cultivation is carried out for 14 days at the cultivation temperature of 22 ℃, then the culture temperature is changed to 22 ℃, the illumination intensity is 3000lx, the illumination length is 12 hours/day, and after the culture is carried out for 40 days, the rooting rate of the 'penguin Bai Momo' root-like stems cultivated in the rooting culture medium 6-2 group is 42.2%, 3.06% and 25.5%, respectively. Wherein the culture medium composition of the rooting culture medium 6 is 1/2MS+6 mu mol/L nerolidol+0.5 mg/L6-BA+2.0 mg/L NAA+25g/L sucrose+7 g/L agar, and the pH value is 6.0; the rooting culture medium 7 has the culture medium components of 1/2MS+0.5 mg/L6-BA+2.0 mg/L NAA+25g/L sucrose+
7g/L agar, pH value is 6.0; the rooting medium 2 has the medium components of 1/2MS+2 mu mol/L nerolidol+0.5 mg/L6-BA+2.0 mg/L NAA+25g/L sucrose+7 g/L agar and the pH value of 6.0.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (5)
1. The application of nerolidol in rooting culture of Mo Langen corm, and the Mo Langen corm is cultured in rooting culture medium for rooting culture, and is characterized in that the rooting culture medium contains nerolidol.
2. The use of nerolidol according to claim 1 in rooting culture of Mo Langen phimosis, wherein the rooting culture medium comprises 2-6 micromoles of nerolidol, 1.0-2.0 milligrams NAA, 0.1-0.5 milligrams 6-BA, 20-30 grams sucrose, 5-7 grams agar powder, the balance being 1/2MS culture medium, and the pH value is 5.4-6.0.
3. The use of nerolidol in Mo Langen like stem rooting culture according to claim 1, wherein components of the rooting medium other than nerolidol are sterilized by high temperature steam, and after the culture is cooled to below 50 ℃, the filtered sterilized nerolidol is added into a sterile operating table.
4. The use of nerolidol according to any one of claims 1-4 in rooting culture of Mo Langen like stems, wherein the rooting culture conditions are: inoculating the Mo Langen stems into a rooting culture medium, performing dark culture at 25-22 ℃ for 7-14 days, transferring to the culture temperature of 25-22 ℃ and the illumination intensity of 2000-3000lx, and culturing for 30-40 days at the illumination length of 12 hours/day.
5. The use of nerolidol according to claim 1 in rooting culture of Mo Langen phimosis, wherein Mo Langen phimosis is the root of cymbidium sinense variety 'penguin Bai Momo cymbidium sinense' bud differentiated.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310448738.3A CN116326483A (en) | 2023-04-24 | 2023-04-24 | Application of nerolidol in rooting culture of Mo Langen-shaped stems |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310448738.3A CN116326483A (en) | 2023-04-24 | 2023-04-24 | Application of nerolidol in rooting culture of Mo Langen-shaped stems |
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| Publication Number | Publication Date |
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| CN116326483A true CN116326483A (en) | 2023-06-27 |
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| CN202310448738.3A Pending CN116326483A (en) | 2023-04-24 | 2023-04-24 | Application of nerolidol in rooting culture of Mo Langen-shaped stems |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102986526A (en) * | 2011-11-02 | 2013-03-27 | 福建省亚热带植物研究所 | Open-type plant tissue culture method |
| US20150173373A1 (en) * | 2013-12-19 | 2015-06-25 | Rhizoflora, Inc. | Plant Activator Composition |
| CN105340757A (en) * | 2015-12-14 | 2016-02-24 | 广东省农业科学院环境园艺研究所 | Tissue culture method for cymbidium tortisepalum and application thereof |
| CN112970588A (en) * | 2021-04-30 | 2021-06-18 | 江西省林业科学院 | Breeding method for efficiently inducing adventitious bud differentiation of sassafras callus |
| CN115518056A (en) * | 2022-10-12 | 2022-12-27 | 中国医学科学院医药生物技术研究所 | Use of nerolidol, nerol and geraniol for antibacterial purpose |
-
2023
- 2023-04-24 CN CN202310448738.3A patent/CN116326483A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102986526A (en) * | 2011-11-02 | 2013-03-27 | 福建省亚热带植物研究所 | Open-type plant tissue culture method |
| US20150173373A1 (en) * | 2013-12-19 | 2015-06-25 | Rhizoflora, Inc. | Plant Activator Composition |
| CN105340757A (en) * | 2015-12-14 | 2016-02-24 | 广东省农业科学院环境园艺研究所 | Tissue culture method for cymbidium tortisepalum and application thereof |
| CN112970588A (en) * | 2021-04-30 | 2021-06-18 | 江西省林业科学院 | Breeding method for efficiently inducing adventitious bud differentiation of sassafras callus |
| CN115518056A (en) * | 2022-10-12 | 2022-12-27 | 中国医学科学院医药生物技术研究所 | Use of nerolidol, nerol and geraniol for antibacterial purpose |
Non-Patent Citations (3)
| Title |
|---|
| 张东旭;李承秀;王长宪;潘银萍;王丰妍;张传芹;: "蕙兰杂交种子的无菌萌发和快速繁殖研究", 中国农学通报, no. 12, pages 159 - 164 * |
| 张志勇 等: "铁皮石斛开放式组织培养体系的建立", 《贵州农业科学》, vol. 46, no. 6, pages 24 - 25 * |
| 许申平;袁秀云;王默霏;崔波;: "墨兰(Cymbiduim sinense)组培快繁技术体系研究", 热带作物学报, no. 05 * |
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