CN106234221A - Oligochitosan is in plant tissue culture and the application of Fast-propagation - Google Patents
Oligochitosan is in plant tissue culture and the application of Fast-propagation Download PDFInfo
- Publication number
- CN106234221A CN106234221A CN201610633896.6A CN201610633896A CN106234221A CN 106234221 A CN106234221 A CN 106234221A CN 201610633896 A CN201610633896 A CN 201610633896A CN 106234221 A CN106234221 A CN 106234221A
- Authority
- CN
- China
- Prior art keywords
- oligochitosan
- culture
- tissue culture
- propagation
- fast
- 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
- 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)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to agrochemical field, especially relate to oligochitosan in plant tissue culture and the application of Fast-propagation.Oligosaccharide adds in culture medium as growth stimulator, and oligochitosan can be dry powder, solution or other water-soluble dosage forms.The present invention utilizes the MS culture medium adding oligochitosan that plant is carried out tissue culture, it is possible to significantly improves callus and sprouts rate, shortens the Callus formation time, shortens subculture cycle.Wherein oligochitosan can be the preparation that other of oligochitosan dry powder, oligochitosan solution or oligochitosan are dissolved in water.The concentration of oligochitosan solution is 5～25ppm.The new application of the oligochitosan of the present invention provides a new method for plant tissue culture Fast-propagation, and what beneficially shortening group cultivated thing breeds the time, increases economic efficiency.
Description
Technical field
The invention belongs to agrochemical field, especially relate to oligochitosan at plant tissue culture and Fast-propagation should
With.
Background technology
Tissue culture be under the artificial aseptic condition created by the isolated organ of life (such as root, stem, leaf, stem section, primary
Plastid), tissue or cell be placed in culture medium, and be placed in suitable environment, cultivate continuously to obtain cell, tissue or
Individual technology.This technology is widely used to agriculture and biological, medical research.With the red palm of economical ornamental flower it is
Example, by tissue culture technique produce red palm seedling mainly have the foundation of regenerating system, enrichment culture, strengthening seedling and rooting, transplanting and
The sport technique segment such as nurse young plants in hothouses.
In tissue culture's production practices of present stage, generally there is on the low side, the successive transfer culture cycle length partially of breeding coefficient etc.
Problem.At present for solving these problems, the hormone combination of variable concentrations the most all can be used to improve breeding coefficient, contract as far as possible
Short culture cycle attaining.Applicant utilizes the MS culture medium adding oligochitosan to carry out tissue culture, forms callus induction, improves
Breeding coefficient, quickening test tube Seedling reproduction speed aspect have extraordinary effect, can be greatly improved plant tissue culture breeding with this
Efficiency.
Summary of the invention
In order to overcome deficiency of the prior art, the present invention provide oligochitosan at plant tissue culture and Fast-propagation should
With, it is possible to significantly improve callus and sprout rate, shorten the Callus formation time, shorten subculture cycle, beneficially shortening group
That cultivates thing breeds the time, increases economic efficiency.
In order to solve above-mentioned technical problem, the technical solution used in the present invention is as follows: oligochitosan adds to as growth stimulator
In culture medium, oligochitosan can be dry powder, solution or other water-soluble dosage forms.
As preferably, above-mentioned oligochitosan solution concentration is 5～25ppm.
As preferably, above-mentioned culture medium is MS culture medium, facilitates the growth of plant callus.
It is required for using in Regeneration System stage, enrichment culture stage and the strengthening seedling and rooting stage of plant tissue culture
MS culture medium, and use add oligochitosan composition modified MS medium can be than the common MS culture medium being not added with oligochitosan composition
Effect is more preferable, and integral cycle is shorter.In such use, the concrete composition of MS culture medium is different because of Object of Development.This is because not
Different to the sensitivity of hormone with plant.But its main component content is constant, simply hormone concentration and media is different.
Additionally, the study find that, utilize the outer implant of the MS culture medium inoculated adding oligochitosan, 26 DEG C ± 2 DEG C, illumination
Cultivating under the conditions of 1500～2000Lx × 12h/d, the callus induction formation time can shorten 1/4th.And continue with
Oligochitosan MS culture medium carries out differentiation culture, and the differentiation culture time also can shorten 1/4th, and differentiation rate is significantly increased.
For general plant, oligochitosan is known has the effect promoting root growth, promoting root system increased number.And
Tissue culture is just referring to that plant parts tissue such as cambium layer, parenchyma, mesophyll tissue, endosperm etc. carry out cultivating and is obtaining regeneration
Plant, is the most all the plant growth regulating substance utilizing variable concentrations proportioning, such as 2, and 4-dichlorphenoxyacetic acid (2,4-D), naphthalene
Acetic acid (naphthalene acetic acid), 6-benzyl purine (6-benzyl aminoadenine) etc. complete moving back of plant parts tissue and are differentiated to form wound healing group
Knitting, then callus breaks up and then grows full plants body.The application finds that oligochitosan can be induced as little molecule
Material, forward regulates each process of tissue culture.
Oligochitosan of the present invention is carried out based on following principle in the application of plant tissue culture and Fast-propagation:
The Principle of Process of plant tissue culture is divided into two parts, one be living cells in outer implant through induction, recover it potential
Totipotency, is changed into meristematic cell, and its derivative cell is divided into parenchyma and forms callus then.Two is to produce
Callus, can induce neomorph or embryoid to form plant under certain condition further.
During Callus formation, outer planting somatic cell needs dedifferentiation after induction, and constantly division, hypertrophy are careful
Born of the same parents.The phytohormone that this derived need is certain, the ms culture medium adding oligochitosan has in terms of callus induction formation
Certain effect.Its principle needs to be studied further, speculates according to available data, it may be possible to owing to oligochitosan is as a kind of little molecule
Saccharide, can interact with the special receptor (glycoprotein) on cellular plasm film, and activation signal conducts, and promotes related gene
Transcribe and express.Its effect of endogenous hormones produced by auto-induction is better than exogenous hormone and adds.
It addition, the phenol pollution in tissue culture procedures can affect culture effect.Phenol pollution level is somatic with outer planting
Polyphenol oxidase activity is relevant.Phenol pollution level in incubation often increases the weight of with the raising of cytokinin concentration.Subculture
Because media environment deteriorates also results in the callus appearance phenomenon such as aging, clayization and abnormal differentiation in incubation.
Oligochitosan is a kind of small-molecule substance with positive charge, can adsorb toxicant, the subculture training of indirect protection callus
The process of supporting.
In the present invention, oligochitosan is the height that the degree of polymerization is 2-30 that glucosamine is formed by connecting by Isosorbide-5-Nitrae-glycosidic bond
The mixture of Molecularly Imprinted Polymer, mean molecule quantity is 1000-3000 dalton.Oligochitosan can be prepared by any means, example
Method as following: deacetylation chitosan more than 70% passes through physical degradation methods, chemical degradation method, enzymatic degradation method, sugar
The mixture of the Chitosan that the degree of polymerization is 2-30 that the methods such as group-transfer method, composite degradation method prepare.Or, pass through
From fungal cell wall extract obtain chitin and pass through physical degradation methods, chemical degradation method, enzymatic degradation method, glycosyl transfer method,
The Chitosan mixture that the degree of polymerization is 2-30 that the methods such as composite degradation method prepare.Or, by by aminoglucose
Sugar is by Chitosan mixture that the 1,4-glycosidic bond chemosynthesis degree of polymerization is 2-30.Oligochitosan is different polymerization at present
The mixture of the high molecular polymer of thing.That is oligochitosan is not the oligosaccharide form of the single degree of polymerization.But every kind
The oligochitosan of the degree of polymerization but soup all there may be.So no matter by which kind of method prepared, or by above-mentioned preparation side
The commercially available prod that method prepares, if its be glucosamine by the degree of polymerization that Isosorbide-5-Nitrae-glycosidic bond is formed by connecting be 2-30,
Mean molecule quantity be the mixture of the daltonian high molecular polymer of 1000-3000 be exactly oligochitosan of the present invention, it can
With containing acetyl group, but the amount that the sugar unit of bonding acetyl group is less than 40%, the amount of preferably more than 30%, particularly preferably
Amount less than 20%.
Oligochitosan is added in MS culture medium by the present invention, and the MS culture medium adding oligochitosan can accelerate induced tissue cultivation
During calli induction, and accelerate test tube Seedling reproduction speed to shorten Subculture Time.Oligochitosan of the present invention is to induction
Callus formation, raising breeding coefficient, quickening test tube Seedling reproduction speed aspect have extraordinary effect, and can be big with this
The big efficiency improving plant tissue culture breeding.
Detailed description of the invention
In order to understand the present invention, further illustrate the present invention with embodiment below, but be not limited to the present invention.
Embodiment
As a example by the red palm of ornamental flower, utilize the stem section of the red palm as initially cultivate induced material (outer implant) carry out with
Lower step.
Being cleaned down by the dust hairbrush of outer planting surface, flowing water is rinsed well, ultra violet lamp 10min.Proceed to ultra-clean
In workbench, the ethanol wash with 75% about 30 seconds, aseptic water washing, then with 0.1% mercuric chloride sterilizing 10-15 minute,
Fully wash with sterilized water, remove sterilized solution.Suck surface moisture with the filter paper through sterilizing, be cut into suitable size.
Outer implant obtained above is inoculated in respectively experimental group oligochitosan modified MS medium and matched group MS culture medium
Middle cultivation.Record induces the average time of available callus (producing the bud of about about 2cm).Wherein, comparison MS training
Foster base is to the addition of 6-benzyl aminoadenine 1.5ml/L, kinetins 1.0mg/L on the basis of regular MS media.Oligochitosan
Modified MS medium be with the addition of on the basis of regular MS media 6-benzyl aminoadenine 1.5ml/L, kinetins 1.0mg/L with
And 5～25ppm oligochitosans.
After forming callus, choose the basically identical red palm test tube Seedling of growing way and be respectively connected to two groups of differences 1/2MS cultivations
Base carries out successive transfer culture.Wherein oligochitosan improvement 1/2MS culture medium is interpolation naphthalene acetic acid on the basis of regular MS media
0.2mg/L, heteroauxing 2.0mg/L, activated carbon 0.2% and 5～25ppm oligochitosans, comparison 1/2MS culture medium is in routine
Naphthalene acetic acid 0.2mg/L, heteroauxing 2.0mg/L, activated carbon 0.2% is added on the basis of MS culture medium.
During after successive transfer culture, can constantly grow sprout, when propagation is to time a certain amount of, it is possible to decrease the amount of hormone is as used
0.1-0.2ml/L 6-benzyl aminoadenine, promotes that sprout is grown up and becomes strong, and grow aerial root.
After strong seedling culture 1-2 generation, the comparison 1/2MS proceeding to additional 0.1 mg/litre heteroauxing or naphthalene acetic acid respectively cultivates
The formation of root is promoted on base and oligochitosan improvement 1/2MS culture medium；Statistics starts to root growth to 3～4 lis from successive transfer culture
Time used during rice.
The time (d) that red palm callus from stem segment is induced by the different MS culture medium of table 1
| MS culture medium mesochite oligosaccharide concentration | Investigation stem section | Callus | Inductivity | Average induction time |
| 5ppm | 50 | 41 | 82% | 37d |
| 15ppm | 50 | 37 | 74% | 39d |
| 25ppm | 50 | 34 | 68% | 44d |
| Comparison MS culture medium | 50 | 28 | 56% | 52d |
Anthurium andraeanum callus enrichment culture extremely can be transplanted time (d) used by the different 1/2MS culture medium of table 2.
| MS culture medium mesochite oligosaccharide concentration | Average time used |
| 5ppm | 39d |
| 15ppm | 41d |
| 25ppm | 41d |
| Comparison 1/2MS culture medium | 49d |
As can be seen from Table 1 and Table 2, the MS culture medium of oligochitosan is used can to significantly improve the effect that plant tissue culture is bred
Rate accelerates manufacturing schedule.
The above, be only presently preferred embodiments of the present invention, and the present invention not makees any pro forma restriction, therefore
Every without departing from technical solution of the present invention content, any simply repaiied according to what above example made by the technical spirit of the present invention
Change, equivalent variations and modification, all still fall within the range of technical solution of the present invention.
Claims (5)
1. oligochitosan is in plant tissue culture and the application of Fast-propagation, it is characterised in that: oligochitosan adds to as growth stimulator
In culture medium, oligochitosan can be dry powder, solution or other water-soluble dosage forms.
Oligochitosan the most according to claim 1 is in plant tissue culture and the application of Fast-propagation, it is characterised in that: shell is few
Sugar juice concentration is 5～25ppm.
Oligochitosan the most according to claim 1 is in plant tissue culture and the application of Fast-propagation, it is characterised in that: cultivate
Base is MS culture medium.
4. oligochitosan as claimed in claim 1 is in plant tissue culture and the application of Fast-propagation, it is characterised in that: plant is
Red palm flowers.
5. oligochitosan as claimed in claim 4 is in plant tissue culture and the application of Fast-propagation, it is characterised in that: include with
Lower step: (1) is clean by the dust brush of red palm plant surface, and flowing water is rinsed well, ultra violet lamp 10min, proceeds to ultra-clean work
In station, with the ethanol wash 30s of 75%, then with aseptic water washing, then with 0.1% mercuric chloride sterilizing 10-15min, continue
Wash with sterilized water, remove sterilized solution；
(2) the outer implant that step (1) obtains is inoculated in oligochitosan modified MS medium, oligochitosan modified MS medium be
The shell widow of 6-benzyl aminoadenine 1.5ml/L, kinetins 1.0mg/L and 5～25ppm is with the addition of on the basis of regular MS media
Sugar；
(3) after step (2) outer planting body forms callus, choose the basically identical red palm test tube Seedling of growing way and be respectively connected to shell widow
Carrying out successive transfer culture in sugar improvement 1/2MS culture medium, oligochitosan improvement 1/2MS culture medium is on the basis of regular MS media
Add naphthalene acetic acid 0.2mg/L, heteroauxing 2.0mg/L, activated carbon 0.2% and 5～25ppm oligochitosans；
(4), during after successive transfer culture, with 0.1-0.2ml/L 6-benzyl aminoadenine, promote that sprout is grown up and become strong, and grow
Aerial root；
(5), after strong seedling culture 1-2 generation, the oligochitosan improvement 1/2MS training of additional 0.1 mg/litre heteroauxing or naphthalene acetic acid is proceeded to
Support the formation promoting root on base.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610633896.6A CN106234221A (en) | 2016-08-03 | 2016-08-03 | Oligochitosan is in plant tissue culture and the application of Fast-propagation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610633896.6A CN106234221A (en) | 2016-08-03 | 2016-08-03 | Oligochitosan is in plant tissue culture and the application of Fast-propagation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106234221A true CN106234221A (en) | 2016-12-21 |
Family
ID=58077756
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610633896.6A Pending CN106234221A (en) | 2016-08-03 | 2016-08-03 | Oligochitosan is in plant tissue culture and the application of Fast-propagation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106234221A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108124768A (en) * | 2017-12-14 | 2018-06-08 | 浙江诚邦园林股份有限公司 | A kind of method suitable for color leafed plants tissue culture seedling rooting |
| CN110810240A (en) * | 2019-11-08 | 2020-02-21 | 甘肃爽口源生态科技股份有限公司 | Stem tissue culture and rapid propagation method for Lanzhou lily plants |
| CN114041421A (en) * | 2021-11-25 | 2022-02-15 | 中国热带农业科学院海口实验站 | Tissue rapid propagation method of avocados |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415384A (en) * | 2011-10-28 | 2012-04-18 | 海南正业中农高科股份有限公司 | Purpose of chitosan oligosaccharide for regulating crop growth |
| CN103444714A (en) * | 2013-09-15 | 2013-12-18 | 海南正业中农高科股份有限公司 | Application of composition containing chitosan oligosaccharide for promoting plant flower organ expansion |
| CN103734009A (en) * | 2013-12-20 | 2014-04-23 | 三明市农业科学研究院 | Tissue culture method of anthurium |
| CN105010141A (en) * | 2015-07-09 | 2015-11-04 | 虞龙 | Anthurium rapid propagation method |
| CN105340415A (en) * | 2015-12-01 | 2016-02-24 | 青岛博智汇力生物科技有限公司 | Application of chitosan oligosaccharide as disease-resistant elicitor to cultivation of plant seedlings |
-
2016
- 2016-08-03 CN CN201610633896.6A patent/CN106234221A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415384A (en) * | 2011-10-28 | 2012-04-18 | 海南正业中农高科股份有限公司 | Purpose of chitosan oligosaccharide for regulating crop growth |
| CN103444714A (en) * | 2013-09-15 | 2013-12-18 | 海南正业中农高科股份有限公司 | Application of composition containing chitosan oligosaccharide for promoting plant flower organ expansion |
| CN103734009A (en) * | 2013-12-20 | 2014-04-23 | 三明市农业科学研究院 | Tissue culture method of anthurium |
| CN105010141A (en) * | 2015-07-09 | 2015-11-04 | 虞龙 | Anthurium rapid propagation method |
| CN105340415A (en) * | 2015-12-01 | 2016-02-24 | 青岛博智汇力生物科技有限公司 | Application of chitosan oligosaccharide as disease-resistant elicitor to cultivation of plant seedlings |
Non-Patent Citations (3)
| Title |
|---|
| 匡银近等: "壳寡糖对秋海棠叶片离体培养的影响", 《安徽农业科学》 * |
| 娄予强等: "壳寡糖及低聚壳聚糖在农业上应用研究综述", 《科技创新导报》 * |
| 黄菊艳等: "壳寡糖对不结球白菜子叶离体培养再生体系的影响", 《植物生理学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108124768A (en) * | 2017-12-14 | 2018-06-08 | 浙江诚邦园林股份有限公司 | A kind of method suitable for color leafed plants tissue culture seedling rooting |
| CN108124768B (en) * | 2017-12-14 | 2020-12-11 | 诚邦生态环境股份有限公司 | Rooting method suitable for tissue culture seedlings of color-leaf plants |
| CN110810240A (en) * | 2019-11-08 | 2020-02-21 | 甘肃爽口源生态科技股份有限公司 | Stem tissue culture and rapid propagation method for Lanzhou lily plants |
| CN114041421A (en) * | 2021-11-25 | 2022-02-15 | 中国热带农业科学院海口实验站 | Tissue rapid propagation method of avocados |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101258835B (en) | Fast reproducing method for high quality seedling of dendrobium officinale | |
| CN102499080B (en) | Plant fast propagating method using fagopyrum tataricum leaf stalks as explants | |
| CN105815213A (en) | Establishing method for in-vitro regeneration system of Kiwi berry | |
| CN102792888B (en) | Method for somatic embryogenesis and plant regeneration of Lijiang spruce | |
| CN102648698A (en) | Pyrus stem tip tissue culture rapid propagation method | |
| CN102771393B (en) | Method for picea balfouriana somatic embryo generation and plant regeneration | |
| CN105766654B (en) | A kind of Nanchuan jackfruit method for tissue culture | |
| CN106234221A (en) | Oligochitosan is in plant tissue culture and the application of Fast-propagation | |
| CN103416304A (en) | Method for cultivating water-saving and drought-resistant rice anther | |
| CN105191794B (en) | Method for rapid propagation of bischofia javanica based on embryogenic callus seedling development | |
| CN110313403B (en) | White birch embryonic callus induction culture medium, method for inducing embryonic callus and culture method of tissue culture seedling | |
| CN108739385A (en) | A kind of method for building up of Chinese pear blades efficient regenerating system and its application | |
| CN105265317B (en) | A kind of For-carrying green onions rapid propagation method | |
| CN105613288B (en) | A kind of construction method of Phnom Penh Chinese littleleaf box rapid propagation system | |
| CN106172008A (en) | A kind of Bowring cattleya fast culture propagation method | |
| CN108142284A (en) | A kind of tissue culture and rapid propagation method of five leaflets maple | |
| CN107646674A (en) | A kind of method that mountain ginseng culture root is produced using bioreactor | |
| CN1305371C (en) | Tufted bamboo tissue culture rapid breeding rooting method | |
| KR101439618B1 (en) | A Method for Mass Propagation of Rhododendron Keiskei var. hypoglaucum by Plant Tissue Culture | |
| CN104686336A (en) | Tissue culture rapid propagation method of ailanthus altissima | |
| CN101904302A (en) | Method for somatic cell embryogeny and plant regeneration of medicinal plant schisandga chinensis baill | |
| JP3877800B2 (en) | Propagation method of aseptic sweet potato seedlings | |
| CN107711501A (en) | A kind of tissue culture and rapid propagation method of radix tylophorae | |
| CN110938549B (en) | Microbial inoculum for promoting growth of euscaphis konishii container seedlings and preparation method and application thereof | |
| CN105815219A (en) | Method for acquiring explants on basis of young shoot pinching in June and July to increase Dangshansu pear tissue culture survival rate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into 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: 20161221 |