CN108056101A - A kind of chitosan oligosaccharide-alginic acid nano particle and its preparation and application - Google Patents
A kind of chitosan oligosaccharide-alginic acid nano particle and its preparation and application Download PDFInfo
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- CN108056101A CN108056101A CN201610978760.9A CN201610978760A CN108056101A CN 108056101 A CN108056101 A CN 108056101A CN 201610978760 A CN201610978760 A CN 201610978760A CN 108056101 A CN108056101 A CN 108056101A
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- chitosan oligosaccharide
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
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Abstract
It prepares and its applies the present invention relates to nano particle, specifically the preparation of chitosan oligosaccharide alginic acid nano particle and its application as plant induced resistance agent and growth accelerator.Chitosan oligosaccharide, alginic acid are carried out polymer precipitation reaction by the present invention, and then ultrasonic, ultracentrifugation, which is separated and dried, prepares chitosan oligosaccharide alginic acid nano particle.Raw material chitosan oligosaccharide and alginic acid of the present invention are natural oligosaccharides and polysaccharide, and raw material is easy to get, cheap；The process for preparing nano particle adopts water as solvent, and preparation process is simple, is easy to amplify；The nano particle good dispersion of synthesis, stability is strong, does not add crosslinking agent and dispersant；The chitosan oligosaccharide alginic acid nano particle that this patent is invented has good bioactivity, while has inducing plant resistance and promote the activity of plant growth, is with a wide range of applications.
Description
Technical field
It prepares and its applies the present invention relates to nano particle, specifically the preparation of chitosan oligosaccharide-alginic acid nano particle
And its application as plant induced resistance agent and growth accelerator.
Background technology
Chitosan oligosaccharide be chitin after deacetylation, the degree of polymerization that hydrolyzes is the oligosaccharides of 2-10, structure for β-(1,
4) -2- deoxidations -2- aminoglucose oligosaccharides.Chitosan oligosaccharide is that a kind of alkalescence for having special physico-chemical property and excellent bioactivity is few
Sugar.With induction plant disease-resistant, degeneration-resistant and antibacterial etc. bioactivity, medicine, health food, cosmetics, agriculture are widely used to
Industry etc..
Alginic acid is a kind of polysaccharide extracted from the sea-plants such as kelp, brown alga and undaria pinnitafida, structure for α-(1 →
4) glycan that the L- guluronic acids (G) of connection and the D-MANNOSE aldehydic acid (M) of β-(1 → 4) connection are formed by connecting.Alginic acid has
Have and promote the bioactivity such as plant growth, drought resisting.
At present, although having alginic acid and itself and ion, such as the nano particle that alginic acid-calcium is formed, as hydrogel or
Person is applied to medical material.Chitosan oligosaccharide has been reported that carrying out precipitation reaction using sodium polyphosphate forms as oligosaccharides, good water solubility
Nano particle, but the nano particle being collectively formed currently without chitosan oligosaccharide and alginic acid is reported.This patent is with chitosan oligosaccharide and brown alga
Acid prepares chitosan oligosaccharide-alginic acid nano particle using simple technique.There is induction plant inducer resistance, alginic acid with reference to chitosan oligosaccharide
With Plant growth-promoting effect effect, nano particle of the present invention can be used as plant induced resistance agent and growth accelerator, and effect simultaneously
It is used alone better than chitosan oligosaccharide and alginic acid.
The content of the invention
It is an object of the invention to provide the preparation of chitosan oligosaccharide-alginic acid nano particle and its in plant induced resistance agent and growth-promoting
Application in long agent.
In order to realize above-mentioned target, the technical solution adopted by the present invention is：
The preparation method of chitosan oligosaccharide-alginic acid nano particle, concretely comprises the following steps:Chitosan oligosaccharide, alginic acid are dissolved respectively
Yu Shuizhong, by chitosan oligosaccharide solution tune pH to neutrality；Chitosan oligosaccharide solution is added dropwise in brown alga acid solution and carries out polymer precipitation instead
Should, the rate of addition of speed of agitator, chitosan oligosaccharide concentration, brown alga acid concentration and chitosan oligosaccharide solution is controlled, treats after completion of the reaction, to surpass
Sound obtains stable nanometer system；Ultracentrifugation is separated and dried to get chitosan oligosaccharide-alginic acid nano particle.
The chitosan oligosaccharide molecular weight is 1000-3000Da, and deacetylation is more than 85%；The concentration of chitosan oligosaccharide solution is
0.1%-10% (g/mL)；The concentration of brown alga acid solution is 0.05-5% (g/mL)；The mass ratio of alginic acid and chitosan oligosaccharide is
0.4-1: 1, when chitosan oligosaccharide solution time for adding is 0.5-3 small.
The rotating speed is 100-1000 revs/min, ultrasonic power 80-180w, and ultrasonic time is 2-60 minutes.
The chitosan oligosaccharide-alginic acid nano particle average grain diameter 50-500nm, Zeta potential 20-50mv.
The chitosan oligosaccharide-alginic acid nano particle can be used as antiseptic or plant induced resistance agent；Or chitosan oligosaccharide-the alginic acid
Nano particle can be used as Plant growth-promoting effect agent, improve yield.
The chitosan oligosaccharide-alginic acid nano particle is 20-200ppm as the concentration of antiseptic and plant induced resistance agent.
Chitosan oligosaccharide-alginic acid nano particle is 10-100ppm as the concentration of Plant growth-promoting effect agent.
The invention has the advantages that：
1. raw material chitosan oligosaccharide of the present invention and alginic acid are natural oligosaccharides and polysaccharide, raw material is easy to get, valency
Lattice are cheap.
2. the process that the present invention prepares nano particle adopts water as solvent, preparation process is simple, is easy to amplify；
3. present invention synthesis nano particle good dispersion, stability is strong, does not add crosslinking agent and dispersant.
4. chitosan oligosaccharide of the present invention-alginic acid nano particle has good bioactivity, while there is induction to plant
Object resistance and the activity for promoting plant growth, are with a wide range of applications.
Specific embodiment
With reference to embodiment, the present invention will be further described：
Embodiment 1:Chitosan oligosaccharide-alginic acid nano particle preparation process
0.01g molecular weight is taken as 1000Da, the chitosan oligosaccharide that deacetylation is 90% is dissolved in 5mL deionized waters, uses ammonium hydroxide
PH to 7 is adjusted, under agitation, is added dropwise in the brown alga acid solution of 4mL 0.25% (w/v), drips off within 30 minutes, mixing speed
For 300 revs/min, reaction 6 it is small when after, 90w power ultrasonics 2 minutes, 12000 revs/min centrifugation 1 it is small when, supernatant discarding, freeze-drying
Up to chitosan oligosaccharide-alginic acid nano particle, average grain diameter is in 200nm.
Embodiment 2:Chitosan oligosaccharide-alginic acid nano particle preparation process
Taking 0.02g molecular weight, deacetylation is dissolved in for 85% chitosan oligosaccharide in 5mL deionized waters, in stirring bar for 1500Da
It under part, is added dropwise in the brown alga acid solution of 4mL 0.5% (w/v), is dripped off in 30 minutes, mixing speed is 400 revs/min, reaction 6
After hour, 120w power ultrasonics 5 minutes, when 12000 revs/min of centrifugations 1 are small, supernatant discarding is freeze-dried up to chitosan oligosaccharide-brown alga
Sour nano particle, average grain diameter is in 100nm.
Embodiment 3:Chitosan oligosaccharide-alginic acid nano particle preparation process
0.05g molecular weight is taken as 3000Da, the chitosan oligosaccharide that deacetylation is 95% is dissolved in 5mL deionized waters, is being stirred
Under the conditions of, it is added dropwise in the brown alga acid solution of 4mL 1% (w/v), is dripped off in 60 minutes, mixing speed is 500 revs/min, reaction 8
After hour, 150w power ultrasonics 30 minutes, when 14000 revs/min of centrifugations 1 are small, supernatant discarding is freeze-dried up to chitosan oligosaccharide-brown
Alginic acid nano particle, average grain diameter is in 300nm.
Embodiment 4:Chitosan oligosaccharide-alginic acid nano particle preparation process
The chitosan oligosaccharide that 0.05g molecular weight is taken to be 3000Da is dissolved in 5mL deionized waters, under agitation, instills 4mL
The brown alga acid solution of 0.5% (w/v), drips off in 40 minutes, mixing speed be 600 revs/min, reaction 8 it is small when after, 180w power surpasses
Sound 10 minutes, when 14000 revs/min of centrifugations 1 are small, supernatant discarding is freeze-dried up to chitosan oligosaccharide-alginic acid nano particle, puts down
Equal grain size is in 270nm.
Embodiment 5：Chitosan oligosaccharide-alginic acid nano particle Rhizoctonia solani Kuhn inhibitory action
Chitosan oligosaccharide is configured to 6 series concentrations of 1000,500,100,50,25,12.5 μ ɡ/ml with sterile water.With nothing
Alginic acid is configured to 6 series concentrations of 5000,2500,1000,500,100,50 μ ɡ/ml by bacterium water, in addition, will with sterile water
6 series that the chitosan oligosaccharide of embodiment 1-alginic acid nano particle is configured to 1000,500,100,50,25,12.5 μ ɡ/ml are dense
Degree.Using inhibition mycelial growth rate method (Pesticide Science journal 2007,9 (4):357-362) processing is measured to inhibit mycelial growth
Rate calculates EC according to various concentration preparation to mycelial growth inhibition rate50Value.
1 chitosan oligosaccharide of table-alginic acid nano particle is to Rhizoctonia solani Kuhn inhibitory activity
As seen from the experiment, chitosan oligosaccharide-alginic acid nano particle has synergistic work to the inhibition of Rhizoctonia solani Kuhn
With EC50Value is far smaller than alginic acid EC50Value, less than chitosan oligosaccharide EC50Value.
Embodiment 6：Chitosan oligosaccharide-alginic acid nano particle is to the field efficacy of rice sheath blight disease
Experiment is carried out in Shenyang City, Liaoning Province new people area rice terrace, and experiment sets 5 processing altogether, made using embodiment 2
Chitosan oligosaccharide-alginic acid nano particle, (1) 50ppm chitosan oligosaccharides-alginic acid nano particle aqua, 30g/ mus；(2) 25ppm shells are few
Sugar-alginic acid nano particle aqua, 40g/ mus；(3) 10ppm chitosan oligosaccharides-alginic acid aqueous nanoparticle powder, 50g/ mus；(4)
50ppm chitosan oligosaccharide aquas, 50g/ mus；(5) clear water compares.3 repetitions are often handled, are applied for the first time in rice sheath blight disease early stage
Medicine, after sprayed 1 time every 7-10 days, continuous spray 3 times, the 7th day investigation rice sheath blight disease incidence after third time dispenser,
Calculate control effect.
2. chitosan oligosaccharides of table-alginic acid nano particle is to the preventive and therapeutic effect of rice sheath blight disease
| Processing | Disease index (%) before medicine | Disease index (%) after medicine | Average preventive effect (%) |
| 1 | 0.92 | 1.28 | 49.68cC |
| 2 | 0.51 | 0.55 | 60.91bB |
| 3 | 1.68 | 0.96 | 79.25AA |
| 4 | 1.33 | 1.38 | 61.44bB |
| CK | 2.21 | 5.97 | ___ |
Oligosaccharides-alginic acid nano particle shows the field control effectiveness test result of rice sheath blight disease 10ppm chitosan oligosaccharides-brown
The processing preventive effect highest of 50g/ mus of alginic acid nano particle is 79.25%.
Embodiment 7：Chitosan oligosaccharide-alginic acid nano particle is to tomato early blight bacterium, cucumber anthracnose, fusarium graminearum
And the germs inhibitory action such as Exserohilum turcicum
Method according to embodiment 5 measures chitosan oligosaccharide-alginic acid nano particle to following nine kinds of germ inhibitory action, calculates
Toxicity.
3. chitosan oligosaccharides of table-alginic acid nano particle is to nine kinds of pathogen inhibitory action measurement results
Embodiment 8：Chitosan oligosaccharide-alginic acid nano particle is to capsicum early blight of tomato field test results
According to the method for embodiment 6, chitosan oligosaccharide-alginic acid nano particle is carried out in Shandong Shouguang to capsicum early blight of tomato
Field trial, result of the test are shown in Table 4.
4. chitosan oligosaccharides of table-alginic acid nano particle is to the preventive effect of early blight of tomato
| Processing | Incidence (%) | Disease index (%) after medicine | Average preventive effect (%) |
| Chitosan oligosaccharide -30g/ mus of alginic acid nano particle | 8.6 | 7.73 | 58.40cC |
| Chitosan oligosaccharide -40g/ mus of alginic acid nano particle | 3.4 | 6.20 | 67.761bB |
| Chitosan oligosaccharide -50g/ mus of alginic acid nano particle | 2.1 | 2.67 | 86.50AA |
| 50g/ mus of chitosan oligosaccharide | 5.0 | 6.83 | 62.20bB |
| CK | 15.7 | 18.3 | ___ |
Result of the test shows that chitosan oligosaccharide-alginic acid nano particle has early blight of tomato preferable preventive effect, and highest preventive effect reaches
86.50%.Chitosan oligosaccharide-alginic acid nano particle has synergistic effect to the preventive effect of early blight of tomato.Embodiment 9：Chitosan oligosaccharide-brown alga
Sour nano particle is to the preventive and therapeutic effect of Strawberry anthracnose
According to the method for embodiment 6, chitosan oligosaccharide-alginic acid nano particle is carried out in Dalian to powdery mildew of strawberry disease field
Between test, result of the test is shown in Table 5.
5. chitosan oligosaccharides of table-alginic acid nano particle is to the preventive effect of Strawberry anthracnose
| Processing | Disease index (%) | Average preventive effect (%) |
| Chitosan oligosaccharide -30g/ mus of alginic acid nano particle | 1.97 | 77.75 |
| Chitosan oligosaccharide -40g/ mus of alginic acid nano particle | 1.84 | 79.34 |
| Chitosan oligosaccharide -50g/ mus of alginic acid nano particle | 0.81 | 90.80 |
| 50g/ mus of chitosan oligosaccharide | 1.72 | 88.70 |
| CK | 8.90 | ___ |
Result of the test shows fine preventive effect of the chitosan oligosaccharide-alginic acid nano particle to Strawberry anthracnose, chitosan oligosaccharide-alginic acid
50g/ mus of processing preventive effects of nano particle are higher than the preventive effect of 50g/ mus of processing of chitosan oligosaccharide.
Embodiment 10：Chitosan oligosaccharide-influence of the alginic acid nano particle to wheat tillering number and grain number per spike
200mg chitosan oligosaccharides-alginic acid nano particle is disperseed in 1L water.Experiment is divided into two groups, is respectively chitosan oligosaccharide-brown alga
The wheat of sour nano particle processing and clear water processing blank control group wheat.Starting sprinkling after into seedling stage, to be diluted to 50mg/L dense
The nanoparticie dispersion agents of degree, every 20 days 1 time, the statistical result after jointing until wheat is turned green.Through statistics obtain the results show that through
The wheat tillering number and grain number per spike of chitosan oligosaccharide-alginic acid nano particle processing significantly improve (table 6).
6. chitosan oligosaccharides of table-influence of the alginic acid nano particle to wheat tillering number and grain number per spike
Visible after chitosan oligosaccharide-alginic acid nano particle processing by result, Plant Height in Wheat increases by 1.98%, and tiller number increases
Add 9.3%.Wheat grains per spike increases by 5.3% simultaneously, and infertile spikelet number reduces 14.0%, chitosan oligosaccharide-alginic acid nanometer
Grain has been obviously promoted the tiller of wheat, adds wheat yield.
Embodiment 11：Chitosan oligosaccharide-alginic acid nano particle Dressing wheat acts on the raising of wheat yield
This experiment uses chitosan oligosaccharide-alginic acid aqueous nanoparticle powder of mass concentration 50mg/L.Wheat is mixed
Kind processing.Subjects are the wheat of western 9871 kind of agriculture.Its mature spike number, grain number per spike and mass of 1000 kernel are carried out in wheat aging time
Measurement, while the check plot of same kind is set, to compare the Application effects of chitosan oligosaccharide-alginic acid nano particle.Experiment finds that shell is few
Wheat mature spike number after the seed dressing of sugar-alginic acid nano particle increases by 6.1% compared with blank control group, and grain number per spike increases by 2.02%, and
And 4.46% and 8.1% (table 7) is respectively increased in mass of 1000 kernel and yield.
The raising of wheat yield is acted on after 7. chitosan oligosaccharides of table-alginic acid nano particle Dressing Wheat Species
Embodiment 12：Chitosan oligosaccharide-alginic acid nano particle improves cherry yield
The cherry that three kinds of different cultivars are preced with using red light, pioneer, dragon carries out yield experiment, by chitosan oligosaccharide-alginic acid nanometer
Particle is made into 25ppm water dispersants, is sprayed before being avenged on April 14th, 2015, and mu applies 200kg liquids, investigates 3 cherry variety productions
Amount, the results show chitosan oligosaccharide-alginic acid nanometer have preferable effect of increasing production, and concrete outcome is as shown in table 3 below.
Table 8. uses chitosan oligosaccharide-influence of the alginic acid nano particle to cherry yield
Embodiment 13：Chitosan oligosaccharide-alginic acid nano particle improves apple production
Yield experiment is carried out for apple in Shaanxi Luochuan Fu County, by chitosan oligosaccharide-alginic acid nanometer on April 10th, 2015
Pellet moisture powder 50ppm, sprays apple tree, and mu applies the liquid of 200kg.Investigate yield and exterior quality, concrete outcome such as table 9
It is shown.
Table 9. uses chitosan oligosaccharide-influence of the alginic acid nano particle to apple production and exterior quality
| Processing | Fruit-setting rate (%) | Single fruit weight (g) | Fruit shape index | Fruit color area (%) | Single plant yield (kg) |
| Control | 36.48 | 248.43 | 0.813 | 64.9 | 20.42 |
| Processing group | 46.12 | 279.52 | 0.865 | 79.7 | 36.54 |
As can be seen that apple production and exterior quality can be significantly improved using chitosan oligosaccharide-alginic acid nano particle.
Claims (7)
1. a kind of preparation method of chitosan oligosaccharide-alginic acid nano particle, it is characterised in that:Chitosan oligosaccharide, alginic acid are dissolved respectively
Yu Shuizhong, by chitosan oligosaccharide solution tune pH to neutrality；Chitosan oligosaccharide solution is added dropwise in brown alga acid solution and carries out polymer precipitation instead
Should, the rate of addition of speed of agitator, chitosan oligosaccharide concentration, brown alga acid concentration and chitosan oligosaccharide solution is controlled, treats after completion of the reaction, to surpass
Sound obtains stable nanometer system；Ultracentrifugation is separated and dried to get chitosan oligosaccharide-alginic acid nano particle.
2. according to the method described in claim 1, it is characterized in that：The chitosan oligosaccharide molecular weight is 1000-3000Da, takes off second
Acyl degree is more than 85%；The concentration of chitosan oligosaccharide solution is 0.1%-10% (g/mL)；The concentration of brown alga acid solution is 0.05-5%
(g/mL)；The mass ratio of alginic acid and chitosan oligosaccharide is 0.4-1: 1, when chitosan oligosaccharide solution time for adding is 0.5-3 small.
3. according to the method described in claim 1, it is characterized in that：The rotating speed is 100-1000 revs/min, and ultrasonic power is
80-180w, ultrasonic time are 2-60 minutes.
4. nano particle prepared by a kind of any methods of claim 1-3, it is characterised in that：Chitosan oligosaccharide-the brown alga
Sour nano particle average grain diameter 50-500nm, Zeta potential 20-50mv.
5. a kind of application of nano particle described in claim 4, it is characterised in that：The chitosan oligosaccharide-alginic acid nano particle can
As antiseptic or plant induced resistance agent；Or the chitosan oligosaccharide-alginic acid nano particle can be used as Plant growth-promoting effect agent, improve production
Amount.
6. application according to claim 5, it is characterised in that：The chitosan oligosaccharide-alginic acid nano particle is as antiseptic
Concentration with plant induced resistance agent is 20-200ppm.
7. application according to claim 5, it is characterised in that：Chitosan oligosaccharide-alginic acid nano particle is as Plant growth-promoting effect
The concentration of agent is 10-100ppm.
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| CN109221286A (en) * | 2018-10-12 | 2019-01-18 | 江西省农业科学院植物保护研究所 | A kind of high-efficiency environment friendly prevents and treats composition and the application of rice disease |
| CN109293438A (en) * | 2018-11-21 | 2019-02-01 | 青岛博智汇力生物科技有限公司 | A kind of flower nutrient solution containing marine oligosaccharide |
| CN114431228A (en) * | 2022-01-18 | 2022-05-06 | 四川海润作物科学技术有限公司 | Method for preventing chitosan oligosaccharide preparation and suspending agent from being mixed to generate flocculation |
| CN115517258A (en) * | 2022-09-15 | 2022-12-27 | 中科荣耀(苏州)生物科技有限公司 | Composition containing brown algae oligosaccharide and chitosan oligosaccharide and preparation method and application thereof |
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| CN109221286A (en) * | 2018-10-12 | 2019-01-18 | 江西省农业科学院植物保护研究所 | A kind of high-efficiency environment friendly prevents and treats composition and the application of rice disease |
| CN109221286B (en) * | 2018-10-12 | 2021-02-26 | 江西省农业科学院植物保护研究所 | Efficient and environment-friendly composition for preventing and treating rice diseases and application thereof |
| CN109293438A (en) * | 2018-11-21 | 2019-02-01 | 青岛博智汇力生物科技有限公司 | A kind of flower nutrient solution containing marine oligosaccharide |
| CN114431228A (en) * | 2022-01-18 | 2022-05-06 | 四川海润作物科学技术有限公司 | Method for preventing chitosan oligosaccharide preparation and suspending agent from being mixed to generate flocculation |
| CN115517258A (en) * | 2022-09-15 | 2022-12-27 | 中科荣耀(苏州)生物科技有限公司 | Composition containing brown algae oligosaccharide and chitosan oligosaccharide and preparation method and application thereof |
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Application publication date: 20180522 |