CN109984130A - A kind of chitosan oligosaccharide composition and application thereof - Google Patents
A kind of chitosan oligosaccharide composition and application thereof Download PDFInfo
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- CN109984130A CN109984130A CN201711494127.3A CN201711494127A CN109984130A CN 109984130 A CN109984130 A CN 109984130A CN 201711494127 A CN201711494127 A CN 201711494127A CN 109984130 A CN109984130 A CN 109984130A
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- chitosan oligosaccharide
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- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 title claims abstract description 81
- 239000000203 mixture Substances 0.000 title claims abstract description 58
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 35
- 201000010099 disease Diseases 0.000 claims abstract description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 18
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 6
- 230000002265 prevention Effects 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 25
- 241000196324 Embryophyta Species 0.000 claims description 22
- 241000219793 Trifolium Species 0.000 claims description 21
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 18
- 235000005881 Calendula officinalis Nutrition 0.000 claims description 13
- 108090000790 Enzymes Proteins 0.000 claims description 13
- 102000004190 Enzymes Human genes 0.000 claims description 13
- 240000000785 Tagetes erecta Species 0.000 claims description 13
- 229940088598 enzyme Drugs 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 11
- 208000036142 Viral infection Diseases 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 241000243785 Meloidogyne javanica Species 0.000 claims description 7
- 238000001514 detection method Methods 0.000 claims description 7
- 241001530056 Athelia rolfsii Species 0.000 claims description 6
- 235000016623 Fragaria vesca Nutrition 0.000 claims description 4
- 240000009088 Fragaria x ananassa Species 0.000 claims description 4
- 235000011363 Fragaria x ananassa Nutrition 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- 244000144730 Amygdalus persica Species 0.000 claims description 3
- 108010059892 Cellulase Proteins 0.000 claims description 3
- 239000004365 Protease Substances 0.000 claims description 3
- 235000006040 Prunus persica var persica Nutrition 0.000 claims description 3
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- 238000005259 measurement Methods 0.000 claims description 3
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- 241000220225 Malus Species 0.000 claims description 2
- 235000011430 Malus pumila Nutrition 0.000 claims description 2
- 235000015103 Malus silvestris Nutrition 0.000 claims description 2
- 235000014826 Mangifera indica Nutrition 0.000 claims description 2
- 240000007228 Mangifera indica Species 0.000 claims description 2
- 235000009811 Momordica charantia Nutrition 0.000 claims description 2
- 235000009812 Momordica cochinchinensis Nutrition 0.000 claims description 2
- 240000001740 Momordica dioica Species 0.000 claims description 2
- 235000018365 Momordica dioica Nutrition 0.000 claims description 2
- 102000016943 Muramidase Human genes 0.000 claims description 2
- 108010014251 Muramidase Proteins 0.000 claims description 2
- 240000005561 Musa balbisiana Species 0.000 claims description 2
- 235000018290 Musa x paradisiaca Nutrition 0.000 claims description 2
- 208000031888 Mycoses Diseases 0.000 claims description 2
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims description 2
- 235000007164 Oryza sativa Nutrition 0.000 claims description 2
- 108090000526 Papain Proteins 0.000 claims description 2
- 235000009827 Prunus armeniaca Nutrition 0.000 claims description 2
- 244000018633 Prunus armeniaca Species 0.000 claims description 2
- 241000220324 Pyrus Species 0.000 claims description 2
- 240000003768 Solanum lycopersicum Species 0.000 claims description 2
- 235000002597 Solanum melongena Nutrition 0.000 claims description 2
- 244000061458 Solanum melongena Species 0.000 claims description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 2
- 244000061456 Solanum tuberosum Species 0.000 claims description 2
- 235000009184 Spondias indica Nutrition 0.000 claims description 2
- 235000021307 Triticum Nutrition 0.000 claims description 2
- 244000098338 Triticum aestivum Species 0.000 claims description 2
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 2
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 2
- 240000006365 Vitis vinifera Species 0.000 claims description 2
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 2
- 240000008042 Zea mays Species 0.000 claims description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 2
- 239000001390 capsicum minimum Substances 0.000 claims description 2
- 235000019693 cherries Nutrition 0.000 claims description 2
- 235000020971 citrus fruits Nutrition 0.000 claims description 2
- 235000005822 corn Nutrition 0.000 claims description 2
- 238000002425 crystallisation Methods 0.000 claims description 2
- 230000008025 crystallization Effects 0.000 claims description 2
- 229960000274 lysozyme Drugs 0.000 claims description 2
- 239000004325 lysozyme Substances 0.000 claims description 2
- 235000010335 lysozyme Nutrition 0.000 claims description 2
- 238000001819 mass spectrum Methods 0.000 claims description 2
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 claims description 2
- 229940055729 papain Drugs 0.000 claims description 2
- 235000019834 papain Nutrition 0.000 claims description 2
- 235000021017 pears Nutrition 0.000 claims description 2
- 235000009566 rice Nutrition 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 244000298715 Actinidia chinensis Species 0.000 claims 1
- 235000009434 Actinidia chinensis Nutrition 0.000 claims 1
- 235000009436 Actinidia deliciosa Nutrition 0.000 claims 1
- 235000021028 berry Nutrition 0.000 claims 1
- 238000003795 desorption Methods 0.000 claims 1
- 230000005611 electricity Effects 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- 241000244206 Nematoda Species 0.000 abstract description 2
- 241000700605 Viruses Species 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 25
- 239000003814 drug Substances 0.000 description 17
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- 241000221785 Erysiphales Species 0.000 description 9
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- 238000006731 degradation reaction Methods 0.000 description 9
- 238000012360 testing method Methods 0.000 description 8
- 239000002689 soil Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 206010039509 Scab Diseases 0.000 description 6
- 238000011835 investigation Methods 0.000 description 6
- 235000007516 Chrysanthemum Nutrition 0.000 description 5
- 244000189548 Chrysanthemum x morifolium Species 0.000 description 5
- 229920001542 oligosaccharide Polymers 0.000 description 5
- 150000002482 oligosaccharides Chemical class 0.000 description 5
- 229920001661 Chitosan Polymers 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 3
- 238000002144 chemical decomposition reaction Methods 0.000 description 3
- 239000002361 compost Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 229960002442 glucosamine Drugs 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 239000004576 sand Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 2
- 239000004367 Lipase Substances 0.000 description 2
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- 241001582888 Lobus Species 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000007123 defense Effects 0.000 description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 125000003147 glycosyl group Chemical group 0.000 description 2
- 235000019421 lipase Nutrition 0.000 description 2
- 230000036244 malformation Effects 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 241000219112 Cucumis Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 1
- 229910021607 Silver chloride Inorganic materials 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 230000003628 erosive effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000001698 laser desorption ionisation Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000008121 plant development Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
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- 239000000047 product Substances 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
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- 210000003462 vein Anatomy 0.000 description 1
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Classifications
-
- 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
Abstract
The present invention is agriculture field, is specifically related to a kind of chitosan oligosaccharide composition, which is made of A and B, and A is the chitosan oligosaccharide of the 2-6 degree of polymerization, and quality accounts for the 40-95% of total amount, and B is the chitosan oligosaccharide and monosaccharide of 7 degree of polymerization or more, and quality accounts for the 5-60% of total amount.Application dose of the chitosan oligosaccharide composition on plant is 0.1-200ppm.Chitosan oligosaccharide composition is suitable for disease Plant nematode, plant epiphyte and the plant virus of prevention and treatment.
Description
Technical field
The invention belongs to agriculture fields, more particularly to a kind of chitosan oligosaccharide composition and application thereof.
Background technique
From organism and the great interest of the good compound of the biodegradability person that caused pesticide research.At this
In a little compounds, have no toxic side effect, the chitosan oligosaccharide of easily biological-degradable has very big potentiality, become the new medicament of control disease.
Chitosan oligosaccharide leads antiviral agents as a kind of important plant inducer, from the research of the oligosaccharide of plant, studies have shown that chitosan oligosaccharide exists
During resisting pathogen erosion, it can induce excitation plant and generate a series of defense reaction, the collaboration of these defense reactions is made
With the propagation and invasion for preventing pathogen, have the function that prevent and cure diseases.Meanwhile chitosan oligosaccharide is also a new class of plant hormone,
Has the effects that regulation and control plant growth and development, it can issue the information for adjusting specified plant function, these functions include
Growth promotion, cold-resistant, drought resisting etc..
However, chitosan oligosaccharide can be prepared by any means at present in chitosan oligosaccharide preparation and application study, such as shell is poly-
Sugar is prepared poly- by the methods of physical degradation methods, chemical degradation method, enzyme edman degradation Edman, glycosyl transfer method, composite degradation method
The mixture of the right chitosan oligomer for 2-20.Alternatively, obtaining crust by extracting from fungal cell wall or insect body wall
Element simultaneously passes through what the methods of physical degradation methods, chemical degradation method, enzyme edman degradation Edman, glycosyl transfer method, composite degradation method were prepared
The degree of polymerization is the mixture of the chitosan oligomer of 2-20.Alternatively, by passing through Isosorbide-5-Nitrae-glycosidic bond chemical synthesis by Glucosamine
The degree of polymerization being polymerized is the mixture of the polymer of 2-20.Chitosan oligosaccharide can be with the oligosaccharides of any degree of polymerization of degradation of chitosan
Form, the chitosan oligosaccharide of a variety of degree of polymerization all there may be.So no matter being prepared by which kind of method, or pass through above-mentioned system
The commercial product that Preparation Method is prepared, as long as it is that Glucosamine passes through the degree of polymerization made of Isosorbide-5-Nitrae-glucosides key connection as 2-
20, it is exactly chitosan oligosaccharide used in existing market that average molecular weight, which is the mixture of the high molecular polymer of 1000-3000 dalton,.
However, the degree of polymerization of β-Isosorbide-5-Nitrae-glucosides key connection 2- Glucosamine composition chitosan oligosaccharide due to the generation of different biodegrading process
It differs greatly, causes its agricultural effect unstable.
The invention proposes the composition of the chitosan oligosaccharide with specific aggregation degree and its effects, for the city of chitosan oligosaccharide pesticide
Field positioning and popularization have positive directive function.
Summary of the invention
To solve the unstable of chitosan oligosaccharide single dose agricultural effect currently on the market, the present invention provides the shells of different polymerization degree
Oligosaccharide composition and application thereof.
The present invention provides a kind of chitosan oligosaccharide compositions, which is characterized in that the chitosan oligosaccharide composition is made of A and B, and A is
The chitosan oligosaccharide of the 2-6 degree of polymerization, quality account for the 10-95% of total amount, and B is the chitosan oligosaccharide of monosaccharide and 7 degree of polymerization or more, and quality accounts for total amount
5-90%.
The present invention provides the application methods of chitosan oligosaccharide composition, are characterized in that application of the chitosan oligosaccharide composition on plant
Dosage is 0.1-200ppm.Preferably, composition application dose is 20-100ppm.
The application method of above-mentioned composition, composition are that the content that the degree of polymerization is 2-6 is higher than 40% chitosan oligosaccharide, the plant
Object is clover, marigold, potato, tomato, citrus, banana, pears, grape, mango, lichee, longan, apple, peach, apricot, Mi
Monkey peach, strawberry, cherry, strawberry, watermelon, cucumber, sponge gourd, balsam pear, wax gourd, capsicum, pimento, eggplant, wheat, rice, corn, tea
Leaf, cotton etc..Preferably, plant is clover, marigold.
It is the application method for chitosan oligosaccharide of the content higher than 40% that the degree of polymerization is 2-6 according to composition, is suitable for the disease of prevention and treatment
Evil has Plant nematode, fungal diseases of plants, the viroses of plant.Preferably, above-mentioned chitosan oligosaccharide composition is white for preventing and treating clover
Thin,tough silk disease and powdery mildew, marigold virosis and root-knot nematode.
The present invention also provides a kind of preparation methods of chitosan oligosaccharide of the invention.Using acetic acid as dissolving medium, the bottom of reaction
Object concentration 10%, the concentration of complex enzyme are 3%, and reaction temperature is 50 DEG C, and mixing speed is 30 revs/min, and the concentration of acetic acid is
3%, the reaction time is 10 hours.Obtained chitosan oligosaccharide solution obtains chitosan oligosaccharide of the invention through separation, crystallization and measurement,
In, the chitosan oligosaccharide of degree of polymerization 2-6 and the mass content of monosaccharide are 74.5%, the quality of more than 7 degree of polymerization chitosan oligosaccharide and monosaccharide
Content is 25.5%.
Complex enzyme used in the preparation of chitosan oligosaccharide of the invention, it is characterised in that complex enzyme is by 20 parts of cellulase, molten
15 parts of bacterium enzyme, 15 parts of lipase, 25 parts of glucolase, 25 parts of papain compositions.Chitosan oligosaccharide composition of the invention can lead to
Physical degradation methods, chemical degradation method, enzyme edman degradation Edman etc. is crossed to prepare.Detection method of the invention, it is characterised in that amino is few
The detection method of sugared element mass fraction includes in the chromatography of ions, and chromatograph is ICS-3000 ion chromatograph, comprising online de-
Quaternary gradient pump, column oven, ED3000 electrochemical detector, leacheate automatic generator, the Chromeleon 6.80 of device of air
Chromatographic work station.Chromatographic condition in the chromatography of ions is analytical column CarboPacTMPA10, and guard column is
CarboPacTMPA10BioLCTM；Using ED3000 Pulse amperometric detection, Au working electrode, Ag/AgCl reference electrode mode,
Four current potential of Standard for Sugars, leacheate are 100mM NaOH and NaAc/H2O=10:90, flow velocity 1.0mL/min.
The detection method of the degree of polymerization of chitosan oligosaccharide composition of the invention, it is characterised in that use sample substrate auxiliary laser
Desorption ionization flight time mass spectrum (MALDI-TOF-MS) method is detected.
The invention has the benefit that composition of the invention can specific aim controlling plant diseases, and agricultural effect is steady
Fixed, effect is higher, while save the cost.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 shows activated oligosaccharide preparation technology flow chart；
Fig. 2 shows oligosaccharide monomer separating technology flow chart.
Specific embodiment
In order to understand the present invention, further illustrated below with embodiment, but be not limited to the present embodiment.
The preparation of 1 chitosan oligosaccharide composition of embodiment and the measurement of the degree of polymerization
The preparation and detection of chitosan oligosaccharide are carried out according to shown in attached drawing 1 and attached drawing 2.Quantitative chitosan is weighed, is with acetic acid
Dissolving medium, degrading enzyme are complex enzyme (by 20 parts of cellulase, 15 parts of lysozyme, 15 parts of lipase, 25 parts of glucolase, wood
25 parts of melon protease compositions) concentration of substrate of enzyme digestion reaction is 8-15%, the concentration of complex enzyme is 1-3%, and the concentration of acetic acid is
3%, reaction temperature is 20-50 DEG C, and mixing speed is 30 revs/min, and the reaction time is 5~10 hours.Suitable control reaction temperature
Degree and reaction time, the chitosan oligosaccharide solution of different batches is obtained, is further separated, crystallized and measured, obtain different polymerization degree
Chitosan oligosaccharide sample.Concrete example is shown in Table 1.
The chitosan oligosaccharide preparation of the different enzymatic hydrolysis conditions of table 1
As can be seen that under different enzymatic hydrolysis conditions, chitosan oligosaccharide sample obtained the 2-6 degree of polymerization mass percentage and
The mass percentage of remaining product is different.
2 chitosan oligosaccharide composition of embodiment is to turfgrass effect
It is mixed into compost with sterilized soil and sterilizing river sand, after mixing well, is mixed into medicament.Test sample concentration is with every thousand
Gram soil contains the milligram number meter of chitosan oligosaccharide, and test sample is made into preparation.Soil is directly mixed with corresponding dosage formulation, is sufficiently mixed
Clear water is added after even makes water content 30%, then is quantitatively packed into basin alms bowl after mixing thoroughly.Clover seed, every basin alms bowl sowing are sowed in basin alms bowl
10,2 weeks after clover germination seedling true leaf expansion, every alms bowl is inoculated with 10 2 age in week southern blight sclerotium, observes three after 15d
Leaf grass disease incidence calculates preventive effect, the results are shown in Table 2.
2 chitosan oligosaccharide composition of table is to Sclerotium rolfsii Causing Southern Blight on Clover effect
| Test sample | Concentration (ppm) | Preventive effect (%) |
| Sample 1 | 40 | 36.7 |
| Sample 2 | 40 | 38.3 |
| Sample 3 | 40 | 48.2 |
| Sample 4 | 40 | 53.3 |
| Sample 5 | 40 | 53.7 |
Note: the degree of polymerization of test sample and content are as shown in table 1 in table 2.
Different chitosan oligosaccharide compositions are shown in Table 2 to Sclerotium rolfsii Causing Southern Blight on Clover effect.As can be seen that under same medicine amount, it is different
Chitosan oligosaccharide sample have a different to the preventive effect of Sclerotium rolfsii Causing Southern Blight on Clover, the preventive effect of sample 1 and sample 2 is lower than sample 3, sample 4
With the preventive effect of sample 5, so, when the chitosan oligosaccharide content that the degree of polymerization is 2-6 is 40% or more, composition is to Sclerotium rolfsii Causing Southern Blight on Clover
Effect is higher than the effect that the chitosan oligosaccharide content that the degree of polymerization is 2-6 is lower than 40%, effect of the chitosan oligosaccharide composition to Sclerotium rolfsii Causing Southern Blight on Clover
Fruit is higher.
Effect of 3 chitosan oligosaccharide composition of embodiment to clover powdery mildew
It is mixed into compost with sterilized soil and sterilizing river sand, clear water, which is added, after mixing well, after mixing well makes water content
30%, then basin alms bowl is quantitatively packed into after mixing thoroughly.5, clover seed are sowed in basin alms bowl, normal watering tube after clover germination
Reason quantify on blade inoculation quantitatively powdery mildew in every alms bowl 1 week after the expansion of seedling true leaf, moves in observation ward and unification moisturizing
It cultivates, investigation disease refers to and sprays after 7d, and 14d continues spray 1 time after medicine.It investigates disease after 20d to refer to, record investigation total number of sheets, sick leaf
Number calculates disease index and preventive effect according to following stage division, the results are shown in Table 3.
Blade grade scale:
0 grade: disease-free
1 grade: the connected area of scab accounts for the 1% or less of entire leaf area；
3 grades: the connected area of scab accounts for the 1%-5% of entire leaf area；
5 grades: the connected area of scab accounts for the 6%-25% of entire leaf area；
7 grades: the connected area of scab accounts for the 26%-50% of entire leaf area；
9 grades: the connected area of scab accounts for 51% or more of entire leaf area；
The different chitosan oligosaccharide composition of table 3 is to clover powdery mildew effect
| Test sample | Concentration (ppm) | Preventive effect (%) |
| Sample 1 | 50 | 40.4 |
| Sample 2 | 50 | 43.5 |
| Sample 3 | 50 | 50.5 |
| Sample 4 | 50 | 55.6 |
| Sample 5 | 50 | 57.5 |
Note: the degree of polymerization of test sample and content are as shown in table 1 in table 3.
Different chitosan oligosaccharide compositions are shown in Table 3 to clover powdery mildew effect.As can be seen that under same medicine amount, it is different
Chitosan oligosaccharide sample have a different to the preventive effect of clover powdery mildew, the preventive effect of sample 1 and sample 2 is lower than sample 3, sample 4
With the preventive effect of sample 5, so, when the chitosan oligosaccharide content that the degree of polymerization is 2-6 is 40% or more, composition is to clover powdery mildew
Preventive effect is 50% or more, and composition is higher to the effect of clover powdery mildew.
Effect of the chitosan oligosaccharide composition of 4 different polymerization degree of embodiment to marigold virosis
Cell is set in marigold flower nursery.Every plot area 5m2, if clear water compares, the cultivation condition of all experimental plots is equal
It is even consistent.With the 1st medication of knapsack sprayer when virosis takes place, it is spaced 1 of 7d medication and shares 3 times.Before medicine and
7d progress disease refers to investigation after medicine, and investigation total number of sheets, the sick number of sheets at different levels calculate control efficiency according to disease index, the results are shown in Table 4.
Blade grade scale
0 grade: asymptomatic:
1 grade: bright arteries and veins, light floral leaf:
3 grades: lobus cardiacus and middle leaf piece floral leaf；
5 grades: lobus cardiacus and middle leaf piece floral leaf, a small number of leaf malformations, shrinkage or plant are slightly downgraded；
7 grades: weight floral leaf, most leaf malformations, shrinkage or plant are downgraded；
9 grades: the obvious deformity of weight floral leaf, blade, leaf line, plant are seriously downgraded, or even dead.
4 chitosan oligosaccharide composition of table is to chrysanthemum virosis effect
Note: the degree of polymerization of test sample and content are as shown in table 1 in table 4.
Different chitosan oligosaccharide compositions are shown in Table 4 to chrysanthemum virosis effect.As can be seen that under same medicine amount, it is different
Chitosan oligosaccharide sample has different to the preventive effect of chrysanthemum virosis, and in terms of investigation result several times, sample 1 is low with the preventive effect of sample 2
In the preventive effect of sample 3, sample 4 and sample 5, so, when the chitosan oligosaccharide content that the degree of polymerization is 2-6 is 40% or more, the 3rd medicine
For the drug effect of 7d up to 70% or more, composition is higher to the effect of chrysanthemum virosis afterwards.
Effect of the different chitosan oligosaccharide composition of embodiment 5 to marigold root-knot nematode
It is mixed into compost with sterilized soil and sterilizing river sand, after mixing well, is mixed into medicament.Drug concentration is with every kilogram of soil
Earth contains the milligram number meter of institute's effective component of the medicament, directly mixes soil with corresponding preparation, and clear water is added after mixing well to be made
Water content 30%, then it is quantitatively packed into basin alms bowl after mixing thoroughly, the almost the same marigold seedling of the plant height, thickness, blade of 20d of emerging is moved
It plants in basin, 7 days after transplanting, every basin accesses 2 instar larvae of chrysanthemum root-knot nematode 400, is placed in greenhouse, Routine Management.After 40d,
Each processing plant root knot number is investigated, and is classified: 0 grade, no root knot；1 grade, 1-2 root knot；2 grades, 3-10 root knot；3 grades,
11-30 root knot；4 grades, 31-100 root knot；5 grades, 100 or more root knots.Count root knot number, the root knot number of different disposal and
Disease scale is shown in Table 5.
The different chitosan oligosaccharide composition of table 5 is to clover powdery mildew effect
| Test medicament | Concentration (mg/Kg) | Root knot number | Disease scale |
| Sample 1 | 60 | 43.2 | 4 |
| Sample 2 | 60 | 39.5 | 4 |
| Sample 3 | 60 | 28.3 | 3 |
| Sample 4 | 60 | 27.4 | 3 |
| Sample 5 | 60 | 26.5 | 3 |
Note: the degree of polymerization and the content that medicament is tested in table 5 are as shown in table 1.
Different chitosan oligosaccharide compositions are shown in Table 5 to the effect of marigold root-knot nematode.As can be seen that under same medicine amount,
Different chitosan oligosaccharide samples have different to the preventive effect of marigold root-knot nematode, in terms of investigation result several times, sample 1 and sample
2 preventive effect is lower than the preventive effect of sample 3, sample 4 and sample 5, so, the chitosan oligosaccharide content that the degree of polymerization is 2-6 is 40% or more
When, disease scale is 3 grades, and composition is higher to the effect of marigold root-knot nematode.
Claims (9)
1. a kind of chitosan oligosaccharide composition, which is characterized in that the chitosan oligosaccharide composition is made of A and B, and A is that the shell of the 2-6 degree of polymerization is few
Sugar, quality account for the 10-95% of total amount, and B is the chitosan oligosaccharide of monosaccharide and 7 degree of polymerization or more, and quality accounts for the 5-90% of total amount.
2. the application method of chitosan oligosaccharide composition according to claim 1, which is characterized in that chitosan oligosaccharide composition is on plant
Application dose is 0.1-200ppm.Preferably, composition application dose is 20-100ppm.
3. application method according to claim 2, composition is that the content that the degree of polymerization is 2-6 is higher than 40% chitosan oligosaccharide, described
Plant for example clover, marigold, potato, tomato, citrus, banana, pears, grape, mango, lichee, longan, apple, peach, apricot,
Kiwi berry, strawberry, cherry, strawberry, watermelon, cucumber, sponge gourd, balsam pear, wax gourd, capsicum, pimento, eggplant, wheat, rice, corn,
Tealeaves, cotton.
4. application method according to claim 3, which is characterized in that plant is clover and marigold.
5. according to the application method of claim 3 or 4, it is characterised in that chitosan oligosaccharide composition is suitable for that the disease of prevention and treatment has plant line
Worm, fungal disease and the viroses of plant.
6. application method according to claim 5, which is characterized in that chitosan oligosaccharide composition is for preventing and treating Sclerotium rolfsii Causing Southern Blight on Clover and white
Powder disease, marigold virosis and root-knot nematode.
7. the preparation method of the chitosan oligosaccharide composition of claim 1, which is characterized in that using acetic acid as dissolving medium, the bottom of reaction
Object concentration is 10%, and the concentration of complex enzyme is 3%, and reaction temperature is 50 DEG C, and mixing speed is 30 revs/min, the concentration of acetic acid
It is 3%, the reaction time is 10 hours, and obtained chitosan oligosaccharide solution obtains chitosan oligosaccharide of the invention through separation, crystallization and measurement,
Wherein, the chitosan oligosaccharide mass content of the 2-6 degree of polymerization is 74.5%, and the mass content of more than 7 degree of polymerization chitosan oligosaccharide and monosaccharide is
25.5%.
8. preparation method according to claim 7, it is characterised in that complex enzyme is by 20 parts of cellulase, 15 parts of lysozyme, fat
15 parts of enzyme, 25 parts of glucolase, 25 parts of papain compositions.
9. the detection method of the chitosan oligosaccharide composition of claim 1, it is characterised in that using sample substrate Assisted Laser Desorption electricity
Degree of polymerization detection is carried out from flight time mass spectrum (MALDI-TOF-MS) method.
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| CN111727971A (en) * | 2020-06-11 | 2020-10-02 | 中科禾一（大连）农业科技有限公司 | Nematode inhibitor containing chitin oligosaccharide and application thereof |
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