CN117461641A - Plant regulator and application thereof in improving antiviral capacity of plants and/or relieving low-temperature stress of plants - Google Patents
Plant regulator and application thereof in improving antiviral capacity of plants and/or relieving low-temperature stress of plants Download PDFInfo
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- CN117461641A CN117461641A CN202311443684.8A CN202311443684A CN117461641A CN 117461641 A CN117461641 A CN 117461641A CN 202311443684 A CN202311443684 A CN 202311443684A CN 117461641 A CN117461641 A CN 117461641A
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- plant
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- chitosan oligosaccharide
- plant regulator
- zinc
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- 230000000840 anti-viral effect Effects 0.000 title claims abstract description 19
- 241000196324 Embryophyta Species 0.000 claims abstract description 182
- 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 claims abstract description 80
- 239000011701 zinc Substances 0.000 claims abstract description 50
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims abstract description 34
- 229910052725 zinc Inorganic materials 0.000 claims abstract description 34
- 229940038879 chelated zinc Drugs 0.000 claims abstract description 29
- 241000208125 Nicotiana Species 0.000 claims description 50
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims description 50
- 241000209140 Triticum Species 0.000 claims description 33
- 235000021307 Triticum Nutrition 0.000 claims description 33
- 239000004094 surface-active agent Substances 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 18
- 239000013543 active substance Substances 0.000 claims description 13
- 239000013522 chelant Substances 0.000 claims description 11
- 238000005507 spraying Methods 0.000 claims description 11
- 238000002791 soaking Methods 0.000 claims description 8
- 229920000136 polysorbate Polymers 0.000 claims description 4
- 238000003973 irrigation Methods 0.000 claims description 3
- 230000002262 irrigation Effects 0.000 claims description 3
- 241000894007 species Species 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 26
- 241000723873 Tobacco mosaic virus Species 0.000 abstract description 16
- 102000004190 Enzymes Human genes 0.000 abstract description 10
- 108090000790 Enzymes Proteins 0.000 abstract description 10
- 230000005764 inhibitory process Effects 0.000 abstract description 10
- 239000003963 antioxidant agent Substances 0.000 abstract description 9
- 230000003078 antioxidant effect Effects 0.000 abstract description 9
- 238000002474 experimental method Methods 0.000 abstract description 9
- 201000010099 disease Diseases 0.000 abstract description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 7
- 230000029553 photosynthesis Effects 0.000 abstract description 7
- 238000010672 photosynthesis Methods 0.000 abstract description 7
- 241000238631 Hexapoda Species 0.000 abstract description 5
- 238000010521 absorption reaction Methods 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 4
- 239000005648 plant growth regulator Substances 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract description 2
- 241000607479 Yersinia pestis Species 0.000 abstract 1
- 230000001276 controlling effect Effects 0.000 abstract 1
- 230000035882 stress Effects 0.000 description 36
- 238000011282 treatment Methods 0.000 description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 27
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 22
- 229960001763 zinc sulfate Drugs 0.000 description 22
- 229910000368 zinc sulfate Inorganic materials 0.000 description 22
- 239000007788 liquid Substances 0.000 description 19
- 239000000243 solution Substances 0.000 description 18
- 239000003153 chemical reaction reagent Substances 0.000 description 14
- 238000002156 mixing Methods 0.000 description 13
- 239000000047 product Substances 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 11
- 239000013642 negative control Substances 0.000 description 11
- 230000000052 comparative effect Effects 0.000 description 10
- 230000012010 growth Effects 0.000 description 10
- 230000009920 chelation Effects 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 7
- 238000000502 dialysis Methods 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 238000001035 drying Methods 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 229930002875 chlorophyll Natural products 0.000 description 5
- 235000019804 chlorophyll Nutrition 0.000 description 5
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 239000011259 mixed solution Substances 0.000 description 5
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- 238000002360 preparation method Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000000967 suction filtration Methods 0.000 description 4
- 239000005723 virus inoculator Substances 0.000 description 4
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- 230000015556 catabolic process Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 3
- 229920000053 polysorbate 80 Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- PUAQLLVFLMYYJJ-UHFFFAOYSA-N 2-aminopropiophenone Chemical group CC(N)C(=O)C1=CC=CC=C1 PUAQLLVFLMYYJJ-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- 239000012295 chemical reaction liquid Substances 0.000 description 2
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- 238000011081 inoculation Methods 0.000 description 2
- 229940118019 malondialdehyde Drugs 0.000 description 2
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- 238000001556 precipitation Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 238000009736 wetting Methods 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 229930192334 Auxin Natural products 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 208000009084 Cold Injury Diseases 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000002595 cold damage Effects 0.000 description 1
- 230000006196 deacetylation Effects 0.000 description 1
- 238000003381 deacetylation reaction Methods 0.000 description 1
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- 230000002708 enhancing effect Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000012844 infrared spectroscopy analysis Methods 0.000 description 1
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- 229910052760 oxygen Inorganic materials 0.000 description 1
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- 230000035790 physiological processes and functions Effects 0.000 description 1
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- 208000024891 symptom Diseases 0.000 description 1
- 150000003752 zinc compounds Chemical class 0.000 description 1
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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/45—Tobacco
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- 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
- A01N55/00—Biocides, pest repellants or attractants, or plant growth regulators, containing organic compounds containing elements other than carbon, hydrogen, halogen, oxygen, nitrogen and sulfur
- A01N55/02—Biocides, pest repellants or attractants, or plant growth regulators, containing organic compounds containing elements other than carbon, hydrogen, halogen, oxygen, nitrogen and sulfur containing metal atoms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
Abstract
The invention belongs to the technical field of plant growth regulators, and particularly relates to a plant regulator and application thereof in improving antiviral capacity of plants and/or relieving low-temperature stress of the plants. The invention enhances the stability of the plant regulator by regulating and controlling the concentration of the chitosan oligosaccharide chelated zinc, is beneficial to the absorption of zinc element by roots and plants, and simultaneously ensures the efficacy of the chitosan oligosaccharide in improving plant resistance to diseases and insect pests and relieving low temperature stress. Experiments show that by adopting the technical scheme provided by the invention, the antioxidant enzyme activity and photosynthesis efficiency of plants in a low-temperature environment are greatly improved, and the highest inhibition rate of the tobacco mosaic virus can reach 71.43%. Therefore, the chitosan oligosaccharide chelated zinc plant regulator provided by the invention not only can improve the stress resistance of plants, but also can realize the high-value utilization of ocean low-value products.
Description
Technical Field
The invention belongs to the technical field of plant growth regulators, and particularly relates to a plant regulator and application thereof in improving antiviral capacity of plants and/or relieving low-temperature stress of the plants.
Background
The low temperature is one of the main natural disasters in agricultural production, and seriously affects the normal growth and development and yield of plants. The low temperature stress can cause peroxidation of the plant material film, and the plasma film structure is damaged; the chlorophyll is degraded, the photosynthesis efficiency is reduced, the growth and development of plants are seriously affected, and even the plants die. Especially, in the seedling raising period and after transplanting of tobacco, the tobacco seedling is often subjected to low-temperature cold injury and virus diseases caused by 'cold in the back spring', so that tobacco seedlings grow slowly, leaves are yellow, the tobacco cannot be quickly developed after transplanting, and the production of tobacco leaves is affected.
The chitosan oligosaccharide is the oligoglucosamine obtained after the degradation of chitosan, mainly comes from the offal of aquatic products such as shrimp and crab shells, is taken as a natural plant exciton, can improve the capability of plants to resist insect diseases and abiotic stress, and has wide application prospect in agricultural production. Zinc can influence photosynthesis of plants by participating in metabolism of auxin, thereby influencing plant growth; zinc can also be used as a metal activator of enzymes, and can improve the activity of antioxidant enzymes in plants and the stress resistance of the plants by participating in important metabolic processes. However, there is currently no report on the use of chitosan oligosaccharide chelated zinc for improving antiviral ability and/or alleviating low temperature stress in plants.
Disclosure of Invention
The invention aims to provide a plant regulator and application thereof in improving plant antiviral ability and/or relieving plant low-temperature stress, and can improve plant antiviral ability while relieving plant low-temperature stress.
The invention provides a plant regulator, wherein the effective components of the plant regulator comprise chitosan oligosaccharide chelated zinc; the concentration of the chitosan oligosaccharide chelated zinc in the plant regulator is 100-1000 mg/L.
Preferably, the mass content of zinc in the chitosan oligosaccharide chelating zinc is 3-4%.
Preferably, the plant regulator further comprises a surfactant; the concentration of the surfactant in the plant regulator is 1-10 mL/L.
Preferably, the surfactant comprises one or more of a tween active agent, a span active agent, an OP active agent and an NP active agent.
The invention provides application of the plant regulator in improving antiviral ability of plants and/or relieving low-temperature stress of plants.
The invention also provides a method for improving the antiviral ability of plants and/or relieving the low-temperature stress of plants, which comprises the following steps: the plant regulator according to the above technical scheme is used for soaking plant seeds and/or applying the plant seed regulator to plants.
Preferably, the mode of administration comprises: spraying and/or root irrigation.
Preferably, the concentration of the plant regulator is 0.1-0.5 g/L when the plant species is soaked; the soaking time is 6-8 h.
Preferably, the concentration of the plant regulator is 0.1-1 g/L and the application amount is 20-80L/mu when the plant regulator is applied to plants.
Preferably, the plant species include: tobacco and/or wheat.
The beneficial effects are that:
the invention provides a plant regulator, wherein the effective components of the plant regulator comprise chitosan oligosaccharide chelated zinc; the concentration of the chitosan oligosaccharide chelated zinc in the plant regulator is 100-1000 mg/L. The stability of the plant regulator is enhanced by using chitosan oligosaccharide chelated zinc with the adaptive concentration, which is beneficial to the absorption of roots and plants to metal zinc, and simultaneously ensures the efficacy of chitosan oligosaccharide in improving plant disease and insect resistance and relieving low temperature stress. Therefore, the chitosan oligosaccharide chelated zinc plant regulator not only can improve the stress resistance of plants, but also can realize the high-value utilization of ocean low-value products.
Based on the technical advantages, the invention also provides a method for improving the antiviral capability of plants and/or relieving the low-temperature stress of plants, which comprises the following steps: the plant regulator according to the above technical scheme is used for soaking plant seeds and/or applying the plant seed regulator to plants. By soaking plant seeds and/or applying to plants, it is beneficial to increase the antiviral ability of the plants and to alleviate the low temperature stress to which the plants are subjected. Experiments show that by adopting the technical scheme provided by the invention, the antioxidant enzyme activity and photosynthesis efficiency of plants in a low-temperature environment are greatly improved, and the highest inhibition rate of the tobacco mosaic virus can reach 71.43%. Therefore, the technical scheme provided by the invention can be used for improving plant antiviral and/or relieving plant low-temperature stress.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.
FIG. 1 is an infrared spectrum of the plant regulator prepared in example 1 and comparative example 1;
FIG. 2 is a graph showing the effect of different treatment groups on tobacco seedling growth in application example 3.
Detailed Description
The invention provides a plant regulator, wherein the effective components of the plant regulator comprise chitosan oligosaccharide chelated zinc; the concentration of the chitosan oligosaccharide chelated zinc in the plant regulator is 100-1000 mg/L.
In the invention, the raw materials and equipment are all purchased conventionally unless otherwise specified.
The concentration of the chitosan oligosaccharide chelate zinc in the plant regulator is 100-1000 mg/L, more preferably 100-500 mg/L or 700-1000 mg/L, and still more preferably 100mg/L. In the present invention, the mass of zinc in the chitosan oligosaccharide chelate zinc is preferably 3% to 4%, more preferably 3%.
The preparation method of the chitosan oligosaccharide chelated zinc preferably comprises the following steps: mixing a chitosan oligosaccharide solution and a zinc compound solution, and then performing a chelation reaction to obtain a chelation reaction solution; and (3) dialyzing, precipitating and drying the chelation reaction liquid to obtain the chitosan oligosaccharide chelated zinc.
The chitosan oligosaccharide solution is preferably obtained by mixing and dissolving chitosan oligosaccharide and water. In the invention, the mass volume ratio of the chitosan oligosaccharide to the water is preferably 10-100 mg:10 to 50mL, more preferably 20mg:10mL; the water is preferably distilled water. The mixing mode of the invention has no special requirement, and the chitosan oligosaccharide is completely dissolved in water. The molecular weight of the chitosan oligosaccharide is preferably 2000-5000 Da, more preferably 3500Da.
The invention also preferably mixes and dissolves the zinc-containing compound with water to obtain a zinc-containing compound solution. In the invention, the mass volume ratio of the zinc-containing compound to water is preferably 10-100 mg:10 to 100mL, more preferably 10mg:10mL; the water is preferably distilled water. The zinc-containing compound of the present invention is preferably zinc sulfate. The manner of mixing according to the invention is not particularly limited, as long as the zinc-containing compound is completely dissolved in water.
After the chitosan oligosaccharide solution and the zinc-containing compound solution are respectively obtained, the chitosan oligosaccharide solution and the zinc-containing compound solution are preferably mixed for chelation reaction to obtain a chelation reaction solution. In the present invention, when the chelation reaction is performed, the mixed solution is preferably heated in a water bath. The pH value of the mixed solution is preferably 5.5-7.0, more preferably 7.0; the temperature of the water bath heating is preferably 35-45 ℃, more preferably 40 ℃; the time of heating in the water bath is preferably 20 to 40 minutes, more preferably 30 minutes.
After the chelate reaction solution is obtained, the present invention preferably dialyzes the chelate reaction solution against distilled water. The dialysis time of the invention is 24-72 hours, more preferably 48 hours; the dialysis is preferably performed in a dialysis bag having a molecular weight of 3500Da. The source of the dialysis bag has no special requirement, and the dialysis bag is purchased conventionally.
After the dialysis, the present invention preferably uses absolute ethanol to carry out precipitation reaction on the trapped fluid to obtain a precipitate. The volume ratio of the absolute ethyl alcohol to the chelation reaction liquid is preferably 3-4:1, more preferably 3:1. In the present invention, the precipitation reaction preferably includes suction filtration, and the suction filtration is not particularly limited, and techniques well known in the art may be employed. The present invention preferably further comprises washing the reaction product obtained by suction filtration to obtain said precipitate, wherein the washing reagent is preferably absolute ethanol, and the washing mode is not particularly required, and the technology well known in the art is adopted.
After the precipitate is obtained, the precipitate is preferably dried to obtain the chitosan oligosaccharide chelated zinc. In the present invention, the temperature of the drying is preferably 45 to 65 ℃, more preferably 60 ℃; the drying equipment is preferably a forced air oven. The invention has no special requirement on the drying time, and the drying is the basis. By the method, fully chelated chitosan oligosaccharide chelated zinc can be prepared, and the stability of zinc element is greatly improved.
In the present invention, the plant regulator further preferably includes a surfactant; the concentration of the surfactant in the plant regulator is 1-10 mL/L, preferably 1-3 mL/L, 3-5 mL/L or 7-10 mL/L; more preferably 1mL/L; the surfactant is preferably one or more of tween active agent, span active agent, OP active agent and NP active agent, more preferably tween active agent. The surface tension of the target solution is reduced by adding the surfactant into the plant regulator, so that the obtained plant regulator can be used as a spraying agent.
The preparation method of the plant regulator preferably comprises the following steps: mixing the chitosan oligosaccharide chelated zinc aqueous solution with a surfactant and fixing the volume to obtain the plant regulator containing the surfactant. In the invention, the chitosan oligosaccharide chelated zinc is mixed with water for dissolution to obtain the chitosan oligosaccharide chelated zinc aqueous solution, the mode of mixing the chitosan oligosaccharide chelated zinc aqueous solution with the surfactant for volume fixation is not required, and the chitosan oligosaccharide chelated zinc aqueous solution and the surfactant can be prepared by adopting the technology well known in the art. The concentration of the surfactant and the chitosan oligosaccharide chelated zinc is accurate by constant volume, so that the zinc is absorbed by roots and plants, and the effects of improving plant disease and insect resistance and relieving low-temperature stress by the chitosan oligosaccharide are ensured.
Based on the technical advantages, the invention also provides a method for improving the antiviral capability of plants and/or relieving the low-temperature stress of plants, which comprises the following steps: the plant regulator according to the above technical scheme is used for soaking plant seeds and/or applying the plant seed.
The present invention preferably provides for the plant regulator to be used to soak plant seeds and to apply to plants. The components and preparation methods of the plant regulator of the present invention are described in detail above, and are not described in detail herein.
In the present invention, the concentration of the plant regulator is preferably 0.1 to 0.5g/L, more preferably 0.1g/L, when the plant species is soaked; the soaking time is preferably 6 to 8 hours, more preferably 6 hours. When seeds are soaked, the plant regulator is used in an amount which is based on that the seeds are completely soaked.
In the present invention, the concentration of the plant regulator is preferably 0.1 to 1g/L, more preferably 0.1g/L, when applied to plants; the application mode is preferably spraying and/or root irrigation, more preferably spraying; the period of application is preferably the seedling stage and/or the maturity stage of the plant. When the plant conditioner is applied to plants, the application amount of the plant conditioner is preferably 20-80L/mu, and more preferably 30L/mu or 60L/mu. In the specific embodiment of the invention, in the tobacco planting process, the application amount of the plant regulator is 30L/mu in the tobacco seedling stage, and the application amount of the plant regulator is 60L/mu in the tobacco maturity stage; in the wheat planting process, the application amount of the plant regulator in the tobacco seedling stage is 30L/mu, and the application amount of the plant regulator in the tobacco maturity stage is 60L/mu.
The invention also provides application of the plant regulator in improving antiviral ability of plants and/or relieving low-temperature stress of plants. In the present invention, the plant species is preferably tobacco and/or wheat, more preferably tobacco and wheat. Experiments show that by adopting the technical scheme provided by the invention, the antioxidant enzyme activity and photosynthesis efficiency of plants in a low-temperature environment are greatly improved, and the antiviral capability of the plants can be effectively improved, and especially the inhibition rate of the tobacco mosaic virus can be up to 71.43%.
For further explanation of the present invention, a plant regulator and its use for improving antiviral ability of plants and/or alleviating low temperature stress of plants, provided by the present invention, will be described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.
In the examples, comparative examples and application examples of the present invention, the methods used are conventional in the art unless otherwise specified.
Example 1
Preparing a plant regulator, which comprises the following steps:
1) 20mg of chitosan oligosaccharide (purchased from Qingdao and Haiotsai biotechnology Co., ltd., average molecular weight is 3500Da, deacetylation degree is not less than 85%) and 10mg of zinc sulfate are weighed, put into two beakers respectively, 10mL of distilled water is added respectively, stirring to dissolve respectively, and then the chitosan oligosaccharide solution and the zinc sulfate solution are mixed and fully stirred to obtain a mixed solution. Regulating the pH value of the mixed solution by using 1% ammonia water solution in parts by weight to slowly rise to 7, placing a beaker filled with the mixed solution in a water bath kettle at 40 ℃, performing chelation reaction for 30min, dialyzing for 48h by using a dialysis bag with the molecular weight cutoff of 3500Da, slowly adding three times of absolute ethyl alcohol into the dialyzate, and washing by using the absolute ethyl alcohol after suction filtration. And then the precipitate is placed in a blast oven at 60 ℃ for drying, and the yellow solid obtained is the chitosan oligosaccharide chelated zinc. The detection shows that the mass fraction of zinc in the chitosan oligosaccharide chelated zinc prepared by the method is 10 percent (zinc sulfate is purchased from pharmaceutical industry Co., ltd., model is analytically pure)
2) Weighing 10mg of chitosan oligosaccharide chelate zinc in the step 1), dissolving in distilled water, adding 100 mu L of tween-80, uniformly stirring, and uniformly stirring to obtain the plant regulator B (the concentration of chitosan oligosaccharide chelate zinc in the plant regulator B is 100 mg/L) after constant volume is reached to 100 mL.
Comparative example 1
Preparing a plant regulator, which comprises the following steps:
weighing 10mg of chitosan oligosaccharide in the step 1) in the example 1, dissolving in 80mL of distilled water, adding 100 mu L of Tween-80, uniformly stirring, fixing the volume to 100mL, and uniformly stirring to obtain the plant regulator A (the concentration of chitosan oligosaccharide chelated zinc in the plant regulator B is 100 mg/L).
Results and analysis
The results of infrared spectroscopic analysis of the plant regulator A and the plant regulator B prepared in comparative example 1 and example 1, respectively, are shown in FIG. 1 (COS represents the preparation of the plant regulator A in comparative example 1; COS-Zn represents the preparation of the plant regulator B in example 1).
As can be seen from FIG. 1, the wave number is about 556.6cm -1 The absorption peak of-NH 2 at the site is obviously weakened, which indicates that the-NH in the chitosan oligosaccharide molecule 2 Participate in the chelation reaction; located at a wavenumber of about 1514.6cm -1 The absorption peak of the-C=O bond is obviously weakened, namely, the-C=O double bond in the chitosan oligosaccharide molecule and zinc ions are chelated to form-C-O-Zn. The infrared spectrum test result shows that chitosan oligosaccharide and zinc ion are chelated to form stable chelate.
Application example 1
Antiviral experiment: the inhibition of Tobacco Mosaic Virus (TMV) by the plant regulator in example 1 and comparative example 1 was determined by potting test and half-leaf spot method (ref. Liu Xiaowei, qin Yuanxia, yuan Lianlian, etc. two strains of rhizosphere growth promoting bacteria were studied for biocontrol effect on TMV [ J ]. Chinese tobacco theory, 2018,24 (06): 78-85.DOI:10.16472/J. Chinatobacco.2018.080).
The experiment consisted of the following steps:
1) Plant regulator: the plant regulator A in comparative example 1 and the plant regulator B in example 1 were diluted 100-fold respectively for use;
control reagent 1: separately preparing a zinc sulfate reagent, and dissolving 100mg of zinc sulfate in 100mL of distilled water to prepare a solution with the final concentration of 1000mg/L for later use;
control reagent 2: directly mixing 100mg of chitosan oligosaccharide with 1000mg of zinc sulfate reagent, and fixing the volume to 1L for later use;
control reagent 3: market products: chitosan oligosaccharide zinc (purchased from Shandong Lloyd biotechnology Co., ltd.)
2) Test crop: the three-stage tobacco producing process includes the steps of removing spot. Planting the three-leaf tobacco with the dead spots in the sterilized seedling raising substances, and placing the seedling raising substances in an insect prevention greenhouse for planting and culturing, wherein the three-leaf tobacco with the dead spots is used when the three-leaf tobacco with the dead spots grows to 7-8 leaf periods (the plants are planted and cultured according to the local planting management habit);
3) Tobacco Mosaic Virus (TMV): the variety of tobacco stored in the insect-proof greenhouse is Yunyan 87.
4) Preparing a test virus inoculation liquid: taking 1g of TMV infected leaf, putting into a mortar to be ground into homogenate, filtering by gauze, transferring filtrate into a centrifuge tube, and mixing with 10mL of water for standby;
5) The treatment group is set: mixing 1mL of the diluted plant regulator A in the step 1) with 1mL of the test virus inoculation liquid in the step 4) (marked as mixed liquid COS), mixing 1mL of the diluted plant regulator B in the step 1) with 1mL of the test virus inoculation liquid in the step 4) (marked as mixed liquid COS-Zn), placing the mixed liquids into 2mL centrifuge tubes respectively, standing for 15min, and respectively inoculating the mixed liquids onto test tobacco leaves (the test tobacco variety is Zhongyan 100) by using a half-leaf method friction, wherein the mixed liquid COS is inoculated onto the right half leaf of the tobacco leaf 1, and the mixed liquid COS-Zn is inoculated onto the right half leaf of the tobacco leaf 2.
Control group settings: mixing 1mL of clean water with 1mL of the virus inoculation liquid to be tested in the step 4) (denoted as mixed liquid CK), and respectively rubbing and inoculating the mixed liquid CK onto the left half leaf which is not treated in the tobacco leaf 1 (namely, the right half of one tobacco leaf is inoculated with COS, the left half is inoculated with CK) and the left half leaf which is not treated in the tobacco leaf 2 (namely, the right half of one tobacco leaf is inoculated with COS-Zn, the left half is inoculated with CK);
inoculating the control reagent 1 to the control reagent 3 by adopting the same half-leaf method; each treatment was repeated in triplicate and the inoculated tobacco was cultivated in a greenhouse.
Results and analysis
The formula for calculating the disease prevention effect is as follows:
disease preventing effect = (number of half leaf blight spots treated-number of half leaf blight spots control)/number of half leaf blight spots control x 100%
Typical occurrence at 48h and 72hCounting the number of dead spots after the dead spot symptoms, wherein the results are shown in table 1, and COS represents tobacco leaves inoculated with CK in the left half leaf and COS in the right half leaf; COS-Zn means tobacco leaves inoculated with CK in the left half and COS-Zn in the right half; znSO (ZnSO) 4 Represents the inoculation of CK on the left half leaf and ZnSO on the right half leaf 4 Is a tobacco leaf of (2); COS+ZnSO 4 Represents the inoculation of CK on the left half leaf and COS+ZnSO on the right half leaf 4 Is a tobacco leaf of (2); the market product represents tobacco leaves inoculated with CK in the left half and with CK in the right half), and the inhibition of Tobacco Mosaic Virus (TMV) by chitosan oligosaccharide and its chelate was studied.
TABLE 1 inhibition of tobacco mosaic Virus by Chitosan oligosaccharide and Chitosan oligosaccharide chelate Zinc
As can be seen from table 1, each treatment group had an inhibitory effect on tobacco mosaic virus compared to the control; the chitosan oligosaccharide or zinc sulfate is singly applied, and the inhibition rate of the chitosan oligosaccharide or zinc sulfate on tobacco mosaic virus is equivalent. However, the chitosan oligosaccharide is mixed with zinc sulfate, so that the inhibition rate of the chitosan oligosaccharide on tobacco mosaic virus is improved; the chitosan oligosaccharide chelated zinc has the best inhibition effect on tobacco mosaic virus, the inhibition rate is as high as 71.43%, and the effect is superior to that of products on the market.
Application example 2
Wheat low temperature resistance experiment: the plant regulators prepared in comparative example 1 and example 1 were sprayed to wheat seedlings in the seedling stage, and the concrete experiments were as follows:
1) Plant regulator: taking the plant regulator A after 100 times dilution, the plant regulator B after 100 times dilution, the control reagent 1 and the control reagent 2 prepared in the step 1) in the application example 1 for standby;
after 100 mu L of Tween 80 surfactant is fixed to 100mL, clean water containing the surfactant is prepared for standby.
2) Test crop: wheat (variety Jimai 22). Sterilizing wheat seeds, soaking, accelerating germination, selecting full and uniform wheat seeds after the seeds are just exposed to white, sowing the seeds into culture dishes, and supplementing 30 grains per dish with HoaglandThe liquid was cultured in a light incubator. The culture conditions are as follows: 25/20 ℃ (day/night), light intensity 800. Mu. Mol/m 2 S, a light cycle of 14/l0h (day/night), a relative humidity of 65.+ -. 5%;
when wheat seedlings grow to the 1-heart stage of 2 leaves, the wheat seedlings are reserved;
3) The treatment group is set: spraying the wheat seedling leaves in the step 2) with the diluted plant regulator A, the diluted plant regulator B, the control reagent 1 and the control reagent 2 in the step 1) (based on the condition that the leaves are completely wetted but the liquid does not flow down), and respectively marking as a COS treatment group, a COS-Zn treatment group and a ZnSO treatment group 4 Treatment group and COS+ZnSO 4 Mixing and assembling;
negative control group 1: spraying the clear water containing the surfactant prepared in the step 1) (based on the condition that the leaves are completely wet but the liquid does not flow down), treating for 24 hours, and culturing in a room temperature environment (about 25 ℃).
Negative control group 2: spraying the surfactant-containing clear water prepared in step 1) (based on the complete wetting of the leaves, but the liquid does not flow down);
comparison of commercial products: chitosan oligosaccharide zinc (purchased from Shandong green Biotechnology Co., ltd.) was sprayed according to the instructions of the product.
Each treatment was repeated 3 times, 30 wheat seedlings each.
4) And (2) respectively treating the treated group, the negative control group 2 and the commercial product control in the step (3) for 24 hours, then placing the treated group, the negative control group 2 and the commercial product control in a low temperature of 4 ℃ for stress for 48 hours, then placing the wheat in a room temperature condition (about 25 ℃) for recovering for 48 hours, and respectively measuring various indexes of wheat seedling leaves in the treated group and the negative control group, wherein the results are shown in tables 2-5.
Results and analysis
TABLE 2 influence of different treatment groups on wheat seedling growth parameters
As can be seen from table 2, wheat seedling growth is inhibited under low temperature stress, and seedling height, root length, fresh weight and dry weight are all significantly reduced; compared with the low-temperature stress group of the negative control group 2, the COS treatment group provided by the comparative example 1 has the advantages that the wheat seedlings are respectively increased by 24.6%,15.3%,35.9% and 56.8% in height, root length, fresh weight and dry weight; after the COS-Zn treatment group provided in example 1 of the present invention was used, the wheat seedlings were raised by 39.0%,21.1%,43.1% and 81.1% in height, root length, fresh weight and dry weight, respectively. After zinc sulfate is singly applied, the height, root length, fresh weight and dry weight of wheat seedlings are respectively improved by 22.2%,11.9%,22.5% and 32.4%; after the chitosan oligosaccharide is mixed with the zinc sulfate, the height, root length, fresh weight and dry weight of wheat seedlings are respectively improved by 28.4%,16.4%,40.2% and 64.9%; after the commercial products are applied, the height, root length, fresh weight and dry weight of wheat seedlings are respectively increased by 28.48%,15.9%,37.9% and 67.6%. Therefore, the plant regulator prepared by the invention can promote the accumulation of plant seedling biomass under low temperature stress, and has better effect than that of independently applying chitosan oligosaccharide or zinc sulfate, and is also better than that of simply compounding chitosan oligosaccharide and zinc sulfate, and is better than that of products on the market.
TABLE 3 influence of different treatment groups on physiological index of wheat seedlings
As can be seen from table 3, the malondialdehyde content of wheat seedling leaves is significantly increased under low temperature stress; the malondialdehyde content in COS-Zn treated groups of application example 1 was reduced by 34.4%,28.9%,34.1% and 62.0%, respectively, by comparison with negative control group 2 by the addition of COS, zinc sulfate, chitosan oligosaccharide alone, and zinc sulfate. Therefore, the peroxidation of the plant material film under low temperature stress can be reduced by different treatments, the structure of the plant material film is protected, and the COS-Zn treatment group effect is obviously superior to other groups. Meanwhile, COS-Zn can also improve the content of proline and soluble sugar in plants under low-temperature stress, maintain the osmotic pressure balance of plant cells and ensure the normal physiological functions of the plants.
In addition, low temperature stress degrades chlorophyll in leaves of wheat seedlings; however, the treatment of chitosan oligosaccharide and the like can relieve chlorophyll degradation caused by low temperature. Compared with control group 2, the chlorophyll content in the COS-Zn treated group of application example 1 was increased by 60.8%,41.9%,59.5% and 63.5%, respectively, by the single application of COS, zinc sulfate, the compounding of chitosan oligosaccharide with zinc sulfate. Therefore, the chitosan oligosaccharide and the chelate thereof can reduce the degradation of plant chlorophyll under low-temperature stress and improve the photosynthesis efficiency of plants, and particularly the plant regulator taking the chitosan oligosaccharide chelated zinc as the main component has more remarkable effect.
TABLE 4 influence of different treatment groups on antioxidant enzyme Activity of wheat leaves
As can be seen from table 4, wheat seedling leaves have enhanced antioxidant enzyme activity under stimulation of low temperature stress; the different treatment groups of COS are capable of further enhancing antioxidant enzyme activity in plants. Compared with control group 2, the activity of SOD, POD and CAT in wheat in COS treatment group is enhanced by 20.0%,18.8% and 56.6%; the activity of SOD, POD and CAT in wheat in COS-Zn treatment group is enhanced by 34.0%,22.7% and 68.9%; the activity of SOD, POD and CAT of wheat is enhanced by 15.8%,9.4% and 41.8% by single application of zinc sulfate; the chitosan oligosaccharide is compounded with zinc sulfate, so that the activities of SOD, POD and CAT of wheat are enhanced by 27.4%,17.1% and 61.8%; the activity of SOD, POD and CAT of wheat is enhanced by 20.0%,18.2% and 65.0% by applying the commercial products. Therefore, although the antioxidant enzyme activity in plants under low-temperature stress can be enhanced by different treatments of chitosan oligosaccharide and zinc element, excessive active oxygen generated by low-temperature stress induction is removed, a plant substance film structure is protected, and the capability of the plants for resisting low-temperature stress is improved, the effect of chelating zinc by chitosan oligosaccharide is better and is better than that of products on the market.
Application example 3
Tobacco low temperature resistance experiment: the invention takes tobacco as an example, and plants regulators prepared in comparative example 1 and example 1 are sprayed on tobacco seedlings, and the concrete experiment is as follows:
1) A plant regulator; the same as in application example 2, step 1);
2) Test for testingCrop: tobacco is medium tobacco 100. Culturing 4-leaf tobacco seedling in illumination incubator under the conditions of 25/20deg.C (day/night) and illumination intensity of 800 μmol/m 2 S, a light cycle of 14/l0h (day/night), a relative humidity of 65.+ -. 5%; when the tobacco seedlings grow to 6 leaf stage, reserve;
3) The treatment group is set: spraying tobacco seedling leaves in the step 2) with the diluted plant regulator A, the diluted plant regulator B, the control reagent 1 and the control reagent 2 in the step 1) (based on the fact that the leaves are completely wetted but the liquid does not flow down), and respectively marking the leaves as a COS treatment group, a COS-Zn treatment group and a ZnSO treatment group 4 Treatment group and COS+ZnSO 4 Mixing and assembling;
negative control group 1: spraying the clear water containing the surfactant prepared in the step 1) (based on the condition that the leaves are completely wet but the liquid does not flow down), treating for 24 hours, and culturing in a room temperature environment (about 25 ℃).
Negative control group 2: spraying the surfactant-containing clear water prepared in step 1) (based on the complete wetting of the leaves, but the liquid does not flow down), and culturing in a low temperature environment (4 ℃).
Each treatment was repeated 3 times, 8 seedlings each.
4) After 24 hours of treatment in the treatment group and the negative control group 2 in the step 3), the tobacco seedlings are subjected to stress for 48 hours at the low temperature of 4 ℃, and then the tobacco is subjected to room temperature (about 25 ℃) for 48 hours, and each index of tobacco seedling leaves in the treatment group and the control group is measured respectively, and the results are shown in Table 5 and FIG. 2.
Results and analysis
TABLE 5 influence of different treatment groups on tobacco seedling growth parameters
As can be seen from table 5 and fig. 2, tobacco seedling growth was inhibited under low temperature stress, and leaf area, above-ground dry weight, root length, and root dry weight were all significantly reduced; compared with the negative control group 2, after being treated by the COS treatment group, the tobacco leaf area, the overground fresh weight, the root length and the root dry weight are respectively improved by 17.9%,36.8%,13.0% and 72.2%; after being treated by COS-Zn treatment group, the leaf area, fresh weight on the ground, root length and root dry weight are respectively improved by 34.0%,68.4%,27.0% and 94.4%. The zinc sulfate or the chitosan oligosaccharide and the zinc sulfate are simply compounded, and the effect on the low temperature resistance of tobacco seedlings is general, because the loss of inorganic zinc is serious, and the inorganic zinc cannot be well absorbed and utilized by plants. Therefore, both the chitosan oligosaccharide and the chitosan oligosaccharide chelating zinc can promote the accumulation of plant seedling biomass under low temperature stress, but the effect of the chitosan oligosaccharide chelating zinc is better than that of the chitosan oligosaccharide, which indicates that the chitosan oligosaccharide chelating zinc not only plays the role of stress resistance and growth promotion of the chitosan oligosaccharide, but also can convert zinc element into an organic state, and better plays the role of growth promotion.
In conclusion, after the plant regulator provided by the invention is applied, the antioxidant enzyme activity and photosynthesis efficiency of plants in a low-temperature environment can be greatly improved, and virus diseases of the plants can be obviously inhibited. Therefore, the chitosan oligosaccharide chelated zinc plant regulator provided by the invention can improve the stress resistance of plants, and lays a foundation for realizing the high-value utilization of ocean low-value products.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (10)
1. The plant regulator is characterized in that the effective components of the plant regulator comprise chitosan oligosaccharide chelated zinc; the concentration of the chitosan oligosaccharide chelated zinc in the plant regulator is 100-1000 mg/L.
2. The plant regulator according to claim 1, wherein the mass content of zinc in the chitosan oligosaccharide chelate zinc is 3% to 4%.
3. The plant regulator of claim 1, further comprising a surfactant; the concentration of the surfactant in the plant regulator is 1-10 mL/L.
4. A plant regulator according to claim 3, wherein the surfactant comprises one or more of tween active agent, span active agent, OP active agent and NP active agent.
5. Use of a plant regulator according to any one of claims 1 to 4 for increasing antiviral ability of plants and/or for alleviating low temperature stress in plants.
6. A method for improving antiviral ability and/or alleviating low temperature stress in a plant comprising the steps of:
plant regulator according to any one of claims 1 to 4 is used to soak plant seeds and/or is applied to plants.
7. The method of claim 6, wherein the means of administration comprises: spraying and/or root irrigation.
8. The method according to claim 6, wherein the concentration of the plant regulator is 0.1 to 0.5g/L when the plant species is soaked; the soaking time is 6-8 h.
9. The method according to claim 6, wherein the plant regulator is applied to the plant at a concentration of 0.1 to 1g/L and at an application rate of 20 to 80L/mu.
10. The method of claim 6, wherein the species of plant comprises: tobacco and/or wheat.
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