CN114916548A - Plant drought-resistant agent and preparation method and application thereof - Google Patents
Plant drought-resistant agent and preparation method and application thereof Download PDFInfo
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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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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- 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
- A01G13/00—Protecting plants
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- 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
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- 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
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- 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
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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
- A01N25/30—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 characterised by the surfactants
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- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
The invention provides a plant drought-resistant agent, a preparation method and application thereof, and belongs to the technical field of plant polypeptides. The plant drought-resistant agent provided by the invention takes a STRPEEP 1 polypeptide mature body as a main component; wherein the amino acid sequence of the mature polypeptide of STRPEEP 1 is as follows: S-Q-V-V-Q-G-S-V-P-S-P-G-V-G-H. The plant drought-resistant agent provided by the invention has the advantages of natural green and pollution-free property, strong biological activity, strong field operability, remarkable drought-resistant effect and no adverse reaction.
Description
Technical Field
The invention belongs to the technical field of plant polypeptides, and relates to a plant drought-resistant agent, and a preparation method and application thereof.
Background
Drought stress is one of the serious challenges facing agriculture in China and even in the world, and is directly related to grain safety and economic safety. Taking wheat as a main food crop as an example, wheat in China is mainly distributed in arid and semiarid regions, and the yield and the quality of wheat are seriously influenced by drought stress.
At present, the molecular mechanism of drought stress resistance of plants is greatly developed, the regulation and control aspects of partial plant hormones and compounds are mainly focused, the excavated genes with drought resistance and drought resistance can be utilized, and new varieties of plant drought resistance germplasm resources are created by combining traditional crossbreeding and modern molecular breeding. In addition, the drought resistance of crops can be improved by improving cultivation and cultivation techniques. However, the method for obtaining drought-resistant plants by the traditional breeding technology is long in time consumption and large in workload; the molecular breeding technology inevitably has ethical problems and is not accepted by the masses; and other chemical agents have production safety problems.
Plant polypeptide hormone is a small secreted peptide whose function is involved in many plant growth and development processes. The polypeptide mature body is a peptide segment with biological activity formed by a series of processing, shearing and modification of a protein precursor coded by a polypeptide gene. The polypeptide mature body is secreted to the outside of the cell, is recognized by a specific membrane receptor thereof, and transduces signals into the cell to further regulate the expression of downstream genes, thereby regulating the growth and development of plants. The mature polypeptide can be obtained by artificial synthesis, and can be directly applied in vitro to regulate and control the growth and development of plants, such as the elongation of roots, the development of lateral roots and the like. However, the plant polypeptide hormone has few reports in plant stress resistance response at present, and the application of the mature polypeptide in plants, especially crops, is not reported.
Disclosure of Invention
The plant drought-resistant agent has the advantages of being natural, green and pollution-free, strong in biological activity, strong in field operability, remarkable in drought-resistant effect and free of adverse reaction.
In order to achieve the aim, the invention provides a plant drought-resistant agent which takes a polypeptide mature body of STRPP 1 as a main component;
wherein the amino acid sequence of the mature polypeptide of STRPEEP 1 is as follows:
S-Q-V-V-Q-G-S-V-P-S-P-G-V-G-H。
preferably, the plant drought-resistant agent further comprises organic silicon or tween-20.
In the scheme, the organic silicon is used as a surfactant, has very low surface tension, can realize quick wetting and dispersion of the STREP 1 drought-resistant agent, and can permeate into pores of plants, so that the absorption of the STREP 1 drought-resistant agent by the plants is promoted, the utilization rate of the drought-resistant agent is improved, and the use cost of the STREP 1 plant drought-resistant agent is reduced. Taking wheat as an example, when the mature polypeptide solution of STRPEEP 1 is sprayed, the solution cannot be uniformly dispersed on the surface of a plant, but forms water drops and then falls off from the surface of the plant; the organic silicon assistant is added into the STRPP 1 mature polypeptide solution and then sprayed, and the solution can be uniformly dispersed on the surface of the plant, so that the surface of the plant is quickly moistened. It will be appreciated that tween-20 may also achieve the above effect as an alternative to silicone, in practice silicone is preferred, but tween-20 may also be used instead.
Preferably, the concentration of the mature polypeptide body of STRPP 1 is 0.5-1.0 mu mol/L; the concentration of the organic silicon or the Tween-20 is 0.5-1.0 mL/L.
In the above protocol, the concentration of the mature polypeptide of strppe 1 may be 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 or any concentration within the above range. Taking wheat as an example, the isolated leaves of the wheat are treated by 0.5 mu mol/L and 1.0 mu mol/L of STRPEEP 1 polypeptide solutions, under drought conditions (20% PEG-6000 simulated drought conditions), the leaves treated by the polypeptide all show a remarkable drought-resistant phenotype, and the chlorophyll content is remarkably higher than that of the leaves not treated by the polypeptide, so that the working concentration of 0.5 mu mol/L of the polypeptide is more preferable from the economic point of view. The concentration of the silicone is not particularly limited as long as the solution can be rapidly dispersed and osmotically wetted on the surface of the plant, and the above range is selected as much as possible from the viewpoint of economy.
The invention provides a preparation method of a plant drought-resistant agent according to any one of the technical schemes, which comprises the following steps:
adding the mature polypeptide mother liquor of STREP 1 into distilled water to prepare a polypeptide solution of 0.5-1.0 mu mol/L, simultaneously adding organic silicon or Tween-20, and uniformly mixing to obtain the plant drought-resistant agent.
Preferably, the strppep 1 mature body polypeptide mother liquor is prepared by the following method:
the STRPEP1 mature polypeptide powder was dissolved in distilled water to prepare a STRPEP1 mature polypeptide mother solution with a concentration of 1 Ag/L for further use.
Preferably, the mother liquid of the mature body polypeptide of STRPEEP 1 is stored at-70 ℃ when being stored for a long time; when used for a short period, it is stored at-20 deg.C.
The invention provides an application of the plant drought-resistant agent according to any one of the technical schemes in improving the drought-resistant capability of plants.
Preferably, the plant drought-resistant agent is directly and uniformly sprayed on the surface of the overground part of the plant, or the plant drought-resistant agent is used for soaking cotton balls and uniformly coating the cotton balls on the overground part of the plant.
Preferably, the plant is wheat or tobacco.
Compared with the prior art, the invention has the advantages and positive effects that:
1. the STRPP 1 mature polypeptide used in the plant drought-resistant agent provided by the invention is a mature polypeptide obtained by artificial synthesis, is convenient to obtain and can be synthesized in a large scale; the nano-molecular compound has the advantages of natural green and no pollution, has strong biological activity, can act at a nano-molar concentration, and greatly reduces the use cost;
2. the preparation and use methods of the plant drought-resistant agent provided by the invention are simple, the field operability is strong, special training is not needed for users, and the operation is simple and practical;
3. the plant drought-resistant agent provided by the invention has a remarkable drought-resistant effect and does not cause adverse reactions.
Drawings
FIG. 1 is a schematic diagram of a result of processing wheat in vitro leaves by an artificially synthesized plant drought resistant agent provided by an embodiment of the invention under a drought condition; wherein, A, wheat in-vitro leaves before treatment; B. wheat in-vitro leaves are treated by drought and plant drought resistance agent for 8 days; C. fig. B shows measurement of chlorophyll content in wheat leaves; D. wheat leaf photosynthetic rate determination in panel B;
FIG. 2 is a schematic diagram of the results of treating Nicotiana benthamiana in vitro leaves with an artificially synthesized plant drought resistant agent provided by an embodiment of the present invention under drought conditions; wherein, A, tobacco leaves in vitro before drought treatment; B. tobacco leaves in vitro after drought and plant drought-resistant agent treatment for 7 days; C. measuring the chlorophyll content of the tobacco leaves in the graph B; D. determination of photosynthetic rate of tobacco leaf in panel B;
FIG. 3 is a schematic diagram of the results of wheat seedling treatment with the artificially synthesized plant drought-resistant agent provided by the embodiment of the invention under drought conditions; a, treating live wheat plants with a plant drought-resistant agent under drought conditions to obtain fruits for 4 days; b, weighing the fresh weight of the wheat plant after drought and plant drought resistance agent treatment; C. measuring the chlorophyll content of the wheat plants after drought and plant drought resistance agent treatment; in the diagram A, bar is 5 cm.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The plant drought-resistant agent comprises 0.5 mu mol/L of STRPEEP 1 mature polypeptide body and 0.5mL/L of organic silicon auxiliary agent.
The preparation method comprises the following steps:
weighing STRPP 1 polypeptide powder, dissolving in distilled water, and preparing into STRPEP1 mature polypeptide mother liquor with concentration of 1 Ag/L for later use;
adding 1.42 mu L of the mature polypeptide mother liquor of STREP 1 into 2mL of distilled water, fully and uniformly mixing, adding 1 mu L of organic silicon, and uniformly mixing again to obtain the mature polypeptide plant drought-resistant agent of STREP 1.
Example 2
The STREP 1 mature polypeptide plant drought-resistant agent comprises 1.0 mu mol/L of STREP 1 mature polypeptide body and 0.5mL/L of organic silicon auxiliary agent.
The preparation method comprises the following steps:
weighing STRPEP1 polypeptide powder, dissolving in distilled water to prepare STRPEP1 mature polypeptide mother solution at concentration 1/L for later use;
and adding 2.84 mu L of the mature polypeptide mother liquor of STRPP 1 into 2mL of distilled water, fully and uniformly mixing, adding 1 mu L of organic silicon, and uniformly mixing again to obtain the mature polypeptide plant drought-resistant agent of STRPP 1.
Example 3
The STREP 1 mature polypeptide plant drought-resistant agent comprises 0.8 mu mol/L of STREP 1 mature polypeptide and 1.0mL/L of Tween-20 auxiliary agent.
The preparation method comprises the following steps:
weighing STRPP 1 polypeptide powder, dissolving in distilled water, and preparing into STRPEP1 mature polypeptide mother liquor with concentration of 1 Ag/L for later use;
and adding 2.27 mu L of the mature polypeptide mother liquor of STREP 1 into 2mL of distilled water, fully and uniformly mixing, adding 202 mu L of Tween, and uniformly mixing again to obtain the mature polypeptide plant drought-resistant agent of STREP 1.
Performance testing
Test for treating in vitro Material
Taking example 1 as an example, the plant drought-resistant agent prepared in example 1 is used for treating the wheat detached leaf and the tobacco detached leaf respectively according to the using method.
The drought stress is realized by 20 percent PEG-6000 solution, and the specific preparation method comprises the following steps:
weighing 20 Ag PEG-6000 solid, dissolving in distilled water, and fixing the volume to 100mL for later use after the solid is completely dissolved.
Treatment 1: treating the in vitro leaves of the wheat: the wheat is grown for two weeks under continuous illumination, the in vitro leaf (the first leaf) at the same leaf position is obtained, the position about 4cm in the middle of the leaf is taken, and the leaf is placed in a culture dish with filter paper laid at the bottom. 2mL of distilled water (blank control), 2mL of STREP 1 drought-resistant agent, 2mL of 20% PEG-6000 (simulated drought stress) and 2mL of 20% PEG-6000 containing STREP 1 drought-resistant agent were added to the petri dish. The phenotype was observed after 8 days of standing.
And (3) treatment 2: treating the tobacco in-vitro leaves: taking the 7 th leaf blade of tobacco in the vigorous growth stage, punching by a puncher with the diameter of 1.0cm, taking a leaf disc, and placing the leaf disc in a culture dish with filter paper laid at the bottom. 2mL of distilled water (blank control), 2mL of STREP 1 drought-resistant agent, 2mL of 20% PEG-6000 (simulated drought stress) and 2mL of 20% PEG-6000 containing STREP 1 drought-resistant agent were added to the petri dish. The phenotype was observed after 7 days of standing.
As shown in FIGS. 1A-B, leaves of wheat with the same leaf position (first piece) and consistent growth are picked by tweezers, and are flatly placed in a culture dish, and the STREP 1 mature polypeptide plant drought-resistant agent is treated in an infiltration manner and is placed under continuous illumination for 8 days.
The result shows that the damage of the wheat detached leaves caused by drought can be obviously inhibited after the STRPP 1 mature polypeptide plant drought-resistant agent is treated. After 8 days, the control group wheat leaves turn yellow from the two ends and the edges of the leaves, while the drought-treated wheat leaves turn yellow in the whole piece, and the yellowing of the wheat leaves caused by drought is obviously delayed after the drought-treated wheat leaves are treated by adding the STREP 1 mature polypeptide plant drought-resistant agent. The chlorophyll content was determined by the ethanol method, as shown in fig. 1C, where the drought treatment resulted in a dramatic decrease in the chlorophyll content of the wheat blades as compared to the control 0.315m Ag/Ag, down to 0.127m Ag/Ag, while the chlorophyll content of the plant drought resistant agent treated blades remained at a higher level, up to 0.341m Ag/Ag, comparable to the blank control. And meanwhile, photosynthetic rate determination is carried out on the leaves in vitro of the wheat, as shown in figure 1D, compared with a control group, the photosynthetic rate of the leaves of the wheat is obviously reduced due to drought treatment, and the photosynthetic rate of the leaves treated by the plant drought-resistant agent is still maintained at a higher level.
As shown in FIGS. 2A-B, the leaves of Nicotiana benthamiana (seventh leaf) with the same leaf position and consistent growth vigor were selected, punched with a punch having a diameter of 1.0cm for sampling, placed in a petri dish regularly, treated with a mature polypeptide plant drought-resistant agent STRPP 1 in an infiltration manner, and left under continuous light for 7 days.
The result shows that the damage of the tobacco leaves in vitro caused by drought can be obviously inhibited after the plant drought-resistant agent of the mature polypeptide of STRPP 1 is treated. After 7 days, the tobacco leaves in the control group almost stay green, while the tobacco leaves subjected to drought treatment almost completely turn yellow, and the tobacco leaves subjected to drought treatment are obviously delayed to turn yellow after being treated by the STRPEEP 1 mature polypeptide plant drought-resistant agent. The chlorophyll content was determined by the ethanol method, as shown in fig. 2C, where the drought treatment resulted in a dramatic decrease in the chlorophyll content of the wheat blades down to 0.160m Ag/Ag compared to the control 0.493m Ag/Ag, while the chlorophyll content of the plant drought resistant agent treated blades remained at a higher level up to 0.420m Ag/Ag. And meanwhile, the photosynthetic rate of the tobacco leaves in vitro is measured, as shown in figure 2D, compared with a control group, the photosynthetic rate of the tobacco leaves is obviously reduced (by 33.8%) due to drought treatment, and the photosynthetic rate of the leaves treated by the plant drought-resistant agent is still maintained at a higher level and is almost not different from that of the control group.
Test for treating living plants
Using example 1 as an example, the mature polypeptide plant drought resistant agent STRPP 1 prepared in example 1 is used for treating live wheat plants according to the using method.
As shown in FIG. 3A, wheat with consistent germination was selected and sown in soil for long-day culture, water-cut and drought-resistant treatment was performed while two cores were kept, and at the same time, spray treatment was performed with STRPEEP 1 mature polypeptide plant drought-resistant agent. The culture was continued under light for 4 days.
The contrast group sprayed with the mature polypeptide plant drought-resistant agent STRPEP1 has the same growth vigor as the wheat seedlings without the mature polypeptide plant drought-resistant agent, has no obvious difference, and after treatment, the wheat seedlings subjected to drought treatment are withered due to severe water loss, and most of the wheat seedlings subjected to spray with the mature polypeptide plant drought-resistant agent STRPEP1 still grow vertically. As shown in fig. 3B, the fresh weight of the overground part of the wheat seedlings shows that the fresh weight of the overground part of the wheat sprayed with the strppep 1 mature polypeptide plant drought-resistant agent in the control group is not significantly different from that of the overground part of the wheat sprayed with the strppep 1 mature polypeptide plant drought-resistant agent in the non-spraying manner, which indicates that the strppep 1 mature polypeptide plant drought-resistant agent has no adverse side effect on plants, and the biomass of the overground part of the wheat is significantly increased (increased by 12.39%) after the plant drought-resistant agent is sprayed under the drought condition. As shown in fig. 3C, the chlorophyll measurement results show that there is no significant difference in chlorophyll content between the control group sprayed and the plant drought resistant agent without spraying the mature polypeptide strppe 1, whereas under drought conditions, the chlorophyll content of wheat sprayed with the plant drought resistant agent is significantly higher than that of wheat not sprayed with the plant drought resistant agent (increased by 68.63%).
Claims (10)
1. The plant drought-resistant agent is characterized in that a polypeptide mature body STRPP 1 is used as a main component; wherein the amino acid sequence of the mature polypeptide of STRPEEP 1 is as follows: S-Q-V-V-Q-G-S-V-P-S-P-G-V-G-H.
2. The plant drought resistant agent of claim 1, further comprising silicone or tween-20.
3. The plant drought resistant agent according to claim 1 or 2, wherein the concentration of the mature polypeptide body of STRPP 1 is 0.5-1.0 μmol/L.
4. The plant drought resistant agent according to claim 1 or 2, wherein the concentration of the organosilicon or tween-20 is 0.5-1.0 mL/L.
5. The method for preparing a plant drought resistant agent according to any of claims 1-4, comprising the steps of:
adding the STRPP 1 mature body polypeptide mother solution into distilled water to prepare 0.5-1.0 mu mol/L polypeptide solution, simultaneously adding organic silicon or Tween-20, and uniformly mixing to obtain the plant drought-resistant agent.
6. The method of claim 5, wherein the mature body polypeptide stock solution of STRPP 1 is prepared by:
the STRPEP1 mature polypeptide powder was dissolved in distilled water and set up as STRPEP1 mature polypeptide mother solution at a concentration of 1 Ag/L for further use.
7. The method according to claim 6, wherein the stock solution of the mature body polypeptide of STRPP 1 is stored at-70 ℃ when stored for a long period; when used in a short period, it is stored at-20 deg.C.
8. Use of a plant drought resistance agent according to any of claims 1-4 for increasing the drought resistance of a plant.
9. The use of claim 8, wherein the drought resistant agent is directly and uniformly sprayed on the surface of the overground part of the plant, or the drought resistant agent is used for soaking cotton balls and uniformly coating the cotton balls on the overground part of the plant.
10. Use according to claim 8 or 9, wherein the plant is wheat or tobacco.
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