CN114916548B - Drought-resistant agent for plants and preparation method and application thereof - Google Patents
Drought-resistant agent for plants and preparation method and application thereof Download PDFInfo
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- CN114916548B CN114916548B CN202210739673.3A CN202210739673A CN114916548B CN 114916548 B CN114916548 B CN 114916548B CN 202210739673 A CN202210739673 A CN 202210739673A CN 114916548 B CN114916548 B CN 114916548B
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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
- A01N37/46—N-acyl derivatives
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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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- Health & Medical Sciences (AREA)
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- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
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- Forests & Forestry (AREA)
- Chemical & Material Sciences (AREA)
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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 STREP 1 polypeptide mature body as a main component; wherein, the amino acid sequence of the STREP 1 polypeptide mature body is: 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 pollution-free property, strong biological activity, strong field operability, obvious 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, a preparation method and application thereof.
Background
Drought stress is one of the serious challenges facing China and even global agriculture, and is directly related to grain safety and economic safety. Taking wheat as a main grain crop as an example, the wheat in China is mainly distributed in arid and semiarid regions, and the drought stress has serious influence on the yield and quality of the wheat.
At present, the molecular mechanism of drought stress resistance of plants has been greatly developed, and the molecular mechanism is mainly concentrated on the aspects of regulation and control of part of plant hormones and compounds, so that the excavated drought-resistant and drought-resistant genes can be utilized, and new varieties of drought-resistant germplasm resources of plants are created by combining traditional crossbreeding and modern molecular breeding. In addition, drought resistance of crops can be improved by improving cultivation and cultivation technology. 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; while other chemicals present production safety issues.
Plant polypeptide hormone is a secreted small peptide whose function involves multiple 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 outside cells and recognized by a specific membrane receptor thereof, and signals are transduced into the cells so as to regulate and control the expression of downstream genes, thereby regulating and controlling the growth and development of plants. Mature polypeptide can be obtained by artificial synthesis, and can be directly applied in vitro to regulate plant growth and development, such as root elongation, lateral root development, etc. However, plant polypeptide hormones are rarely reported in plant stress resistance response, and application of mature polypeptides to plants, especially crops is not reported.
Disclosure of Invention
The invention provides a plant drought-resistant agent, a preparation method and application thereof, and the obtained plant drought-resistant agent has the advantages of natural green pollution-free property, strong biological activity, strong field operability, obvious drought-resistant effect and no adverse reaction.
In order to achieve the aim, the invention provides a plant drought-resistant agent, which takes STRP P1 polypeptide mature body as a main component;
wherein, the amino acid sequence of the STREP 1 polypeptide mature body is:
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 organosilicon is used as the surfactant, has very low surface tension, can realize rapid wetting and dispersion of the STRP 1 drought-resistant agent and permeates into plant pores, thereby promoting the absorption of the STRP 1 drought-resistant agent by plants, improving the utilization rate of the STRP 1 drought-resistant agent and reducing the use cost of the STRP 1 plant drought-resistant agent. Taking wheat as an example, when the solution of the mature polypeptide of the STRP 1 is sprayed, the solution cannot be uniformly dispersed on the surface of the plant, but rolls off from the surface of the plant after forming water drops; and the organosilicon auxiliary agent is added into the STREP 1 mature polypeptide solution and then sprayed, so that the solution can be uniformly dispersed on the surface of the plant, and the surface of the plant is quickly wetted. It will be appreciated that as an alternative to silicone, tween-20 may also achieve the above effect, in practice silicone is preferred, but tween-20 may also be used instead.
Preferably, the concentration of the STREP 1 mature polypeptide body is 0.5-1.0 mu mol/L; the concentration of the organosilicon or tween-20 is 0.5-1.0mL/L.
In the above scheme, in actual use, the concentration of the STREP 1 mature polypeptide can 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, in vitro wheat leaves are treated by 0.5 mu mol/L and 1.0 mu mol/L STRP 1 polypeptide solution, and under drought conditions (20% PEG-6000 simulates drought conditions), the leaves treated by the polypeptide show a remarkable drought-resistant phenotype, and the chlorophyll content is remarkably higher than that of the leaves treated by non-polypeptide, so that the concentration of the polypeptide of 0.5 mu mol/L is more preferable as the working concentration from the economic point of view. The concentration of the organosilicon is not particularly limited as long as the solution can be rapidly dispersed and infiltrated into the plant surface, and the above range is selected as much as possible from the viewpoint of economy.
The invention provides a preparation method of the plant drought-resistant agent according to any one of the technical schemes, which comprises the following steps:
adding STRP 1 mature polypeptide mother liquor into distilled water to prepare 0.5-1.0 mu mol/L polypeptide solution, adding organic silicon or tween-20, and uniformly mixing to obtain the plant drought-resistant agent.
Preferably, the STRPEP1 mature polypeptide mother liquor is formulated by the following method:
the STRP 1 mature polypeptide powder is dissolved in distilled water to prepare STRP 1 mature polypeptide mother liquor with the concentration of 1-fold/L for standby.
Preferably, the STREP 1 mature polypeptide mother liquor is stored at-70 ℃ when stored for a long time; when used for a short period, the product is stored at-20 ℃.
The invention provides an application of the plant drought-resistant agent according to any one of the technical schemes in improving drought resistance of plants.
Preferably, the drought-resistant agent is directly and uniformly sprayed on the surface of the overground part of the plant, or the cotton balls soaked by the drought-resistant agent are uniformly coated 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 STRP 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 preparation method has the advantages of natural green and pollution-free properties, high biological activity and the effect of nanomolar concentration, and greatly reduces the use cost;
2. the plant drought-resistant agent provided by the invention has the advantages of simple preparation and use method, strong field operability, no special training for users, and simple and practical operation;
3. the drought-resistant agent for plants provided by the invention has remarkable drought-resistant effect and does not cause adverse reaction.
Drawings
FIG. 1 is a schematic diagram of the results of processing wheat in vitro leaves with the artificially synthesized plant drought-resistant agent provided by the embodiment of the invention under drought conditions; wherein, A, wheat in-vitro leaves before treatment; B. wheat in-vitro leaves after drought and plant drought resistance agent treatment for 8 days; C. measuring chlorophyll content of wheat leaves in a drawing B; D. wheat leaf photosynthetic rate determination in panel B;
FIG. 2 is a schematic diagram of the results of processing in vitro leaves of Nicotiana benthamiana with the artificially synthesized plant drought-resistant agent provided by the embodiment of the invention under drought conditions; wherein, A, tobacco leaves in vitro before drought treatment; B. tobacco leaves in vitro after drought and plant drought resistance agent treatment for 7 days; C. measuring chlorophyll content of tobacco leaves in a graph B; D. leaf photosynthetic rate determination of tobacco in panel B;
FIG. 3 is a schematic diagram of the result of wheat seedlings treated with the artificially synthesized plant drought-resistant agent provided by the embodiment of the invention under drought conditions; wherein, the plant drought-resistant agent is used for treating the wheat living plant result for 4 days under the drought condition; b, fresh weight of wheat plants after drought and plant drought-resistant agent treatment is weighed; C. measuring chlorophyll content of wheat plants after drought and plant drought-resistant agent treatment; bar=5 cm in panel a.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The plant drought-resistant agent comprises 0.5 mu mol/L STRPP 1 mature polypeptide body and 0.5mL/L organosilicon auxiliary agent.
The preparation method comprises the following steps:
weighing STREP 1 polypeptide powder, dissolving in distilled water, and preparing STREP 1 mature polypeptide mother liquor with concentration of 1 g/L for standby;
1.42 mu L of STREP 1 mature polypeptide mother liquor is taken and added into 2mL of distilled water, fully and uniformly mixed, then 1 mu L of organic silicon is added, and uniformly mixed again, thus obtaining the STREP 1 mature polypeptide plant drought-resistant agent.
Example 2
A plant drought-resistant agent for STRP 1 mature polypeptide comprises 1.0 mu mol/L STRP 1 mature polypeptide body and 0.5mL/L organosilicon auxiliary agent.
The preparation method comprises the following steps:
weighing STREP 1 polypeptide powder, dissolving in distilled water, and preparing STREP 1 mature polypeptide mother liquor with concentration of 1 g/L for standby;
2.84 mu L of STREP 1 mature polypeptide mother liquor is taken and added into 2mL of distilled water, fully and uniformly mixed, then 1 mu L of organic silicon is added, and uniformly mixed again, thus obtaining the STREP 1 mature polypeptide plant drought-resistant agent.
Example 3
The plant drought-resistant agent for STRP 1 mature polypeptide comprises 0.8 mu mol/L STRP 1 mature polypeptide and 1.0mL/L Tween-20 auxiliary agent.
The preparation method comprises the following steps:
weighing STREP 1 polypeptide powder, dissolving in distilled water, and preparing STREP 1 mature polypeptide mother liquor with concentration of 1 g/L for standby;
adding 2.27 mu L of STREP 1 mature polypeptide mother liquor into 2mL of distilled water, fully and uniformly mixing, adding 2 mu L of Tween-20 mu L, and uniformly mixing again to obtain the STREP 1 mature polypeptide plant drought-resistant agent.
Performance testing
Test for in vitro Material handling
Taking example 1 as an example, the plant drought-resistant agent prepared in example 1 is used for respectively treating wheat in-vitro leaves and tobacco in-vitro leaves according to a using method.
Drought stress was achieved with a 20% peg-6000 solution, specifically formulated as follows:
weighing 20 g PEG-6000 solid, dissolving in distilled water, and fixing volume to 100mL after complete dissolution for standby.
Treatment 1: treatment of wheat ex-vivo leaves: the wheat grows by continuous illumination for two weeks, the in-vitro leaf (first leaf) at the same leaf position is obtained, the middle of the leaf is taken to be about 4cm, and the leaf is placed in a culture dish with filter paper paved at the bottom. 2mL of distilled water (blank control), 2mL of STRP 1 drought-resistant agent, 2mL of 20% PEG-6000 (simulating drought stress) and 2mL of 20% PEG-6000 containing the STRP 1 drought-resistant agent are respectively added into a culture dish. Phenotypes were observed after 8 days of standing.
Treatment 2: treatment of tobacco ex-vivo leaves: taking 7 th leaf leaves of tobacco in the vigorous growth period, punching by using a puncher with the diameter of 1.0cm, and placing a leaf taking tray in a culture dish with filter paper paved at the bottom. 2mL of distilled water (blank control), 2mL of STRP 1 drought-resistant agent, 2mL of 20% PEG-6000 (simulating drought stress) and 2mL of 20% PEG-6000 containing the STRP 1 drought-resistant agent are respectively added into a culture dish. Phenotypes were observed after 7 days of standing.
As shown in fig. 1A-B, the wheat leaves with identical growth vigor and the same leaf position (first leaf) are selected by forceps, placed in a culture dish flatly, the STRPEP1 mature polypeptide plant drought-resistant agent is treated in a soaking manner, and placed under continuous illumination for 8 days.
The result shows that the STREP 1 mature polypeptide plant drought-resistant agent can obviously inhibit the damage of wheat in vitro leaves caused by drought after being treated. After 8 days, the wheat leaves of the control group begin to turn yellow from the two ends and the edges of the leaf segments, and the wheat leaves subjected to drought treatment are turned yellow in whole, and after the STRP 1 mature polypeptide plant drought-resistant agent is added for treatment, the wheat leaves caused by drought are obviously delayed to turn yellow. As shown in FIG. 1C, compared with the chlorophyll content of 0.315m of the control group per gram, the chlorophyll content of the wheat leaves is sharply reduced by drought treatment to 0.127m of the total chlorophyll content per gram, while the chlorophyll content of the leaves treated by the plant drought-resistant agent is still kept at a higher level to 0.341m of the total chlorophyll content per gram, which is not different from that of the blank control group. At the same time, the photosynthetic rate of the wheat in-vitro leaf is measured, as shown in fig. 1D, compared with a control group, the photosynthetic rate of the wheat leaf is obviously reduced by drought treatment, and the photosynthetic rate of the leaf treated by the plant drought-resistant agent is still maintained at a higher level.
As shown in fig. 2A-B, leaf position (seventh leaf) of benshi tobacco leaf with consistent growth vigor is selected, perforated and sampled by a puncher with the diameter of 1.0cm, and the leaf position is placed in a culture dish regularly, the STRP 1 mature polypeptide plant drought-resistant agent is treated in a soaking manner, and the leaf position is placed under continuous illumination for 7 days.
The result shows that the STREP 1 mature polypeptide plant drought-resistant agent can obviously inhibit tobacco in-vitro leaf damage caused by drought after being treated. After 7 days, the tobacco leaves of the control group are almost green, and the tobacco leaves subjected to drought treatment are almost all yellow, and the yellow of the tobacco leaves caused by drought is obviously delayed after being treated by adding the STRP 1 mature polypeptide plant drought resistance agent. As shown in FIG. 2C, compared with the chlorophyll content of the control group with 0.493 mBar/bar, the chlorophyll content of the wheat leaves is sharply reduced by drought treatment to 0.160 mbar/bar, and the chlorophyll content of the leaves treated by the plant drought-resistant agent is still kept at a higher level to reach 0.420 mbar/bar. At the same time, the photosynthetic rate of the tobacco leaves in vitro is measured, as shown in fig. 2D, compared with a control group, the photosynthetic rate of the tobacco leaves is obviously reduced (reduced by 33.8%) by drought treatment, and the photosynthetic rate of the leaves treated by the plant drought-resistant agent still maintains a higher level and has almost no difference from the control group.
Treatment of living plants test
Taking example 1 as an example, the STREP 1 mature polypeptide plant drought-resistant agent prepared in example 1 is used for treating wheat living plants according to a using method.
As shown in fig. 3A, wheat with consistent germination is selected and sown in soil for long-day cultivation, water cut-off and drought treatment is carried out when two leaves and one core are adopted, and meanwhile, the STRP P1 mature polypeptide plant drought-resistant agent is adopted for spraying treatment. Culturing under continuous illumination for 4 days.
The growth vigor of wheat seedlings of the control group sprayed with the STRP 1 mature polypeptide plant drought-resistant agent is consistent with that of wheat seedlings of the control group not sprayed with the STRP 1 mature polypeptide plant drought-resistant agent, no obvious difference exists, and after the control group is treated, the wheat seedlings subjected to drought treatment seriously lose water and wilt, and most of the wheat seedlings still keep upright growth after being sprayed with the STRP 1 mature polypeptide plant drought-resistant agent. As shown in fig. 3B, the fresh weight of the aerial parts of wheat seedlings is measured, and the fresh weight of the aerial parts of wheat is not significantly different between the control group sprayed and the plant drought-resistant agent without the STRPEP1 mature polypeptide, which indicates that the plant drought-resistant agent with the STRPEP1 mature polypeptide has no adverse side effect on plants, and the biomass of the aerial parts of wheat is significantly increased (increased by 12.39%) after the plant drought-resistant agent is sprayed under drought conditions. As shown in fig. 3C, the chlorophyll measurement result shows that the chlorophyll content of the drought-resistant agent of the plant of the control group is not significantly different from that of the drought-resistant agent of the plant without STRPEP1, and the chlorophyll content of the wheat with the drought-resistant agent of the plant is significantly higher than that of the wheat without the drought-resistant agent of the plant under drought conditions (the improvement of 68.63%).
Claims (9)
1. The plant drought-resistant agent is characterized by taking STREP 1 polypeptide mature body as a main component; wherein, the amino acid sequence of the STREP 1 polypeptide mature body is: S-Q-V-V-Q-G-S-V-P-S-P-G-V-G-H; the plant is wheat or tobacco.
2. The plant drought-resistance agent according to claim 1, further comprising a silicone or tween-20.
3. The plant drought-resistance agent according to claim 1, characterized in that the concentration of the STRPEP1 mature polypeptide body is 0.5-1.0 μmol/L.
4. The plant drought-resistance agent according to claim 2, wherein the concentration of said organic silicon or tween-20 is 0.5-1.0mL/L.
5. The method for preparing a plant drought-resistant agent according to any one of claims 1 to 4, comprising the steps of:
adding STRP 1 mature polypeptide mother liquor 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.
6. The method of claim 5, wherein the STRPEP1 mature polypeptide mother liquor is formulated by:
the STRP 1 mature polypeptide powder is dissolved in distilled water to prepare STRP 1 mature polypeptide mother liquor with the concentration of 1-fold/L for standby.
7. The method of claim 6, wherein the STRPEP1 mature polypeptide mother liquor is stored at-70 ℃ when stored for long periods of time; when used for a short period, the product is stored at-20 ℃.
8. Use of a plant drought-resistance agent according to any one of claims 1-4 for increasing drought resistance of plants, characterized in that the plants are wheat or tobacco.
9. The use according to claim 8, wherein the drought-resistant agent is sprayed directly and uniformly on the surface of the plant aerial parts or cotton balls soaked with the drought-resistant agent are uniformly coated on the plant aerial parts.
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| CN113841700A (en) * | 2021-09-30 | 2021-12-28 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | SENPP 1-3 mature polypeptide plant senescence promoter, preparation method and application thereof |
| CN113907077A (en) * | 2021-09-30 | 2022-01-11 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | PSY1 mature polypeptide plant senescence inhibitor, preparation method and application thereof |
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