CN114176083A - Application of chlorogenic acid in improving plant salt stress resistance - Google Patents

Application of chlorogenic acid in improving plant salt stress resistance Download PDF

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CN114176083A
CN114176083A CN202111464074.7A CN202111464074A CN114176083A CN 114176083 A CN114176083 A CN 114176083A CN 202111464074 A CN202111464074 A CN 202111464074A CN 114176083 A CN114176083 A CN 114176083A
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chlorogenic acid
salt stress
tobacco seedlings
tobacco
plant
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CN114176083B (en
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陈帅
杨爱国
吴风燕
佟英
任民
张银超
武秀明
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Qingzhou Tobacco Research Institute of China National Tobacco Corp of Institute of Tobacco Research of CAAS
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Qingzhou Tobacco Research Institute of China National Tobacco Corp of Institute of Tobacco Research of CAAS
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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
    • A01N37/36—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 singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
    • A01N37/38—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 singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
    • A01B79/00—Methods for working soil
    • 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
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/002—Culture media for tissue culture
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00—Soil-conditioning materials or soil-stabilising materials
    • C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2109/00—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE pH regulation

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  • Organic Chemistry (AREA)
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  • Agronomy & Crop Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
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Abstract

The invention discloses application of chlorogenic acid in improving plant salt stress resistance, and belongs to the technical field of improving plant salt stress capability. The invention determines the effect of chlorogenic acid on increasing the salt stress tolerance of plants by measuring the growth condition of the root length of tobacco seedlings and the change of the levels of hydrogen peroxide and superoxide anion in the tobacco seedlings, finds that the chlorogenic acid can improve the growth rate of the tobacco seedlings in the salt stress process, reduce the levels of hydrogen peroxide and superoxide anion, relieve the toxic action of oxidative stress (hydrogen peroxide and superoxide anion) on plant cells under the salt stress, and enable the tobacco to normally grow under the high salt stress, thereby providing a new method for relieving the salt stress of plants in saline-alkali soil.

Description

Application of chlorogenic acid in improving plant salt stress resistance
Technical Field
The invention relates to the technical field of improving plant salt stress capability, in particular to application of chlorogenic acid in improving plant salt stress resistance.
Background
The salinization of soil affects the sustainable development of agriculture in China. Due to the conditions of improper climate and agricultural technical measures and the like, the secondary salinization harm of cultivated land is increased year by year. Most crops are sensitive to salt stress, and the salt stress becomes one of the important factors limiting the yield and quality of crops with the increasing salinization area of cultivated land. Salt stress can induce plant cells to generate excessive active oxygen, so that the oxidative damage of biomacromolecules is caused, and the normal metabolic process of the plant cells is interfered. The plant growth can be inhibited when the soil contains too much salt, and the harm of the soil salinization to the plants is reflected in that: na in cells+The excessive accumulation can break the ion balance in cells, inhibit the normal metabolism in plant cells, reduce the photosynthesis capability of plants, and lead the growth of the plants to lack nutrients and then be inhibited and even die finally; the salinity of the saline-alkali soil enables the osmotic pressure of the soil to be higher than that of a plant root system, so that the water absorption of the plant is difficult, and the plant is dehydrated and dried up; the salinization of the land can destroy the balance of generation and elimination of active oxygen in plants, cause the accumulation of a large amount of free radicals, generate membrane lipid peroxidation, destroy cell membranes and enable organic matters of cells to permeate outwards, thereby inactivating the cells.
Researches find that the polyphenol substances are important secondary metabolites in plants, have strong antioxidant capacity, and play an important role in resisting oxidative stress damage under the growth and development and abiotic stress of the plants as important antioxidants. Accumulation of flavonoids such as quercetin, anthocyanin and kaempferol can increase tolerance of plants to salt stress. In addition, moderate salt stress treatment can cause accumulation of polyphenol substances in plants, and improve the product quality of crops and medicinal plants. The above studies indicate that polyphenols play an important role in plant salt stress resistance, but the specific regulatory mechanisms are not clear.
Chlorogenic acid belongs to polyphenol substances, has strong oxidation resistance and very rich pharmacological activity, and has wide application in the fields of medicines, chemical industry, foods and the like. However, in the field of botany, no patent and related report that chlorogenic acid directly influences the stress resistance of plants are seen at present.
Disclosure of Invention
The invention aims to provide application of chlorogenic acid in improving salt stress resistance of plants so as to solve the problems in the prior art.
Adding NaCl with a certain concentration into an MS culture medium to be treated by salt stress, taking MS as control treatment, and observing the root length elongation condition of the tobacco seedlings on the MS culture medium subjected to salt stress treatment and not subjected to salt stress treatment. It was found that the growth of tobacco roots was significantly inhibited after high concentration salt stress (200mM) treatment, and the length was significantly shorter than the control treatment with MS medium. The content of chlorogenic acid in the tobacco seedlings after the salt stress treatment is obviously increased.
In order to achieve the purpose, the invention provides the following scheme: application of chlorogenic acid in improving plant salt stress resistance is provided.
Further, the chlorogenic acid is administered at a concentration of 100 μ M.
Further, the method of applying comprises: chlorogenic acid is applied to the young plants stressed by salt.
Further, the method of applying comprises: adding exogenous chlorogenic acid into plant culture environment or adding chlorogenic acid into fertilizer for use; the fertilizer comprises any one of organic fertilizer and biological fertilizer.
Further, the method of applying comprises: chlorogenic acid is prepared into a saline-alkali soil improver for use.
Chlorogenic acid can remarkably reduce the content of hydrogen peroxide and superoxide anion in plants, remarkably increase the growth of plant roots and improve the salt tolerance of plants.
The invention discloses the following technical effects:
according to the invention, through exogenous addition of chlorogenic acid, the chlorogenic acid is found to be capable of improving the growth rate of tobacco seedlings in a salt stress process, reducing the content of hydrogen peroxide and the level of superoxide anions, and relieving the toxic action of oxidative stress (hydrogen peroxide and superoxide anions) on tobacco cells under salt stress, so that the tobacco can normally grow under high salt stress, and the chlorogenic acid is proved to have the capability of remarkably improving the plant salt stress resistance, thereby providing a new method for relieving the salt stress of plants in saline-alkali soil.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a graph of the growth of tobacco seedlings according to different treatments of example 2 of the present invention, wherein a is the root length change and b is the tobacco seedling;
FIG. 2 is a graph of hydrogen peroxide levels in tobacco seedlings from different treatments of example 3 of the present invention;
FIG. 3 is a graph of superoxide anion levels in tobacco seedlings from different treatments of example 4 of the present invention.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The proper salt stress can effectively improve the content of chlorogenic acid in leaves of sea fennel, artichoke, honeysuckle, artichoke and the like, and further analysis shows that the transcription levels of chlorogenic acid synthesis pathway key enzyme genes HQT and PAL1, PAL2 and PAL3 are obviously improved under the salt stress. Chlorogenic acid is widely existed in plants, and the content of chlorogenic acid in tobacco reaches about 80% of the total polyphenol substances, so that tobacco is taken as a model plant.
Example 1
Root length determination experiment:
(1) and (3) after the tobacco seeds are sterilized in a clean bench (the method is the same as aseptic seedling culture), respectively dibbling the seeds in an MS culture medium, dibbling 20 seeds in each dish for culture, and culturing for 15d to obtain tobacco seedlings.
(2) Transferring tobacco seedlings obtained by culturing in the step (1) to square culture dishes of MS culture media respectively containing 0mM NaCl (blank control CK), 200mM NaCl and 200mM NaCl plus 100 mu M chlorogenic acid (CHA), transferring 10 seedlings to each dish, linearly arranging the seedlings at the upper end of the culture dish, repeating the steps for 3 times, vertically placing and culturing, observing and measuring the growth condition of seedling roots after 15 days, wherein the growth condition of the tobacco seedlings is shown in figure 1.
As can be seen from the figure 1, the growth of the tobacco seedlings is severely inhibited under the stress action of 200mM NaCl, when 100 mu M chlorogenic acid is added to the tobacco seedlings under the stress action of 200mM NaCl, the stress action of NaCl can be effectively resisted, the salt stress resistance of the tobacco seedlings is improved, the growth vigor of the tobacco seedlings is equivalent to that of the tobacco seedlings of a blank control group, and the chlorogenic acid has a remarkable effect on improving the salt stress capability of the tobacco seedlings.
Example 2
Measurement of hydrogen peroxide level by DAB staining method:
(1) preparation of plant tissue: taking tobacco seedlings cultured for 15 days in the step (2) in the embodiment 2 to obtain tobacco stems and leaves (pretreated plant tissues) of a blank control group, a 200mM NaCl group and a 200mM NaCl +100 mu M chlorogenic acid group;
(2) taking a proper amount of DAB dye solution (1 mg.mL)-1pH3.8), adjusting pH to 5.8 with NaOH, adding the pretreated plant tissue, and storing at 28 deg.C in dark for 8 hr. And then sucking off the dye solution, adding 80% ethanol by volume fraction, boiling the water bath for several minutes, sucking out the liquid, adding absolute ethanol, boiling the water bath until leaves are completely green, sucking out the liquid again, adding the absolute ethanol, storing the liquid in a refrigerator at 4 ℃ for a period of time, observing, and displaying the hydrogen peroxide levels in the tobacco seedlings subjected to different treatments in a graph 2.
Stress can cause plant cells to produce excessive reactive oxygen species, which can lead to oxidative damage of cells to interfere with normal metabolic processes of plant cells, and the like. As can be seen from FIG. 2, the tobacco seedlings which were not subjected to salt treatment (blank control) were colorless in staining, and the tobacco cultured in 200mM NaCl medium was darkened to a dark brown color, indicating that the hydrogen peroxide content of the tobacco seedlings was increased after salt treatment; the dyeing color becomes light obviously after the exogenous addition of the chlorogenic acid, which shows that the exogenous addition of the chlorogenic acid can obviously reduce the hydrogen peroxide content in the tobacco seedlings caused by salt stress treatment.
Example 3
Determination of superoxide anion levels by NBT staining:
(1) preparation of plant tissue: taking the tobacco seedlings cultured for 15 days in the step (2) in the embodiment 2, removing roots to obtain tobacco leaves of a blank control group, a 200mM NaCl group and a 200mM NaCl +100 mu M chlorogenic acid group;
(2) the leaves were placed on 12-well plates, and 2mL of NBT staining solution was added to immerse the leaves. Adding Na into the leaves2HPO4For comparison. And (3) putting the 12-hole plate in a vacuum environment, carrying out vacuum infiltration for 5min, taking out, covering an aluminum foil, oscillating on an oscillator for 2h, and pouring the NBT dyeing solution at the rotating speed of 80-100 rpm. Adding ethanol with the volume fraction of 95%, and carrying out water bath at 90-95 ℃ for 5 min; adding anhydrous ethanol (bleaching solution), and boiling in water bath until leaves are removed in green. When observing, a small amount of water is added to prevent rapid dehydration and wilting (the decolorized transparent leaves can also be stored in 50% glycerol for observation under a microscope), and the observation result is shown in FIG. 3.
As can be seen from FIG. 3, the tobacco seedlings which were not subjected to salt treatment (blank control) were colored in light blue, and the tobacco seedlings cultured in 200mM NaCl medium were colored more deeply and turned into dark blue, indicating that the superoxide anion content of the ordinary tobacco seedlings was increased after salt treatment. After exogenous addition of chlorogenic acid, the dyeing color becomes remarkably light, which shows that the exogenous addition of chlorogenic acid can remarkably reduce the content of superoxide anions in tobacco seedlings caused by salt stress treatment.
In conclusion, chlorogenic acid plays an important role in plant salt stress resistance. Under the condition of salt stress, the exogenous addition of chlorogenic acid can obviously reduce the content of hydrogen peroxide and superoxide anions in the plant, relieve oxidative stress toxicity, obviously increase the growth of plant roots and improve the salt tolerance of the plant.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.

Claims (5)

1. Application of chlorogenic acid in improving plant salt stress resistance is provided.
2. The use according to claim 1, wherein the chlorogenic acid is administered at a concentration of 100 μ M.
3. The application according to claim 1, wherein the method of applying comprises: chlorogenic acid is applied to the young plants stressed by salt.
4. The application according to claim 1, wherein the method of applying comprises: adding exogenous chlorogenic acid into plant culture environment or adding chlorogenic acid into fertilizer for use; the fertilizer comprises any one of organic fertilizer and biological fertilizer.
5. The application according to claim 1, wherein the method of applying comprises: chlorogenic acid is prepared into a saline-alkali soil improver for use.
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Cited By (2)

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Publication number Priority date Publication date Assignee Title
CN116735320A (en) * 2023-06-26 2023-09-12 中国水稻研究所 Determination method for activity of rice tillering buds and detection method for active oxygen level in tillering buds
CN117296595A (en) * 2023-10-25 2023-12-29 玉林师范学院 Application of 2,4-epibrassinolide in promoting growth of Yulan in salt stress and method for relieving Yulan salt stress toxicity

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116735320A (en) * 2023-06-26 2023-09-12 中国水稻研究所 Determination method for activity of rice tillering buds and detection method for active oxygen level in tillering buds
CN117296595A (en) * 2023-10-25 2023-12-29 玉林师范学院 Application of 2,4-epibrassinolide in promoting growth of Yulan in salt stress and method for relieving Yulan salt stress toxicity

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