CN114176083B - Application of chlorogenic acid in improving salt stress resistance of plants - Google Patents
Application of chlorogenic acid in improving salt stress resistance of plants Download PDFInfo
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- CN114176083B CN114176083B CN202111464074.7A CN202111464074A CN114176083B CN 114176083 B CN114176083 B CN 114176083B CN 202111464074 A CN202111464074 A CN 202111464074A CN 114176083 B CN114176083 B CN 114176083B
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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
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- 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
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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
- 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
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- 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
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- 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
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 detects the effect of chlorogenic acid in increasing plant salt stress tolerance by measuring the growth condition of tobacco seedling root length and the change of hydrogen peroxide and superoxide anion level in tobacco seedlings, and discovers that chlorogenic acid can improve the tobacco seedling growth rate in the salt stress process, reduce the levels of hydrogen peroxide and superoxide anion, and relieve the toxic effect of oxidative stress (hydrogen peroxide and superoxide anion) on plant cells under salt stress, so that tobacco can normally grow under high salt stress, thereby providing a novel method for relieving the salt stress of saline-alkali soil plants.
Description
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
Soil salinization affects the sustainable development of agriculture in China. The secondary salinization hazard of cultivated lands caused by improper use of climate and agricultural technical measures is aggravated year by year. Most crops are relatively sensitive to salt stress, which has become one of the important factors limiting crop yield and quality as the salinized area of cultivated lands increases. Salt stress can induce plant cells to produce excessive active oxygen, thereby causing oxidative damage of biological macromolecules and interfering with the normal metabolic process of the plant cells. Excessive soil salinization can inhibit plant growth, and the damage caused by soil salinization to plants is shown in the following steps: na in cells + The excessive accumulation of (2) can break the ion balance in the cells, inhibit the normal metabolism in the cells of the plant, reduce the photosynthesis capacity of the plant, and the plant growth can lack nutrients and be inhibited or even finally dead; the salt in the saline-alkali soil makes the osmotic pressure of the soil higher than that of the plant root system, so that the plant is difficult to absorb water, and the plant is dehydrated and dried; land salinization can destroy the balance of active oxygen generation and scavenging in plants, cause free radical accumulation in large quantity, generate membrane lipid peroxidation, destroy cell membranes and extravasate cell organic matters, thereby inactivating cells.
The research shows that the polyphenols are important secondary metabolic substances in plants, have strong antioxidant capacity, and play an important role in resisting oxidative stress damage under the growth and development of plants and abiotic stress 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 yet clear.
Chlorogenic acid belongs to polyphenols, has strong antioxidant capacity and very rich pharmacological activity, and has wide application in the fields of medicine, chemical industry, food and the like. However, in the field of botanic technology, patents and related reports of chlorogenic acid directly affecting plant stress resistance are not 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 of the prior art.
NaCl with a certain concentration is added into an MS culture medium to serve as salt stress treatment, MS serves as control treatment, and root length elongation of tobacco seedlings on the MS culture medium subjected to salt stress treatment and not subjected to salt stress treatment is observed. It was found that tobacco root growth was significantly inhibited following high concentration salt stress (200 mM) treatment, with a significantly shorter length than control treatment of MS medium. Chlorogenic acid content of tobacco seedlings after salt stress treatment is obviously increased.
In order to achieve the above object, the present invention provides the following solutions: use of chlorogenic acid for increasing salt stress resistance of plants.
Further, the chlorogenic acid is administered at a concentration of 100 μm.
Further, the method of application includes: chlorogenic acid is applied to young plants that are subject to salt stress.
Further, the method of application includes: adding chlorogenic acid exogenously into a plant culture environment or adding chlorogenic acid exogenously into a fertilizer for use; the fertilizer comprises any one of an organic fertilizer and a biological fertilizer.
Further, the method of application includes: chlorogenic acid is prepared into a saline-alkali soil modifier for use.
Chlorogenic acid can obviously reduce the content of hydrogen peroxide and superoxide anions in plants, obviously increase the growth of plant root systems 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 can improve the growth rate of tobacco seedlings in the salt stress process, reduce the content of hydrogen peroxide and the level of superoxide anions, and relieve the toxic effect of oxidative stress (hydrogen peroxide and superoxide anions) on tobacco cells under salt stress, so that tobacco can grow normally under high salt stress, and the chlorogenic acid has the capability of obviously improving the resistance of plants to salt stress, so that a novel method is provided for relieving the salt stress of saline-alkali soil plants.
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 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 other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the growth of tobacco seedlings treated differently in example 2 of the present invention, a is root length variation, b is tobacco seedlings;
FIG. 2 is a graph showing hydrogen peroxide levels in various treated tobacco seedlings according to example 3 of the present invention;
FIG. 3 is a graph showing the levels of superoxide anions in various treated tobacco seedlings according to example 4 of the present invention.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions 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. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, 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 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 invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
The proper salt stress can effectively improve the content of chlorogenic acid in the leaves of sea fennel, artichoke, honeysuckle, artichoke and the like, and further analysis shows that the transcription level of key enzyme genes HQT and PAL1, PAL2 and PAL3 of chlorogenic acid synthesis paths is obviously improved under the salt stress. Chlorogenic acid is widely present in plants, and the content in tobacco is more about 80% of the total amount of polyphenol substances, so tobacco is used as model plant.
Example 1
Root length measurement experiment:
(1) After tobacco seeds are sterilized in an ultra-clean workbench (the method is the same as aseptic seedling culture), the seeds are respectively planted in an MS culture medium, 20 seeds are planted in each dish for culture, and tobacco seedlings are obtained after 15 days of culture.
(2) The tobacco seedlings obtained by culturing in the step (1) were transferred to square dishes containing MS medium of 0mM NaCl (blank CK), 200mM NaCl, 200mM NaCl+100. Mu.M chlorogenic acid (CHA), 10 seedlings were transferred to each dish, and the seedlings were placed in line at the upper end of the dish, and were repeated 3 times, and were vertically placed for culturing, and after 15 days, the growth of the roots of the seedlings was observed and measured, and the growth of the tobacco seedlings was shown in FIG. 1.
As can be seen from the graph 1, the tobacco seedlings are severely inhibited from growing under the stress action of 200mM NaCl, and when the tobacco seedlings are added with 100 mu M chlorogenic acid while being stressed by 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 in a blank control group, and the chlorogenic acid has a remarkable effect of improving the salt stress capability of the tobacco seedlings.
Example 2
DAB staining method to determine hydrogen peroxide levels:
(1) Preparation of plant tissue: taking tobacco seedlings after 15d cultivation in the step (2) of the example 2, and obtaining a blank control group, a 200mM NaCl group, a 200mM NaCl+100 mu M chlorogenic acid group tobacco stem leaves (pretreated plant tissues);
(2) A suitable amount of DAB dye solution (1 mg. ML) was taken -1 pH 3.8), adjusting pH to 5.8 with NaOH, adding pretreated plant tissue, and storing at 28deg.C in dark place for 8 hr. Then the dye liquor is sucked off, 80% ethanol is added, the liquor is sucked off in a boiling water bath for a few minutes, absolute ethanol is added and the boiling water bath is added until the green leaves are completely removed, the liquor is sucked off again, the absolute ethanol is added, the tobacco seedlings are stored in a refrigerator at the temperature of 4 ℃ for a period of time and then are observed, and the hydrogen peroxide levels in the tobacco seedlings are treated differently, wherein the hydrogen peroxide levels are shown in figure 2.
Adversity stress can cause plant cells to produce excessive active oxygen, so that cell oxidative damage interferes with normal metabolic processes of the plant cells and the like. As can be seen from FIG. 2, the staining result of the tobacco seedlings which were not subjected to salt treatment (blank control) was colorless, and the tobacco cultured in the 200mM NaCl medium was deeply stained to become dark brown, indicating that the hydrogen peroxide content of the tobacco seedlings was increased after salt treatment; after the chlorogenic acid is externally added, the dyeing color is obviously lightened, which indicates that the content of hydrogen peroxide in tobacco seedlings caused by salt stress treatment can be obviously reduced by the externally added chlorogenic acid.
Example 3
NBT staining assay for superoxide anion levels:
(1) Preparation of plant tissue: taking tobacco seedlings after 15d cultivation in the step (2) of the example 2, and 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, respectively, and 2mL of NBT staining solution was added to submerge the leaves. In addition, na is added into the blade 2 HPO 4 As a control. Putting the 12-pore plate into a vacuum environment, vacuum infiltration for 5min, taking out, covering an aluminum foil, vibrating for 2h on a vibrator, and pouring out NBT dyeing liquid at a rotating speed of 80-100 rpm. Adding 95% ethanol by volume fraction, and carrying out water bath at 90-95 ℃ for 5min; adding absolute ethyl alcohol (bleaching liquid) and then boiling water bath until the leaves are green and removed. A small amount of water was added during observation to prevent rapid dehydration and wilting (decolored and transparent leaves can also be stored in 50% glycerol for observation under a microscope), and the observation results are shown in FIG. 3.
As can be seen from FIG. 3, the staining result of the tobacco seedlings which were not subjected to the salt treatment (blank control) was light blue, and the staining of the tobacco seedlings which were cultured in the 200mM NaCl medium was deepened to dark blue, indicating that the superoxide anion content of the ordinary tobacco seedlings was increased after the salt treatment. After the chlorogenic acid is externally added, the dyeing color is obviously lightened, which indicates that the content of superoxide anions in tobacco seedlings caused by salt stress treatment can be obviously reduced by the externally added chlorogenic acid.
In summary, 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 plants, relieve oxidative stress toxicity, obviously increase the growth of plant root systems and improve the salt tolerance of the plants.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (5)
1. Use of chlorogenic acid for reducing hydrogen peroxide content and superoxide anion level in tobacco.
2. The use according to claim 1, wherein chlorogenic acid is administered at a concentration of 100 μm.
3. The application according to claim 1, wherein the method of application comprises: chlorogenic acid is applied to tobacco seedlings that are subjected to salt stress.
4. The application according to claim 1, wherein the method of application comprises: adding chlorogenic acid exogenously into tobacco culture environment or adding chlorogenic acid into fertilizer; the fertilizer comprises any one of an organic fertilizer and a biological fertilizer.
5. The application according to claim 1, wherein the method of application comprises: chlorogenic acid is prepared into a saline-alkali soil modifier for use.
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| 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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| CN106187577A (en) * | 2016-06-30 | 2016-12-07 | 山东胜伟园林科技有限公司 | A kind of anti-salt alkali composition containing Fructus Arctii and preparation method thereof |
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| A peroxidase/phenolics/ascorbate system can scavenge hydrogen peroxide in plant cells;Umeo Takahama and Takayuki Oniki;PHYSIOLOGIA PLANTARUM;第101卷;845-852 * |
| 肉桂酸及其衍生物对活性氧H2O2清除作用;赵春贵 等;化学研究与应用;第16卷(第5期);685-687 * |
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