CN111436437A - Reagent for antagonizing physiological action of n-butyl alcohol - Google Patents
Reagent for antagonizing physiological action of n-butyl alcohol Download PDFInfo
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- CN111436437A CN111436437A CN202010332541.XA CN202010332541A CN111436437A CN 111436437 A CN111436437 A CN 111436437A CN 202010332541 A CN202010332541 A CN 202010332541A CN 111436437 A CN111436437 A CN 111436437A
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
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- 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
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
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- A—HUMAN NECESSITIES
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- A01N45/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds having three or more carbocyclic rings condensed among themselves, at least one ring not being a six-membered ring
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Abstract
The invention provides a reagent for antagonizing the physiological action of n-butanol, and relates to the technical field of hormone function research. In the embodiment of the invention, 10 mu M Fluridone can effectively antagonize the effect of n-butanol on inhibiting seed germination; gibberellin can improve the seed germination rate and the cotyledon extension rate under the condition of n-butanol inhibition to different degrees, and the higher the concentration is, the more obvious the effect is, and the antagonistic effect of 30 mu M gibberellin is optimal; high concentrations of GA (30. mu.M) restore to a large extent the dividing capacity of root tip stem cells and eventually partially the main root growth.
Description
Technical Field
The invention belongs to the technical field of hormone function research, and particularly relates to a reagent for antagonizing physiological effects of n-butanol.
Background
Gibberellin (Gibberellins, GAs), diterpene phytohormones, mainly promote vegetative growth of plants, including elongation of internodes, elongation of roots and expansion of leaves, gibberellin signals are transferred to pathways, DE LL A protein serving as a plant growth inhibitor can be counteracted by active gibberellin, the gibberellin signals start degradation of DE LL A protein, expression of gibberellin response genes is regulated, and therefore growth of the plants is promoted, and DE LL A protein plays a role in negative feedback regulation on growth and development of the plants.
Transphosphatidylation refers to the reaction of phosphodiester bonds of glycerophospholipids to break and transfer the phosphatidyl group of phospholipids to primary alcohols or water under the catalytic action of Phospholipase D (P L D), thereby producing phosphatidylols or phosphatidic acids, which was first discovered in the early 60 th century by Ferrari and Sastry et al when studying the biosynthesis and metabolism of plant phospholipids, Yang et al (1967) demonstrated that the enzymes catalyzing this reaction are Phospholipase D (phophospase D, P L D), while glycerophospholipids and hydroxyl-containing compounds, respectively, as donor and acceptor for this reaction, when water molecules are used as substrate, the products are phosphatidic acids (phophatidicacid, PA) and organic bases, this reaction is referred to as hydrolysis of P L D, when reacting with other hydroxyl-containing compounds, forming new phospholipids and new hydroxyl compounds, this reaction is referred to as phosphatidic acid (phophatidicacid, PA) and organic bases, this reaction is known as a reaction, when the reaction with water molecules is carried out as substrate, the general, the hydrolysis of n-butanol, the n-butanol, and n-butanol, which are involved in the general processes that the reactions of phosphosphingomythophatidicacid reactions, the physiological processes, the substrate and n-butanol, the alcohol-butanol, which have been found to affect the development of plants.
Disclosure of Invention
In view of the above, the present invention aims to provide a reagent for gibberellin to antagonize the physiological action of n-butanol, which can antagonize the physiological action of n-butanol in plants, animals and microorganisms, and provide a new theoretical basis for studying signal transduction of gibberellin and abscisic acid.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a reagent for antagonizing the physiological action of n-butanol, and the active component of the reagent comprises an abscisic acid synthesis inhibitor or gibberellin.
Preferably, the physiological effects of n-butanol include inhibition of seed germination, inhibition of root growth, and induction of accumulation of an inhibitor of DE LL a protein GA 1-3.
Preferably, the abscisic acid synthesis inhibitor comprises fluazinone.
Preferably, the working concentration of the abscisic acid synthesis inhibitor is 1-10 mu mol/L.
Preferably, the working concentration of the gibberellin is 10-100 mu mol/L.
Preferably, the working concentration of gibberellin is 30. mu. mol/L.
The invention also provides a kit for antagonizing the physiological action of n-butanol, which comprises the reagent.
In the embodiment of the invention, 1 mu M, 10 mu M and 20 mu MABA synthetic inhibitor Fluridone (Fluridone) are simultaneously added into a culture medium containing 0.1% of n-butanol to improve the germination rate and the cotyledon elongation rate of seeds inhibited by n-butanol to a certain extent, and the germination rate and the cotyledon elongation rate of the arabidopsis seeds are both not obviously different by utilizing 1 mu M and 10 mu M Fluridone, when 10 mu M, 30 mu M, 50 mu M and 100 mu M gibberellin are simultaneously added into the culture medium containing 0.1% of n-butanol to improve the germination rate and the cotyledon elongation rate under the inhibition of n-butanol to a different extent, the effect difference between 50 mu M and 100 mu M is not obvious when the concentration is higher, the antagonistic effect of 30 mu M gibberellin is optimal when the concentration is not higher, the n-butanol can expand cells, the root stem apex cell division cell is obviously inhibited, the number of the root apex cell is obviously reduced, the root apex cell division region is not obviously inhibited, the root tuber growth inhibitor is finally recovered by adding other RGA (RGGA) and the root tuber growth inhibitor LL A, the accumulation of the RGA is induced, the root apex protein (RGA) is not greatly reduced, and the accumulation of the butanol accumulation of the root apex protein (RGA) is induced, the RGA, the accumulation of the RGGA 3, and the RGGA (RGGA 3 is induced by the butanol concentration of the RGA, the RGGA 3.
Drawings
FIG. 1 is a comparison of ABA synthesis inhibitors and n-butanol on germination of Arabidopsis seeds;
FIG. 2 is a graph showing the influence of ABA synthesis inhibitor and n-butanol on the germination rate of Arabidopsis seeds;
FIG. 3 is a graph showing the influence of ABA synthesis inhibitors and n-butanol on the cotyledon formation rate of Arabidopsis seeds;
FIG. 4 is a comparison of gibberellin and n-butanol for germination of Arabidopsis thaliana seeds;
FIG. 5 is a graph showing the effect of gibberellin and n-butanol on the germination rate of Arabidopsis thaliana seeds;
FIG. 6 is a graph showing the effect of gibberellin and n-butanol on the cotyledon formation rate of Arabidopsis seeds;
FIG. 7 is a graph showing the effect of gibberellin and n-butanol on root length of Arabidopsis seedlings;
FIG. 8 is a graph comparing the effect of gibberellin and n-butanol on Arabidopsis thaliana root cells;
FIG. 9 is a graph showing the effect of gibberellin and n-butanol on the number of cells in the meristematic region of the root tip of Arabidopsis thaliana;
FIG. 10 is a graph showing the accumulation of RGA in DE LL A protein induced by n-butanol;
FIG. 11 is a graph of the effect of different butanol isomers on Arabidopsis germination and seedling growth, wherein: MS; b. n-butanol; c. sec-butyl alcohol; d. tert-butyl alcohol; e. isobutanol; ms +30 μ M GA; g. n-butanol +30 μ M GA; h. sec-butanol +30 μ MGA; i. tert-butanol +30 μ M GA; j. isobutanol +30 μ M GA;
FIG. 12 is a graph of the cumulative effect of different butanol isomers on RGA, a member of the DE LL A protein family.
Detailed Description
The invention provides a reagent for antagonizing the physiological action of n-butanol, and the active component of the reagent comprises an abscisic acid synthesis inhibitor or gibberellin.
The physiological effects of n-butanol according to the present invention preferably include physiological effects that can be produced against plants, animals or microorganisms, and more preferably include inhibition of seed germination, inhibition of root growth and induction of accumulation of an inhibitor (RGA) of DE LL A protein GA 1-3.
The abscisic acid synthesis inhibitor preferably comprises fluazinone (Fluridone), and the working concentration of the abscisic acid synthesis inhibitor is preferably 1-10 mu mol/L, more preferably 10 mu mol/L.
The working concentration of the gibberellin is preferably 10-100 mu mol/L, and more preferably 30 mu mol/L.
The invention also provides a kit for antagonizing the physiological action of n-butanol, which comprises the reagent.
The agents antagonistic to the physiological action of n-butanol provided by the present invention will be described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
ABA synthetic inhibitor antagonizes n-butanol to inhibit germination of arabidopsis thaliana seeds
N-butanol inhibits germination of arabidopsis seeds and induces cotyledon modification. The seed germination rate and cotyledon extension rate of n-butanol inhibition can be improved to a certain extent by adding 1 mu M, 10 mu M and 20 mu MABA synthesis inhibitor Fluridone into a culture medium containing 0.1% of n-butanol. When arabidopsis thaliana seeds were grown on MS medium and medium containing 0.1% n-butanol, 0.1% n-butanol and Fluridone under illumination for 3 days, the effect on germination was as shown in fig. 1 and 2, the germination rate of arabidopsis thaliana seeds in MS culture was about 95%, the germination rate of arabidopsis thaliana seeds treated with 0.1% n-butanol was about 25%, the germination rate of arabidopsis thaliana seeds treated with 0.1% n-butanol and 10 μ M Fluridone was about 80%, and the germination rates of arabidopsis thaliana seeds treated with both 0 μ M and 10 μ M MFluridone were not significantly improved. The result shows that 10 mu M Fluridone can effectively antagonize n-butanol to inhibit seed germination, and no significant effect exists when the concentration is too high or too low.
The effect on cotyledon occurrence of arabidopsis thaliana seeds grown for 7 days on MS medium and on medium containing 0.1% n-butanol, 0.1% n-butanol and Fluridone under light irradiation is shown in fig. 3, where the cotyledon protrusion rate of arabidopsis thaliana in MS culture was about 98%, while the cotyledon protrusion rate of arabidopsis thaliana treated with 0.1% n-butanol was about 0%, the arabidopsis thaliana protrusion rate of arabidopsis thaliana treated with 0.1% n-butanol and 10 μ M Fluridone simultaneously was about 60%, and the arabidopsis thaliana protrusion rate of arabidopsis thaliana treated with 0.1% n-butanol and 20 μ M Fluridone simultaneously was about 27%, whereas the cotyledon protrusion rates of arabidopsis thaliana seeds were not significantly improved by adding 1 μ M and 5 μ M flurido. The 10 mu M Fluridone can effectively antagonize the inhibition effect of n-butanol on the growth of the germinated seeds, but the effect is reduced when the concentration is too high, and no significant effect is generated when the concentration is too low.
Example 2
High-concentration gibberellin antagonism n-butanol inhibition arabidopsis seed germination aspect effect
The seed germination rate and cotyledon extension rate under the n-butanol inhibition can be improved to different degrees by adding 10 mu M, 30 mu M, 50 mu M and 100 mu M gibberellin into the culture medium containing 0.1 percent of n-butanol. As a result of growing Arabidopsis seeds on MS medium and medium containing 0.1% n-butanol, 0.1% n-butanol and gibberellin at various concentrations under light for 3 days, as shown in FIGS. 4 and 5, the germination rate of Arabidopsis seeds in MS culture was about 95%, while the germination rate of Arabidopsis seeds treated with the addition of 0.1% n-butanol was about 12%, the germination rate of Arabidopsis seeds treated with the addition of 0.1% n-butanol and 10. mu.M gibberellin medium was about 26%, the germination rate of Arabidopsis seeds treated with the addition of 0.1% n-butanol and 30. mu.M gibberellin medium was about 40%, the germination rate of Arabidopsis seeds treated with the addition of 0.1% n-butanol and 50. mu.M gibberellin medium was about 60%, and the germination rate of Arabidopsis seeds treated with the addition of 0.1% n-butanol and 100. mu.M gibberellin medium was about 60%.
When Arabidopsis seeds were grown for 7 days under the light irradiation of MS medium and media containing 0.1% n-butanol, 0.1% n-butanol and gibberellin at various concentrations, the results of counting the cotyledon protrusion rate were found as shown in FIG. 6, the Arabidopsis seed protrusion rate in MS culture was about 100%, the seed germination rate was almost 0% under the treatment of adding 0.1% n-butanol, the Arabidopsis seed protrusion rate was about 25% under the treatment of adding 0.1% n-butanol and 10. mu.M gibberellin medium simultaneously, the Arabidopsis seed protrusion rate was about 27% under the treatment of adding 0.1% n-butanol and 20. mu.M gibberellin medium simultaneously, the Arabidopsis seed protrusion rate was about 80% under the treatment of adding 0.1% n-butanol and 30. mu.M gibberellin medium simultaneously, and the Arabidopsis seed protrusion rate was 70% under the treatment of adding 0.1% n-butanol and 50. mu.M or 100. mu.M gibberellin simultaneously. The gibberellin can effectively antagonize n-butanol to inhibit seed germination, the higher the concentration is, the more obvious the effect is, the effect difference between 50 mu M and 100 mu M is not obvious, and the antagonistic effect of 30 mu M gibberellin is optimal.
Example 3
Gibberellin antagonizing n-butanol to inhibit growth of main root of arabidopsis thaliana
Arabidopsis seeds were cultured in MS medium for 3 days, and then the seedlings were transferred to MS normal medium, medium containing 30. mu.M gibberellin, medium containing 0.2% n-butanol and 30. mu.M gibberellin, medium containing 0.4% n-butanol and 30. mu.M gibberellin, respectively, and the cultivation was continued vertically for 5 days under light. Determination of Primary root Length As shown in FIG. 7, the primary root length of seedlings grown on MS and a culture containing 30. mu.M gibberellin was about 35mm, the primary root length of seedlings grown on a medium containing 0.2% n-butanol and a medium containing 0.2% n-butanol plus 30. mu.M gibberellin was 22mm and 31mm, respectively, and the primary root length of seedlings grown on a medium containing 0.2% n-butanol and a medium containing 0.2% n-butanol plus 30. mu.M gibberellin was 4mm and 29mm, respectively. The results show that 30 mu M gibberellin can remarkably relieve the inhibition effect of n-butyl alcohol on the elongation of the main roots of seedlings.
Through observation of the number of the root tip stem cells, the static center of the root tip (figure 8) and the cell number of the root tip meristematic region (figure 9), the n-butanol can obviously inhibit the division of the root tip stem cells, the deformity of the static center cells and the reduction of the cell number of the root tip meristematic region. The number of cells of the primary root meristematic region of seedlings grown on MS and on a culture containing 30. mu.M gibberellin was 27, the number of cells of the primary root tip of seedlings grown on a medium containing 0.2% n-butanol and a medium containing 0.2% n-butanol plus 30. mu.M gibberellin was 13 and 27, respectively, and the number of cells of the primary root tip of seedlings grown on a medium containing 0.4% n-butanol and a medium containing 0.4% n-butanol plus 30. mu.M gibberellin was 3 and 23, respectively. The results show that the n-butyl alcohol can expand root system cells, remarkably inhibit division of root tip stem cells, remarkably reduce the number of cells in a root tip meristematic region and finally cause the growth of a main root to be inhibited. However, high concentrations of GA can restore the dividing capacity of root tip stem cells to a large extent and eventually partially restore primary root growth.
Example 4
Application of n-butanol to induce accumulation of DE LL A protein RGA
Arabidopsis seedlings germinating for two days on a normal MS culture medium are respectively transferred to a culture medium of MS, MS +50 mu M GA, MS + 0.2% n-butanol and MS + 0.2% n-butanol +50 mu M GA, and are cultured in an acclimatization room with the relative humidity of 60% at 22 ℃ and the photoperiod of 12h, the accumulation condition of the fusion protein is observed and processed by a laser confocal microscope for three days, the RGA-GFP overexpression Arabidopsis root tip is used for expressing the accumulation condition of the fusion protein, the result is shown in figure 10, and the accumulation of the RGA of a DE LL A protein family member is detected by confocal detection after the normal MS culture medium is processed by the n-butanol.
Example 5
The other isomers of n-butanol did not affect plant growth nor induce the accumulation of RGA, a member of the DE LL A protein family
Seeds of Arabidopsis thaliana wild type Arabidopsis thaliana were sown on a medium of MS, MS + 30. mu.M GA, MS + 0.1% n-butanol (1-butanol) and MS + 0.1% n-butanol + 30. mu.M GA, MS + 0.1% sec-butanol (sec-butanol) and MS + 0.1% sec-butanol + 30. mu.M GA, MS + 0.1% tert-butanol (tert-butanol) and MS + 0.1% tert-butanol + 30. mu.M GA, MS + 0.1% isobutanol (iso-butanol) and MS + 0.1% isobutanol + 30. mu.M GA, and cultured in a acclimation room at 22 ℃ with a relative humidity of 60% and a photoperiod of 12 hours for 7 days. As shown in FIG. 11, the isomers of n-butanol, sec-butanol, tert-butanol, isobutanol (iso-butanol) did not inhibit seed germination and post-germination growth, while the addition of gibberellin did not significantly affect the growth thereof.
Seeds of an RGA-GFP mutant strain were sown in a normal MS medium for two days and the seedlings were transferred to a medium of MS, MS + 30. mu.M GA, MS + 0.1% n-butanol and MS + 0.1% n-butanol + 30. mu.M GA, MS + 0.1% sec-butanol and MS + 0.1% sec-butanol + 30. mu.M GA, MS + 0.1% tert-butanol and MS + 0.1% tert-butanol + 30. mu.M GA, MS + 0.1% iso-butanol and MS + 0.1% iso-butanol + 30. mu.M GA, respectively, and incubated at 22 ℃ under a relative humidity of 60% and a light period of 12 hours, and the accumulation of the fusion protein was observed in an acclimatization room treated with a confocal laser microscope for three days in which the root tip of RGA-GFP overexpressing Arabidopsis thaliana was treated, and the results were shown in FIG. 12, and sec-butanol, did not induce the accumulation of RGA of the family RGA protein family RGA of DE LL A.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (7)
1. An agent for antagonizing the physiological action of n-butanol, wherein the active ingredient of said agent comprises an abscisic acid synthesis inhibitor or gibberellin.
2. The reagent of claim 1, wherein the physiological effects of n-butanol include inhibition of seed germination, inhibition of root growth, and induction of accumulation of an inhibitor of the DE LL a protein GA 1-3.
3. The reagent of claim 1, wherein the inhibitor of abscisic acid synthesis comprises fluazinone.
4. The reagent according to claim 1 or 3, wherein the abscisic acid synthesis inhibitor has a working concentration of 1-10 μmol/L.
5. The reagent of claim 1, wherein the working concentration of gibberellin is 10-100 μmol/L.
6. The reagent of claim 5, wherein the working concentration of gibberellin is 30 μmol/L.
7. A kit for antagonizing the physiological effects of n-butanol, comprising the agent of any one of claims 1 to 6.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113243372A (en) * | 2021-04-15 | 2021-08-13 | 湖南农业大学 | Application of fluazinone in promotion of rice shoot differentiation and ratoon rice planting |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107810958A (en) * | 2017-10-19 | 2018-03-20 | 中国农业大学 | A kind of method for promoting Yunnan Paris polyphylla seed to sprout |
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| DANICA E GOGGIN等: "Fluridone: a combination germination stimulant and herbicide for problem fields?", 《PEST MANAG SCI》 * |
| JOHN GARDINER等: "The Effects of the Phospholipase D-Antagonist 1-Butanol on Seedling Development and Microtubule Organisation in Arabidopsis", 《PLANT CELL PHYSIOL.》 * |
| 王华磊等: "氟啶酮对管花肉苁蓉种子萌发影响的研究", 《中国中药杂志》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113243372A (en) * | 2021-04-15 | 2021-08-13 | 湖南农业大学 | Application of fluazinone in promotion of rice shoot differentiation and ratoon rice planting |
| CN113243372B (en) * | 2021-04-15 | 2022-07-15 | 湖南农业大学 | Application of fluazinone in promotion of rice shoot differentiation and ratoon rice planting |
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