KR101993612B1 - Composition for promoting germination and enhancing salt stress tolerance of plant comprising polymer of naphthalene or phenolic material as effective component and uses thereof - Google Patents

Abstract

The present invention relates to a composition containing naphthalene or a polymer of a phenolic substance as an active ingredient for promoting germination and enhancing resistance to salt stress, and a use thereof. The polymer of the present invention is developed as artificial humic acid, It is expected to be replaced.

Description

TECHNICAL FIELD The present invention relates to a composition for promoting germination and resistance to salt stress in a plant containing a polymer of naphthalene or a phenolic substance as an active ingredient and a use thereof for a stress tolerance of a plant, uses thereof}

The present invention relates to a composition for promoting germination and for enhancing resistance to salt stress of a plant containing a polymer of naphthalene or a phenolic substance as an active ingredient, and a use thereof.

A natural humic substance is a complex substance in the form of a colloidal substance which is decomposed by a variety of microorganisms to destroy the organic matter applied to the soil and has no altered original structure or newly synthesized brown or dark brown color. It is found in sediments, coal, peat, etc. and includes polymeric phenol materials that can chelate metals and minerals. In addition, natural humic substances play an important role in linking inorganic and organic compounds in a natural environment, and it is possible to add aromatic, hydroxyl group, carbonyl group, carboxy group, it is presumed that these compounds have various properties and structures because they include carboxyl group, amine group and sulfhydryl group. Natural humic substances can be classified into fulvic acid and humic acid depending on their solubility in acid and alkali solutions. Fulvic acid dissolves in acid solution, but humic acid does not dissolve in acid solution.

Humic acid can be obtained from sediments that have undergone the processes of fermentation and mass transfer by microorganisms, enzymes, fungi, or protozoa for tens of millions of years after plant residues have been deposited. Humic acid acts as a source and a repository for nutrients by converting insoluble minerals into a receptive form that plants can absorb and slowly dissociating reversibly bound minerals. Humic acid can also improve plant nutrition by promoting plant growth and seed germination, increasing the vitamin content of plants, and promoting absorption of inorganic elements through the leaves.

Humic acid is generally in the form of a humic acid salt, distributed in less than 2% of the soil and 20 to 80% or more of the combustible minerals.

Until now, humic acid salts have been extracted from flammable minerals such as peat soil or leaf mold distributed in lignite, peat or swamp or forest which have been buried in the ground for a long time as an alkali solution And purified. However, in the conventional extraction method of humic acid salt from combustible minerals, an excess alkali solution is contained in the solution of the extracted humic acid salt, or the yield is very low, which is inefficient. Also, there is a problem in that the quality and quantity of humic acid extracted according to the organic material as a raw material are significantly different. Therefore, it is required to develop a candidate substance capable of high-quality mass production that can replace natural humic acid.

Korean Patent No. 1041345 discloses a method for culturing bean sprouts containing polyphenol components and Chinese Patent Publication No. 105638470 discloses a culture composition for germination of ginseng seeds, There is no description of a composition for promoting germination and for enhancing resistance to salt stress of a plant containing a polymer of naphthalene or a phenolic substance as an effective ingredient and its use.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and the present inventors have found that a homopolymer of 2,7-dihydroxynaphthalene; Copolymers of catechol, vanillylic acid and 3-pentadecyl phenol; And copolymers of catechol, vanillyric acid and L-3,4-dihydroxyphenylalanine were prepared and the polymers were treated with the plants. As a result, the polymers of the present invention showed not only the seed germination improving effect of plants, The present inventors have completed the present invention.

In order to solve the above problems, the present invention provides a composition for promoting germination and for enhancing resistance to salt stress of a plant containing a polymer of naphthalene or a phenolic substance as an effective ingredient.

The present invention also provides a method for promoting germination and salt stress tolerance of a plant, comprising the step of treating the composition with a plant, a seed of a plant, or a plant cultivated soil.

The polymer of the naphthalene or phenolic material of the present invention exhibits a seed germination promoting effect similar to that of humic acid. Therefore, it is developed as artificial humic acid and can be effectively used as an effective ingredient for growth regulators, fertilizers and the like. In addition, the polymer of the naphthalene or phenolic substance of the present invention can enhance the salt stress tolerance of the plant, and thus it is expected that the polymer can be usefully used for plants that are grown in high salt and coastal areas.

Fig. 1 shows the result of treating the polymer material (DHN, CV-3P-P, CV-L-DOPA) of the present invention with Arabidopsis thaliana as a model plant and confirming the seed germination rate of the plant. HA; Humic acid, DHN; Homopolymer of 2,7-dihydroxynaphthalene, CV-3P-P; Copolymers of 3-pentadecylphenol, catechol and vanillyric acid mixtures, CV-L-DOPA; L-3,4-dihydroxyphenylalanine, copolymers of catechol and vanillyric acid mixtures.
Figure 2 shows the results of analysis of the chlorophyll content of plants grown by treating the Arabidopsis thaliana as model plants with high salt and polymeric materials of the present invention (DHN, CV-3P-P, and CV-L-DOPA). MS only; Plant growth medium, MS + Lac; MS medium supplemented with laccase. The y-axis salt / control means that the same number of Arabidopsis is incubated for the same time in salt-free medium and salt-free medium, respectively, and chloroplasts are extracted from the plants. Therefore, the higher the ratio, the higher the salt resistance, and the above value can not exceed 1.

In order to achieve the object of the present invention, the present invention provides a composition for promoting germination and for enhancing resistance to salt stress of a plant containing a polymer of naphthalene or a phenolic substance as an active ingredient.

In the composition for promoting germination and enhancing resistance to salt stress according to an embodiment of the present invention, the polymer of naphthalene is prepared by polymerizing 2,7-dihydroxynaphthalene monomer with an oxidizing agent But are not limited to, homopolymers.

In addition, in the composition for promoting germination and salt stress tolerance of plants of the present invention, the polymer of the phenolic substance may be catechol; Vanillic acid; And a copolymer prepared by polymerizing a phenol monomer having an amine group with an oxidizing agent, and the phenol monomer having the amine group may be L-3,4-dihydroxyphenylalanine (L-3,4-dihydroxyphenylalanine) But is not limited thereto.

The polymer of the phenolic substance according to an embodiment of the present invention may be a copolymer of L-3,4-dihydroxyphenylalanine, catechol and vanillyric acid, but is not limited thereto.

Also, the copolymer of L-3,4-dihydroxyphenylalanine, catechol, and vanillic acid according to an embodiment of the present invention is obtained by copolymerizing L-3,4-dihydroxyphenylalanine, catechol, and vanillyric acid monomer More preferably L-3,4-dihydroxyphenylalanine, catechol and vanillylic acid monomer in a weight ratio of 0.1: 0.3 to 0.7: 0.9: 0.9: 1.2, But not limited to, those prepared by mixing lactic acid in a weight ratio of 0.8: 1.

In the production of polymers of the naphthalene or phenolic materials of the present invention, the catechol, vanillic acid, 2,7-dihydroxynaphthalene and L-3,4-dihydroxyphenylalanine are commercially synthesized / But is not limited to.

In the production of the polymer of the naphthalene or phenolic material of the present invention, the oxidant may be at least one selected from the group consisting of oxidase, copper (I) chloride, ammonium peroxodisulfate, sodium periodate and potassium chloride. Do not. In addition, the oxidizing enzyme may be, but is not limited to, laccase, peroxidase, or a mixture thereof. The lacquer is a Trametes < RTI ID = 0.0 > versicolor), Po Po scalpel menta Julius (Fomes fomentarius), sat hatred Thermo fillets (Chaetomium the four thermophile , Neurospora crassa , Colorius < RTI ID = 0.0 > versicol , Botrytis cinerea , Rigidoporus lignosus , Ganoderma lucidum , Coriolus hirsutus , Russula delica ( Russula delica) ), Pleurotos ( Pleurotos ostreatus), Aspergillus nidul lance (Aspergillus nidulans , and the like, but the present invention is not limited thereto.

The polymer of the naphthalene or phenolic material of the present invention has a weight average molecular weight of 5,000 or more. The term "weight average molecular weight" may refer to a conversion value for standard polystyrene measured by Gel Permeation Chromatograph (GPC). In this specification, unless otherwise specified, the term "molecular weight" means "weight average molecular weight ".

The homopolymers or copolymers of the present invention can be prepared by a process for producing oxidized polymers of conventional phenolic compounds known in the art.

The present invention also provides a method for promoting germination of a plant, comprising the step of treating a plant, a seed of a plant or a planting material soil with a composition for promoting germination and salt stress tolerance of the plant of the present invention.

The present invention also provides a method for enhancing the salt stress tolerance of a plant, comprising the step of treating a plant, a plant seed or a plant food soil with a composition for promoting germination and salt stress tolerance of a plant of the present invention.

As a method for promoting germination and salt stress tolerance of the plant, the plant may be immersed in a composition containing the polymer of the naphthalene or phenolic substance according to the present invention as an active ingredient, or the composition according to the present invention may be applied to a plant, For example. For the method of soaking, the composition of the present invention may be poured into the soil around the plant, or the seed may be immersed in the composition, but is not limited thereto.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Production Example 1. Production of homopolymer (DHN) of 2,7-dihydroxynaphthalene

2 g of 2,7-dihydroxynaphthalene (Sigma, USA) was added to 192 mL of a 100 mM sodium acetate buffer containing 48 mL of 100% ethanol and mixed. 20 mg of laccase (Sigma) was added to the mixed solution, and the mixture was stirred at 36 ° C and 400 rpm for 24 hours to induce synthesis of oxidized polymers. The crosslinking solution was centrifuged at 13,000 rpm to obtain a polymer precipitate, and the obtained polymer precipitate was washed with distilled water. The precipitate-free solution was subjected to desalting using an Amicon ultrafilter (Millipore, Germany) with a molecular weight cut-off of 5 kDa to separate the polymer in the solution and then centrifuged The obtained polymer was mixed with the polymer separated through the ultrafilter to prepare a homopolymer. The polymer concentration of the mixed solution was confirmed by measuring the solid weight by completely drying 0.5 mL of the mixed solution at 90 ° C. and diluting the mixed polymer solution to various concentrations in the plant growth medium (MS agar medium) Germination test was performed. In the case of the control (untreated), only the enzyme catalyst was added without using distilled water or phenol monomer as the polymer solution, and the ultrafiltered solution was diluted.

Preparation Example 2. Copolymer of 3-pentadecyl phenol, catechol and vanillyric acid mixture (CV-3P-P) manufacture

1 g of catechol, 0.8 g of vanillylic acid and 0.2 g of 3-pentadecylphenol were added to 120 mL of a 100 mM sodium acetate buffer containing 120 mL of 100% ethanol and mixed. Thereafter, an oxidant is added to induce the synthesis of the polymer, and the process of separating and purifying the polymer is the same as described above. Through the above procedure, a copolymer of monomeric phenol having an alkyl group was prepared.

Production Example 3. Copolymer (CV-L-DOPA) of L-3,4-dihydroxyphenylalanine, catechol and vanillyric acid mixture

0.8 g of catechol, 1 g of vanillylic acid and 0.2 g of L-3,4-dihydroxyphenylalanine were added to 192 mL of 100 mM sodium acetate buffer containing 48 mL of 100% ethanol . Thereafter, an oxidant is added to induce the synthesis of the polymer, and the process of separating and purifying the polymer is the same as described above. Through the above procedure, a copolymer of monomeric phenol having amine groups was prepared.

EXAMPLES Example 1: Identification of polymer germination promoting effect

In order to confirm the plant germination promoting effect of the polymer produced through Production Example 1 to Production Example 3, seed germination rate was measured using Arabidopsis thaliana as a model plant.

Arabidopsis seeds were prepared by surface sterilization by immersing 1.5% sodium hypochlorite solution containing 0.02% triton X-100 in a solution diluted with distilled water at a ratio of 3: 7 (v: v) Washed with distilled water and subjected to cancer treatment at 4 ° C for two days. The disinfected Arabidopsis seeds were MS 4.3 g L ^-1 , sugar 30 g L ^-1 , 0.6% agar, and 8.6, 86 or 430 mg L ^-1 concentration of polymer (DHN, CV-3P-P or CV (Murashige and Skoog), pH 5.8, which was prepared containing the L-DOPA (-L-DOPA) and incubated for 2 days in a growth chamber set at 23 ° C and 16 hours: 8 hours (female). 50 seeds per group were repeated 3 times and germination rate was measured based on the emergence of young seeds of seeds.

As a result, it was confirmed that the germination rate of Arabidopsis seeds was increased depending on the treatment concentration, similar to the case of treatment with natural humic acid (HA) at 8.6 and 86 mg · L ^-1 concentration treatment of DHN and CV-L-DOPA (Fig. 1). In particular, CV-L-DOPA showed higher germination rate than the humic acid treatment at the same concentration of 86 mg · L ^-1 . However, in both DHN and CV-L-DOPA, the seed germination rate was reduced at the 430 mg · L ^-1 concentration. When compared to the degree of reduction, the CV-L-DOPA substance showed the highest seed germination rate similar to humic acid at high concentration, indicating that the plant stimulation ability was changed by the presence of amine groups. In the case of CV-3P-P, the seed germination rate did not increase at the treatment at 8.6 and 86 mg · L ^-1 , and in particular, the seed germination was not observed at the treatment concentration of 430 mg · L ^-1 .

From the above results, it can be inferred that the polymer of the naphthalene or phenolic material according to the present invention can be used as a material of a similar organic polymer fertilizer that can replace commercial humic acid.

Example 2. Confirmation of enhancing stress tolerance of plant salt of polymer

In order to confirm the stress tolerance enhancing effect of the polymer prepared from the preparation examples 1 to 3, the salt resistance of the plant was analyzed using the Arabidopsis thaliana as a model plant.

Arabidopsis seeds were planted in MS medium and cultured for 13 days in a growth chamber set at 23 ° C and 16 hours: 8 hours (female) photoperiod condition. The grown Arabidopsis seedlings were grown in 250 mM sodium chloride (NaCl) and 106 mg The cells were cultured in MS medium containing L- ¹ concentration of the polymer (DHN, CV-3P-P or CV-L-DOPA). After 6 days of incubation, the Arabidopsis was harvested and chloroplast was extracted with 80% (v / v) acetone and the chlorophyll content of the plant was measured using a spectrophotometer. Three groups of 30 seeds per group were used and Arabidopsis thaliana grown on MS medium without sodium chloride and polymer was used as a control. The amount of chloroplast extracted from the Arabidopsis thaliana grown in the control and the amount of chloroplast extracted from the Arabidopsis thaliana grown at each experimental treatment were expressed as a ratio.

As a result, the content of chlorophyll was higher in DHN, CV-3P-P and CV-L-DOPA treatments than MS + Lac treated with distilled water only or diluted with oxidation catalyst only. It was also observed that the levels of chlorophyll were higher in DHN and CV-L-DOPA polymer treatments of the present invention than in natural humic acid (HA) treatments (FIG. 2).

From the above results, it was found that the polymer of the present invention has an effect of enhancing the salt resistance of the plant, so that the polymer of the present invention is considered to be useful for plant growth of high salt and coastal area.

Claims (9)

A composition for promoting germination and for improving salt stress tolerance of a plant containing a polymer of naphthalene or a phenolic substance as an active ingredient,
The polymer of naphthalene is a homopolymer of 2,7-dihydroxynaphthalene,
The polymer of said phenolic material is selected from the group consisting of catechol; Vanillic acid; And L-3,4-dihydroxyphenylalanine (L-3,4-dihydroxyphenylalanine); and a composition for enhancing germination and enhancing resistance to salt stress. delete delete delete The composition according to claim 1, wherein the polymer is polymerized using an oxidizing agent. 6. The method according to claim 5, wherein the oxidizing agent comprises at least one member selected from the group consisting of oxidase, copper (I) chloride, ammonium peroxodisulfate, sodium periodate and potassium chloride. / RTI > [Claim 7] The composition according to claim 6, wherein the oxidase is lactase, peroxidase or a mixture thereof. A method for promoting germination of a plant comprising the step of treating plant, plant seed or plant material soil with the composition for promoting plant germination and salt stress tolerance according to claim 1. A method for promoting salt stress tolerance of a plant, comprising the step of treating the plant germination promoting and salt stress tolerance enhancing composition according to claim 1 to a plant, a plant seed or a plant cultivation soil.

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