KR101384470B1 - Methods of preparation of plant-growth-promoter by using solid fermenation of chitosan - Google Patents

Abstract

The present invention relates to a method for preparing a plant growth promoter by solid fermentation of chitosan and a method of promoting plant growth using the method. The plant growth promoter according to the present invention is prepared by solid fermentation of chitin using Rhizopus oligosporus strain and has good preservation and plant growth promoting effect. The chitosan useful to a human body may be absorbed into plants.

Description

Method for preparation of plant growth promoter by chitosan solid fermentation {Methods of preparation of plant-growth-promoter by using solid fermenation of chitosan}

The present invention relates to a plant growth promoter through solid fermentation of chitosan.

Chitin is the second most abundant resource after cellulose, and is a natural macromolecular substance present in the shells of shellfish, crabs, shrimps, and higher plants such as fungi and algae. Chitosan is prepared by deacetylating chitin, and its solubility in water varies depending on the degree of deacetylation. Recently, chitosan has been used in various fields because of its high toxicity, environmental friendliness and biocompatibility. Chitosan is known to activate cells, inhibit aging, strengthen immunity and prevent disease, and regulate rhythm with the function of activating the body's natural healing ability. In addition, it is known to exhibit the adsorption, excretion and anticancer action of harmful cholesterol, and by inhibiting the absorption in the intestine by adsorbing chloride ions that cause the increase in blood pressure is discharged to the outside of the body to suppress blood pressure rise.

Conventionally, strong acids (hydrochloric acid) and strong bases (caustic soda solution) were used for shelling of shellfish such as crabs, crawfish, and shrimps. However, strong acids and strong bases have a problem that adversely affects the environment. Therefore, there is a need for a method of producing chitosan at low cost without adversely affecting the environment.

In order to supply chitosan useful to the human body, anchovy salt containing chitosan, kimchi containing chitosan, bread containing chitosan, rice cake containing chitosan, and miso containing chitosan were prepared. In another attempt, a study was conducted to absorb chitosan from fruit vegetables and to supply them through chitosan-containing fruit vegetables.

However, there have been no reports of successful absorption of chitosan into plants by decomposing chitosan through solid culture and then supplying it to plants. The present inventors have found that the strain Rhizopus oligosporus can produce chitosan by decomposing the chitin component of the shellfish through solid culture, and can be absorbed into the plant when the solid culture fermentation product is treated to the plant, thereby completing the present invention. .

Korean Patent Application No. 2009-0135168 discloses (a) washing, drying, and grinding shellfish shells; (b) a decolorizing step of extracting astaxanthin contained in the shellfish crust crushed in (a) as a spirit; (c) a deliming step of extracting and separating ash by putting the decolored crustacean shell in (b) into an extraction tank containing lactic acid and boiling; (d) The method of obtaining a chitin is provided by deshelling the shellfish, which has been subjected to the deliming step, into an extraction tank containing potassium carbonate (K2CO3) and boiling to extract and separate proteins to obtain chitin. Korean Patent Application No. 2007-0096386 discloses a method for producing anchovy salted fish, mainly based on chitosan-containing ocher aqueous extract. Patent No. 0880603 provides a water treatment plant sludge treatment method using chitosan and chitosan degrading enzyme, and a fertilizer manufacturing method using the same. Patent No. 0992213 relates to a method for producing low molecular chitosan using Ryzopus and a method for producing chitosan-containing plants using the same, the method comprising: dissolving chitosan in a lactic acid solution; Mixing low-cost chitosan having a molecular weight of 3600 or less by mixing the culture of Lyzopus oligosporus KCCM 10624 with the chitosan-dissolved solution; And it provides a low-molecular chitosan-containing tomato production method comprising the step of administering the low-molecular chitosan to the tomato. US Patent Application 2002-161187 discloses a method for preparing chitosan using Rhizopus azygosporus. In this patent, however, liquid culture was used to culture Rhizopus azygosporus. In addition, US Patent No. US 6255085 B1 discloses a method for producing chitosan by culturing Actinomucor taiwanensis fungi in medium about 5 to 60 g / L corn steep liquor.

Zhang et al. Disclose a method of producing chitosan using Rhizopus japonicus in Carbohydrate Polymers (Volume 101, Jan. 30, 2014, pp. 57-67). Chatterjee et al. Studied the effects of plant hormones on the production of chitosan using Rhizopus orizae in the International Journal of Biological Macromolecules (Volume 42, Issue 2, pp. 120-126, March 1, 2008). It is reported. Mahata et al. Reported a method for producing chitosan using Rhizopus oligosporus in Carbohydrate Research (In Press, Accepted Manuscript, Available online 13 June 2013).

An object of the present invention is to provide a method for producing a plant growth promoter containing chitosan through solid culture of chitin using the Rhizopus oligosporus strain.

The present invention is chitin Rhizopus An object of the present invention is to provide a method for increasing the content of chitosan in a plant, characterized by treating a solid culture of an oligosporus strain.

A first aspect of the present invention for this purpose comprises the steps of preparing a grain: adsorbing a chitin material to the grain; Grinding the grains to which the chitin is adsorbed; And Rhizopus in the ground mill Chitosan-containing plant growth promoter manufacturing method comprising the step of inoculating the oligosporus strain. The grain may be any type of grain available to the Rhizopus oligosporus strain. Preferably, the grains may be any one or more grains selected from the group consisting of soybean meal, corn, wheat flour.

A second aspect of the invention provides a method of preparing a grain comprising: adsorbing a chitin material to the grain; Grinding the grains to which the chitin is adsorbed; Rhizopus on the pulverized ground A method of increasing the amount of chitosan uptake in a plant, comprising inoculating an oligosporus strain and fermenting the same, and treating the plant with the fermentation product. The plant may be a vegetable such as melons, tomatoes, and the like. More preferably lettuce. The grain may be any type of grain available to the Rhizopus oligosporus strain. Preferably, the grains may be any one or more grains selected from the group consisting of soybean meal, corn, wheat flour.

In the present invention, when chitosan is prepared by decomposing chitin using Rhizopus oligosporus strain, a solid culture method is used without using a buffer solution such as lactic acid for dissolving chitin, and using no strong acid or strong base. It is cost effective because it can reduce the contamination and reduce the cost of removing the buffer solution.

The present invention is also characterized in that the grains used can be used to promote plant growth, and in the process of the present invention, chitin is crushed by physical force, so that the chitosan content in the form that the plant can absorb is increased.

Since the composition according to the present invention is not a liquid phase but a solid phase, there is little contamination by microorganisms, and thus, the shelf life is almost semipermanent.

1 is a result of requesting the detection of chitosan in the Korea Food Research Institute for lettuce containing chitosan.

The invention is explained in more detail through the following examples. However, the following examples are only for illustrating the present invention and do not limit the scope of the present invention. The scope of the present invention is to be interpreted by the claims, to the extent that can be easily substituted by those skilled in the art.

Example 1 Method for Preparing Chitosan-Containing Plant Growth Accelerator
Chitin was degraded and plant growth promoters containing chitonic acid were prepared by the following steps.
(a) preparing 25 kg of grain (main malt flour for animal diet) and adsorbing 200 g of chitin powder
(b) adding 30 ml of microbial culture (Ryzopus oligosporus (1 species of white downy mildew)) and 3000 ml of water (containing minerals) to the grains adsorbed with the chitin powder;
(c) mixing the composition prepared in step (b) into a stirrer and mixing;
(d) fermenting the composition prepared in step (c) in an airy container for 4 days;
(e) drying the fermented product of step (d);

(f) The dried fermented product was prepared by grinding at 250 mesh.

Example  2. Identification of Chitosan in Lettuce by Chitosan-Containing Plant Growth Accelerator

1. Method

After spraying the chitosan-containing microbial agent in the soil and mixed, foliar fertilization or irrigation at intervals of 15 to 20 days.

2. Results

As a result of requesting the detection of chitosan to the Korea Food Research Institute, 0.06 mg / g of chitosan was detected (FIG. 1). These results show that chitosan can be contained by the method according to the invention for lettuce which cannot synthesize chitosan.

Example 3. Confirmation of the effect of chitosan-containing plant growth promoter (Power Green-F)

1. Purpose of test

This study intends to find out the effect of Power Green-F prepared according to Example 1 on crop growth.

2. Test method

end. Start: Leaf Vegetable-Yeolmu [Everyone]

I. Survey content: height, leaf width, number of leaves, chlorophyll content (SPAD), basement length, fresh weight (ground, basement)

All. Processing contents

Test Materials Growth survey test Non-experimental test request
company Dilution factor and
usage When and how to process Standard amount Ship Power Green-F 1000 times dilution of volume treatment 500 times dilution of volume treatment Dilute the test materials immediately before the seedlings (heatless radish) by 1000 times dilution and 500 times the dilution. After soil irrigation (12/17), two leaves were treated twice every two weeks in the 3 ~ 4 season (2/1, 2). / 15) 500-fold dilution 250-fold dilution Wells Farm Co., Ltd. No treatment - - - - -

la. Seedling overview: The radish seeds were sown at 30 intervals on December 17, and no pesticides or fertilizers were treated during the cultivation period. Soil hydration was irrigated with groundwater whenever needed. Others are based on conventional cultivation.

hemp. Test Arrangement: 3 times of ingot method

division Test area Treated water Number of repeats Muzzle count Gudang area Area required gun
Area required Ineffective 2 2 3 12 20 m ² 240 m ² 300 m ² Than 2 2 3 12 5 m ² 60 m ²

3. Survey Method

division Survey item Number of investigations Date of investigation Investigation method Growth survey test Growth survey One 02/22 Published by the Rural Development Administration
Agricultural Research Standards Comparison test Compared with 3 02/04, 02/06, 02/08 Investigation of the presence or absence of plants

4. Test score

end. Growth Test: Crop Growth Effect of 'Power Green-F'

Table 4 shows the effect of Power Green-F treatment on the ultra long.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Extra long (cm) Indices(%) Extra long (cm) Indices(%) Power Green-F Appropriate 14.6a ^{2 )} 104.3 16.4a 104.5 Power Green-F Displacement 15.6b 111.4 17.1b 108.9 No treatment 14.0a - 15.7 -

¹⁾ Appropriate treatment: 1000 times dilution treatment, Dose treatment: 500 times dilution

^{2) There was} no significant difference between DMRT results at the 5% significance level.

Table 5 shows the effect of Power Green-F treatment on the leaf width.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Leaf width (cm) Indices(%) Leaf width (cm) Indices(%) Power Green-F Appropriate 5.3a ^{2 )} 100.0 5.3a 106.0 Power Green-F Displacement 5.4a 101.9 5.5a 110.0 No treatment 5.4a - 5.0a -

¹⁾ Appropriate treatment: 1000 times dilution treatment, Dose treatment: 500 times dilution

^{2) There is} no significant difference between DMRT results at the 5% level of similar characters.

Table 6 shows the effect of Power Green-F treatment on the leaves.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Leaves () Indices(%) Leaves () Indices(%) Power Green-F Appropriate 8.8a ^{2 )} 123.9 9.3b 127.4 Power Green-F Displacement 9.3b 131.0 9.7b 132.9 No treatment 7.1a - 7.3a -

¹⁾ Appropriate amount treatment: 1000 times dilution treatment, dosing treatment: 500 times dilution

^{2) There is} no significant difference between DMRT results at the 5% level of similar characters.

Table 7 shows the effect of Power Green-F treatment on chlorophyll content.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Chlorophyll content
(SPAD) Indices(%) Chlorophyll content
(SPAD) Indices(%) Power Green-F Appropriate 36.9a ^{2 )} 102.8 37.8a 103.8 Power Green-F Displacement 36.2a 100.8 37.3a 102.5 No treatment 35.9a - 36.4a -

¹⁾ Appropriate amount treatment: 1000 times dilution treatment, dosing treatment: 500 times dilution

^{2) There is} no significant difference between DMRT results at the 5% level of similar characters.

Table 8 shows the impact of Power Green-F treatment on the ground load.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Ground weight (g) Indices(%) Ground weight (g) Indices(%) Power Green-F Appropriate 13.9b ^{2 )} 113.9 14.0b 120.7 Power Green-F Displacement 16.1b 132.0 16.6b 143.1 No treatment 12.2a - 11.6a -

¹⁾ Appropriate amount treatment: 1000 times dilution treatment, dosing treatment: 500 times dilution

^{2) There is} no significant difference between DMRT results at the 5% level of similar characters.

Table 9 shows the effect of PowerGreen-F treatment on the underground length.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Underground length (cm) Indices(%) Underground length (cm) Indices(%) Power Green-F Appropriate 14.1b ^{2 )} 121.6 14.4b 128.6 Power Green-F Displacement 16.1b 138.8 16.1b 143.8 No treatment 11.6a - 11.2a -

¹⁾ Appropriate amount treatment: 1000 times dilution treatment, dosing treatment: 500 times dilution

^{2) There is} no significant difference between DMRT results at the 5% level of similar characters.

Table 10 shows the effect of Power Green-F treatment on the underground load.

Treatment District ^{1 )} Chungnam National University Boeun, Chungbuk Underground weight (g) Indices(%) Underground weight (g) Indices(%) Power Green-F Appropriate 0.9a ^{2 )} 112.5 0.9a 128.6 Power Green-F Displacement 0.9a 112.5 0.9a 128.6 No treatment 0.8a - 0.7a -

¹⁾ Appropriate amount treatment: 1000 times dilution treatment, dosing treatment: 500 times dilution

^{2) There is} no significant difference between DMRT results at the 5% level of similar characters.

I. Comparison test

Table 11 shows the test results after the Power Green-F treatment.

Test Materials Test crop
(kind) Comparative accuracy (0 ~ 5) Remarks Standard amount Quantity Power Green-F Yeolmu
(everyone) 0 0 No comparison

All. Hazardous Ingredient Analysis

Table 12 shows the harmful component analysis of the test sample. The unit is mg / kg.

Yu Sun component chrome lead cadmium nickel Mercury zinc arsenic Copper Test Sample Content 2.8467 ND 0.0933 ND ND 19.7933 0.0849 2.2333 Ingredient Standard ^* <300 <150 <5 <50 <2 <50 <5 <50

^* Standard: Rural Development Administration Notice No. 2009-30 Fertilizer Process Specification

5. Summary of Results

end. Growth promoting effect

In summary, the results of investigating the growth-promoting effect of thermal radish by treating 'Power Green-F' according to Example 1 of the present invention are as follows.

1) When Power Green-F was treated, grass height increased by at least 4.3% (Chungnam National University), and up to 11.4% (Chungnam National University).

2) When Power Green-F was treated, the leaf width increased by 1.9% (Chungnam National University) and 10.0% (Boeun).

3) When Power Green-F was treated, the number of leaves increased by 23.9% (Chungnam National University) and 32.9% (Boeun).

4) When Power Green-F was treated, the chlorophyll content was increased by 0.8% (Chungnam University drainage treatment) and 3.8% (Boeun appropriate treatment).

5) When Power Green-F was treated, the ground load increased by at least 13.9% (Chungnam National University), and up to 43.1% (Chungnam National University).

6) Under the treatment of Power Green-F, the underground length increased by 21.6% (Chungnam National University) and 43.8% (Boeun).

7) When Power Green-F was treated, the underground load increased by at least 12.5% (Chungnam National Treatment Zone) and up to 28.6% (Boeun Treatment Zone).

I. Than

The test materials showed no symptoms compared to the heat crops, which were the target crops, in the standard and batch treatments.

All. Hazardous Ingredient Analysis

As a result of analyzing the harmful components of 'Power Green-F' according to Example 1 of the present invention, it was not detected exceeding the component standards of the establishment and designation of the fertilizer process standard No. 2009-30 RDA.

6. Conclusion

The composition according to the present invention has an effect on promoting the growth of plants, and did not appear compared to the thermal radish which is the test crop, it could be utilized as an environmentally friendly growth material.

Claims (5)

(a) preparing 25 kg of grain and adsorbing 200 g of chitin powder to the grain;
(b) injecting 30 ml of Ryzopus oligosporus culture medium and 3000 ml of water into the grains to which the chitin powder is adsorbed;
(c) mixing the composition prepared in step (b) into a stirrer and mixing;
(d) fermenting the composition prepared in step (c) in an airy container for 4 days;
(e) drying the fermented product of step (d); And
(F) a method for producing a plant growth accelerator composition comprising chitosan made by decomposing chitin consisting of grinding the dried fermented product at 250 mesh. The method of claim 1, wherein the grain is at least one grain selected from the group consisting of soybean meal, corn, and wheat flour. (a) preparing 25 kg of grain and adsorbing 200 g of chitin powder to the grain;
(b) injecting 30 ml of Ryzopus oligosporus culture medium and 3000 ml of water into the grains to which the chitin powder is adsorbed;
(c) mixing the composition prepared in step (b) into a stirrer and mixing;
(d) fermenting the composition prepared in step (c) in an airy container for 4 days;
(e) drying the fermented product of step (d); And
(f) increasing the amount of chitosan uptake in lettuce, comprising treating the lettuce with a plant growth promoter composition comprising chitosan produced by decomposing chitin, which is composed of grinding the dried fermented product at 250 mesh Way. delete The method of claim 3, wherein the grains are any one or more grains selected from the group consisting of soybean meal, corn, and wheat bran.

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