KR102634297B1 - agent for promoting growth of crop and cultivating method - Google Patents

Abstract

The present invention relates to a crop growth promoter and a crop cultivation method using the same. More specifically, it relates to a growth promoter that can effectively promote crop growth using microorganisms of the Saccabacillus genus and a crop cultivation method using the same.

Description

Crop growth promoter using microorganisms of the Saccabacillus genus and crop cultivation method using the same {agent for promoting growth of crop and cultivating method}

The present invention relates to a crop growth promoter and a crop cultivation method using the same. More specifically, it relates to a growth promoter that can effectively promote crop growth using microorganisms of the Saccabacillus genus and a crop cultivation method using the same.

In general, in order to improve the quality and maximize the yield when growing crops, there are methods of applying various types of fertilizers as irrigation or foliar application.

When nutrients cannot be absorbed in a balanced manner due to imbalance or antagonism of each nutrient in the soil, a foliar application method is used in which fertilizer is diluted in water and sprayed on stems, leaves, and fruits. This is used when it is necessary to quickly replenish nutrients to address deficiencies in specific nutrients, slow growth, or improve quality. These nutrients include minerals such as nitrogen, phosphoric acid, potassium, kaolin, manganese, boron, iron, molybdenum, zinc, copper, and calcium, as well as urea, high molecular weight nitrogen compounds, amino acid fertilizers, organic phosphoric acid, organic acid potassium, organic phosphorus, carbohydrate derivatives, Organic compounds such as vitamins, sugars, seaweed extracts, and plant extracts are used in crops.

Conventional farming methods using chemical fertilizers or chemical pesticides result in a decrease in soil strength and acidification of the soil. Therefore, there is a transition from inorganic farming using chemical fertilizers to environmentally friendly organic farming.

In particular, interest in eco-friendly farming using microorganisms has recently increased significantly. There are many types of microorganisms that promote crop growth, but among them, the crop growth promotion effect of Bacillus has been well identified.

Plant hormones produced by microorganisms as metabolites are effective in promoting the growth of crops. These plant hormones include auxin, gibberellin (GA), cytokinin, and ethylene.

Korean Patent Publication No. 10-2017-0136081, Korean Patent Publication No. 10-2008-0090803, etc. disclose the growth promotion of crops using microorganisms. However, the genus Saccharibacillus is still sp.) There is no known effect of using microorganisms to promote crop growth.

1. Republic of Korea Patent Publication No. 10-2017-0136081: Bacillus amyloliqueficiens IM1 strain, plant disease control and growth promotion composition containing the same 2. Republic of Korea Patent Publication No. 10-2008-0090803: Crop growth promoting microorganism Bulkholderia cepacia SV-4 and method for producing gibberellin using the same

The present invention was created to improve the above problems, and its purpose is to provide a growth promoter that can effectively promote crop growth using microorganisms of the genus Saccabacillus and a crop cultivation method using the same.

The crop growth promoter using microorganisms of the genus Saccharibacillus of the present invention to achieve the above object is Saccharibacillus genus (Saccharibacillus sp.) Contains secondary metabolites of microorganisms.

The microorganism is Saccharibacillus brassicae .

The secondary metabolites include indole acetic acid.

The growth promoter is the culture medium of the microorganism or its dry powder.

The culture medium contains yeast extract, dextrose, NaCl, K ₂ HPO ₄ , The microorganism was cultured in a medium containing Na ₂ CO ₃ , MgSO ₄ , and L-Trytophan.

And in the crop cultivation method of the present invention to achieve the above object, crops are cultivated by fertilizing the above growth promoter by foliar spraying or irrigation spraying.

The present invention suggests that the culture medium of microorganisms of the genus Sacacillus contains indole acetic acid as a plant hormone. Therefore, it is possible to provide a growth promoter that can effectively promote the growth of crops by using the culture medium of microorganisms of the Saccabacillus genus.

The crop growth promoter of the present invention can increase disease resistance and promote crop growth by fertilizing the soil or the leaves and roots of growing crops by foliar spraying or irrigation spraying.

Figure 1 is a graph showing the results of HPLC analysis of microbial culture medium,
Figure 2 is a graph showing the HPLC analysis results of prototype SP20,
Figures 3 to 5 are graphs showing the measured values of leaf length, leaf width, and fresh weight as a result of lettuce growth experiments, respectively;
Figure 6 is a photograph showing the results of a lettuce growth experiment,
Figure 7 is a photograph showing the results of a tomato growth experiment,
Figure 8 is a photograph showing the results of a tomato growth experiment,
Figure 9 is a photograph showing the results of a tomato toxicity test.

Hereinafter, a crop growth promoter using microorganisms of the genus Saccabacillus according to a preferred embodiment of the present invention and a crop cultivation method using the same will be described in detail.

The present invention relates to the genus Saccharibacillus sp.) By containing secondary metabolites of microorganisms, it is possible to provide a natural growth promoter that is harmless to crops instead of conventional artificially synthesized artificial hormones.

Saccharibacillus genus sp.) Saccharibacillus brassicae can be used as a microorganism.

Secondary metabolites can be obtained by culturing microorganisms.

Unlike primary metabolites, such as amino acids, nucleic acids, proteins, lipids, carbohydrates, etc., which are essential for the growth of microorganisms, secondary metabolites are products of secondary metabolism of microorganisms and are directly involved in the growth of microorganisms. It means a substance that is not involved.

Indole-3-acetic acid (IAA) is a secondary metabolite of the Saccabacillus brassicae microorganism.

Indole acetic acid is a natural auxin, a type of plant hormone that regulates plant growth. Indole acetic acid is a synthetic auxin that is artificially synthesized, such as 2,4-dichlorophenoxyacetic acid, 1-naphthaleneacetic acid, and 2-naphthoxyacetic acid. Likewise, it is known to induce cell growth and cell division.

Since the present invention contains indole acetic acid, a natural auxin, as a secondary metabolite of microorganisms, it is harmless compared to using artificially synthesized synthetic auxin, and allows organic certification of crops.

Various types of liquid media can be used as a medium for cultivating microorganisms of the Saccabacillus genus.

Examples of liquid media include distilled water, yeast extract, dextrose, NaCl, K ₂ HPO ₄ , Those with added Na ₂ CO ₃ and MgSO ₄ can be used. In this case, the liquid medium contains 0.6 to 1.0% by weight of yeast extract, 0.2 to 0.8% by weight of dextrose, 0.1 to 0.2% by weight of NaCl, K ₂ HPO ₄ , It may contain 0.1 to 0.5% by weight, Na ₂ CO ₃ , 0.01 to 0.1% by weight, MgSO ₄ , 0.05 to 0.15% by weight.

In order to increase the content of indole acetic acid, a secondary metabolite of microorganisms, a precursor of indole acetic acid can be further added to the medium used for culturing microorganisms. L-Tryptophan can be used as a precursor for indole acetic acid. By adding L-tryptophan to the medium, the content of indole acetic acid can be increased when culturing microorganisms. Examples of such media include 0.6 to 1.0% by weight of yeast extract, 0.2 to 0.8% by weight of dextrose, 0.1 to 0.2% by weight of NaCl, K ₂ HPO ₄ , It may contain 0.1 to 0.5% by weight, Na ₂ CO ₃ , 0.01 to 0.1% by weight, MgSO ₄ , 0.05 to 0.15% by weight, and 0.05 to 0.15% by weight of L-tryptophan.

The crop growth promoter of the present invention is the culture solution itself obtained by culturing microorganisms in a medium or is separated from the culture solution. Additionally, it may be a dry powder obtained by drying the culture medium.

Additionally, it can be used as a crop growth promoter by removing bacterial cells from the culture medium. A typical solid-liquid separation method can be used to remove bacteria from the culture medium. For example, the culture medium can be centrifuged to obtain a supernatant from which the bacteria have been removed. The supernatant obtained through centrifugation is used as a crop growth promoter.

In addition, of course, the crop growth promoter of the present invention can be manufactured in the form of a wettable powder, granule or encapsulation by adding known formulation materials for the purpose of stable formulation. In addition, of course, known additives such as anti-freezing agents, preservatives, dispersants, pH adjusters, etc. may be added. In this case, the crop growth promoter of the present invention may contain 1 to 90% by weight of culture medium or dry powder.

The crop growth promoter of the present invention can increase disease resistance and promote crop growth by fertilizing the soil or the leaves and roots of growing crops by foliar spraying or irrigation spraying.

Hereinafter, the present invention will be described through examples. However, the following examples are for illustrating the present invention in detail, and the scope of the present invention is not limited to the following examples.

(Example)

1. Culture

Yeast extract, dextrose, NaCl, K ₂ HPO ₄ in distilled water, A liquid medium was prepared by adding Na ₂ CO ₃ , MgSO ₄ , and L-tryptophan. At this time, each component in the liquid medium is 0.8% by weight of yeast extract, 0.5% by weight of Dextrose, 0.15% by weight of Nacl, 0.25% by weight of K ₂ HPO ₄ , 0.05% by weight of Na ₂ CO ₃ , 0.1% by weight of MgSO ₄ , and 0.1% by weight of L-Trytophan. It was added to adjust to %.

Saccharibacillus brassicae KCTC 43072 was cultured in the prepared liquid medium at 30°C and 150 rpm for 72 hours to obtain a microbial culture medium. The number of strains in the microbial culture medium was 8.8×10 ⁸ cfu/ml.

And the dry powder was obtained by spray drying the microbial culture solution using a spray dryer. The number of strains in the dry powder was 1.6×10 ¹⁰ cfu/ml.

2. Prototype manufacturing

Four types of prototypes were manufactured using the above microbial culture medium and dry powder. The mixing ratio (% by weight) of each component is shown in Table 1 below.

Sample Microbial culture medium dry powder Distilled water SS10 10 - 90 SS20 20 - 80 SP10 - 10 90 SP20 - 20 80

<Analysis of microbial secondary metabolites>

To confirm the components of secondary metabolites produced by Sacacillus brassicae, microbial culture medium and prototype SP20 were used as samples.

The microbial culture medium and prototype SP20 were centrifuged at 4000 rpm for 15 minutes to obtain a supernatant from which the bacteria were separated. After pretreatment of the supernatant using the QuEChERS Extraction method, the separated material was analyzed using Agilent 1100 series, API 3200 Q TRAP LC MSMS. Column used was X-Bridge _{( 2.1} The material was analyzed under the following conditions: % TFA, Flow rate: 0.2mL/min, Injection volume: 5μL.

The analysis results are shown in Table 2 below. Each sample was measured three times.

division Calculated Concentration(ng/ml) Average(ng/ml) SD
Microbial culture medium 18.7
19.65
1.67 22 18.26
SP20 50
50.20
3.51 54.6 46

Referring to the results in Table 2 above, indole acetic acid (IAA), known as a growth promoting substance, was detected in the microbial culture medium and prototype. The content of IAA was detected at 19.65 ng/ml in the microbial culture medium, and 50.20 ng/ml in the prototype using dried material with the number of strains concentrated about 100 times, so the content of IAA was about 2.5 times more in the prototype compared to the culture solution. I was able to confirm that it was high.

And the HPLC analysis results of the microbial culture medium and prototype SP20 are shown in Figures 1 and 2, respectively. Compared to the IAA standard material (Sigma's Indole-3-Acetic Acid, CAS No. 87-51-4), a peak was confirmed at the same retention time (RT: 7.71 min). Therefore, it was confirmed that the microbial culture medium and prototype SP20 contained indole acetic acid (IAA), a plant hormone.

<Lettuce growth experiment>

To test the growth promotion effect of the prototype in-flight, lettuce seeds (Cheongchima, Nonghyup Seedlings) were sown and germinated at 26℃ for one week, then transplanted into 9cm Afterwards, each prototype was diluted 500 times and irrigated twice at 7-day intervals.

The appearance of each lettuce after 7 days of treatment is shown in Figure 6. And the measurement results of leaf length, leaf width, and live weight are shown in Table 3 and Figures 3 to 5 below.

division leaf print leaf width live weight cm Increase/decrease rate (%) cm Increase/decrease rate (%) g Increase/decrease rate (%) Untreated 13.2 - 5.1 - 13.7 - SS10 14.5 9.8 5.2 2.5 14.4 4.9 SS20 14.5 9.9 5.2 1.0 14.2 3.7 SP10 14.9 12.3 5.2 2.5 14.6 6.3 SP20 14.8 12.5 5.3 3.7 14.7 7.2

As a result of the experiment, it was found that the leaf length, leaf width, and live weight of all prototypes were increased compared to the untreated ones. In particular, in the SP20 prototype containing 20% of dry powder, it was confirmed that the leaf length increased by 12.5%, the leaf width increased by 3.7%, and the fresh weight increased by 7.2% compared to the untreated product.

<Tomato growth experiment>

In the above experiment, the prototype SP20, which had the best growth promotion effect, was diluted 500 times and a tomato growth experiment was performed.

In a field test for tomato growth promotion conducted in Gokseong-gun, Jeollanam-do, the experiment was divided into an irrigation treatment group 7 days after planting, an irrigation treatment group 14 days after planting, an irrigation treatment group 21 days after planting, and a non-treatment group. The first harvest was conducted on November 2nd, about 3 months after planting the tomatoes, and the second harvest was conducted on November 9th, and then the yield was investigated.

The actual appearance of the untreated section during the first harvest season and the irrigated section 14 days after planting are shown in Figure 7. When observed with the naked eye, it was found that there was a difference in the quantity of fruit on the tomatoes.

And Figure 8 shows a comparison of the first harvested tomato fruits. Referring to Figure 8, there was a clear difference in the quantity of fruit in the untreated group and the irrigated group.

The yield and increase rate of the untreated and irrigated groups after the first harvest are shown in Table 4 below.

treatment area Yield (g/week) Increase rate (%) ⅠRepeat ⅡRepeat ⅢRepeat average Official 7-day post-drench treatment 1098.0 1119.0 1151.5 1122.8 23.2 Official 14-day post-drench treatment 1137.0 1104.0 1171.0 1137.3 24.8 Official 21-day post-drench treatment 1130.5 1098.5 1146.3 1125.1 23.5 Untreated 924 899 911 911.3 -

Referring to the results in Table 4 above, the yield of the first harvest period on November 2 was investigated, and the average was 1122.8g/week in the irrigation treatment group 7 days after planting, 1137.3g/week in the irrigation treatment group 14 days after planting, and 1125.1g/week in the irrigation treatment group 21 days after planting. /Indicates the harvest of the week. It was confirmed that the yield increased by 23.2%, 24.8%, and 23.5%, respectively, compared to 911.3g/week in the untreated area.

The yield and increase rate of the untreated and irrigated groups after the second harvest are shown in Table 5 below.

treatment area Yield (g/week) Increase rate (%) ⅠRepeat ⅡRepeat ⅢRepeat average Official 7-day post-drench treatment 1156.4 1152.2 1157.1 1155.2 17.3 Official 14-day post-drench treatment 1163.9 1177.5 1187.8 1176.4 19.4 Official 21-day post-drench treatment 1188.4 1193.7 1195.5 1192.5 21.1 Untreated 956.7 1004.8 993.4 984.9 -

Referring to the results in Table 5 above, as a result of investigating the yield during the second harvest on November 9, the yield was 1155.2g/week in the irrigation treatment group 7 days after planting, 1176.4g/week in the irrigation treatment group 14 days after planting, and 1176.4g/week in the irrigation treatment group 21 days after planting. 1192.5g/week was measured. As a result, it was confirmed that there was an increase of 17.3%, 19.4%, and 21.1%, respectively, compared to 984.4g/week in the untreated group.

According to the results of the tomato growth experiment, it was confirmed that treatment of the prototype affected the initial harvest and increased yield compared to the untreated group.

<Weakness test>

Prototype SP20 was diluted 500 times (quantity) and 250 times (double volume) to confirm the occurrence of damage 7 days, 14 days, and 21 days after planting.

As a result of examining a total of 21 treatment areas of 5 m ² per area, it was confirmed that no damage occurred. Figure 9 is a photograph showing the appearance of tomatoes 21 days after planting.

Above, the present invention has been described with reference to embodiments, but these are merely illustrative, and those skilled in the art will understand that various modifications and equivalent embodiments are possible therefrom. Therefore, the true scope of protection of the present invention should be determined only by the appended claims.

Claims (6)

It is a dried powder of the culture broth of Saccharibacillus brassicae microorganisms and contains secondary metabolites of the Saccharibacillus brassicae,
The secondary metabolites include indole acetic acid,
The culture medium contains 0.8% by weight of yeast extract, 0.5% by weight of dextrose, 0.15% by weight of Nacl, 0.25% by weight of K ₂ HPO ₄ , 0.05% by weight of Na ₂ CO ₃ , 0.1% by weight of MgSO ₄ , and 0.1% by weight of L-Trytophan. A crop growth promoter using microorganisms of the Saccabacillus genus, characterized in that the microorganisms are cultured for 72 hours at 30° C. and 150 rpm in a medium. delete delete delete delete A crop cultivation method characterized in that crops are grown by applying the growth promoter of claim 1 through foliar spraying or irrigation spraying.