KR102571461B1 - Antibacterial composition for cultivating ginger and the use thereof - Google Patents

Abstract

The present invention relates to an antibacterial extract against ginger growth and quality-inhibiting microorganisms and its use. According to the composition for controlling microorganisms that inhibit the growth and quality of ginger according to the present invention and the method for controlling microorganisms using the same, since it has the effect of killing soft rot-causing bacteria, it can be preferably used for controlling these microorganisms, and the initial stage of the ginger cultivation process It is thought that bacteria that cause soft rot of ginger can be preferentially controlled in the cultivation area composition, which is the stage, and natural control effects can be seen when early symptoms occur during the growth process. In addition, it is considered that it can serve as a natural bactericidal preservative that can prevent quality-degrading bacteria from entering from the outside by using it for storage after harvesting ginger.

Description

Antibacterial extract for cultivating ginger growth and quality-inhibiting microorganisms and its use {Antibacterial composition for cultivating ginger and the use thereof}

The present invention relates to an antibacterial extract against ginger growth and quality-inhibiting microorganisms and its use.

Ginger is an important crop along with garlic as a seasoning vegetable. Its origin is in subtropical regions with hot and humid environments such as India and Malaysia. In Korea, it is sown from mid-April to early May and harvested before frost in October. It is a long crop.

In general, crops have an adverse effect on yield and quality of crops if the cultivation method is not suitable with the growing environment and soil, and ultimately causes a decrease in the income of farmers who grow crops. Insecticides, etc. are used, but there is a problem that the soil quality deteriorates or the soil is devastated due to excessive use.

As a technology for controlling pests and diseases for ginger cultivation, Korean Publication No. 10-2005-0014286 (February 7, 2005) describes a method for cultivating ginger or garlic, which is a root vegetable. Seed preparation step; A land leveling step of making ditches and furrows while evenly plowing the ground so that roots grow well after applying manure such as compost and fertilizer; a seeding step of planting the seeds on the ridge; A paper coating step of coating paper over the ridge where seeds are sown in order to prevent weeds from occurring; A second coating step of coating the paper coated on the ridge with rice straw or pine needles; After the sprouts of ginger or garlic pierce through the paper coating, a method of cultivating ginger or garlic using paper coating is disclosed, comprising a watering step, a manure step, a pest control step, etc. in a conventional manner. there is.

In addition, Korean Publication No. 10-2016-0068366 (June 15, 2016) discloses a method of cultivating ginger, which sprays a soil insecticide when the sprouts rise, and when the sprouts rise 70 to 90% It includes a pest control step of spraying a fungicide when

However, in general farms, there are problems such as management of pollutants in the case of using chemicals, so economical and environmentally friendly methods of using natural materials are still continuously required.

Korean Publication No. 10-2016-0068366 (June 15, 2016) Korea Publication No. 10-2005-0014286 (February 7, 2005)

As a result of intensive research to solve the above problems according to the prior art and develop an eco-friendly antibacterial composition, the present inventors found that the natural extracts described below can be preferably used for the above purpose, and completed the present invention. I came to do it.

Therefore, an object of the present invention, in one aspect, to provide a composition containing an environmentally friendly natural extract for controlling the microorganisms for inhibiting the growth and quality of ginger.

The above object of the present invention can be achieved by providing a composition for controlling microorganisms that inhibit the growth and quality of ginger, characterized in that it comprises hibiscus extract, ginkgo extract, persimmon leaf extract, or a mixture thereof as an active ingredient. .

According to the composition for controlling microorganisms that inhibit the growth and quality of ginger according to the present invention and the method for controlling microorganisms using the same, since it has the effect of killing soft rot-causing bacteria, it can be preferably used for controlling these microorganisms, and the initial stage of the ginger cultivation process It is thought that bacteria that cause soft rot of ginger can be preferentially controlled in the cultivation area composition, which is the stage, and natural control effects can be seen when early symptoms occur during the growth process. In addition, it is considered that it can serve as a natural bactericidal preservative that can prevent quality-degrading bacteria from entering from the outside by using it for storage after harvesting ginger.

1 is a manufacturing process diagram of an antibacterial extract for ginger growth and quality-inhibiting microorganisms.
Figure 2 is a separation process diagram of microorganisms inhibiting the growth of ginger quality.
Figure 3 is an antibacterial test process chart for ginger quality and growth-inhibiting microorganisms by the paper disk method.
Figure 4 is an antibacterial test process chart for ginger quality and growth-inhibiting microorganisms by measuring the number of bacteria.
5 and 6 are sequence comparison diagrams showing the results of identifying Pseudomonas chlororaphis strains by gene sequencing.
7 and 8 are sequence comparison diagrams showing the results of identifying Pseudomonas plecoglossicida strains by gene sequencing.
9 and 10 are sequence comparison diagrams showing the results of identifying Pseudomonas peli strains by gene sequencing.
11 and 12 are sequence comparisons showing the results of Bacillus aerius strain identification by gene sequencing.
13 and 14 are sequence comparison diagrams showing the results of identification of Pectobacterium carotovorum strains by gene sequencing.
15 and 16 are sequence comparison diagrams showing results of identification of Paenibacillus mobilis strains by genetic sequencing.
17 and 18 are sequence comparisons showing the results of Oerskovia enterophila strain identification by gene sequencing.

In one aspect, the present invention provides a composition for controlling microorganisms that inhibit the growth and quality of ginger, comprising a hibiscus extract, a ginkgo extract, a persimmon leaf extract, or a mixture thereof as an active ingredient.

In a further aspect of the present invention, the extract is prepared by washing the raw material, drying the washed raw material naturally or using a dryer, mixing 70% fermented alcohol and the raw material at a weight ratio of 9: 1 It provides a composition for controlling microorganisms that inhibit the growth and quality of ginger, characterized in that it is prepared by extracting at room temperature for 22 to 25 hours, and filtering each extract obtained by extraction.

In another further aspect of the present invention, the composition is a composition for controlling microorganisms that inhibit the growth and quality of ginger, characterized in that the composition comprises a hibiscus extract and a persimmon leaf extract in a ratio of 1:0.5 to 2 based on weight. provides

In another further aspect of the present invention,

Preparing a hibiscus extract, ginkgo biloba extract or persimmon leaf extract or a mixture thereof; and

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It provides a method for controlling microorganisms that inhibit the growth and quality of ginger, comprising the step of applying the extract or mixture to ginger planted or stored in a harvested state.

Hereinafter, a composition for controlling microorganisms that inhibit the growth and quality of ginger according to the present invention and a method for controlling microorganisms using the same will be described in more detail with reference to the accompanying drawings.

Terms or words used in this specification and claims should not be construed as being limited to ordinary or dictionary meanings, and the inventor may appropriately define the concept of terms in order to explain his or her invention in the best way. It should be interpreted as a meaning and concept consistent with the technical idea of the present invention based on the principle that there is. Therefore, the embodiments described in this specification and the configurations described in the drawings are only the most preferred examples of embodiments, and do not represent all of the technical spirit of the present invention, so various equivalents and It should be understood that variations may exist.

According to the composition for controlling microorganisms that inhibit the growth and quality of ginger according to the present invention and the method for controlling microorganisms using the same, as confirmed in the analysis results described later, when using a hibiscus extract at a high concentration of 100,000 ppm, it is effective in killing soft rot-producing bacteria. It can be preferably used for the control of these microorganisms.

The composition may more preferably include a hibiscus extract and a persimmon leaf extract in a ratio of 1:0.5 to 2 based on weight for efficient control of microorganisms affecting the quality of ginger.

In particular, it is believed that bacteria that cause soft rot of ginger can be preferentially controlled in the plantation composition, which is the initial stage of the ginger cultivation process, and natural control effects can be seen when early symptoms occur during the growth process.

In addition, it is considered that it can serve as a natural bactericidal preservative that can prevent quality-degrading bacteria introduced from the outside by using it for storage after harvesting ginger.

Figure 1 is a manufacturing process diagram of an antibacterial extract against ginger growth and quality-inhibiting microorganisms, Figure 2 is a separation process diagram of ginger quality growth-inhibiting microorganisms, Figure 3 is an antibacterial test process diagram for ginger quality and growth-inhibiting microorganisms by the paper disk method 4 is an antibacterial test process diagram for ginger quality and growth-inhibiting microorganisms by measuring the number of bacteria.

As shown in Figure 1, the antibacterial extract according to the present invention can be prepared by washing, drying, extracting and filtering raw materials such as hibiscus, ginkgo leaves, and persimmon leaves.

First, in a1) washing step, the raw material is washed with clean water. The washing removes foreign substances such as dust, soil, insects, etc. from the raw material in a manner such as spraying purified water, and the quality of the finally manufactured fruit tablets can be prevented from deterioration by bacteria inhabiting the foreign substances in the subsequent process. .

Then, in the drying step a2), the washed raw materials are naturally dried or dried using a dryer.

In the case of natural drying, for example, the raw material is spread evenly to a thickness of 1 to 2 cm in a well-ventilated room, placed on a shelf or a net, turned over intermittently for 3 to 30 hours at room temperature, and dried so that moisture evaporates evenly. It is better to leave it in a state where it is properly oxidized.

On the other hand, the drying time when drying under direct sunlight is 0.2 to 1 hour depending on the amount of sunlight, preferably 0.5 to 1 hour.

In addition, when using a dryer, it may be preferable to dry at a temperature of 50 ° C. to 65 ° C. or less for 2 hours to 48 hours.

a3) Next, in the extraction step, it may be desirable to mix 70% fermented alcohol and raw materials in a weight ratio of 9:1 and extract for 22 to 25 hours at room temperature (eg, 28 to 32 ° C, preferably 30 ° C). there is.

In the natural product extraction process, the destruction of active ingredients is minimized, and in the alcohol extraction process, the alcohol component is volatilized during high-temperature extraction, reducing the capacity and preventing the additional step of dilution by the reduced amount, and also reducing the risk of fire and explosion during high-temperature extraction. room temperature extraction was performed.

a4) filtration step

Each extract obtained by extraction can be filtered by filtering out solid foreign substances and particles using, for example, a filter membrane, paper filter paper, hemp, nonwoven fabric, or cotton, or using ultrafiltration, cryofiltration, centrifugation, etc. . In addition, it may be preferable to perform filtration by first filtering using gauze and then performing secondary filtering using Whatman paper and then refrigerating for subsequent treatment.

The extract extracted as above can be preferably used as a composition for improving the antibacterial or growth environment of ginger by mixing in an appropriate mixing ratio or combining with additives known in the field of ginger cultivation or growth.

<Example>

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention.

Example 1: Preparation of antibacterial extract

As shown in FIG. 1, individual extracts were prepared by washing, drying, extracting, and filtering raw materials such as hibiscus, ginkgo leaves, and persimmon leaves.

After washing each raw material with clean water, spread it evenly to a thickness of 1 to 2 cm in a well-ventilated room, place it on a shelf, and dry it by turning it over intermittently for 24 hours at room temperature so that moisture evaporates evenly. 70% fermented alcohol and raw materials were mixed at a weight ratio of 9: 1, extracted at room temperature (30 ° C) for 24 hours, filtered, and the supernatant was used in subsequent tests.

Example 2: Isolation of microorganisms inhibiting the growth of ginger quality

After taking a sample from ginger and diluting it 10-fold with sterile water, 1% peptone water was used to separate and count microorganisms from the diluted solution to 10 ^-1 , 10 ^-2 , and 10 ^-8 Diluted to concentrations , Take 1 ml of the diluted solution, spread it on a plate medium (PCA), incubate at 30 ° C. for 48 hours, isolate genomic DNA from colonies that appear, perform 16S rDNA sequencing to identify microorganisms, and the remaining diluted samples are liquid medium ( MRS broth) at 30 ° C for 48 hours and then stored frozen (-70 ° C).

The strains isolated in the above example were subjected to comparative analysis of sequences with the following strains as targets for comparison, and the results are shown in Table 1 and FIGS. 5 to 18.

Table 1 shows the identification results of microorganisms. 5 and 6 are sequence comparison diagrams showing results of identification of Pseudomonas chlororaphis strains by gene sequencing, and FIGS. 7 and 8 are sequence comparisons showing results of identification of Pseudomonas plecoglossicida strains by gene sequencing, and FIGS. 9 and 10 are gene sequence comparisons. A sequence comparison diagram showing the results of Pseudomonas peli strain identification by sequencing, Figures 11 and 12 are sequence comparison diagrams showing the results of Bacillus aerius strain identification by gene sequencing, and Figures 13 and 14 are Pectobacterium carotovorum by gene sequencing. 15 and 16 are sequence comparison diagrams showing results of identification of Paenibacillus mobilis strains by genetic sequencing, and FIGS. 17 and 18 are sequences showing results of identification of Oerskovia enterophila strains by genetic sequencing. is a comparison.

The strains used for gene sequencing as public strains are Pseudomonas chlororaphis KCCM 11363 strain (ATCC 9446), Pseudomonas plecoglossicida strain (ATCC 700383), Pseudomonas peli strain (KCTC 22660), Bacillus aerius strain (KCTC 13384), Pectobacterium carotovorum subsp. carotovorum KCCM 11319 (ATCC 15390) strain, Paenibacillus mobilis strain (KCTC 33848), and Oerskovia enterophila strain (KCTC 19998) can be used.

As a result of microbial identification, a total of 7 microorganisms were analyzed, and one bacterium causing soft rot was detected.

As a result of sequence comparison, it was confirmed that the strains had 98% or more homology with the published strains.

Example 3: Antibacterial test for ginger quality and growth-inhibiting microorganisms by paper disk method

Antibacterial tests for ginger quality and growth-inhibiting microorganisms by the paper disk method were performed on the diluted samples as shown in FIG.

Diameter of the clear zone formed on each medium after smearing microorganisms on a sterilized plate medium, loading the sample (100, 1000, 10,000, 100,000) for each concentration using a paper disk and incubating it in an incubator at 30 ° C for 48 hours The antibacterial activity was compared by measuring. The results are shown in Tables 2 and 3. Persimmon: Persimmon leaf extract, Silver: Ginkgo biloba extract, Hi: Hibiscus extract.

division persimmon silver he Pseudomonas chlororaphis 1.1 2 Pseudomonas plecoglossicida 1.7 Pseudomonas peli 1.6 2.8 2.8 Bacillus aerius 1.6 2.3 Pectobacterium carotovorum 1.2 1.7 Paenibacillus mobilis 2.8 2.9 Oerskovia enterophila 1.5 1.4

division persimmon + silver silver + he hi + feeling Persimmon + Eun + Hee Pseudomonas chlororaphis 1.8 1.8 1.7 Pseudomonas plecoglossicida 1.7 1.8 1.5 Pseudomonas peli 1.2 1.4 One 2.1 Bacillus aerius 1.1 1.3 1.4 Pectobacterium carotovorum 1.5 1.8 1.4 Paenibacillus mobilis 2.5 2.5 2.5 2.8 Oerskovia enterophila 1.8 1.8 1.9 2

As a result of the antibacterial activity test, the hibiscus extract alone showed antibacterial efficacy against all 7 types of microorganisms, the persimmon leaf extract showed antibacterial efficacy against 5 types and the ginkgo leaf extract showed antibacterial efficacy against 2 types.

Example 4: Antibacterial test for ginger quality and growth-inhibiting microorganisms by bacterial count measurement

Antibacterial tests for ginger quality and growth-inhibiting microorganisms by measuring the number of bacteria were performed as shown in FIG. 4.

To this end, after inoculating the microorganism in a liquid medium, each of the hibiscus, ginkgo leaf, and persimmon leaf extracts prepared in Example 1 was mixed and loaded at 10% or 5%, cultured in an incubator at 30 ° C. for 48 hours, and then peptone After diluting with water, it was spread on a plate medium, and again incubated at 30 ° C. for 48 hours to measure the number of bacteria, and the results are shown in Tables 4 to 6. Table 6 shows the antibacterial efficacy evaluation results for each hibiscus concentration.

division common persimmon leaves ginkgo leaves hibiscus Pseudomonas chlororaphis 1.0×10 ⁸ 1.0×10 ⁸ 1.0×10 ⁸ 7.0×10 ⁵ Pseudomonas plecoglossicida 1.2×10 ¹¹ 1.2×10 ¹¹ 1.2×10 ¹¹ 1.0×10 ⁵ Pseudomonas peli 3.8×10 ¹⁰ 1.0×10 ⁵ 3.8×10 ¹⁰ 1.0×10 ⁵ Bacillus aerius 3.8×10 ¹⁰ 1.0×10 ⁷ 1.8×10 ⁸ 3.0 × 10 ⁶ Pectobacterium carotovorum 7.3×10 ¹⁰ 7.3×10 ¹⁰ 1.5×10 ⁸ 2.0×10 ⁶ Paenibacillus mobilis 7.0×10 ¹¹ 4.0×10 ⁶ 1.2×10 ⁸ 1.0×10 ⁵ Oerskovia enterophila 4.6×10 ¹⁰ 1.0×10 ⁷ 3.0×10 ⁸ 2.0×10 ⁶

division winding dare Eunhi dare Pseudomonas chlororaphis 1.5×10 ⁹ 3.0×10 ⁶ 2.0×10 ⁷ 2.0×10 ⁷ Pseudomonas plecoglossicida 2.1×10 ⁸ 2.0×10 ⁶ 0 1.0×10 ⁶ Pseudomonas peli 2.0×10 ⁶ 2.0×10 ⁷ 0 2.0×10 ⁶ Bacillus aerius 1.0×10 ⁷ 2.0×10 ⁶ 1.0×10 ⁷ 2.0×10 ⁶ Pectobacterium carotovorum 3.1×10 ⁷ 4.0×10 ⁵ 1.3×10 ⁶ 3.0×10 ⁶ Paenibacillus mobilis 0 1.0×10 ⁷ 1.0×10 ⁶ 1.0×10 ⁶ Oerskovia enterophila 3.0×10 ⁷ 1.0×10 ⁶ 1.0×10 ⁶ 1.0×10 ⁶

division 100ppm 1,000ppm 10,000ppm 100,000ppm Pseudomonas chlororaphis 2.7 × 10 ⁸ 1.6 × 10 ⁸ 7.3 × 10 ⁸ 1.0 × 10 ⁶ Pseudomonas plecoglossicida 1.6 × 10 ⁹ 1.1 × 10 ⁹ 6.2 × 10 ⁹ 1.0 × 10 ⁷ Pseudomonas peli 7.0 × 10 ⁵ 1.0 × 10 ⁵ 1.0 × 10 ⁵ 3.0 × 10 ⁵ Bacillus aerius 5.2 × 10 ⁷ 2.8 × 10 ⁸ 4.0 × 10 ⁸ 3.0 × 10 ⁶ Pectobacterium carotovorum 8.4 × 10 ⁷ 3.8 × 10 ⁷ 1.6 × 10 ⁷ 0 Paenibacillus mobilis 3.0 × 10 ⁵ 1.0 × 10 ⁵ 0 0 Oerskovia enterophila 2.5 × 10 ⁸ 9.0 × 10 ⁶ 5.8 × 10 ⁷ 0

As a result of the antibacterial test, in the growth inhibition result using natural (hibiscus, persimmon leaf, ginkgo leaf) extracts, it was analyzed that the growth of 7 microorganisms was inhibited when hibiscus was used alone.

In addition, as a result of the growth inhibition test by mixing each natural product, the inhibition rate was lower than that of hibiscus alone, but higher than that of persimmon leaf and ginkgo leaf alone. On the other hand, when persimmon leaves and hibiscus were mixed, the inhibitory effect of Pectobacterium carotovorum , a bacterium causing soft rot, was increased. Therefore, it is thought that the use of hibiscus in combination with incision rather than alone will be effective in inhibiting soft rot-producing bacteria.

Regarding the concentration, it was analyzed that the hibiscus extract at a high concentration of 100,000 ppm was effective in killing soft rot-producing bacteria.

According to this test, it is considered that the priority is to control bacteria that cause soft rot of ginger in the plantation composition, which is the initial stage of the ginger cultivation process, and natural control effects can be seen when early symptoms occur during the growth process.

In addition, it is thought that it can serve as a natural sterilization preservative that can prevent quality-degrading bacteria from entering from the outside by using it to store ginger harvest water.

As described above, the present invention has been described by the limited embodiments and drawings, but the present invention is not limited to the above embodiments, and those skilled in the art in the field to which the present invention belongs can make various modifications and transformation is possible Therefore, the spirit of the present invention should be grasped only by the claims described below, and all equivalent or equivalent modifications thereof will be said to belong to the scope of the present invention.

Claims (4)

Contains hibiscus extract and persimmon leaf extract as active ingredients, wherein the extract includes washing raw materials, naturally drying the washed raw materials or drying them using a dryer, 70% fermented alcohol and raw materials at a weight ratio of 9:1 It is prepared by mixing and extracting at room temperature for 22 to 25 hours, and filtering each extract obtained by extraction, wherein the hibiscus extract and persimmon leaf extract are included in a ratio of 1: 1 based on weight, A composition for controlling microorganisms that inhibit the growth and quality of ginger, characterized in that the concentration of the hibiscus extract is 100,000 ppm. delete delete delete