KR20150049136A - Plant disease comprising and method for manufacturing the same - Google Patents

Abstract

The present invention relates to a composition for preventing plant diseases and a manufacturing method thereof wherein the composition comprises Bacillus amyloliquefaciens M27 whose accession number is KACC91208P strain culture medium or residual culture medium except for mycobint from the culture medium as an active ingredient, thereby having effects of preventing powdery mildew or inhibiting phytopathogenic germ and facilitating plant growth for at least one among Sclerotinia nivalis, Alternariasp._ and Colletotrichum panacicola.

Description

[0001] The present invention relates to a composition for controlling plant diseases and a method for producing the same,

The present invention relates to a composition for controlling a plant disease and a method for producing the same, and more particularly, to a plant disease control composition which exhibits an effect of controlling powdery mildew disease or controlling a plant bacterium, Compositions and methods for their preparation.

Control of plant diseases has typically relied on extensive use of toxic chemicals (pesticides).

These organic synthetic pesticide control methods often require toxic chemicals in the soil as well as the target plants and plant products, resulting in plant residue and soil residual toxicity problems, It is toxic to microorganisms and animals, and is known to be harmful to humans.

Therefore, much of the recent research on plant disease control is aimed at finding an environmentally friendly plant disease control method, and one of them is the development of biopesticides using bio-resources that are antagonistic to plant pathogenic microorganisms.

An antagonistic microorganism is a microorganism that inhibits the growth of another microorganism when cultured with another kind of microorganism. Antagonistic microorganisms are known to cause parasitism, predation, and antibiotic action.

Korean prior art 10-1260170 entitled " Pseudomonas CP236, a novel strain having antimicrobial activity, and a plant disease control composition containing the same, " or Korean Patent No. 10-1166853 " have.

An object of the present invention is to provide a composition for controlling plant diseases and a method for producing the same, which can improve the control effect of powdery mildew disease, inhibit bacterial pathogen growth, and control plant bacterium, will be.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

In order to achieve the above object, the present invention provides a plant disease controlling composition comprising Bacillus amyloliquefaciens M27 KACC91208P strain culture medium or a culture filtrate excluding the bacterial cells as an active ingredient, effects or scale Klee Loti Chania you balises (Sclerotinia nivalis), Alter and Leah spp (Alternaria sp.), Colle sat tree Colchicum waves Nicholas fungus (Colletotrichum panacicola), Shirin draw carboxylic phones des True Tansu (Cylindrocarpon destructans), Fish helpful Pseudomonas spp . , Pythium sp . , Pectobacterium carotovora subsp. Cartovorum , Acidovorax citrulli , Agrobacterium tumefacienes , At least one selected from Burkholderia glumae , Ralstonia solanacearum , And has the effect of inhibiting plant pathogenic bacteria and promoting plant growth.

The culture solution is characterized by being cultured in LB medium.

The LB culture broth is characterized in that the Bacillus amyloliquefaciens M27 KACC91208P strain is cultured at 26 to 33 ° C for 2 to 3 days.

The culturing solution or the culturing solution is characterized by being diluted 2 to 10 times with the culture filtrate excluding the cells.

The culture filtrate or the culture filtrate for the culture broth excluding the microbial cells contains 10 to 50% by weight based on the total weight of the composition.

The powdery mildew is grapes spa as fuse car (Podosphaera fusca), Spy Rotterdam car sick varnish (Sphaerotheca aphanis), Spy Rotterdam car plate labor (Sphaerotheca pannosa), Erie Shirai pesok beginning serum (Erysiphe cichoracearum), L'first base la tau Rica ( Leveillula taurica ). ≪ / RTI >

The composition may further include 0.01 to 0.05% by weight of an electrodeposition agent based on the total weight of the composition.

The plant is characterized in that it exhibits the growth promoting effect of tomato or pepper.

The method for producing a plant disease controlling composition of the present invention comprises the steps of: preparing a strain of Bacillus amyloliquefaciens M27 KACC91208P; preparing the strain by incubating the strain in an LB medium at 26 to 33 DEG C for 2 to 3 days; A second step of preparing a culture solution or a culturing filtrate excluding the cells for the culture solution, and a second step of culturing the culture solution prepared in step 2 or the culture solution excluding 0.01-2.0 wt% And a third step of preparing a composition for preventing plant diseases, which comprises the step of controlling a powdery mildew disease or a method of controlling a disease preventing effect of Sclerotinia nivalis , Alternaria sp . , Colletotrichum panana Nicholas fungus (Colletotrichum panacicola), Shirin von draw carboxylic True death Tansu (Cylindrocarpon destructans), Fish Umm spp. (Pythium sp.), Peck tobak Te Karo Solarium sat Fora variants Carl Topo Bacteria (Pectobacterium carotovora subsp. Cartovorum), walk Locks sheet rules rigyun (Acidovorax citrulli) know, Agrobacterium-to-mepa sieon seugyun (Agrobacterium tumefacienes), Burkina kolde Ria glue megyun (Burkholderia glumae), Central Stony Oh Solana Asia rumgyun ( Ralstonia solanacearum ) against plant pathogenic bacteria and plant growth promoting effect.

In the third step, the culture filtrate or the culture filtrate for the culture broth excluding the microbial cells contains 10 to 50% by weight based on the total weight of the composition.

The plant growth promotion is characterized by being tomato or pepper.

The plant disease control method of the present invention is characterized in that an effective amount of the plant disease control composition prepared by the above-described production method is sprayed on the host plants.

The host plant is characterized in that it is at least one selected from cucumber, amber, melon, watermelon, melon, pak, strawberry, rose, tomato and pepper.

According to the present invention, there is provided a plant disease controlling composition capable of enhancing the control effect against powdery mildew, at the same time inhibiting bacterial pathogen growth, preventing plant bacterium disease, promoting plant growth, and enabling development of microbial fertilizer.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a diagram showing the nucleotide sequence of Bacillus amyloliquefaciens M27 KACC91208P of the present invention.
2 is a graph showing the effect of B. amyloliquefaciens strain M27 on the control of cucumber powdery mildew according to the culture medium and the culture period.
FIG. 3 is a graph showing the comparison of the activity of controlling the powderiness of cucumber powdery mildew according to the dilution ratios of the culture medium of B. amyloliquefaciens M27 strain
4 is a graph showing the activity of powdery mildew according to the culture temperature and incubation period in the selection medium.
FIG. 5 is a graph showing the inhibitory effect of B. amyloliquefaciens strain M27 on LB broth medium and the occurrence of cucumber powdery mildew by culture temperature and culture days.
FIG. 6 is a graph showing the activity of controlling the powdery mildew of the culture solution, culture filtrate and cell suspension. FIG.
Fig. 7 is a graph showing the activity of controlling the cucumber powdery mildew in the culture broth, culture filtrate, and cell suspension cultured in LB medium of B. amyloliquefaciens M27 strain.
FIG. 8 is a graph showing the effect of B. amyloliquefaciens strain M27 on cucumber powdery mildew control according to the treatment time.
Fig. 9 is a graph showing the effect of B. amyloliquefaciens M27 on mycelial growth of plant pathogenic bacteria.
(1) Collectotrichum panacicola
(2) Cylindrocarpon destructans
(3) Pythium sp.
(4) Sclerotinia nivalis
(5) Alternaria sp .

The terms, techniques, and the like described in this specification are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified.

The present invention relates to a composition for controlling a plant disease.

The active ingredient contained in the plant disease controlling composition is preferably a culture solution of a strain of Bacillus amyloliquefaciens M27 KACC91208P, or a culture filtrate excluding the cells for the culture solution.

The Bacillus amyloliquefaciens M27 KACC91208P strain of the present invention can be used in the present invention to obtain a suitable optimum medium such as a suitable medium type and a dilution factor for obtaining a powdery mildew control effect, Cultivation conditions and the like, it is possible to provide a composition for controlling plant diseases, which is capable of controlling bacterial pathogens by inhibiting growth of bacterial pathogens, enabling the development of microbial fertilizers by promoting plant growth, I will.

In other words, to, as shown in Table 4. The powdery mildew is grapes spa as fuse car (Podosphaera fusca), Spy Rotterdam car sick varnish (Sphaerotheca aphanis), Spy Rotterdam car plate labor (Sphaerotheca pannosa), Erie when pesok beginning La serum (Erysiphe cichoracearum ), Leveillula taurica , and the grape spa Podosphaera fusca is commonly used for cucumber, amber, melon, watermelon, melon and foil. , the spy Lanzarote car sick Nice (Sphaerotheca aphanis) is in the subject tomatoes intended for strawberries, the spy Rotterdam car plates labor (Sphaerotheca pannosa) intended for roses, Eri Shirai pesok beginning Serum (Erysiphe cichoracearum), the Leveillula taurica ( Leveillula taurica ) is a strain causing powdery mildew in red pepper. When the antimicrobial activity was confirmed, all of the antimicrobial activities were confirmed.

In addition, as shown in the following Tables 5, 6 and 9, it is possible to obtain a plant disease-causing fungus Sclerotinia nivalis , Alternaria sp . , Colletotrichum pneumoniae Colletotrichum panacicola , Cylindrocarpon destructans , Pythium sp . Pectobacterium carotovora subsp. Cartovorum , which exhibits antimicrobial activity against at least one phytopathogenic fungus selected from the group consisting of Pectobacterium carotovora subsp. Cartovorum causing fruit rot of the vegetable , Ashobloxacum causing fruit rot of watermelon Acidovorax citrulli , which causes root canal disease of vegetables and fruits Agrobacterium tumefacienes , Burkholderia glumae causing bacterial rice blight of rice , Ralstonia solanacearum causing root blight of branches and plants, Which is an antimicrobial activity against at least one plant pathogenic bacterium.

In addition, as shown in Table 7 below, it was confirmed that tomato and pepper also exhibited plant growth promoting effect, and thus it was confirmed that it could be used as a microbial fertilizer.

Hereinafter, the method of preparing the composition for controlling plant diseases according to the present invention will be described in detail as follows.

1. First step; Strain preparation

A strain of Bacillus amyloliquefaciens M27 KACC91208P is prepared.

In order to isolate microorganisms from lettuce rootstocks in the lettuce cultivation package, the strain is heat treated at 80 ° C for 7 minutes, plated on 10% TSA medium, and cultured.

Identification of the isolates was carried out by examining the size, shape and spore position of the isolates. The physiological examination was carried out by temperature reaction, pH, and various biochemical experiments. Molecular biology was carried out by isolating the DNA and determining the base sequence of 16S rDNA It was identified as Bacillus subtilis M. 27 in December 2006 and was deposited with KACC 91208P on December 6, 2006.

However, on May 21, 2013, a record of its scientific nature and taxonomic location marking and modifications was found to be applicable to Bacillus amyloliquefaciens .

Therefore, the strain used hereafter is referred to as " Bacillus amyloliquefaciens M27 KACC91208P strain" represented by the nucleotide sequence of Fig.

2. The second step; culture

The prepared strains are cultured in LB medium at 26 to 33 DEG C for 2 to 3 days.

Bacillus amyloliquefaciens M27 KACC91208P, a newly named strain, was prepared by using tryptic soy broth, NB (nutrient broth), LB (Luria- Bertani broth), KB (King's B broth), and MHB (Mueller hinton broth) medium. As a result, it was confirmed that LB medium was most suitable (as shown in FIG. 2)

Thus, the Bacillus amyloliquefaciens M27 KACC91208P strain prepared above is preferably cultured in LB medium.

Further, it was confirmed that culturing at 26 to 33 ° C for 2 to 3 days was most suitable for culturing temperature and culture period, which showed optimization for the plant disease control effect even in the LB medium (FIG. 3, 4)

In other words, it is difficult to obtain the desired control effect of the present invention when the strain is cultured for less than 2 days in the LB medium, and it is difficult to expect a further increase effect when culturing for more than 3 days. Further, It is difficult to obtain the desired effect of the present invention and it is difficult to expect a further increased effect when cultivation is conducted at a temperature higher than 33 ° C.

Thus, the optimum control effect of the strain on the plant disease varies according to conditions such as the kind of medium, culture temperature, culture period, and the like.

In addition, the culture solution or the culture solution excluding the microbial cells may be used as it is or diluted 2 to 10 times. Described below is that the number of microorganisms contained in the culture medium itself is about 1 × 10 ⁸ cfu / ml, and the dilution thereof or the culture filtrate thereof (a filtrate obtained by filtering microbes by filtration in a liquid medium in which microorganisms have been cultured) It was found that the use of the composition as an active ingredient of the composition for controlling plant diseases was the most suitable.

At this time, when the dilution ratio is too high, it is rather difficult to obtain the desired control effect of the present invention because the amount of the bacteria present is rather small, so it is best to dilute the dilution to 2 to 10 times at the dilution (FIG. 7, )

3. The third step; Preparation of composition for plant disease control

The culture solution cultured in step 2 or the cultured solution is added to the culture filtrate except the cells for 0.01 to 0.05% by weight based on the total weight of the composition to prepare a composition for controlling plant disease.

Here, the electrodeposition agent is a kind of auxiliaries used for spreading the pesticide spraying solution on the surface of plants or pests, and it is possible to further improve the effect of controlling plant diseases by adding a complexing agent (Table 3).

At this time, the electrodeposition agent is preferably contained in an amount of 0.01 to 0.05% by weight based on the total weight of the composition. If the electrodeposit is contained in an amount of less than 0.01% by weight, it is difficult to obtain a synergistic effect with respect to the effect of controlling only the culture filtrate except for the bacterial cells in the culture solution. If the concentration exceeds 0.05% by weight, It is not elevated in the disease control effect and is abused.

In addition, it is preferable that the culture filtrate or the culture filtrate for the culture fluid excluding the microbial cells contains 10 to 50% by weight based on the total weight of the composition. This is because it is difficult to obtain the desired plant disease control effect of the present invention when the cultured solution or culture filtrate except for the cells is contained in an amount of less than 10% by weight based on the total weight of the composition. If the culture filtrate contains more than 50% by weight, The mixing ratio with other materials to be used is not matched and it becomes difficult to commercialize them.

The thus prepared plant disease control composition of the present invention not only exhibits excellent control effect against powdery mildew but also exhibits an inhibitory effect against specific fungi of plant hospitals and plant pathogens and promotes growth of plants such as tomatoes and peppers .

Accordingly, the present invention provides a plant disease controlling composition as described above, which is used by spraying an effective amount on a host plant, wherein the host plant is selected from the group consisting of cucumber, pumpkin, melon, watermelon, melon, , And red pepper.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following examples and experimental examples are provided only for the understanding of the present invention, and the scope of the present invention is not limited to the following examples and experimental examples.

<Example 1> Preparation of composition for controlling plant diseases of the present invention

A strain of Bacillus amyloliquefaciens M27 KACC91208P having the nucleotide sequence shown in Figure 1 was prepared

The prepared strains were cultured in LB medium for 2 to 3 days at 26 to 33 ° C.

The cultured culture solution was diluted 10-fold with water, and a plant disease control composition of the present invention containing 50% by weight of the diluted culture solution and 0.03% by weight of an electrodeposition agent (purge) was prepared.

Experimental Example 1 Effect of Cultivation Medium and Culture Period on Control of Cucumber Powdery Mildew

1. Experimental Method

(Tryptic soy broth), NB (nutrient broth), LB (Luria-Bertani broth), KB (King's B) and the like, which are commonly used for bacterial culture for the selection of the medium for optimizing the powdery mildew control activity of B. amyloliquefaciens M27 strain of the present invention, broth) and MHB (Mueller hinton broth), respectively.

Each of the sterilized media was inoculated with M27 strain and incubated at 26 ° C in an incubator at 150 rpm for 7 days with shaking.

The cultures were centrifuged (8,000 rpm, 30 minutes) for 1 day to 7 days, and the culture filtrate was diluted 2-fold. The cucumber (crispy whitefish) cultivated in the greenhouse was sprayed twice at 7-day intervals 7 days after the final treatment, the area ratio of powdery mildew disease was examined.

At this time, the composition of the used medium is as follows.

TSB (Tryptic soy broth) medium; Casein 17 g, Soybean 3 g, Dextrose 2.5 g, NaCl 5 g, K ₂ HPO ₄ 2.5 g

NB (Nutrient broth) medium; Beef extract 3g, Peptone 5g

MHB (Mueller hinton broth) medium; Beef extract 2.0g, Acid digest of casein 17.5g, Soluble starch 1.5g

KB (King's broth) medium; Proteose peptone 10 g, Glycerol 15 g, K ₂ HPO ₄ 1.5 g, MgSO ₄ 5 g

LB (Luria Bertani broth) medium; 10 g of NaCl, 10 g of Tryptone, 5 g of yeast extract

2. Experimental results

As shown in FIG. 2, when the LB medium was used, it was confirmed that the B. amyloliquefaciens M27 strain of the present invention had the best effect for controlling powdery mildew in a short period of time.

<Experimental Example 2> Determination of the activity of controlling cucumber powdery mildew by the dilution ratio of the culture medium

1. Experimental Method

In order to select the medium most suitable for the cultivation of strain M27 among the TSB medium, LB medium and MHB medium showing excellent effect of the powdery mildew control activity shown in Experimental Example 1, the culture filtrate diluted by each medium was treated, Respectively.

These media were inoculated with M27 bacteria and cultured in a shaking incubator (150 rpm / 26 DEG C) for 3 days. The cultures were centrifuged, and the culture filtrate was diluted 10 times and 20 times, and then sprayed twice at 7 days intervals in a cucumber (crispy white bag) of the second leaf stage of the main leaf, and the area ratio of powdery mildew disease was measured 7 days after the final spraying. As a control, a chemical agent phenolic emulsion (4,000 times) was used.

2. Experimental results

As shown in FIG. 3, it was found that, when the culture medium in which the LB medium was diluted 10 times in the medium was used, the area ratio of the cucumber powdery mildew disease was the best, and similar to the phenarymine emulsion which is a control chemical commonly used as a plant disease control agent Cucumber powdery mildew disease area ratio.

<Experimental Example 3> Determination of the activity of powdery mildew according to the culture temperature and incubation period in the selection medium

1. Experimental Method

The inhibitory effect of cucumber powdery mildew disease on the cultured filtrate was investigated by culturing the culture temperature of 23 ℃, 26 ℃ and 33 ℃ for 1 day to 3 days using the LB medium with excellent control of powdery mildew.

The cultures were centrifuged, and the culture filtrate was diluted 10 times and 20 times, and then sprayed twice at 7 days intervals in a cucumber (crispy white bag) of the second leaf stage of the main leaf, and the area ratio of powdery mildew disease was measured 7 days after the final spraying.

2. Experimental results

As shown in FIGS. 4 and 5, even when the LB medium was used, it was confirmed that the M27 strain exhibited excellent powdery mildew disease area when cultured at 26 ° C and 33 ° C for 2 days and 3 days.

<Experimental Example 4> B. amyloliquefaciens M27 inhibitory activity against powdery mildew

1. Experimental Method

(1 × 10 ⁸ cfu / ml) of a 10-fold dilution of the culture, a 10-fold dilution of the culture filtrate and a suspension of the culture (1 × 10 ⁸ cfu / ml) were cultured in LB medium at 26 ° C. and 150 rpm for 3 days with shaking After 2 days of spraying and 7 days of final spraying, the lesion area was examined.

2. Experimental results

As shown in FIGS. 6 and 7, it was confirmed that even in the case of using LB medium, the dilution of the culture broth or culture filtrate thereof by 10 times dilution showed better powdery mildew disease area than the bacterial suspension.

<Experimental Example 5> Effect of B. amyloliquefaciens M27 culture filtrate on the control of cucumber powdery mildew by dilution multiples

1. Experimental Method

To determine the effective dilution factor for cucumber powdery mildew disease, cultivate in LB medium at 26 ° C and 150 rpm for 3 days with shaking. Dilute the culture solution to 2, 5, 10, 20, 50, and 100 times, The leaf area of the freshly caught white cucumber was examined by 7 days after the last spraying. In this experiment, 2 repetitions of cucumber seedlings were performed for 5 weeks, and a chemical agent phenaryol emulsion (4,000 times) was used as a control group.

2. Experimental results

Dilution factor (x) Percentage of lesion area (%) Control (%) 2 0.0 ± 0.00 100 5 0.0 ± 0.00 100 10 0.0 ± 0.00 100 20 1.3 ± 0.81 97.8 50 5.0 ± 1.31 91.7 100 40.0 + - 6.54 33.3 Fenarimol EC 12.5 ± 3.78 79.2 No treatment 60.0 + - 5.34 -

As shown in Table 1, it was confirmed that the culture filtrate of the M27 strain of the present invention exhibited a powdery mildew area percentage of 100% when the diluted solution was diluted 2 to 10 times.

<Experimental Example 6> Effect of B. amyloliquefaciens strain M27 on the prevention and treatment of cucumber powdery mildew

1. Experimental Method

검정 Prevention of cucumber powdery mildew in M27 strain

To prevent powdery mildew of the M27 strain, cucumber (crispy white dough) of the second leaf of the main stem was used, and the LB culture solution for 3 days was diluted 10 times, The area ratio of powdery mildew disease caused by spraying and 7 days after the final spraying was examined. In this experiment, 2 repetitions of cucumber seedlings were performed for 5 weeks, and a chemical agent phenaryol emulsion (4,000 times) was used as a control group.

② Treatment effect of cucumber powdery mildew of M27 strain

In order to investigate the effect of M27 strain on powdery mildew, LB culture filtrate was diluted 10 times and sprayed twice in 7-day intervals at 4-leaf cucumber (crispy white) leaves with 50% or more area of powdery mildew in cucumber. The area ratio of powdery mildew disease was investigated. In this experiment, 2 repetitions of cucumber seedlings were performed for 5 weeks, and a chemical agent phenaryol emulsion (4,000 times) was used as a control group.

2. Experimental results

Treatment Area percentage of powdery mildew of cucumber powdery mildew (%) Pre-treatment Post-onset treatment Culture filtrate 10 times 0 ± 0.0 2 ± 1.22 Fenarimol EC 0 ± 0.0 8 ± 1.22 No treatment 47 ± 4.90 92 ± 3.74

As shown in Table 2, the area ratio of powdery mildew disease before and after onset of the LB culture filtrate of strain M27 was shown to be both preventive or therapeutic against powdery mildew.

<Experimental Example 7> Effect of treatment time on control of powdery mildew of cucumber

1. Experimental Method

In order to compare seasonal disease inhibition effects, the culture filtrate of M27 was diluted 10 times and 20 times in July, October and November using cucumber cultivated in the greenhouse to investigate disease inhibition.

The M27 strain was inoculated on LB medium, cultured in a 26 ° C incubator at 150 rpm for 3 days with shaking, and centrifuged. Cucumbers were cucumbers with more than 30% of powdery mildew in the second leaf of the fresh leaf. A control chemical agent phenalmol emulsion (4,000 times) was used.

2. Experimental results

As shown in FIG. 8, it was confirmed that the diluted solution of the culture filtrate of the M27 bacterium of the present invention diluted by 10 times exhibited excellent powdery mildew lesion area ratio regardless of the season.

<Experimental Example 8> Effect of the dilution of the electrodeposition solution on the LB culture of M27 strain to control cucumber powdery mildew

1. Experimental Method

M27 strain was inoculated into LB medium and cultured in a 26 ° C incubator at 150 rpm for 24 hours. Then, the strain was mixed with the culture medium (5 times to 100 times), the culture medium (5 times to 100 times) and the electrodeposition agent The area ratio of cucumber powdery mildew disease was investigated by spraying three times with dilution multiples.

2. Experimental results

Processing contents Area of powdery mildew disease (%) Control (%) Culture medium × 5 1.3 97.5 Culture solution × 10 6.0 88.9 Culture × 20 11.6 78.6 Culture × 30 18.4 66.0 Culture × 40 23.7 56.1 Culture × 50 29.4 45.4 Culture solution × 100 30.8 42.9 Culture medium × 5 + fuzzy (× 3,000) 1.0 98.1 Culture medium × 10 + fuzzy (× 3,000) 2.7 95.1 Culture medium × 20 + fuzzy (× 3,000) 4.0 93.0 Culture medium × 30 + fuzzy (× 3,000) 4.9 90.9 Culture medium × 40 + fuzzy (× 3,000) 8.9 83.5 Culture medium × 50 + Fuzzy medium (× 3,000) 10.2 81.0 Culture medium × 100 + Fuzzy medium (× 3,000) 15.7 70.9 No treatment 53.9 -

As shown in Table 3, it was confirmed that the use of the electrodepositive agent showed an increased control effect against powdery mildew than that using only the culture medium.

Especially, it was confirmed that the use of the electrodeposition agent in the diluted solution diluted 5 ~ 10 times with the culture liquid showed the control effect against the best powdery mildew disease of 95% or more.

<Experimental Example 9> Antimicrobial activity test of M27 strain against powdery mildews of plants

1. Experimental Method

Powdery mildew occurs in the greenhouse while growing cucumber, pumpkin, melon, watermelon, melon, pak, strawberry, rose, tomato and pepper in the greenhouse. At the beginning, the plant disease control composition of the present invention prepared in Example 1 is spray- And to suppress the occurrence of powdery mildew.

2. Experimental results

Powdery mildew Host plant Antimicrobial activity Grapes Spa Ilafuska
( Podosphaera fusca )
(= Sphaertheca fuligenea ) Cucumber, pumpkin, melon
Watermelon, melon, night + Spyroteka Afanis
( Sphaertheca aphanis ) Strawberry + Spyroteka
( Sphaertheca pannosa ) rose + Elixir Perfumed Primrose Serum
( Erysiphe cichoracearum ) tomato + Levile Lauta Rica
( Leveillula taurica ) pepper +

As shown in Table 4, the composition for controlling plant diseases according to the present invention was sprayed on a host plant such as cucumber, pumpkin, melon, watermelon, melon, pak, strawberry, rose, Powdery mildew).

<Experimental Example 10> Inhibitory effect of B. amyloliquefaciens strain M27 on mycelial growth of plant pathogenic bacteria

1. Experimental Method

검정 Antimicrobial activity test of M27 strain against fungus in phytosanitary hospital

Sclerotinia nivalis and Botrytis cinerea in a potato dextrose agar medium (PDA) of a plant hospital fungus were cultured at 20 ° C, Alterna Ria spp (Alternaria sp.), Alterna Ria Panax (Alternaria panax), Collet sat tree glutamicum waves Nicholas fungus (Colletotrichum panacicola), ache draw carboxylic von des true Tansu (Cylindrocarpon destructans), Fusarium oxy's forum bacteria ( Fusarium oxysporum , Fusarium sp . , Pythium sp . , Rhizoctonia solani were cultivated at 25 ° C for 7 to 15 days and cultured in a 5 mm diameter cork box Was inoculated at the center of the PDA medium, and the M27 strain was inoculated into the LB medium at a distance of 3 cm therefrom. The cells were cultured for 24 hours at 28 ° C and 150 rpm for inoculation. After that, each low support was cultured for 8 ~ 15 days.

② Antimicrobial activity test of M27 strain against botanical bacterium

A plant pathogenic bacterium, such as Pectobacterium carotovora subsp. Cartovorum , Acidovorax citrulli , Agrobacterium tumefacienes , Bourcolieria glue, megyun (Burkholderia glumae), LAL Stony Oh Solana cyano rumgyun (Ralstonia solanacearum) and then cultured with shaking for 24 hours in 28 ℃, 150rpm in TSB medium and dispensed into sterile this TSA medium, respectively.

The antimicrobial activity was examined after 5 days of incubation at 28 ° C by treating M27 culture medium in a vacant space separated from TSA medium by 3 mm diameter corckborer.

2. Experimental results

Plant pathogen Mycelial growth inhibition (nm) Sclerotinia nivalis 6.03 ± 0.91 Botrytis cinerea  0.00 ± 0.00 Alternaria sp . 14.17 ± 0.58 Alternaria panax  0.00 ± 0.00 Collectotrichum panacicola 10.28 + - 0.36 Cylindrocarpon destructans 13.68 ± 0.37 Fusarium oxysporum (lettuce)  0.00 ± 0.00 Fusarium sp.  0.00 ± 0.00 Pythium sp.  3.39 ± 0.15 Rhizoctonia solani  0.00 ± 0.00

Plant hospital bacteria Plants and pathology Growth inhibition
(Growth inhibitory ability) Pectobacterium carotovora
subsp. cartovorum Vegetable ripeness + Acidovorax citrulli Fruit rot of watermelon + Agrobacterium tumefacienes A sprinkling of vegetables and edible flowers + Burkholderia glumae Bacterial rice grain blight of rice + Ralstonia solanacearum Foot blight of branches and plants +

As shown in Tables 5, 6 and 9, among the various phytopathological fungi, Sclerotinia nivalis , Alternaria sp . , Colletotrichum panacicola ) , Cylindrocarpon destructans , and Pythium sp . , And among the plant pathogenic bacteria, Pectobacterium carotovora variant Kaltopoleum, which causes the rotting of vegetables, ( Pectobacterium carotovora subsp. Cartovorum ) , Acidovorax citrulli causing watermelon fruit rot, Acidovorax citrulli causing fruit and vegetable root and throat diseases Agrobacterium tumefacienes , Burkholderia glumae causing bacterial rice blight of rice , Ralstonia solanacearum causing root blight of crops and crops, Antibacterial Respectively.

<Experimental Example 11> Plant growth promotion

1. Experimental Method

After sprouting pepper (Manithe) and tomato (Seokwon) seeds, strain M27 was cultured in TSB medium for 2 days at 28 ° C with shaking at 150 rpm, centrifuged at 5,000 rpm for 30 minutes, collected spores and adjusted to 1 × 10 ⁸ / ㎖ And 10 ml each of them were treated 6 times at 7 days intervals to measure plant height, root length and fresh weight. At this time, the measured exponent value is converted to% in terms of non-treatment, and it is processed immediately after sowing.

2. Experimental results

plant Exponent (% compared to no treatment) Plant height Rootstock Fresh weight tomato 115.1 117.0 121.3 pepper 120.1 158.7 148.8

As shown in Table 7, it was confirmed that the composition for controlling plant diseases of tomato and pepper showed the effect of promoting the growth of the plant, and therefore, it could be used as a microbial fertilizer.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (13)

Bacillus amyloliquefaciens M27 KACC91208P) culture broth or a culture filtrate excluding the cells of the culture broth as an active ingredient,
A preventive effect against powdery mildew, or an effective amount of the extract of Sclerotinia nivalis , Alternaria sp . , Colletotrichum panacicola , Cylindrocarpon destructans , Pseudomonas aeruginosa , Pythium sp . , Pectobacterium carotovora subsp. Cartovorum , Acidovorax citrulli , Agrobacterium tumefacienes , Agrobacterium tumefacienes , , Burkholderia glumae , and Ralstonia solanacearum . The method according to claim 1, wherein the plant growth promoting effect is a plant pathogenic bacterium inhibiting effect and the plant growth promoting effect is at least one selected from the group consisting of Burkholderia glumae , Ralstonia solanacearum ,
A composition for controlling plant diseases.
The method according to claim 1,
Wherein the culture medium is cultured in an LB medium.
A composition for controlling plant diseases.
3. The method of claim 2,
Wherein the LB culture medium is obtained by culturing the strain of Bacillus amyloliquefaciens M27 KACC91208P at 26 to 33 DEG C for 2 to 3 days.
A composition for controlling plant diseases.
The method according to claim 1,
Characterized in that the culture solution or the culture filtrate excluding the cells is diluted 2 to 10 times with respect to the culture solution.
A composition for controlling plant diseases.
The method according to claim 1,
Characterized in that the culture filtrate or the culture filtrate excluding the microbial cells for the culture broth contains 10 to 50% by weight based on the total weight of the composition.
A composition for controlling plant diseases.
The method according to claim 1,
The powdery mildew is grapes spa as fuse car (Podosphaera fusca), Spy Rotterdam car sick varnish (Sphaerotheca aphanis), Spy Rotterdam car plate labor (Sphaerotheca pannosa), Erie Shirai pesok beginning serum (Erysiphe cichoracearum), L'first base la tau Rica ( &Lt; / RTI &gt; Leveillula taurica ).
A composition for controlling plant diseases.
The method according to claim 1,
Wherein the composition further comprises 0.01 to 0.05% by weight of an electrodeposition agent based on the total weight of the composition.
A composition for controlling plant diseases.
The method according to claim 1,
Wherein the plant exhibits the growth promoting effect of tomato or pepper.
A composition for controlling plant diseases.
A first step of preparing a strain of Bacillus amyloliquefaciens M27 KACC91208P;
Culturing the prepared strain at 26 to 33 ° C for 2 to 3 days in an LB medium to prepare a culture solution or culture filtrate excluding the cells for the culture solution,
A third step of adding 0.01 to 0.05% by weight of an electrodeposit to the total weight of the composition of the culture solution prepared in step 2 or culturing the culture solution,
A preventive effect against powdery mildew, or an effective amount of the extract of Sclerotinia nivalis , Alternaria sp . , Colletotrichum panacicola , Cylindrocarpon destructans , Pseudomonas aeruginosa , Pythium sp . , Pectobacterium carotovora subsp. Cartovorum , Acidovorax citrulli , Agrobacterium tumefacienes , Agrobacterium tumefacienes , , Burkholderia glumae , and Ralstonia solanacearum . The method according to claim 1, wherein the plant growth promoting effect is a plant pathogenic bacterium inhibiting effect and the plant growth promoting effect is at least one selected from the group consisting of Burkholderia glumae , Ralstonia solanacearum ,
A method for producing a composition for controlling a plant disease.
10. The method of claim 9,
In the third step, the culture filtrate or the culture filtrate for the culture broth, excluding the cells, contains 10 to 50% by weight based on the total weight of the composition.
A method for producing a composition for controlling a plant disease.
10. The method of claim 9,
Wherein the plant growth promotion is tomato or red pepper.
A method for producing a composition for controlling a plant disease.
A plant disease control method, comprising spraying an effective amount of a plant disease control composition prepared by the method of any one of claims 9 to 11 on a host plant.
13. The method of claim 12,
Wherein the host plant is at least one selected from cucumber, pumpkin, melon, watermelon, melon, foil, strawberry, rose, tomato, and red pepper.

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