KR100574277B1 - Antagonistic mold for protection of damping-off disease and bed soil comprising the mold and method thereof - Google Patents

Abstract

The present invention relates to a novel antagonist and a method for producing the antimicrobial soil using the same, and to a method for producing the same, the antimicrobial soil of Tricodeha Hamanum KF-119, KCCM 10582, the antibacterial for the control of mozzarlock disease of the present invention (A) culturing the antagonist Trichoderma hazianum KF-119 (KF-119), KCCM 10582 for the production of antagonistic bacteria cultures for the control of Mozolox disease ; (b) drying and stabilizing the antagonistic culture to produce an antagonistic culture dried product; And (c) dividing the antagonist culture dry matter with the normal soil evenly, and diluting the antagonist culture medium to spray on the soil. It is effective.

Mozlox disease, antimicrobial soil, antagonistic bacteria, Trichoderma hagianum KF-119 (Trichoderma harzianum KF-119), KCCM 10582

Description

Antagonists for the control of mozzarlock disease and antimicrobial clay using the same and manufacturing method thereof {ANTAGONISTIC MOLD FOR PROTECTION OF DAMPING-OFF DISEASE AND BED SOIL COMPRISING THE MOLD AND METHOD THEREOF}

1 is a PDA medium culture photograph of the antagonist Trichoderma hajianum KF-119 ( Trichoderma harzianum KF-119), KCCM 10582 of the present invention.

The present invention relates to an antagonist for the control of mozzarlock disease , and an antibacterial soil using the same and a method for producing the same. More specifically, the novel antagonist Trichoderma hagianum KF-119 ( trichoderma harzianum KF-119), KCCM 10582 and the antagonist It relates to an antimicrobial clay soil for the control of mozzarella disease and a method for producing the same.

Vegetable cultivation of eggplant, tomatoes, cucumbers, etc., first germinate seeds in bed soil, and then grow them by transplanting them into the main fabric after a certain amount of growth.

Top soil is the soil of the bed to grow well, you need to prepare a good top soil that is not contaminated with various pathogens. Good soils should be soft soil with good water retention and drainage, full of organic matter, breathable and good root extension. In addition, the topsoil should be a source of water and various inorganic nutrients during the growth of seedlings, and it should not interfere with the nutrition supply even without feeding during the seedling period.

Damping off is a condition in which young plants become totally dead, and they may be transferred to the main follicles by parasitic young seedlings before or after germination on the soil, and even during transplantation. These mozallocs are found not only in the ground but also in the basement. In the case of the underground, roots, fungi, and late blight are developed and become mozaloc.

Mozalok disease is the most contagious soil infectious disease in eggplant, tomato, and cucumber cultivation, which causes enormous damage in seedlings and early stages because pathogens survive disease in the soil for a long time. According to the 2001 survey on the damage caused by mozzarella disease in the cucumber jujube complex, it is 23% in seedlings and 16% in mainpo.

In order to control such mozzarlock disease, conventional farmhouses have controlled pesticides such as edazole, metasildong, tachimil, dachigaren, geopan, or the like by diluting the solution with soil or diluting the liquid to soil. However, this method of spraying pesticides is somewhat effective when the appropriate amount of spraying at the appropriate time, but it is insufficient in the control of certain pathogens, in particular, it is not very effective for soil diseases, such as mozalok disease. In addition, especially when the seedlings are softly sprayed immediately after germination, or when the onset of cotyledon or main leaf perennial disease is confirmed, misuse of the drug and high concentration spraying may cause weakness. In addition, problems such as soil pollution, environmental pollution, residual toxicity caused by spraying pesticides are also raised.

Recently, a method of using antagonists against pathogens has been proposed in order to solve the problem effectively.

In order to effectively control soil diseases using antagonists, the soil (or soil) settlement rate and growth rate of antagonists should be excellent. These soil settlements are effectively settled and propagated along the roots at the same time as seed germination, and antagonists living in the rhizomes of the seedlings are transferred to the main cell along with the seedlings so that they can be reliably settled in the crop roots of the main cell without the addition of antagonists. Sufficient control should be available.

However, in the conventional method of controlling antagonistic bacteria, the antagonistic bacteria are grown in nutrient media and then produced and marketed as soil microbial agents (antagonist microbial cultures themselves). ) By spraying or spraying.

However, in the conventional method for controlling antagonists, the antagonists are sprayed only on the upper surface of the soil, and when they are sprayed on the seedlings, they are settled in the roots of the seedlings sufficiently in competition with the pathogens or other microorganisms already formed in the root zone of the crop. There is a problem that the control effect is insufficient due to being lost or killed.

 Thus, the present inventors have found a novel antagonist having excellent antagonistic ability against Mozalok disease, and when the antagonist is mixed with the seedling soil to prepare antimicrobial soil and using the same, the seed germination and antagonist bacteria at the same time Therefore, it is effective to settle and breed, and antagonists living in the root of the seedlings are transferred to the main cell along with the seedlings when the main transplant is transplanted, so that the antagonists are firmly settled in the root of crops without additional injection of antagonists. It became.

Therefore, the present invention is to solve the above problems, the present invention provides a novel Trichoderma harzianum KF-119, KCCM 10582 .

In addition, the present invention provides antimicrobial soil for the control of mozzarella disease comprising the dried antagonist Trichoderma hazianum KF-119 ( Kchoophylloma harzianum KF-119), KCCM 10582 .

In addition, the present invention comprises the steps of (a) culturing the antagonistic bacteria Trichoderma hazianum KF-119 ( Tchochoderma harzianum KF-119), KCCM 10582 for the production of antagonistic bacteria ; (b) drying and stabilizing the antagonistic culture to produce an antagonistic culture dried product; And (c) provides a method for producing antimicrobial clay for controlling Mozal rock disease comprising the step of evenly mixing the antagonist cultured dry with normal clay.

Hereinafter, the present invention will be described in detail.

The present inventors have made efforts to isolate and identify novel antagonists with excellent antagonistic activity against Mozallock disease. As a result, the present inventors have found an antagonistic bacterium excellent in the control of Mozallock disease and completed the present invention based on these findings.

The present inventors collected the soil of farmers cultivating pesticide-free cucumbers in Ochang area, Chungbuk, and separated them into sterile distilled water to separate various microorganisms, and then to Rhizoctonia sp., Pythium sp. The antimicrobial activity was evaluated and the final selection of the antagonistic fungus with the best growth inhibitory effect of Mozalloc pathogen was selected.

The main mycological characteristics of the selected fungi are as follows.

[Characteristics in PDA (potato dextrose agar) medium]

1 is antagonistic trichoderma hajianum KF-119 ( Trichoderma) harzianum KF-119), PDA culture of KCCM 10582 .

At the beginning of the culture, it grows into white mycelium, and as time passes, dark green asexual spores are formed, and the whole colony turns green. As it grows, it secretes a yellow pigment, making the back of the colony yellow. The fungus grows radially from the point of inoculation, forming a limbal. The surface of the colony has a green dry spores lumps, and is easily scattered by a slight wind. Dark green oval (obovoid, ellipsoid) conidia (3.2-4.0 X 2.0 X 3.2 μm) and conidia spore diameter of about 3 μm X 12-15 μm . Conidia speculum has long, short secondary branches but secondary branches do not branch again. Mycelia have a diaphragm and form 1-3 branches to form asexual spores

[Physiological characteristics]

Optimal growth pH of the present invention is 4 to 8, the optimum pH of growth is 4.5. In addition, the optimum growth temperature of the present invention bacteria is 20 ~ 32 ℃, the optimum growth temperature is 26 ℃. Arabitol, Starch, Inulin are the best among various carbon sources

In view of the above physiological characteristics, the present inventors named the present invention Trichoderma harzianum KF-119.

Trichoderma harzianum KF-119 ( antagonist Trichoderma harzianum KF-119) for the control of the present invention has been deposited with the accession number of KCCM 10582 to the Korea Microbiological Conservation Center on June 19, 2004.

The antibacterial activity of the Trichoderma harzianum KF-119, KCCM 10582 is shown in Table 1. Antimicrobial activity was measured by dual culture method and inoculated with Mozolox pathogen on one side of PDA medium, Trichoderma harzianum KF-119, KCCM 10582 on the other side for 5 days. After incubation, the bacterial total area of the pathogen was measured.

Trikoderma Haji Anum KF-119 (Trichoderma                                                                          harzianum                                      KF-119), Antimicrobial Activity against Rhizoctonia solani of KCCM 10582 process Size of the Mozalok pathogen mycelial growth ring (㎠) Trikoderma Haji Anum KF-119 6 Trikoderma Haji Anum KF-65 28 No treatment 580

The method for producing antimicrobial clay for controlling mozzarella disease of the present invention,

(a) culturing the antagonistic Trichoderma hazianum KF-119, KCCM 10582 for the production of antagonistic bacteria for the control of Mozolox disease ;

(b) drying and stabilizing the antagonistic culture to produce an antagonistic culture dried product; And

(c) comprising the step of evenly mixing the antagonist cultured dry with normal soil.

Antagonist culture in step (a) is Trichoderma harzianum KF-119, KCCM 10582 solid food medium (rice bran 30%, peat moss 70%, water content 65%) for 7-8 days It can be obtained by culturing at.

In the step (b), the antagonist culture dry can be prepared by drying the antagonist culture at about 50 degrees Celsius or less about 2 days.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are provided only to more easily understand the present invention, and the present invention is not limited by the following examples.

Example 1

The present inventors collected soil of farmers cultivating pesticide-free cucumbers in Ochang area, Chungbuk, and transported them to the laboratory to dilute the microorganisms by continuous dilution 10 times in sterile distilled water. The isolated microorganisms were evaluated for antimicrobial activity against Rhizoctonia sp. And Pythium sp., Which are Mozlox pathogens, in PDA medium by replacement culture method. Was selected as one of the best antagonistic molds. As a result of investigating various microbiological and culture characteristics of the final selection bacteria and the shape, size and color of spores, the present invention was finally named Trichoderma harzianum .

The main mycological characteristics of the selected fungi are as follows.

[Characteristics on PDA Medium]

At the beginning of the culture, it grows into white mycelium, and as time passes, dark green asexual spores are formed, and the whole colony turns green. As it grows, it secretes a yellow pigment, making the back of the colony yellow. The fungus grows radially from the point of inoculation, forming a limbal. The surface of the colony has a green dry spores lumps, and is easily scattered by a slight wind. Dark green oval (obovoid, ellipsoid) conidia (3.2-4.0 X 2.0X3.2㎛) and condensed spore diameter of about 3㎛ X 12-15㎛ Have Conidia speculum has long, short secondary branches but secondary branches do not branch again. Mycelia have a diaphragm and form 1-3 branches to form asexual spores.

[Physiological characteristics]

Optimal growth pH of the present invention is 4 to 8, the optimum pH of growth is 4.5. In addition, the optimum growth temperature of the present invention bacteria is 20 ~ 32 ℃, the optimum growth temperature is 26 ℃. Arabitol, Starch, Inulin are the best among various carbon sources

Example 2

a. Tricoderma Haji Anum KF-119, KCCM 10582 Preparation of Cultured Dried

Trichoderma Hajianum KF-119, KCCM 10582 The preparation of the culture dried is as follows. Antagonist cultures are prepared by culturing Trichoderma hajianum KF-119 and KCCM 10582, which have excellent antimicrobial activity, against 7-day nutrient medium. The antagonist culture is dried at a temperature of 50 degrees Celsius or less to prepare an antagonist culture dry.

b. Mix with normal soil

Trichoderma Hajianum KF-119, KCCM 10582 prepared as described above is added to the general soil and mixed evenly to prepare antimicrobial soil for the control of mozolox disease.

Example 3

Trichoderma Hajianum KF-119 and KCCM 10582 were selected as the best microorganisms with antagonistic resistance against cucumber Rhizoctonia solani . Trichoderma Hajianum KF-119, KCCM 10582 were incubated in solid nutrient medium (30% rice bran, 70% peat moss, 65% water content) for 7 days, dried at 45 degrees Celsius for 2 days, and added to normal soil and mixed evenly. The antimicrobial clay was prepared.

Experimental Example 1 Density Change of Trichoderma Hajianum KF-119, KCCM 10582 in Antimicrobial Soil

The results of analyzing the density of Trichoderma hagia Kum-Kan, KCCM 10582 in the antimicrobial soil prepared in Example 3 are shown in Table 2 below, Trichoderma Hajianum KF-119, KCCM 10582 is 1 in the antimicrobial soil It did not appear to die at all for years.

Analysis results of antagonistic bacteria density in antimicrobial soils over time Days Since Elapsed Early 10 30 60 120 180 360 Trichoderma Hajianum KF-119, KCCM 10582 Density (× 1,000cfu / g) 298 295 297 299 296 295 296

Experimental Example 2 Density Changes of Trichoderma Hagianum KF-119 and KCCM 10582 in the Roots of Seedlings in Seedlings

Cucumber seedlings were carried out using the antimicrobial soil prepared in the same manner as in Example 3, and during the seedling period, the topsoil of the rhizome was collected from time to time to examine the density of Trichoderma hagianum KF-119, KCCM 10582 The antagonistic density of was investigated. As a control, Trichoderma Hajianum KF-65 was treated early in seedlings and compared with Tricoderma Hagianum KF-119 and KCCM 10582. The analysis results are shown in Table 3 below. In the antimicrobial soil treatment group, the density of Trichoderma hagianum KF-119 and KCCM 10582 increased with time, but the Trichoderma hagianum KF-65 treatment did not increase in density. .

Investigation of the Changes of Antagonistic Bacteria Density in Cucumber Seedling Roots Density of Bacillus subtilis KB-6 (× 100 cfu / g) Elapsed Days (Days) Early 2 5 10 20 Trichoderma Hajianum KF-119, KCCM 10582 (Treatment) 38 118 254 371 368 Trikoderma Haji Anum KF-65 (Comparative District) 39 41 45 43 41  Note: Density of antagonist is the density of Trichoderma hagiaumum surviving within 1 g of root zone soil.

[Experimental Example 3] Effect of Mozalok Disease Control in Pot

The antimicrobial soil prepared in the same manner as in Example 3 was used to evaluate the effect of controlling cucumber disease in the pot, and the results of the incidence at the end of seedlings are shown in Table 4 below. Pathogens were mixed with antimicrobial soil, seeded and seeded cucumber seed was treated as treatment, pathogen was mixed with normal soil and seeded and seeded cucumber seeded as comparison. In addition, seedlings and seedlings of cucumber seeds were mixed with antimicrobial soil without seedlings and seedlings without seedlings, and seedlings and seedlings of seedlings without seedlings with seedlings with seedlings were added to control soils. In other words, when seedlings were inoculated with Rhizoctonia solani in antimicrobial soils, the incidence of mozalococcal disease was only 12%. appear.

Antimicrobial Effect of Cucumber Wear Disease division % Of mozzarlock disease Treatment 12 Comparison 88 Control 1 4 Control 2 28  [Note] Treatment: Seeding and raising seedlings of cucumber seeds by mixing pathogens in antimicrobial soil Comparative group: Seeding and raising seedlings of cucumber seeds by mixing pathogens in normal soil Control 1: Controlling seedlings and seedlings on antibacterial soil Seeding and seeding cucumber seeds on topsoil

[Experimental Example 4] Effect of Mozalok Disease Control in Nursery

The results of 25 days of seedling growth in the actual cucumber nursery using the antimicrobial clay prepared in the same manner as in Example 3 are shown in Table 5 below. The height and dry weight were averaged by randomly picking 10 bags after the completion of seedlings. In the case of treatment with antimicrobial soils, the treatment of other seedlings is similar to that of the control soils treated with normal soils, but it shows that the control effect of Mozalok disease is excellent.

Results of Cucumber Seedling Results at the Nursery Site division Germination rate (%) Extra long (cm) Dry weight (g) % Of mozzarlock disease Treatment 97 17.2 0.29 4 Comparison 97 15.6 0.24 18  [Note] Treatment: seedlings using antibacterial soils Comparative section: seedlings using normal soils

[Experimental Example 5] Test for Control of Mozal Disease in Main Fabric

Using the antimicrobial clay prepared in the same manner as in Example 3, the seedlings of seedlings were grown and transplanted in the main body, and the results of examining the onset and cultivation results of Mozal disease are shown in Table 6 below. The height and dry weight were averaged by randomly picking 10 bags at 20 days of planting. The height and dry weight of the treatment group treated with antimicrobial soil were similar to those of the control group treated with normal soil, but it showed that the effect of days and final incidence until the initial onset of Mozalok disease was much better than that of the control. It is believed that this is because the antagonist itself has excellent antagonistic power, has excellent antagonistic bacteria fixability and proliferation ability to the soil, and reliably inhibited the growth of pathogens present in the soil even after the transplantation.

Investigation on the effects of warehousing and cultivation results at the headquarters Days to First Development (Days) Final Incidence (%) Extra long (cm) Dry weight (g) Treatment 19 4.9 68.3 4.9 Comparison 5 22.1 62.8 4.4  [Note] Treatment: transplanted seedlings grown using antimicrobial soil on the main field.Compared: transplanted seedlings grown using normal soil on the main field.

As can be seen in the above Examples and Experimental Examples, the seeding and seeding of seeds using the antimicrobial clay soil control according to the present invention and seedling resulted in the density of Trichoderma hagianum KF-119, KCCM 10582 in the root zone, that is, antagonist density Increased and the incidence of mozzarlock disease in the nursery and main gun was significantly reduced. Thus, seeding and seeding the seed using the antimicrobial soil for control of mozzarella disease according to the present invention, when the seedlings are grown and settled along the roots of seedlings, trichoderma hagianum KF-119, KCCM 10582, which are antagonists in the antimicrobial soil according to the present invention It can be seen that an antagonistic protective film was formed on the substrate. In addition, when seedlings are transplanted as the answer, the antagonists living in the roots of the seedlings are transferred to the main cell, and as the crop matures, the antagonists in the root zones continue to multiply to form antagonist populations around the roots, and root infestation of soil infectious pathogens occurs. It can be seen that by inhibiting soil infectious diseases such as mozzarlock disease.

As described above, when sowing and raising seedlings using antimicrobial soil containing antagonistic trichoderma hagianum KF-119 ( Trichoderma harzianum KF-119), KCCM 10582 according to the present invention, the environment caused by the use of pesticides, etc. It can solve the problems of pollution, residual toxicity, promote the formation of effective biomass in the soil, and the long-term control effect can improve agricultural productivity.

Claims (3)

Trichoderma hagianum KF-119 (Trichoderma harzianum KF-119), KCCM 10582. Antimicrobial clay for the control of mozzarlock disease comprising Trichoderma hajianum KF-119, KCCM 10582 of claim 1. (A) culturing the antagonist Trichoderma Hagianum KF-119 (Krichorma harzianum KF-119), KCCM 10582 for the control of Mozolox disease to produce an antagonistic culture; (b) drying and stabilizing the antagonistic culture to produce an antagonistic culture dried product; And (c) a method for producing antimicrobial clay for controlling mozzarella disease, comprising the step of uniformly mixing the antagonist cultured dry matter with normal clay.

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