KR20100121208A - Soil gas bioassay kit and uses of the same - Google Patents

Abstract

PURPOSE: A soil gas bioassay kit(SGB kit) is provided to quickly and simply measure physical chemical and to maximize use amount and efficiency of fumigant. CONSTITUTION: A soil gas bio assay kit comprises: a gas injection device having a injection tube for injecting gaseous sample or reagent, a moving tube through which injected gas moves to a sealed box; and a control device for controlling the amount of gas; the sealed box which is connected to a exhaust line(exhaust pipe) for discharging soil gas; a gas collection device having the exhaust line and control device.

Description

Soil gas bioassay kit and uses of the same

The present invention relates to a soil gas bioassay kit (SGB kit) and a method for determining the potency and effective amount of a soil fumigant using the same.

Korean ginseng is the best medicine and health food of oriental medicine with international reputation as a national image product of Korea. With the development of communication and transportation, the world market is becoming one and agricultural products are no exception. Korea's agriculture, which has narrow agricultural land for main grain food production, has no choice but to go to export farming. Although the economic status of 1.4 billion Chinese consumers who have used Korean ginseng for more than 5,000 years is improving, Korean ginseng, which is essential for their prescription, is still not available due to the increase in domestic ginseng prices.

The high price of Korean ginseng is attributable to the increase in domestic consumption, but the main reason is the increase in production cost by six years of cultivation. Ginseng has 22% colony in three years of root crops due to root diseases, and an average of 50% of alveolar alveolar alveoli in six years of age. In recent years, the cultivation area has been expanded, resulting in an insufficient shortage of planting sites for ginseng. Ginseng plants once planted even after 20 years of root disease is severe and can not be planted. These ginseng disturbances are caused by the growth of other species of soil-derived pathogenic bacteria, mainly Cylindrocarpon destructans . After six years of rice farming, it can be replanted due to the reduction of pathogens, but there are few rice paddies that can follow the field conditions, and it is difficult to improve the physical properties of the paddy soils as the field soils.

Naturally, the limit of the planned resources is to plant a lot of ginseng in the unsuitable land, resulting in worse crops and a lot of pesticides. For this reason, land rent is the highest at 28% of ginseng production cost, and resources for planned site is the biggest factor for raw ginseng production. Especially for the production of clean raw ginseng, the planned resources without pathogens are the most important. When the roots of ginseng begin to rot, the roots are in the soil, so there is no way to control them. However, while recommending dazomet (trade name bassamide) for the most important pathogen sterilization, there is no specific treatment in the RDA (Ginseng Cultivation, Rural Development Administration, 2000).

Soil pathogen disinfection has used a number of chemical fumigants that produce poisonous gases. Methyl bromide has been banned due to global environmental problems, and recently, bassamide, which has a relatively low residual problem, is also allowed in ginseng. However, the use of bassamide is rarely used because it is not reliable because the soil conditions are different from place to place, rather than the complexity of the work.

Lime nitrogen is more eco-friendly than bassamide, so it is an eco-friendly soil fumigation and fertilizer in Japan (Environmental Conservation Agricultural Prediction 2, Comprehensive Preventive Soil Disease Countermeasures, 2005). In Korea, research has been conducted on the effects of Chinese cabbage root disease. Lime nitrogen is significantly more effective than the 77% incidence of commercial drugs (fursulfamide), but is still 33 to 48% (Kim, Soon-Soon et al., 2: 3, 278-283, 2006). Since 2015, the use of methyl bromide has been banned in China, and the effectiveness of lime nitrogen as a strawberry releasing agent as an alternative was tested.The amount of appropriate fumigant differed according to the pathogen, and the appropriate soil moisture was different for each pathogen (Wang Lijing et al., Frontiers of Agriculture in China 1: 183-187, 2007).

The above results suggest that it is difficult to find the selection of the fumigant for each pathogen and the determination of the proper amount of use by the field experiments with the change of factors. Most soil fumigants enter the soil and react with moisture to generate toxic gases and sterilize them.

Therefore, the present inventors have developed a small soil gas bioassay kit and its use that can efficiently select the appropriate soil fumigant and determine the amount of use by pathogens and pavement in the field test process for selecting eco-friendly soil fumigants using the ginseng farmland rehabilitation method. This invention was completed by demonstrating the effect by several experiments.

It is an object of the present invention to provide a Soil Gas Bioassay Kit (SGB kit), and a method of using the kit for various uses, such as selecting a soil fumigant and determining an appropriate amount for each soil.

In order to achieve the above object, the present invention provides a soil gas bioassay kit (SGB kit) configured by connecting the gas injection device, a small closed box for generating soil gas and the generated gas recovery device is connected to each other.

The present invention also provides a method for screening soil fumigant candidates using the soil gas bioassay kit.

In addition, the present invention provides a method for measuring the potency (sterilization effect, herbicidal effect, insecticidal effect) of the soil fumigant using the soil gas bioassay kit.

In addition, the present invention provides a method for determining the effective dose per package of soil fumigant using the soil gas bioassay kit.

Soil Gas Bioassay Kit (SGB kit) of the present invention can quickly and simply measure the physical and chemical conditions of the soil to maximize the use of the soil fumigant for each pathogen and the fumigation efficiency, ginseng and other crops It can be useful for disinfecting soil of cropland.

Hereinafter, terms used in the present invention are defined.

As used herein, the term 'bioassay' is a method of assaying a substance by administering a bioactive compound to an organism and observing the response thereof, and using a small amount of the substance to examine a biological effect on the development or function of the organism. When testing, means the method used.

The term 'soil fumigant' used in the present invention refers to a material that is easily evaporated at room temperature and the gas is injected into the soil as a pesticide having bactericidal and insecticidal properties to control harmful animals and plants in the soil.

The term 'dazomet' used in the present invention is known under the trade name 'basamide' as a fungicide pesticide, and means a material having a nematode and herbicidal effect by sterilizing various pathogens in the soil by soil fumigation.

The term 'lime nitrogen (CaN ₂ )' used in the present invention is a nitrogen fertilizer containing calcium cyanide as a main component, and it has a pesticidal effect and sterilization effect of soil by soil fumigation and neutralizes soil acidity. It also means sanctions used.

Hereinafter, the present invention will be described in detail.

The present invention provides a soil gas bioassay kit (SGB kit) in which a gas injection device, a small sealed box for generating soil gas, and a generated gas recovery device are connected to each other.

Specifically, the soil gas bioassay kit of the present invention (Soil Gas Bioassay Kit, SGB kit) is an injection tube (intake) into which a gaseous sample or reagent is injected, a moving tube connected to a sealed box and the injected gas is moved to a sealed box. And a gas injection device having a control device capable of adjusting the amount of gas moved to the moving tube. An airtight box connected to the inlet gas device through a moving tube, having an opening and closing device, including soil, and connected to a discharge pipe (exhaust) for discharging the generated soil gas; And a gas recovery device connected to the closed box and having a discharge pipe (exhaust pipe) for discharging soil gas and a control device for controlling the amount of discharge gas discharged to the discharge pipe, but not limited thereto. Do not.

The gas injection device preferably further includes an injection gas filter device including an air cleaning solution or an injection gas filter device including a filter, but is not limited thereto.

The closed box is preferably a transparent box, the shape can be any shape. Preferably, the closed box has a length, a width, and a height of 10 to 50 cm, more preferably 20 to 30 cm, respectively, but is not limited thereto. The sealed box has an opening and closing lid, it is preferable that the inside of the box is sealed when the lid is closed to block the air inside and outside. The sealed box is preferably able to put the target soil sample 1 to 10 kg in weight, more preferably 2 to 5 kg in weight is not limited thereto. The sealed box may further include a pad or a petri dish for culturing soil pathogens when measuring the bactericidal power of the soil fumigant against soil pathogens, but is not limited thereto.

The gas recovery device preferably further includes a gas collection device including a gas collection liquid or a chemical reaction device including a gas detoxification compound, but is not limited thereto.

The gas recovery device is preferably used for measuring gas generation, collecting gas for safe disposal of reactive gas, or detoxification by chemical reaction, but is not limited thereto. The generated gas collecting device is preferably used by additionally connecting a suction pump, but is not limited thereto. The generated gas collecting device may be measured by putting a chemical reacting appropriately into the collecting container or collecting the gas with a gas collecting syringe and injecting it into gas chromatography.

In the present invention, the present invention is an injection gas washing apparatus including an air cleaning solution (I of FIG. 1), a sealed box for generating soil gas (C of FIG. 1), and a generated gas collecting device including a gas collecting liquid (FIG. 1). Soil Gas Bioassay Kit (SGB kit) consisting of the details of O) was prepared (see Fig. 1).

In addition,

1) injecting a test sample into the soil gas bioassay kit of the present invention comprising a test soil sample and mixing it with the soil sample;

2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions;

3) measuring the survival rate of the soil pathogens; And

4) provides a method for screening candidates for soil fumigant, comprising selecting a test sample that significantly reduces the survival rate of the soil pathogen compared to a control which is not injected with the test sample.

In the above method, the test sample of step 1) is preferably a granule, a powder or a gas, but is not limited thereto. Any formulation that can be used as a soil fumigation agent is possible. The test sample is preferably a composition or a compound, but is not limited thereto.

In the above method, the soil sample of step 2) preferably uses the soil of ginseng cultivation, and the soil pathogen is preferably Rhizoctonia solani or Pythium sp. .

In the above method, the soil pathogens of step 2) are directly cultured and mixed with the culture bacteria in the soil sample, the method of inoculating the soil pathogens and incubated on the pad, the method of burying the pad on the soil sample or on top, and soil pathogens It is preferable to leave the petri dish in any one method selected from the group consisting of a method of inoculating the petri dish after incubation in a petri dish, but is not limited thereto. The leaving is preferably left for 5 days to 10 days, and more preferably left for 6 to 8 days, but is not limited thereto. Specifically, the use of the soil gas bioassay kit of the present invention, after the test material is injected through the intake port or directly processed into the soil and mixed with the soil, the soil pathogens are put in or inside the soil sample to seal the sealed box, After being left for about a week, it is preferable to carry out by collecting the generated soil gas in a collecting device, but is not limited thereto.

In the above method, the survival rate of the soil pathogen of step 3) is determined by colony forming unit (CFU) assay, Most Probable Number (MPN), Phospholipid Fatty Acid (PLFA) assay and fluorescent antibody. It is preferable to use any one selected from the group consisting of fluorescent antibody probes (FA), but is not limited thereto.

The present inventors collected a specific soil sample and put it in the sealed box area of the soil gas bioassay kit of the present invention and injected the soil fumigant candidate material, and then put the soil pathogen and closed the lid and left it at room temperature for 7 to 10 days in a sealed condition. By examining the sterilization degree, it was confirmed that the fumigant candidates could be selected for specific pavement soil.

In addition,

1) adjusting the amount of soil fumigant to be injected into the soil gas bioassay kit of the present invention comprising the sample to be mixed with the soil sample;

2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions;

3) measuring the survival rate of the soil pathogens; And

4) It provides a method for determining the effective amount of the soil fumigant for the test soil, comprising the step of determining the amount of soil fumigant significantly reducing the survival rate of the soil pathogen.

In the above method, the soil fumigant of step 1) preferably uses dazomet or lime nitrogen (CaCN ₂ ), but is not limited thereto, and all soil fumigants may be used. The soil fumigant is preferably granulated, powdered or gaseous, but is not limited thereto. Soil fumigants of all formulations may be used.

In the above method, the soil sample of step 2) preferably uses the soil of ginseng cultivation, and the soil pathogen is preferably Rhizoctonia solani or Pythium sp. .

In the above method, the soil pathogens of step 2) are directly cultured and mixed with the culture bacteria in the soil sample, the method of inoculating the soil pathogens and incubated on the pad, the method of burying the pad on the soil sample or on top, and soil pathogens It is preferable to leave the petri dish in any one method selected from the group consisting of a method of inoculating the petri dish after incubation in a petri dish, but is not limited thereto. The leaving is preferably left for 5 days to 10 days, and more preferably left for 6 to 8 days, but is not limited thereto. Specifically, the use of the soil gas bioassay kit of the present invention, after injecting the soil fumigant through the inlet (adjust the injection amount using the injection control device) or by directly treating the soil (treated in various amounts), and mixed with the soil, Soil pathogens are put in or at the top of the soil sample, the sealed box is sealed, and then left to stand for about a week, it is preferable to carry out by collecting the collected soil gas in the collecting device, but is not limited thereto.

In the above method, the survival rate of the soil pathogen of step 3) is determined by colony forming unit (CFU), Most Probable Number (MPN), Phospholipid Fatty Acid (PLFA) assay and fluorescent antibody electrode method. (fluorescent antibody probes, FA) is preferably used, but not limited to any one selected from the group consisting of.

The present inventors take the target soil sample and put it in the closed box of the soil gas bioassay kit of the present invention, and inject the soil fumigant by varying the amount, and then put the soil pathogen and close the lid for 7 to 10 days under the sealed condition. It was confirmed that the optimum amount of fumigant for a specific pavement soil could be investigated by leaving it at room temperature and examining the degree of sterilization.

In addition,

1) injecting a soil fumigant into the soil gas bioassay kit of the present invention comprising a sample to be mixed with the soil sample;

2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions;

3) measuring the survival rate of the soil pathogens; And

4) It provides a method for determining soil conditions (soil, temperature, moisture content or organic matter content) to increase the effect of the soil fumigant comprising the step of selecting a test soil sample that significantly reduces the survival rate of the soil pathogens. .

In the above method, the soil fumigant of step 1) preferably uses dazomet or lime nitrogen (CaCN ₂ ), but is not limited thereto, and all soil fumigants may be used. The soil fumigant is preferably granulated, powdered or gaseous, but is not limited thereto. Soil fumigants of all formulations may be used.

In the above method, the soil sample of step 2) preferably uses the soil of ginseng cultivation, and the soil pathogen is preferably Rhizoctonia solani or Pythium sp. .

In the above method, the soil pathogens of step 2) are directly cultured and mixed with the culture bacteria in the soil sample, the method of inoculating the soil pathogens and incubated on the pad, the method of burying the pad on the soil sample or on top, and soil pathogens It is preferable to leave the petri dish in any one method selected from the group consisting of a method of inoculating the petri dish after incubation in a petri dish, but is not limited thereto. The leaving is preferably left for 5 days to 10 days, and more preferably left for 6 to 8 days, but is not limited thereto. Specifically, the use of the soil gas bioassay kit of the present invention, first, after putting soil samples of various conditions (different in soil, temperature, moisture content or organic matter content) into the sealed box in the kit, and then injected the soil fumigant through the intake vent Or by directly treating the soil and mixing it with the soil sample, putting the soil pathogen in or above the soil sample and sealing the sealed box, and left for about a week, and then discharges the generated soil gas and collects it in the collecting device. It is preferable to carry out as, but is not limited thereto.

In the above method, the survival rate of the soil pathogen of step 3) is determined by colony forming unit (CFU), Most Probable Number (MPN), Phospholipid Fatty Acid (PLFA) assay and fluorescent antibody electrode method. (fluorescent antibody probes, FA) is preferably used, but not limited to any one selected from the group consisting of.

The present inventors take the target soil sample and change only the target properties of the soil, put it in the sealed box of the soil gas bioassay kit of the present invention, inject the soil fumigant, put the soil pathogens and close the lid 7 It is possible to determine soil conditions (soil temperature, chemical and physical properties, clay content, organic matter content or soil moisture content) that increase the effectiveness of soil fumigants by leaving them at room temperature for 10 days and then examining the sterilization degree. .

In addition,

1) injecting a soil fumigant into the soil gas bioassay kit of the present invention comprising a sample to be mixed with the soil sample;

2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions; And

3) provides a method for measuring the bactericidal effect of the soil fumigant comprising the step of measuring the survival rate of the soil pathogen.

In the above method, the soil fumigant of step 1) preferably uses dazomet or lime nitrogen (CaCN ₂ ), but is not limited thereto, and all soil fumigants may be used. The soil fumigant is preferably granulated, powdered or gaseous, but is not limited thereto. Soil fumigants of all formulations may be used.

In the above method, the soil sample of step 2) preferably uses the soil of ginseng cultivation, and the soil pathogen is preferably Rhizoctonia solani or Pythium sp. .

In the above method, the soil pathogens of step 2) are directly cultured and mixed with the culture bacteria in the soil sample, the method of inoculating the soil pathogens and incubated on the pad, the method of burying the pad on the soil sample or on top, and soil pathogens It is preferable to leave the petri dish in any one method selected from the group consisting of a method of inoculating the petri dish after incubation in a petri dish, but is not limited thereto. The leaving is preferably left for 5 days to 10 days, and more preferably left for 6 to 8 days, but is not limited thereto. Specifically, the use of the soil gas bioassay kit of the present invention, after injecting a soil fumigant through the intake port or directly treated in the soil and mixed with the soil, and put the soil pathogens in or inside the soil sample to seal the sealed box, After being left for about a week, it is preferable to carry out by collecting the generated soil gas in a collecting device, but is not limited thereto.

In the above method, the survival rate of the soil pathogen of step 3) is determined by colony forming unit (CFU), Most Probable Number (MPN), Phospholipid Fatty Acid (PLFA) assay and fluorescent antibody electrode method. (fluorescent antibody probes, FA) is preferably used, but not limited to any one selected from the group consisting of.

The present inventors collect a specific soil sample and put it in the sealed box of the soil gas bioassay kit of the present invention, inject the test soil fumigant, insert the soil pathogen, close the lid, and leave it at room temperature for 7 to 10 days in a sealed condition. By examining the bactericidal power, it was confirmed that the bactericidal effect of soil pathogens (spore or mycelium) of soil pathogen of the tested soil fumigant could be investigated. In addition, in order to measure the effect of soil fumigant on the soil microbial ecology, after treating the soil fumigant can be investigated by classifying the microorganisms of the soil into fungi, bacteria or actinomycetes.

In addition,

1) injecting a soil fumigant into the soil gas bioassay kit of the present invention comprising a sample to be mixed with the soil sample;

2) placing seeds in the soil sample included in the kit and then leaving them under sealed conditions; And

3) It provides a method for measuring the herbicidal effect of soil fumigant comprising the step of measuring the seed germination rate.

In the above method, the soil fumigant of step 1) preferably uses dazomet or lime nitrogen (CaCN ₂ ), but is not limited thereto, and all soil fumigants may be used. The soil fumigant is preferably granulated, powdered or gaseous, but is not limited thereto. Soil fumigants of all formulations may be used.

In the above method, the soil sample of step 2) is preferably to use the soil of ginseng cultivation medium, but is not limited thereto.

In the above method, the seed of step 2) is preferably Sudangrass seed, radish or weed seed, more preferably Sudangrass seed, but is not limited thereto. The seed is preferably placed in a semantic yarn and placed in a soil sample or placed in a small bowl and placed on the soil sample, but is not limited thereto.

The present inventors collect a specific soil sample, put it in the sealed box of the soil gas bioassay kit of the present invention, inject the test soil fumigant, insert the seed, close the lid, and leave the seed at room temperature for 7 to 10 days under the sealed condition. It was confirmed that the herbicidal effect of soil fumigation agent (seed or nutrient growth agent) by the growth stage of the tested soil fumigation agent could be investigated.

In addition,

1) injecting a soil fumigant into the soil gas bioassay kit of the present invention comprising a sample to be mixed with the soil sample;

2) putting pests or eggs into the soil samples included in the kit and leaving them under sealed conditions; And

3) provides a method for measuring the pesticidal effect of the soil fumigant comprising the step of measuring the survival rate of the pest.

In the above method, the soil fumigant of step 1) preferably uses dazomet or lime nitrogen (CaCN ₂ ), but is not limited thereto, and all soil fumigants may be used. The soil fumigant is preferably a granule, powder or gas, but is not limited thereto. Soil fumigants of all formulations may be used.

In the above method, the soil sample of step 2) is preferably to use the soil of ginseng cultivation medium, but is not limited thereto.

The present inventors collected a specific soil sample and put it in the sealed box area of the soil gas bioassay kit of the present invention, injected the test soil fumigant, and then put the pest or eggs and close the lid, and left it at room temperature for 7 to 10 days in a sealed condition. In addition, it was confirmed that the pesticidal effect of the soil pests of the tested soil fumigant (eg eggs, larvae, adults, pupa, etc.) could be examined by examining the insecticidal power.

In addition,

1) injecting a soil fumigant into the soil gas bioassay kit of the present invention, including the soil sample to be mixed with the soil sample, and leaving it under sealed condition; And

2) provides a method for quantifying the amount of gas generated by the soil fumigant comprising the step of measuring the amount of gas collected by the kit.

In the above method, the soil fumigant of step 1) preferably uses dazomet or lime nitrogen (CaCN ₂ ), but is not limited thereto, and all soil fumigants may be used. The soil fumigant is preferably granulated, powdered or gaseous, but is not limited thereto. Soil fumigants of all formulations may be used.

In the above method, the soil sample of step 2) is preferably to use the soil of ginseng cultivation medium, but is not limited thereto.

 The present inventors collect a specific soil sample and put it in a sealed box of the soil gas bioassay kit of the present invention, inject a test soil fumigant, close the lid and leave it at room temperature for 7 to 10 days in a sealed condition, and then collect the collected gas. The analysis confirmed that the respiratory volume of the gas or soil itself generated by the test soil fumigant can be quantified.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the present invention, but the content of the present invention is not limited to the following examples.

Example 1 Preparation of Soil Gas Bioassay Kit (SGB kit)

To manufacture the SGB kit, the inventors have provided a transparent sealed box with a lid (FIG. 1C), a gas washing bottle (I in FIG. 1) capable of washing gas when needed to inject gas from the outside, and soil and After the reaction, it was prepared to consist of three parts of the gas washing bottle (O of FIG. 1) that can perform its function when it is necessary to collect the gas in the closed box. The sealed box was a transparent acrylic plate (1 cm thick) was made horizontal, vertical, height all 20 cm. The air washer was purchased from a chemical device dealer.

<Example 2> Ginseng Soil Pathogen Lyraitononia solani ( Rhizoctonia solani ) And Pygeum ( Pythium  disinfection by soil fumigant for

The present inventors used two soil fumigants, commercially available, Dazomet (Bassamide, Dongbu Fine Chemical) and Lime Nitrogen (CaCN ₂ ) (Pungong, Korea). Ginseng cultivation soil (Gildo, Anjuk-ri, Iljuk-myeon, 6-year-old ginseng mined soil 4 kg) was mixed evenly. Rhizoctonia solani and Pythium sp., Soil pathogens and causative agents of ginseng, were isolated and isolated from ginseng seedlings and ginseng seedlings showing mozzarella disease symptoms and PDA (Potato dextrose) agar, Difco) The bacterial flora of pathogens cultured in plate media for 7 days was cut to 4 mm in diameter using sterile platinum teeth and then inoculated onto new PDA plate media. After incubation for 1 week on the soil of the SGB kit prepared inoculated PDA flat medium, colony forming unit (CFU) was used to examine the bactericidal effect.

As a result, as shown in Table 1, Figure 2 and 3, the two fumigants increased sterilization potency with the increase in the amount of fumigant, the degree of sterilization potency showed a significant difference between the two fumigants. That is, dazometh (Vassamide) was 100% sterilization at the recommended amount of use, but lime nitrogen was not completely sterilized even if more than 10 times the recommended amount (see Tables 1, 2 and 3).

Ginseng Wearing Bacteria by Fumigation Agents ( Pythium  sp.) bactericidal power (%) Bassamide Lime Nitrogen Capacity (ppm) Sterilization (%) Capacity (ppm) Sterilization (%) Formulation  10  14 5,000  0 Granulation  20  36 10,000 44 Granulation  50  92 5,000  0 Powder 100 100 10,000 80 Powder

Example 3 Change in Potency of Soil Fumigation Agent by Soil Moisture Content

After varying the soil moisture content and mixing 100 ppm of Dazomet (Vassamide) with the soil sample, the inventors used the SGB kit for 7 days in a PDA plate medium for incubation of Rhizoctonia solani . After incubation, the bacterial flora of the pathogen was cut to 4 mm in diameter using sterile platinum teeth and inoculated on a new PDA plate medium. The inoculated PDA plate medium was left in the upper or side space of the soil layer for 7 days and then tested for bactericidal effect by using colony forming unit (CFU).

As a result, as shown in FIG. 4, the maximum sterilizing effect was obtained when the relative pavement moisture content of the soil moisture was 30% or more (see FIG. 4).

Example 4 Effect of Fumigation Agent Dose on Soil Microbial Density

The present inventors mixed Dazomet (Vassamide) with soil samples up to the recommended amount of 100 ppm by varying the amount, and left for 7 days in the SGB kit, and then examined the density of fungi, bacteria, actinomycetes in the soil. As the density of microorganisms in soil, mold was used as PDA (Potato dextrose agar, Difco), bacteria as TSA (Tryptic soy agar, Difco), and actinomycetes as AIA (Actinomycete isolation agar, Difco) medium. It was.

As a result, as shown in Table 2, bacteria and actinomycetes showed little bactericidal effect, but showed a significant bactericidal effect against mold (see Table 2).

Changes in Soil Microbial Density by Application of Bathamide (cfu / g) Capacity (ppm) Total germ Total actinomycetes Total fungus 10 8.65 x 10 ⁵ 3.70 x 10 ⁵ 5.1 x 104 20 9.60 x 10 ⁵ 3.45 x 10 ⁵ 5.2 x 104 50 8.40 x 10 ⁵ 1.75 x 10 ⁵ 1.4 x 103 100 7.55 x 10 ⁵ 2.95 x 10 ⁵ 5.0 x 101

Example 5 Investigation of Herbicidal Strength of Soil Fumigation Agent

The inventors mixed 100 ppm Dazomet (Vassamide) (Eastern Agrochemical, Korea), 500 ppm Lime Nitrogen (Pungong, Korea), and 100 ppm Lime Nitrogen (Alfa Aesar) to the soil, In the SGB kit, Sudangrass seeds purchased from Anseong Agricultural Cooperatives were placed in Semangsa, buried in the soil, left for 7 days, and the seeds were taken out.

As a result, as shown in Fig. 5, Dazomet (Vassamide), all seeds died at the recommended amount of 100 ppm, but lime nitrogen (manufactured by PN) 70% germinated at 500 ppm higher than the recommended amount of 138 ppm, lime 100% germination was carried out at 100 ppm of nitrogen (manufactured by AA).

Through the above results, it can be seen that the SGB kit of the present invention can determine the soil conditions and the application amount of the fumigant for each package to increase the sterilization power of the soil fumigant to the maximum.

Through the present invention, it is possible to quickly and easily select the soil fumigant and determine the appropriate amount of use, thereby effectively disinfecting the soil, thereby significantly increasing the yield of ginseng and other crops, and in developing new soil fumigants and studying soil conditions. It can be usefully used.

1 is a soil gas consisting of the injection gas washing device of the present invention (I of Figure 1), a sealed box for generating soil gas (C of Figure 1), and the generated gas collecting device (O of Figure 1) This figure shows the schematic diagram of the Soil Gas Bioassay Kit (SGB kit).

Figure 2 shows the degree of colony formation in PDA (Potato dextrose agar) medium showing the bactericidal effect of Pythium sp. According to the recommended amount of soil fumigant, measured using the soil gas bioassay kit of the present invention Picture is:

A: no treatment;

B: treated with 100 ppm of bassamide;

C: 10,000 ppm lime nitrogen powder treatment; And

D: 10,000 ppm lime quenching agent.

Figure 3 is the degree of colony formation in PDA (Potato dextrose agar) medium showing the bactericidal effect of Rhizoctonia solani according to the recommended amount of soil fumigant, measured using the soil gas bioassay kit of the present invention Here is a picture that shows:

A: no treatment;

B: 100 ppm bassamide;

C: 5,000 ppm lime nitrogen powder; And

D: lime nitrogen scavenger 5,000 ppm.

Figure 4 is a graph showing the bactericidal effect on the Pythium sp. Of dazomet (Vassamide) by soil moisture content, measured using the soil gas bioassay kit of the present invention.

Figure 5 is a diagram showing the germination rate of the seed showing the killing effect of the suicide grease by the fumigant according to the recommended amount of soil fumigant, measured using the soil gas bioassay kit of the present invention:

A: 100 ppm bassamide;

B: 500 ppm lime nitrogen [manufactured by PN Corporation]; And

C: lime nitrogen [manufactured by AA] 100 ppm.

Claims (16)

It is characterized in that it has an injection tube (intake) through which a gaseous sample or reagent is injected, a moving tube connected to a sealed box, and a gas injected into the sealed box, and an adjusting device for adjusting the amount of gas moved to the moving tube. A gas injection device; An airtight box connected to the inlet gas device through a moving tube, having an opening and closing device, including soil, and connected to a discharge pipe (exhaust) for discharging the generated soil gas; Soil gas bioassay kit consisting of a gas recovery device characterized in that it has a discharge device (exhaust pipe) is connected to the closed box to discharge the soil gas and the control device for controlling the amount of the discharge gas discharged to the discharge pipe ( Soil Gas Bioassay Kit, SGB kit). The kit as claimed in claim 1, wherein the gas injection device further includes an injection gas cleaning device including an air cleaning solution or an injection gas filter device including a filter. The kit as claimed in claim 1, wherein the closed box has a length, a width, and a height of 10 to 50 cm, respectively. The kit of claim 1, wherein the gas recovery device further comprises a gas collection device including a gas collection liquid or a chemical reaction device including a gas detoxification compound. The kit as claimed in claim 1, wherein the gas recovery device further comprises an air suction pump. 1) injecting a test sample into the soil gas bioassay kit of the present invention comprising a test soil sample and mixing it with the soil sample; 2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions; 3) measuring the survival rate of the soil pathogens; And 4) screening a test sample that significantly reduces the survival rate of the soil pathogens compared to the control group is not injected with the test sample, soil fumigation candidate screening method. 1) mixing the soil sample with the soil sample by adjusting the amount of soil fumigant to the soil gas bioassay kit of claim 1 comprising the sample to be tested; 2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions; 3) measuring the survival rate of the soil pathogens; And 4) determining the effective amount of the soil fumigant for the test soil, comprising the step of determining the amount of soil fumigant that significantly reduces the survival rate of the soil pathogen. 1) injecting a soil fumigant into the soil gas bioassay kit of claim 1 comprising a soil sample to be mixed with the soil sample; 2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions; 3) measuring the survival rate of the soil pathogens; And 4) selecting a soil sample to significantly reduce the survival rate of the pathogen, soil conditions for increasing the effect of the soil fumigant. 1) injecting a soil fumigation agent into the soil gas bioassay kit of claim 1 comprising a sample to be mixed with the soil sample; 2) putting soil pathogens in the soil samples included in the kit and leaving under sealed conditions; And 3) comprising the step of measuring the survival rate of the soil pathogen, the method of measuring the bactericidal effect of the soil fumigant. 10. The method according to any one of claims 6 to 9, wherein the soil pathogens of step 2) are directly cultured to mix the culture bacteria in the soil sample, and the pads are inoculated onto the soil samples after inoculation with the soil pathogens. Buried or placed on the top, and inoculating the soil pathogens in petri dishes and incubating the petri dishes are placed in any one method selected from the group consisting of the top or side of the soil sample. . 1) injecting a soil fumigant into the soil gas bioassay kit of claim 1 comprising a sample to be mixed with the soil sample; 2) placing seeds in the soil sample included in the kit and then leaving them under sealed conditions; And 3) A method of measuring the herbicidal effect of the soil fumigant comprising the step of measuring the seed germination rate. 12. The method of claim 11, wherein the seed is placed in a semantic yarn and placed in a soil sample or placed in a bowl and placed on the soil sample. 1) injecting a soil fumigant into the soil gas bioassay kit of claim 1 comprising a sample to be mixed with the soil sample; 2) putting pests or eggs into the soil samples included in the kit and leaving them under sealed conditions; And 3) comprising the step of measuring the survival rate of the pest, pesticide effect measurement method of soil fumigant. 1) injecting a soil fumigant into the soil gas bioassay kit of claim 1 comprising a sample to be tested and mixed with the soil sample, and then leaving it under sealed condition; And 2) comprising the step of measuring the amount of gas collected by the kit, the amount of gas generated by the soil fumigant. 15. The method of any one of claims 6-9, 11, 13 or 14, wherein the neglect is left for 5-10 days. The method according to any one of claims 7 to 9, 11, 13 or 14, wherein the soil fumigant is dazomet or lime nitrogen (CaCN ₂ ).

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