WO2012121527A2 - Seed treatment method for inducing induced systemic resistance in a plant while minimising effects on plant growth - Google Patents

Seed treatment method for inducing induced systemic resistance in a plant while minimising effects on plant growth Download PDF

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WO2012121527A2
WO2012121527A2 PCT/KR2012/001616 KR2012001616W WO2012121527A2 WO 2012121527 A2 WO2012121527 A2 WO 2012121527A2 KR 2012001616 W KR2012001616 W KR 2012001616W WO 2012121527 A2 WO2012121527 A2 WO 2012121527A2
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bth
growth
plant
seeds
soybean
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PCT/KR2012/001616
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French (fr)
Korean (ko)
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WO2012121527A3 (en
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류충민
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한국생명공학연구원
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • A01C1/08—Immunising seed
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
    • A01N43/74—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms five-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,3
    • A01N43/78—1,3-Thiazoles; Hydrogenated 1,3-thiazoles
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
    • A01N43/82—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms five-membered rings with three ring hetero atoms

Definitions

  • the present invention relates to a seed treatment method for controlling various plant diseases during seedlings of plants by treating BTH (benzothiadiazole), which is a substance inducing plant whole body resistance, to seeds.
  • BTH benzothiadiazole
  • the present invention has been made in accordance with the above requirements, the inventors of the present invention to know the immersion time and concentration of BTH to minimize the effect on plant growth inhibition using BTH (benzothiadiazole), an induction resistant material, three crops To find out the most effective conditions for seed treatment.
  • BTH benzothiadiazole
  • the present invention provides a method for controlling plant diseases by treating BTH (benzothiadiazole) on the seeds of the plant without minimizing the growth of the plant or minimizing the effect of plant growth.
  • BTH benzothiadiazole
  • the present invention also provides a seed treatment composition for controlling plant diseases affecting pepper, soybean and cucumber seedlings without minimizing the growth of pepper, soybean and cucumber or minimizing the effects of growth.
  • the present invention also provides pepper seed, soybean seed and cucumber seed coated with BTH prepared by immersion in a certain concentration of BTH solution.
  • the present invention also provides a seed treatment method for controlling plant diseases affecting seedlings of peppers, soybeans and cucumbers without minimizing the growth of peppers, soybeans and cucumbers or minimizing the effects of growth.
  • the method of the present invention can control plant diseases from seedlings, and to control plant diseases through derivation of optimal treatment conditions for each seed. It can be used to develop seed treatments.
  • Figure 2 shows the effect of plant induction resistance and its growth on soybean bacterial spot pathogens ( Pseudomonas syringae pv. Syringae ) with the same treatment as in Figure 1 using another seed, soybean. Stem length (top) and disease resistance (bottom) for each treatment were measured. The control plants were water, and the incidence was measured by the number of the indicative parts, and 7 days after the pathogen inoculation. The data represents the mean.
  • Figure 3 shows the effect of plant induction resistance and growth according to cucumber seed spot pathogens ( Pseudomonas syringae pv. Lachrymans ) with the treatment as shown in Figs. Stem length (top) and disease resistance (bottom) for each treatment were measured. The control plants were water, and the incidence was measured by the number of the indicative parts, and 7 days after the pathogen inoculation. The data represents the mean.
  • the present invention provides a method for controlling plant diseases without disrupting the growth of the plant or minimizing the effect of the growth of the plant.
  • the method may include treating BTH (benzothiadiazole) with the seed of the plant, and inducing induced systemic resistance of the treated plant.
  • Plant disease according to the present invention may be the onset of plant seedlings, but is not limited thereto.
  • the plant disease may preferably be a bacterial disease, and more preferably, may be pepper bacterial spot pattern disease, soybean bacterial spot pattern disease or cucumber bacterial spot pattern disease, but is not limited thereto.
  • the concentration of BTH and the treatment time for the seed of the plant may vary depending on the seed of the plant.
  • 2-50 ⁇ M BTH solution is applied for 1 second to 1 hour, or 0.02 to 50
  • the ⁇ M BTH solution may be treated for 4-8 hours, more preferably the 10 ⁇ M BTH solution may be treated for 1-10 seconds and the 0.1 ⁇ M or 10 ⁇ M BTH solution for 6 hours.
  • preferably 0.02-50 ⁇ M BTH solution may be treated for 4-8 hours, more preferably 0.1 ⁇ M or 10 ⁇ M BTH solution may be treated for 6 hours.
  • preferably 2-50 ⁇ M BTH solution may be treated for 1 second to 1 hour, and more preferably 10 ⁇ M BTH solution may be treated for 1-10 seconds.
  • the present invention provides a seed treatment composition for controlling plant diseases affecting plant seedlings without minimizing the growth of plants or minimizing the effects of plant growth.
  • Red pepper or cucumber seed treatment composition according to an embodiment of the present invention may preferably contain 2 to 50 ⁇ M BTH as an active ingredient, more preferably 10 ⁇ M BTH as an active ingredient.
  • the soybean seed treatment composition may preferably contain 0.02 to 50 ⁇ M BTH as an active ingredient, and more preferably 0.1 ⁇ M or 10 ⁇ M BTH as an active ingredient.
  • compositions according to one embodiment of the invention may comprise agrochemically acceptable buffers, carriers, adjuvants or excipients, which are well known in the art.
  • the composition of the present invention may be lyophilized through lyophilization, spray drying or spray cooling.
  • buffer is intended to mean an aqueous solution containing an acid-base mixture for the purpose of stabilizing pH.
  • Buffers may include tris, phosphate, carbonate, acetate, citrate, glycolate, lactate, borate, tartrate, cacodylate, ethanolamine, glycine, imidazole, imidazolactic acid, and the like.
  • diluent is intended to mean an aqueous or non-aqueous solution for the purpose of diluting BTH.
  • the diluent can be one or more saline, water, polyethylene glycol, propylene glycol, ethanol or oils such as corn oil, peanut oil, cottonseed oil or sesame oil.
  • adjuvant is intended to mean a specific compound added to the formulation to increase the biological effect of BTH.
  • Excipients can be one or more carbohydrates, polymers, lipids and minerals.
  • carbohydrates include, for example, lactose, sucrose, mannitol and cyclodextrins added to the composition to facilitate lyophilization.
  • polymers examples include starch, cellulose ether, cellulose carboxymethyl cellulose, alginate, carrageenan, hyaluronic acid, polyacrylic acid, polysulfonate, polyethyleneglycol / polyethylene oxide, polyvinylalcohol / polyvinylacetate with different degrees of hydrolysis, and polyvinylpi There is rolidone (all different in molecular weight).
  • composition of the present invention can make formulations such as emulsions, emulsions, hydrates, powders, granules, tablets, aerosols, ointments, and the like. If necessary, an emulsifier, a suspending agent, an electrodeposition agent, a penetrating agent, a humectant, a thickening agent (muscilage, etc.), a stabilizer and the like can be added. These formulations can be prepared by methods of preparation known in the art.
  • the present invention provides a plant seed coated with BTH prepared by immersing the plant seed in a BTH solution and then drying.
  • Pepper seeds coated with BTH are preferably pepper seeds soaked in 2-50 ⁇ M BTH solution for 1 second to 1 hour, or 0.02-50 ⁇ M BTH solution for 4-8 hours. After that, it may be prepared by drying, more preferably, it may be prepared by immersing in 10 ⁇ M BTH solution for 1 to 10 seconds, or 0.1 ⁇ M or 10 ⁇ M BTH solution for 6 hours, and then dried.
  • the soybean seed coated with BTH may be prepared by soaking the soybean seed, preferably, dipped in 0.02-50 ⁇ M BTH solution for 4-8 hours, and then drying, and more preferably, Soybean seeds can be prepared by soaking in 0.1 ⁇ M or 10 ⁇ M BTH solution for 6 hours and then drying.
  • Cu seeds coated with BTH may be prepared by immersing the cucumber seeds in 2-50 ⁇ M BTH solution for 1 second to 1 hour, and then drying them, more preferably.
  • Cucumber seeds can be prepared by immersing in 10 ⁇ M BTH solution for 1 to 10 seconds, and then dried.
  • the present invention provides a plant disease to the seedlings of the plant without disturbing the growth of the plant or minimizing the effect of the growth of the plant comprising the step of treating the BTH solution to the plant seed
  • a seed treatment method for controlling Provided is a seed treatment method for controlling.
  • Treatment of the BTH solution to the plant seeds is preferably pepper seed, 2 to 50 ⁇ M BTH solution can be treated for 1 second to 1 hour, or 0.02 to 50 ⁇ M BTH solution for 4 to 8 hours, more preferably The 10 ⁇ M BTH solution may be treated for 1-10 seconds, or 0.1 ⁇ M or 10 ⁇ M BTH solution for 6 hours. Soybean seeds may preferably be treated with 0.02-50 ⁇ M BTH solution for 4-8 hours, more preferably 0.1 ⁇ M or 10 ⁇ M BTH solution for 6 hours. Cucumber seeds may preferably be treated with 2-50 ⁇ M BTH solution for 1 second to 1 hour, and more preferably 10 ⁇ M BTH solution for 1-10 seconds.
  • red pepper Heungnong seedling, Bugang pepper
  • cucumber Nongwoo bio, Baekbong tea
  • soybean soybean were used.
  • Pepper, cucumber and soybean seeds were used to determine the optimal BTH soaking time and concentration for each seed. Each seed was immersed in BTH solutions at concentrations of 0.1 ⁇ M, 10 ⁇ M and 1 mM, and the immersion time was divided again into 0, 6 and 12 hours. The 0 hour immersion is a brief immersion (1-10 seconds) in BTH at each concentration. After soaking, a 36-hole pot (28 ⁇ 54 ⁇ 6) filled with sterile filter paper (Whatman) was kept at room temperature for 3 days and filled with horticultural soil (Punong Co., Ltd, Korea). cm), pepper seeds were grown for 6 weeks, cucumber seeds for 3 weeks, and soybean seeds for 4 weeks in the greenhouse at 20-30 °C.
  • Pepper crops were injected into the leaves at 10 7 CFU / ml Bacterial spot pathogen ( Xanthomonas axonopodis pv.vesicatoria ). After 7 days, the disease degree was measured by 1 to 5, and the length of the stem was also measured. Bacterial inoculum was incubated at 30 ° C. for 48 hours by adding 100 ⁇ g / ml of antibiotic Rifampicin to LB (Luria-Bertani) solid medium.
  • Soybean crops were sprayed onto the leaves at 10 7 CFU / ml of Psuedomonas syringae pv. Syringae. After 7 days, the disease degree was measured by 1 to 5, and the length of the stem was also measured. Bacterial inoculum was incubated at 30 ° C. for 48 hours with the addition of 100 ⁇ g / ml of antibiotic Rifampicin in King's B solid medium.
  • Cucumber crops were sprayed onto the leaves at a concentration of 10 7 CFU / ml B. psuedomonas syringae pv. Lachrymans . After 7 days, the disease degree was measured by 1 to 5, and the length of the stem was also measured. Bacterial inoculum was incubated at 30 ° C. for 48 hours with the addition of 100 ⁇ g / ml of antibiotic Rifampicin in King's B solid medium.
  • Example 1 Induction resistance assay and plant growth measurement in red pepper
  • the severity of disease was decreased statistically in all treatments at 0.1 ⁇ M, 10 ⁇ M, and 1 mM BTH after 6 hours immersion, and 6 hours In the immersion treatment, except for 1 mM BTH, the degree of disease was significantly reduced at two BTH concentrations. Contrary to the results of the 6-hour soaking treatment, the 12-hour soaking treatment showed resistance to disease only in the 1 mM BTH soaking treatment.
  • the 0.1 ⁇ M BTH treatment showed resistance to disease at 0 h and 6 h, and no difference was observed at 12 h.
  • the 10 ⁇ M BTH treatment showed the same results as the 0.1 ⁇ M BTH treatment and showed the best disease resistance at 0 and 6 hours immersion among all treatments.
  • 1 mM BTH treatment showed the best disease resistance after immersion for 0 hours, but no disease resistance after 6 hours immersion, and resistance to 12 hours immersion.
  • the above-described two concentrations were different from the tendency of disease resistance to disappear after 12 hours of soaking.
  • the optimal treatment method is to immerse 10 ⁇ M BTH in red pepper seeds for 0 hours in order to develop a seed treatment using low concentration and minimum immersion time.
  • the resistance to disease was statistically different at 0.1 ⁇ M, 10 ⁇ M, and 1 mM BTH concentrations, respectively, and at 0 and 12 h, no resistance to disease was observed. 6 hours of soaking did not show the effect of plant growth inhibition compared to the 0 hours of soaking treatment. However, the comparison between BTH concentrations showed that 0.1 ⁇ M and 10 ⁇ M BTH treatments showed better growth than 1 mM BTH treatments. Based on the above facts, in order to develop a seed treatment using a low concentration and minimal soaking time, it is considered that the optimal treatment method is to soak 0.1 ⁇ M BTH for 6 hours in soybean seed.
  • Example 3 Induction resistance assay and plant growth measurement of cucumber

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Dentistry (AREA)
  • Plant Pathology (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pest Control & Pesticides (AREA)
  • General Health & Medical Sciences (AREA)
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Abstract

The present invention clarifies the concentration and time for treatment with BTH (benzothiadiazole), which induces induced systemic resistance in plants, so as to minimise the plant growth suppressing effect that appears when BTH is used to achieve sustained disease resistance in three crops, namely pepper, soybean and cucumber.

Description

식물 생장의 영향을 최소화하면서 식물 전신유도저항성을 유도하는 종자처리 방법Seed treatment method to induce plant whole body resistance while minimizing the effect of plant growth
본 발명은 식물 전신유도저항성을 유도하는 물질인 BTH (benzothiadiazole)를 종자에 처리하여 식물의 유묘기에 다양한 식물병을 방제하기 위한 종자처리 방법에 관한 것이다.The present invention relates to a seed treatment method for controlling various plant diseases during seedlings of plants by treating BTH (benzothiadiazole), which is a substance inducing plant whole body resistance, to seeds.
기존 식물 병원균의 발생 억제 및 방제를 위한 방법은 주로 유묘기에서 연구가 되었으며, 화학적 합성 농약을 주로 사용하였다. 그러나 이러한 합성 농약으로 인한 여러 문제점이 야기되어 그 사용이 제한되고 있고, 최근 종자에서부터의 방제에 관심이 집중되어 튼튼한 종자 개발에 관한 연구가 활발히 진행되고 있는 실정이다. The methods for suppressing and controlling the occurrence of existing plant pathogens were mainly studied in seedlings, and chemical synthetic pesticides were mainly used. However, due to the various problems caused by the synthetic pesticides are limited in use, and the recent focus on the control of the seed has been actively researched on the development of strong seeds.
식물은 한번 정착하면 움직일 수가 없기 때문에 외부 환경의 변화에 효과적으로 대응하기 위하여 내부와의 신호전달체계를 발전시켜왔다. 특히 병원균의 다양한 공격에 대항하는 면역시스템을 가지고 있는데, 이러한 저항성 반응 중 병원균이 침입된 잎뿐만 아니라 침입되지 않은 식물체 전신에서 저항성 반응이 일어나게 되어, 뒤이어 침입하는 다양한 병원균에 대한 저항성을 나타내게 되는 전신 획득 저항성 (systemic acquired resistance: SAR)이 있다. 여러 SAR 유도체 중에서 BTH (benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester)라는 물질이 가장 강력한 효과를 보이고 있다. 하지만 화학농약의 문제점을 극복할 것 같았던 이러한 SAR 유도물질에도 심각한 문제점이 발견되었는데, 실제 포장상태에서 농작물에 처리 시 식물체의 생육이 억제되거나 처리하지 않은 대조구에 비해 수확량이 현저히 줄어드는 현상이 관찰되었다. 이러한 현상을 생태학적 측면으로 보면, 병원균이 없는데도 많은 병원성 유전자를 발현시킴으로써 식물의 에너지를 과다하게 소비하여 생장에 사용하여야 할 부분을 병 저항성에 사용하기에 이런 일이 일어난다고 보고, 이런 현상을 "allocation fitness cost"라 한다.Since plants cannot move once settled, they have developed a signaling system with the interior to effectively respond to changes in the external environment. In particular, it has an immune system against various attacks of pathogens. Among these resistance reactions, resistance reactions occur not only in the leaves of the pathogens but also in the entire invasive plant, resulting in resistance to various invading pathogens. There is systemic acquired resistance (SAR). Among the various SAR derivatives, BTH (benzo (1,2,3) -thiadiazole-7-carbothioic acid S-methyl ester) has the strongest effect. However, serious problems were found in these SAR-induced substances, which were likely to overcome the problems of chemical pesticides, and when the crops were treated in actual packaging, the growth of plants was suppressed or the yield was significantly reduced compared to the untreated control. The ecological aspect of this phenomenon is that this phenomenon occurs because it expresses many pathogenic genes without pathogens and consumes excessive amounts of plant energy and uses parts for disease resistance. allocation fitness cost ".
본 발명자들은 식물에 유도저항성을 유도하면서 이러한 allocation fitness cost를 극복할 수 있는 처리방법을 종자학에서 말하는 "프라이밍(priming)"이라는 개념으로 극복하고자 했다. "프라이밍"은 종자 발아를 촉진시키기 위해 종자에 수분을 처리하고 뿌리가 발육되기 직전에 다시 건조시킴으로써 다시 종자가 수분에 접하게 되었을 때 그렇지 않은 종자에 비해 종자의 발아를 더욱 촉진시키기 위한 기술로서, SAR 유도물질을 종자에 직접 처리하여 식물 생장 억제에 미치는 영향을 최소화하면서 식물 전신유도저항성을 유도할 수 있는지를 조사하였다.The present inventors have attempted to overcome the treatment method that can overcome this allocation fitness cost while inducing resistance to plants with the concept of "priming" in seed science. "Priming" is a technique for further promoting germination of seeds compared to those that do not, when the seed comes into contact with moisture again by treating the seed with moisture to promote seed germination and drying again just before root development. We investigated whether plant-derived resistance could be induced by minimizing the effects of inducers directly on seeds.
본 발명은 상기와 같은 요구에 의해 안출된 것으로서, 본 발명자들은 유도저항성 물질인 BTH (benzothiadiazole)를 이용하여 식물 생장억제에 미치는 영향을 최소화하는 BTH의 침지 시간 및 농도를 알기 위해, 3가지 작물을 이용하여 종자처리제로서 가장 효과적인 조건을 알아보고자 한다.The present invention has been made in accordance with the above requirements, the inventors of the present invention to know the immersion time and concentration of BTH to minimize the effect on plant growth inhibition using BTH (benzothiadiazole), an induction resistant material, three crops To find out the most effective conditions for seed treatment.
상기 과제를 해결하기 위해, 본 발명은 식물체의 종자에 BTH (benzothiadiazole)를 처리하여 식물의 생장 장애 없이 또는 식물의 생장의 영향을 최소화하면서 식물병을 방제하는 방법을 제공한다.In order to solve the above problems, the present invention provides a method for controlling plant diseases by treating BTH (benzothiadiazole) on the seeds of the plant without minimizing the growth of the plant or minimizing the effect of plant growth.
본 발명은 또한, 고추, 대두 및 오이의 생장 장애 없이 또는 생장의 영향을 최소화하면서 고추, 대두 및 오이 유묘에 발병하는 식물병을 방제하기 위한 종자처리 조성물을 제공한다.The present invention also provides a seed treatment composition for controlling plant diseases affecting pepper, soybean and cucumber seedlings without minimizing the growth of pepper, soybean and cucumber or minimizing the effects of growth.
본 발명은 또한, 특정 농도의 BTH 용액에 침지한 후, 건조시켜 제조한 BTH가 도포된 고추 종자, 대두 종자 및 오이 종자를 제공한다.The present invention also provides pepper seed, soybean seed and cucumber seed coated with BTH prepared by immersion in a certain concentration of BTH solution.
본 발명은 또한, 고추, 대두 및 오이의 생장 장애 없이 또는 생장의 영향을 최소화하면서 고추, 대두 및 오이의 유묘기에 발병하는 식물병을 방제하기 위한 종자 처리 방법을 제공한다.The present invention also provides a seed treatment method for controlling plant diseases affecting seedlings of peppers, soybeans and cucumbers without minimizing the growth of peppers, soybeans and cucumbers or minimizing the effects of growth.
BTH를 유효성분으로 함유하는 조성물을 종자에 처리하여 병에 대한 저항성을 유도하는 본 발명의 방법으로 유묘기부터 식물병을 방제할 수 있으며, 각 종자별 최적 처리조건의 도출을 통해 식물병 방제용 종자처리제 개발에 활용할 수 있다.By treating the seed containing a composition containing BTH as an active ingredient to the seed, the method of the present invention can control plant diseases from seedlings, and to control plant diseases through derivation of optimal treatment conditions for each seed. It can be used to develop seed treatments.
도 1은 온실 조건에서 고추 종자를 SAR 유도 물질인 BTH를 농도별로 각각 0.1 μM, 10 μM, 및 1 mM로 나누고, 이 처리구를 다시 침지 시간별로 0, 6 및 12시간으로 처리하였을 때, 고추 세균점무늬병원균(Xanthomonas axonopodis pv. vesicatoria)에 대한 식물의 유도저항성과 그에 따른 생장의 효과를 나타낸 것이다. 각 처리구에 대한 줄기의 길이 (위) 및 병에 대한 저항성 (아래)을 측정하였다. 대조군 식물에는 물을 처리하였고, 발병도 (0 ~ 5)를 병원균 접종 후 7일째에 측정하였다 (0: 증상 없음, 5: 심한 괴사 증상). 데이터는 평균을 나타낸다. 모든 데이터는 JMP 소프트웨어 ver. 4.0을 이용하여 통계분석을 하였다 (SAS institute Inc., Cary, NC, USA). 처리구간 통계적 차이는 피셔(Fisher)의 최소유의차 검정법(LSD)에 의해 유의수준 P = 0.05에서 F-값의 등급으로 결정했고, 통계적 차이가 있는 처리구를 각각 a, b, c, d로 나타내었다.1 shows pepper seeds under greenhouse conditions when BTH, a SAR-inducing substance, is divided into concentrations of 0.1 μM, 10 μM, and 1 mM, respectively, and the treatment group is treated with 0, 6 and 12 hours again by soaking time. It shows the effect of plant induction resistance and its growth on the spotted pathogen ( Xanthomonas axonopodis pv.vesicatoria ). Stem length (top) and disease resistance (bottom) for each treatment were measured. Control plants were treated with water and the incidence (0-5) was measured 7 days after pathogen inoculation (0: no symptoms, 5: severe necrosis). The data represents the mean. All data is JMP software ver. Statistical analysis was performed using 4.0 (SAS institute Inc., Cary, NC, USA). Statistical differences in treatment intervals were determined by Fisher's least significant difference test (LSD) as a rating of F -value at the significance level P = 0.05, and the treatments with statistical differences were represented as a, b, c, and d, respectively. It was.
도 2는 다른 종자인 대두를 이용하여 도 1과 같은 처리구를 가지고 대두 세균점무늬병원균(Pseudomonas syringae pv. syringae)에 대한 식물의 유도저항성과 그에 따른 생장의 효과를 나타낸 것이다. 각 처리구에 대한 줄기의 길이 (위) 및 병에 대한 저항성 (아래)을 측정하였다. 대조군 식물은 물을 사용하였고, 발병정도는 병징을 나타내는 부분의 수를 측정하였고, 병원균 접종 후 7일째에 측정되었다. 데이터는 평균을 나타낸다.Figure 2 shows the effect of plant induction resistance and its growth on soybean bacterial spot pathogens ( Pseudomonas syringae pv. Syringae ) with the same treatment as in Figure 1 using another seed, soybean. Stem length (top) and disease resistance (bottom) for each treatment were measured. The control plants were water, and the incidence was measured by the number of the indicative parts, and 7 days after the pathogen inoculation. The data represents the mean.
도 3은 오이 종자를 이용하여 도 1 및 2와 같은 처리구를 가지고 오이 세균점무늬병원균 (Pseudomonas syringae pv. lachrymans)에 대한 식물의 유도저항성과 그에 따른 생장의 효과를 나타낸 것이다. 각 처리구에 대한 줄기의 길이 (위) 및 병에 대한 저항성 (아래)을 측정하였다. 대조군 식물은 물을 사용하였고, 발병정도는 병징을 나타내는 부분의 수를 측정하였고, 병원균 접종 후 7일째에 측정되었다. 데이터는 평균을 나타낸다.Figure 3 shows the effect of plant induction resistance and growth according to cucumber seed spot pathogens ( Pseudomonas syringae pv. Lachrymans ) with the treatment as shown in Figs. Stem length (top) and disease resistance (bottom) for each treatment were measured. The control plants were water, and the incidence was measured by the number of the indicative parts, and 7 days after the pathogen inoculation. The data represents the mean.
본 발명의 목적을 달성하기 위하여, 본 발명은 식물의 생장 장애 없이 또는 식물의 생장의 영향을 최소화하면서 식물병을 방제하는 방법을 제공한다. 구체적으로는, 상기 방법은 식물체의 종자에 BTH (benzothiadiazole)를 처리하는 단계, 및 처리된 식물체의 전신유도저항성(induced systemic resistance)을 유도하는 단계를 포함할 수 있다. In order to achieve the object of the present invention, the present invention provides a method for controlling plant diseases without disrupting the growth of the plant or minimizing the effect of the growth of the plant. Specifically, the method may include treating BTH (benzothiadiazole) with the seed of the plant, and inducing induced systemic resistance of the treated plant.
본 발명에 따른 식물병은 식물의 유묘기에 발병하는 것일 수 있으나, 이에 제한되지 않는다. 상기 식물병은 바람직하게는, 세균성 병일 수 있으며, 더욱 바람직하게는, 고추 세균성 점무늬병, 대두 세균성 점무늬병 또는 오이 세균성 점무늬병일 수 있으나, 이에 제한되지 않는다.Plant disease according to the present invention may be the onset of plant seedlings, but is not limited thereto. The plant disease may preferably be a bacterial disease, and more preferably, may be pepper bacterial spot pattern disease, soybean bacterial spot pattern disease or cucumber bacterial spot pattern disease, but is not limited thereto.
식물체의 종자에 처리하는 BTH의 농도 및 처리 시간은 식물체의 종자에 따라 상이할 수 있는데, 고추 종자의 경우에는, 바람직하게는 2~50 μM BTH 용액을 1초 내지 1시간 동안, 또는 0.02~50 μM BTH 용액을 4~8시간 동안 처리할 수 있으며, 더욱 바람직하게는, 10 μM BTH 용액을 1~10초 동안, 0.1 μM 또는 10 μM BTH 용액을 6시간 동안 처리할 수 있다.The concentration of BTH and the treatment time for the seed of the plant may vary depending on the seed of the plant. In the case of red pepper seeds, preferably 2-50 μM BTH solution is applied for 1 second to 1 hour, or 0.02 to 50 The μM BTH solution may be treated for 4-8 hours, more preferably the 10 μM BTH solution may be treated for 1-10 seconds and the 0.1 μM or 10 μM BTH solution for 6 hours.
대두 종자의 경우에는, 바람직하게는 0.02~50 μM BTH 용액을 4~8시간 동안 처리할 수 있으며, 더욱 바람직하게는 0.1 μM 또는 10 μM BTH 용액을 6시간 동안 처리할 수 있다.In the case of soybean seeds, preferably 0.02-50 μM BTH solution may be treated for 4-8 hours, more preferably 0.1 μM or 10 μM BTH solution may be treated for 6 hours.
오이 종자의 경우에는, 바람직하게는 2~50 μM BTH 용액을 1초 내지 1시간 동안 처리할 수 있으며, 더욱 바람직하게는 10 μM BTH 용액을 1~10초 동안 처리할 수 있다.In the case of cucumber seeds, preferably 2-50 μM BTH solution may be treated for 1 second to 1 hour, and more preferably 10 μM BTH solution may be treated for 1-10 seconds.
본 발명의 또 다른 목적을 달성하기 위하여, 본 발명은 식물의 생장 장애 없이 또는 식물의 생장의 영향을 최소화하면서 식물 유묘에 발병하는 식물병을 방제하기 위한 종자처리 조성물을 제공한다. 본 발명의 일 구현예에 따른 고추 또는 오이 종자 처리용 조성물은 바람직하게는 2~50 μM BTH를 유효성분으로 함유할 수 있으며, 더욱 바람직하게는 10 μM BTH를 유효성분으로 함유할 수 있다. 대두 종자 처리용 조성물은 바람직하게는 0.02~50 μM BTH를 유효성분으로 함유할 수 있으며, 더욱 바람직하게는 0.1 μM 또는 10 μM BTH를 유효성분으로 함유할 수 있다. In order to achieve still another object of the present invention, the present invention provides a seed treatment composition for controlling plant diseases affecting plant seedlings without minimizing the growth of plants or minimizing the effects of plant growth. Red pepper or cucumber seed treatment composition according to an embodiment of the present invention may preferably contain 2 to 50 μM BTH as an active ingredient, more preferably 10 μM BTH as an active ingredient. The soybean seed treatment composition may preferably contain 0.02 to 50 μM BTH as an active ingredient, and more preferably 0.1 μM or 10 μM BTH as an active ingredient.
본 발명의 일 구현예에 따른 조성물은 농약학적으로 허용되는 완충제, 담체, 보조제 또는 부형제를 포함할 수 있으며, 이는 당업계에 주지되어 있다. 본 발명의 조성물은 동결건조, 분무건조 또는 분무냉각을 통해 동결건조시킬 수 있다.Compositions according to one embodiment of the invention may comprise agrochemically acceptable buffers, carriers, adjuvants or excipients, which are well known in the art. The composition of the present invention may be lyophilized through lyophilization, spray drying or spray cooling.
용어 "완충제"는 pH를 안정화시킬 목적의 산-염기 혼합물을 함유하는 수용액을 의미하는 것으로 의도된다. 완충제는 트리스, 포스페이트, 카보네이트, 아세테이트, 시트레이트, 글리콜레이트, 락테이트, 보레이트, 타르트레이트, 카코딜레이트, 에탄올아민, 글리신, 이미다졸, 이미다졸락트산 등을 이용할 수 있다.The term "buffer" is intended to mean an aqueous solution containing an acid-base mixture for the purpose of stabilizing pH. Buffers may include tris, phosphate, carbonate, acetate, citrate, glycolate, lactate, borate, tartrate, cacodylate, ethanolamine, glycine, imidazole, imidazolactic acid, and the like.
용어 "희석제(또는 담체)"는 BTH를 희석시킬 목적의 수성 또는 비-수성 용액을 의미하는 것으로 의도된다. 희석제는 하나 이상의 염수, 물, 폴리에틸렌 글리콜, 프로필렌 글리콜, 에탄올 또는 오일(예: 옥수수 오일, 땅콩 오일, 면화씨 오일 또는 참깨 오일)일 수 있다.The term "diluent (or carrier)" is intended to mean an aqueous or non-aqueous solution for the purpose of diluting BTH. The diluent can be one or more saline, water, polyethylene glycol, propylene glycol, ethanol or oils such as corn oil, peanut oil, cottonseed oil or sesame oil.
용어 "보조제"는 BTH의 생물학적 효과를 증가시키기 위해 제형에 가해진 특정 화합물을 의미하는 것으로 의도된다.The term "adjuvant" is intended to mean a specific compound added to the formulation to increase the biological effect of BTH.
부형제는 하나 이상의 탄수화물, 중합체, 지질 및 무기물일 수 있다. 탄수화물의 예는 예를 들면, 동결건조를 용이하게 하기 위해 조성물에 첨가되는 락토즈, 슈크로즈, 만니톨 및 사이클로덱스트린을 포함한다.Excipients can be one or more carbohydrates, polymers, lipids and minerals. Examples of carbohydrates include, for example, lactose, sucrose, mannitol and cyclodextrins added to the composition to facilitate lyophilization.
중합체의 예로는 전분, 셀룰로즈 에테르, 셀룰로즈 카복실메틸셀룰로즈, 알기네이트, 카라기난, 하이알루론산, 폴리아크릴산, 폴리설포네이트, 폴리에틸렌글리콜/폴리에틸렌 옥사이드, 가수분해도가 상이한 폴리비닐알콜/폴리비닐아세테이트, 및 폴리비닐피롤리돈 (분자량이 상이한 것 모두)이 있다.Examples of polymers include starch, cellulose ether, cellulose carboxymethyl cellulose, alginate, carrageenan, hyaluronic acid, polyacrylic acid, polysulfonate, polyethyleneglycol / polyethylene oxide, polyvinylalcohol / polyvinylacetate with different degrees of hydrolysis, and polyvinylpi There is rolidone (all different in molecular weight).
본 발명의 조성물은 유제(乳劑), 유제(油劑), 수화제(水和劑), 분제(粉劑), 입제, 정제, 에어로졸제, 연고제 등과 같은 제형을 만들 수 있다. 필요하다면, 유화제, 현탁제, 전착제(展着劑), 침투제, 습윤제, 농후제(muscilage 등), 안정제 등을 배합시킬 수 있다. 이들 제형은 당업계에 공지된 제조 방법에 의하여 제조할 수 있다.The composition of the present invention can make formulations such as emulsions, emulsions, hydrates, powders, granules, tablets, aerosols, ointments, and the like. If necessary, an emulsifier, a suspending agent, an electrodeposition agent, a penetrating agent, a humectant, a thickening agent (muscilage, etc.), a stabilizer and the like can be added. These formulations can be prepared by methods of preparation known in the art.
본 발명의 또 다른 목적을 달성하기 위하여, 본 발명은 식물 종자를 BTH 용액에 침지한 후, 건조시켜 제조한 BTH가 도포된 식물 종자를 제공한다. 본 발명의 일 구현예에 따른 BTH가 도포된 고추 종자는 고추 종자를 바람직하게는, 1초 내지 1시간 동안 2~50 μM BTH 용액에, 또는 4~8시간 동안 0.02~50 μM BTH 용액에 침지한 후, 건조시켜 제조할 수 있으며, 더욱 바람직하게는, 1~10초 동안 10 μM BTH 용액에, 또는 6시간 동안 0.1 μM 또는 10 μM BTH 용액에 침지한 후, 건조시켜 제조할 수 있다.In order to achieve another object of the present invention, the present invention provides a plant seed coated with BTH prepared by immersing the plant seed in a BTH solution and then drying. Pepper seeds coated with BTH according to one embodiment of the present invention are preferably pepper seeds soaked in 2-50 μM BTH solution for 1 second to 1 hour, or 0.02-50 μM BTH solution for 4-8 hours. After that, it may be prepared by drying, more preferably, it may be prepared by immersing in 10 μM BTH solution for 1 to 10 seconds, or 0.1 μM or 10 μM BTH solution for 6 hours, and then dried.
본 발명의 일 구현예에 따른 BTH가 도포된 대두 종자는 대두 종자를 바람직하게는, 4~8시간 동안 0.02~50 μM BTH 용액에 침지한 후, 건조시켜 제조할 수 있으며, 더욱 바람직하게는, 대두 종자를 6시간 동안 0.1 μM 또는 10 μM BTH 용액에 침지한 후, 건조시켜 제조할 수 있다. The soybean seed coated with BTH according to one embodiment of the present invention may be prepared by soaking the soybean seed, preferably, dipped in 0.02-50 μM BTH solution for 4-8 hours, and then drying, and more preferably, Soybean seeds can be prepared by soaking in 0.1 μM or 10 μM BTH solution for 6 hours and then drying.
본 발명의 일 구현예에 따른 BTH가 도포된 오이 종자는 오이 종자를 바람직하게는, 1초 내지 1시간 동안 2~50 μM BTH 용액에 침지한 후, 건조시켜 제조할 수 있으며, 더욱 바람직하게는, 오이 종자를 1~10초 동안 10 μM BTH 용액에 침지한 후, 건조시켜 제조할 수 있다. Cu seeds coated with BTH according to an embodiment of the present invention may be prepared by immersing the cucumber seeds in 2-50 μM BTH solution for 1 second to 1 hour, and then drying them, more preferably. , Cucumber seeds can be prepared by immersing in 10 μM BTH solution for 1 to 10 seconds, and then dried.
본 발명의 또 다른 목적을 달성하기 위하여, 본 발명은 식물 종자에 BTH 용액을 처리하는 단계를 포함하는 식물의 생장 장애 없이 또는 식물의 생장의 영향을 최소화하면서 식물의 유묘기에 발병하는 식물병을 방제하기 위한 종자 처리 방법을 제공한다. In order to achieve another object of the present invention, the present invention provides a plant disease to the seedlings of the plant without disturbing the growth of the plant or minimizing the effect of the growth of the plant comprising the step of treating the BTH solution to the plant seed Provided is a seed treatment method for controlling.
상기 식물 종자에 BTH 용액 처리는 고추 종자에 바람직하게는, 2~50 μM BTH 용액을 1초 내지 1시간 동안, 또는 0.02~50 μM BTH 용액을 4~8시간 동안 처리할 수 있으며, 더욱 바람직하게는, 10 μM BTH 용액을 1~10초 동안, 또는 0.1 μM 또는 10 μM BTH 용액을 6시간 동안 처리할 수 있다. 대두 종자에 바람직하게는, 0.02~50 μM BTH 용액을 4~8시간 동안 처리할 수 있으며, 더욱 바람직하게는, 0.1 μM 또는 10 μM BTH 용액을 6시간 동안 처리할 수 있다. 오이 종자에 바람직하게는, 2~50 μM BTH 용액을 1초 내지 1시간 동안 처리할 수 있으며, 더욱 바람직하게는, 10 μM BTH 용액을 1~10초 동안 처리할 수 있다. Treatment of the BTH solution to the plant seeds is preferably pepper seed, 2 to 50 μM BTH solution can be treated for 1 second to 1 hour, or 0.02 to 50 μM BTH solution for 4 to 8 hours, more preferably The 10 μM BTH solution may be treated for 1-10 seconds, or 0.1 μM or 10 μM BTH solution for 6 hours. Soybean seeds may preferably be treated with 0.02-50 μM BTH solution for 4-8 hours, more preferably 0.1 μM or 10 μM BTH solution for 6 hours. Cucumber seeds may preferably be treated with 2-50 μM BTH solution for 1 second to 1 hour, and more preferably 10 μM BTH solution for 1-10 seconds.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.
1. 재료 및 방법1. Materials and Methods
식물 생장에 미치는 영향을 최소화하면서 식물 전신유도저항성을 유도하는 종자처리 방법을 알아보기 위해 고추 (㈜흥농종묘, 부강고추), 오이 (㈜농우바이오, 백봉다다기), 및 대두를 사용하였다.In order to investigate the seed treatment method that induces the systemic induction resistance while minimizing the effect on the plant growth, red pepper (Heungnong seedling, Bugang pepper), cucumber (Nongwoo bio, Baekbong tea) and soybean were used.
2.2. BTH 침지 시간 및 농도BTH Immersion Time and Concentration
각 종자에 대한 최적의 BTH 침지 시간 및 농도를 알아보기 위해 고추, 오이 및 대두 종자를 사용하였다. 각 종자를 0.1 μM, 10μM 및 1mM 농도의 BTH 용액에 침지하고, 침지 시간을 다시 0, 6 및 12시간으로 나누었다. 0 시간 침지는 각 농도의 BTH에 잠깐(1~10초) 침지한 것이다. 침지 후 종자를 건조시키기 위해 멸균 여과지 (Whatman)를 깐 페트리디쉬(petri dish)에 3일간 상온에 두었다가, 원예용 상토 (Punong Co., Ltd, Korea)를 채운 36공 포트 (28 × 54 × 6 cm)로 옮겨 심어, 고추 종자는 6주간, 오이 종자는 3주간, 그리고 대두 종자는 4주간 온실에서 20 ~ 30℃로 유지하며 재배하였다. Pepper, cucumber and soybean seeds were used to determine the optimal BTH soaking time and concentration for each seed. Each seed was immersed in BTH solutions at concentrations of 0.1 μM, 10 μM and 1 mM, and the immersion time was divided again into 0, 6 and 12 hours. The 0 hour immersion is a brief immersion (1-10 seconds) in BTH at each concentration. After soaking, a 36-hole pot (28 × 54 × 6) filled with sterile filter paper (Whatman) was kept at room temperature for 3 days and filled with horticultural soil (Punong Co., Ltd, Korea). cm), pepper seeds were grown for 6 weeks, cucumber seeds for 3 weeks, and soybean seeds for 4 weeks in the greenhouse at 20-30 ℃.
3. 병원균 접종3. Pathogen inoculation
고추작물은 세균점무늬병원균(Xanthomonas axonopodis pv. vesicatoria)을 107 CFU/ml 농도로 잎에 주입하였다. 7일 후에 병징을 관찰하면서 병 정도를 1 ~ 5로 측정하였고, 동시에 줄기의 길이도 측정하였다. 세균 접종원은 LB (Luria-Bertani) 고체 배지에 항생제 리팜피신(Rifampicin) 100 ㎍/㎖를 첨가하여 48시간 동안 30℃에서 배양하였다.Pepper crops were injected into the leaves at 10 7 CFU / ml Bacterial spot pathogen ( Xanthomonas axonopodis pv.vesicatoria ). After 7 days, the disease degree was measured by 1 to 5, and the length of the stem was also measured. Bacterial inoculum was incubated at 30 ° C. for 48 hours by adding 100 μg / ml of antibiotic Rifampicin to LB (Luria-Bertani) solid medium.
대두작물은 세균점무늬병원균(Psuedomonas syringae pv. syringae)을 107 CFU/ml 농도로 잎에 분무하였다. 7일 후에 병징을 관찰하면서 병 정도를 1 ~ 5로 측정하였고, 동시에 줄기의 길이도 측정하였다. 세균 접종원은 King's B 고체 배지에서 항생제 리팜피신(Rifampicin) 100 ㎍/㎖를 첨가하여 48시간 동안 30℃에서 배양하였다. Soybean crops were sprayed onto the leaves at 10 7 CFU / ml of Psuedomonas syringae pv. Syringae. After 7 days, the disease degree was measured by 1 to 5, and the length of the stem was also measured. Bacterial inoculum was incubated at 30 ° C. for 48 hours with the addition of 100 μg / ml of antibiotic Rifampicin in King's B solid medium.
오이작물은 세균점무늬병원균(Psuedomonas syringae pv. lachrymans)을 107 CFU/ml 농도로 잎에 분무하였다. 7일 후에 병징을 관찰하면서 병 정도를 1 ~ 5로 측정하였고, 동시에 줄기의 길이도 측정하였다. 세균 접종원은 King's B 고체 배지에서 항생제 리팜피신(Rifampicin) 100 ㎍/㎖를 첨가하여 48시간 동안 30℃에서 배양하였다.Cucumber crops were sprayed onto the leaves at a concentration of 10 7 CFU / ml B. psuedomonas syringae pv. Lachrymans . After 7 days, the disease degree was measured by 1 to 5, and the length of the stem was also measured. Bacterial inoculum was incubated at 30 ° C. for 48 hours with the addition of 100 μg / ml of antibiotic Rifampicin in King's B solid medium.
병징의 병 정도: Disease degree of symptom:
0 = 증상 없음, 1 = 약한 백화 증상, 2 = 백화 증상, 3 = 백화 증상과 약한 괴사, 4 = 괴사, 5 = 심한 괴사 증상 0 = no symptoms, 1 = mild bleaching, 2 = bleaching, 3 = bleaching and mild necrosis, 4 = necrosis, 5 = severe necrosis
실시예 1: 고추에서의 유도저항성 검정 및 식물 생장 측정Example 1: Induction resistance assay and plant growth measurement in red pepper
고추종자의 각 처리구를 대조군 0시간 침지 물 처리구와 비교해 보면 0시간 침지 했을 경우 0.1 μM, 10 μM, 그리고 1 mM BTH 모든 처리구에서 병의 정도가 통계적으로 차이가 나게 감소하는 것을 확인하였고, 6시간 침지 처리구에서는 1 mM BTH를 제외하고 두 가지 BTH 농도에서 병의 정도가 통계적으로 차이가 나게 감소하는 것을 확인하였다. 6시간 침지 처리구의 결과와는 반대로 12시간 침지 처리구에서는 1 mM BTH 침지 처리구에서만 병에 대한 저항성이 나타났다. 0.1 μM BTH 처리구는 0시간과 6시간에서 병에 대한 저항성이 나타났고, 12시간 침지 했을 때는 대조군과 비교했을 때 차이를 확인할 수 없었다. 10 μM BTH 처리구는 0.1 μM BTH 처리구와 같은 결과를 보였고, 모든 처리구들 중 0시간 및 6시간 침지에서 최고의 병저항성을 보였다. 1 mM BTH 처리구에서는 0시간 침지 시 가장 뛰어난 병저항성을 보였으나 6시간 침지 시에는 병저항성이 나타나지 않았고, 다시 12시간 침지 시 병저항성이 관찰되었다. 1 mM BTH 처리구에서는 상기 설명한 두 가지 농도가 12시간 침지했을 때 병저항성이 사라지는 경향과 다른 양상을 보였다. Comparing the treatment of pepper seeds with the control 0 hour immersion water treatment group, the severity of disease was decreased statistically in all treatments at 0.1 μM, 10 μM, and 1 mM BTH after 6 hours immersion, and 6 hours In the immersion treatment, except for 1 mM BTH, the degree of disease was significantly reduced at two BTH concentrations. Contrary to the results of the 6-hour soaking treatment, the 12-hour soaking treatment showed resistance to disease only in the 1 mM BTH soaking treatment. The 0.1 μM BTH treatment showed resistance to disease at 0 h and 6 h, and no difference was observed at 12 h. The 10 μM BTH treatment showed the same results as the 0.1 μM BTH treatment and showed the best disease resistance at 0 and 6 hours immersion among all treatments. 1 mM BTH treatment showed the best disease resistance after immersion for 0 hours, but no disease resistance after 6 hours immersion, and resistance to 12 hours immersion. In the 1 mM BTH treatment, the above-described two concentrations were different from the tendency of disease resistance to disappear after 12 hours of soaking.
각 처리구를 식물의 생장과 비교해 보았을 때, 1 mM BTH 처리구를 6시간 침지 시 식물생장이 저해되는 것을 제외하고는, 고추종자에서 침지 방법에 의해 병에 대한 저항성 유발로 인한 생장의 저해 현상은 나타나지 않는다는 것을 확인하였다. 상기 사실을 바탕으로 저농도 및 최소의 침지시간을 이용한 종자처리제를 개발하기 위해서는 고추종자에 10 μM BTH를 0시간 침지하는 것이 최적의 처리방법인 것으로 사료된다. Comparing each treatment with the growth of plants, the inhibition of growth due to induction of resistance to disease was not observed by the soaking method in pepper seeds, except that plant growth was inhibited after 6 hours of 1 mM BTH treatment. It was confirmed. Based on the above facts, it is considered that the optimal treatment method is to immerse 10 μM BTH in red pepper seeds for 0 hours in order to develop a seed treatment using low concentration and minimum immersion time.
실시예 2: 대두의 유도저항성 검정 및 식물 생장 측정Example 2: Induction Resistance Assay of Soybean and Measurement of Plant Growth
대두 종자의 각 처리구를 대조군 0시간 침지 물 처리구와 비교해 보면, 특이하게 BTH의 농도에 연관되어 저항성을 보이는 것이 아니라 침지 시간에 연관되어 저항성이 나타나는 것을 확인하였다. Comparing each treatment group of soybean seed with the control group 0 hours immersion treatment, it was confirmed that the resistance is not related to the concentration of BTH specifically, but to the immersion time.
6시간 침지 시에는 0.1 μM, 10 μM, 그리고 1 mM BTH 농도 각각 병에 대한 저항성이 통계적으로 차이가 나게 나타났고, 0시간 및 12시간 침지 시에는 병에 대한 저항성이 나타나지 않았다. 6시간 침지 시 0시간 물 침지 처리구와 비교해 보면 식물 생장억제 영향도 나타나지 않았다. 다만 BTH 농도 간 비교를 해보면 0.1 μM과 10 μM BTH 처리구가 1 mM BTH 처리구보다 생장이 더 잘 되는 것을 확인할 수 있었다. 상기 사실을 바탕으로 저농도, 최소의 침지시간을 이용한 종자처리제를 개발하기 위해서는 대두종자에 경우 0.1 μM BTH를 6시간 침지하는 것이 최적의 처리방법인 것으로 사료된다.At 6 h immersion, the resistance to disease was statistically different at 0.1 μM, 10 μM, and 1 mM BTH concentrations, respectively, and at 0 and 12 h, no resistance to disease was observed. 6 hours of soaking did not show the effect of plant growth inhibition compared to the 0 hours of soaking treatment. However, the comparison between BTH concentrations showed that 0.1 μM and 10 μM BTH treatments showed better growth than 1 mM BTH treatments. Based on the above facts, in order to develop a seed treatment using a low concentration and minimal soaking time, it is considered that the optimal treatment method is to soak 0.1 μM BTH for 6 hours in soybean seed.
실시예 3: 오이의 유도저항성 검정 및 식물 생장 측정Example 3: Induction resistance assay and plant growth measurement of cucumber
오이종자의 각 처리구를 대조군 0시간 침지 물 처리구와 비교해 보면 0시간 침지 시 10 μM와 1 mM BTH 처리구에서 병에 대한 저항성이 나타나는 것을 확인할 수 있었고, 0.1 μM BTH 처리구에서는 저항성이 나타나지 않았다. 6시간 침지 시에는 모든 농도에서 대조군보다 더 많은 병증상이 나타났다. 12시간 침지 시 10 μM BTH 처리구는 병증상을 대조군과 비교했을 때 더 심하게 나타났고, 다른 처리구는 통계적으로 차이가 나타나지 않았다. 0.1 μM BTH 처리구는 침지 시간에 따른 병에 대한 저항성이 나타나지 않았고, 10 μM BTH 처리구의 경우는 0시간 침지 시 병 저항성이 나타났으나 6시간과 12시간 침지 시에는 오히려 대조군보다 높은 병저항성이 나타났고, 1 mM BTH 처리구는 0시간 침지 시에 나타나는 병 저항성이 6시간 및 12시간 침지 시에는 사라지는 현상을 관찰할 수 있었다. Comparing each treatment group of cucumber seeds with the control group 0 hours immersion water treatment it was confirmed that the resistance to the disease at 10 μM and 1 mM BTH treatment at 0 hours soaking, the resistance did not appear in the 0.1 μM BTH treatment. After soaking for 6 hours, all symptoms showed more symptoms than the control group. At 12 h immersion, 10 μM BTH treatment showed more severe symptoms when compared to the control, and the other treatments showed no statistical difference. The 0.1 μM BTH treatment showed no resistance to disease according to the soaking time, and the 10 μM BTH treatment showed resistance to disease at 0 h immersion, but higher resistance at 6 h and 12 h than the control. In the 1 mM BTH treatment group, the disease resistance that appeared at 0 hours immersion was observed disappearing at 6 hours and 12 hours immersion.
오이종자의 경우, 대조군인 물 처리구를 처리 시간에 따라 비교해 보면 침지 시간이 길수록 병에 대한 저항성이 낮아지는 현상을 관찰할 수 있었는데 이러한 현상이 10 μM와 1 mM BTH 처리구에서도 나타나는 것을 발견할 수 있었다. 각 처리구를 식물의 생장과 비교해 보았을 때, 6시간 침지 시와 같이 병에 대한 저항성이 나타나지 않았을 때 생장이 더 잘 된다는 것을 확인할 수 있었다. 병에 대한 저항성이 나타났던 10 μM와 1 mM BTH 처리구에서는 1 mM BTH 처리구에서 생장의 저해가 일어나는 것을 관찰할 수 있었지만, 10 μM BTH 0 시간 처리구에서는 병에 대한 저항성이 나타날 뿐만 아니라 생장의 억제현상도 나타나지 않는 것으로 보아 오이에서 종자처리제로서 최적의 처리구인 것으로 확인되었다. In the case of cucumber seeds, the control of water treatment group was compared according to the treatment time, and the longer the soaking time, the lower the resistance to disease was observed. This phenomenon was also found in the 10 μM and 1 mM BTH treatment groups. . Comparing each treatment with the growth of the plant, it was confirmed that the growth was better when the disease resistance did not appear, such as when soaked for 6 hours. In 10 μM and 1 mM BTH treatments, growth inhibition was observed in 1 mM BTH treatment, but disease resistance and growth inhibition were observed in 10 μM BTH 0 hours treatment. In addition, it was confirmed that it is an optimal treatment as a seed treatment agent in cucumber.

Claims (16)

  1. 식물체의 종자에 BTH (benzothiadiazole)를 처리하는 단계; 및Treating BTH (benzothiadiazole) on the seed of the plant; And
    처리된 식물체의 전신유도저항성(induced systemic resistance)을 유도하는 단계를 포함하는 식물의 생장 장애 없이 또는 식물의 생장의 영향을 최소화하면서 식물병을 방제하는 방법.A method of controlling plant diseases without disrupting plant growth or minimizing the effects of plant growth, comprising inducing induced systemic resistance of the treated plant.
  2. 제1항에 있어서, 상기 식물병은 식물의 유묘기에 발병하는 것을 특징으로 하는 방법.The method of claim 1, wherein the plant disease occurs during the seedling period of the plant.
  3. 제1항에 있어서, 상기 식물병은 세균성 병인 것을 특징으로 하는 방법.The method of claim 1, wherein the plant disease is a bacterial disease.
  4. 제3항에 있어서, 상기 세균성 병은 고추 세균성 점무늬병, 대두 세균성 점무늬병 또는 오이 세균성 점무늬병인 것을 특징으로 하는 방법.The method of claim 3, wherein the bacterial disease is pepper bacterial spot pattern disease, soybean bacterial spot pattern disease or cucumber bacterial spot pattern disease.
  5. 제1항에 있어서, 고추 종자에 2~50 μM BTH 용액을 1초 내지 1시간 동안, 또는 0.02~50 μM BTH 용액을 4~8시간 동안 처리하는 것을 특징으로 하는 방법.The method of claim 1 wherein the pepper seeds are treated with 2-50 μM BTH solution for 1 second to 1 hour, or 0.02-50 μM BTH solution for 4-8 hours.
  6. 제1항에 있어서, 대두 종자에 0.02~50 μM BTH 용액을 4~8시간 동안 처리하는 것을 특징으로 하는 방법.The method of claim 1 wherein the soybean seed is treated with 0.02-50 μM BTH solution for 4-8 hours.
  7. 제1항에 있어서, 오이 종자에 2~50 μM BTH 용액을 1초 내지 1시간 동안 처리하는 것을 특징으로 하는 방법.The method of claim 1, wherein the cucumber seed is treated with a 2-50 μM BTH solution for 1 second to 1 hour.
  8. 2~50 μM BTH를 유효성분으로 함유하는, 고추의 생장 장애 없이 또는 고추의 생장의 영향을 최소화하면서 고추 유묘에 발병하는 식물병을 방제하기 위한 종자처리 조성물.A seed treatment composition containing 2-50 μM BTH as an active ingredient for controlling plant diseases affecting pepper seedlings without minimizing the growth of peppers or minimizing the effects of pepper growth.
  9. 0.02~50 μM BTH를 유효성분으로 함유하는, 대두의 생장 장애 없이 또는 대두의 생장의 영향을 최소화하면서 대두 유묘에 발병하는 식물병을 방제하기 위한 종자처리 조성물.A seed treatment composition containing 0.02-50 μM BTH as an active ingredient for controlling plant diseases affecting soybean seedlings without minimizing soybean growth or minimizing the effects of soybean growth.
  10. 2~50 μM BTH를 유효성분으로 함유하는, 오이의 생장 장애 없이 또는 오이의 생장의 영향을 최소화하면서 오이 유묘에 발병하는 식물병을 방제하기 위한 종자처리 조성물.A seed treatment composition containing 2-50 μM BTH as an active ingredient for controlling plant diseases affecting cucumber seedlings without minimizing the growth of cucumbers or minimizing the effects of cucumber growth.
  11. 고추 종자를 1초 내지 1시간 동안 2~50 μM BTH 용액에, 또는 4~8시간 동안 0.02~50 μM BTH 용액에 침지한 후, 건조시켜 제조한 BTH가 도포된 고추 종자.Pepper seeds coated with BTH prepared by immersing red pepper seeds in a 2-50 μM BTH solution for 1 second to 1 hour or in a 0.02-50 μM BTH solution for 4-8 hours.
  12. 대두 종자를 4~8시간 동안 0.02~50 μM BTH 용액에 침지한 후, 건조시켜 제조한 BTH가 도포된 대두 종자.BTH-coated soybean seeds prepared by soaking soybean seeds in 0.02-50 μM BTH solution for 4-8 hours and then drying.
  13. 오이 종자를 1초 내지 1시간 동안 2~50 μM BTH 용액에 침지한 후, 건조시켜 제조한 BTH가 도포된 오이 종자.Cucumber seeds coated with BTH prepared by immersing the cucumber seeds in a 2-50 μM BTH solution for 1 second to 1 hour and then drying.
  14. 고추 종자에 2~50 μM BTH 용액을 1초 내지 1시간 동안, 또는 0.02~50 μM BTH 용액을 4~8시간 동안 처리하는 것을 특징으로 하는 고추의 생장 장애 없이 또는 고추의 생장의 영향을 최소화하면서 고추의 유묘기에 발병하는 식물병을 방제하기 위한 종자 처리 방법.Pepper seeds are treated with 2-50 μM BTH solution for 1 second to 1 hour, or 0.02-50 μM BTH solution for 4 to 8 hours without pepper growth disorders or while minimizing the effects of pepper growth. Seed treatment method for controlling plant disease that occurs in pepper seedlings.
  15. 대두 종자에 0.02~50 μM BTH 용액을 4~8시간 동안 처리하는 것을 특징으로 하는 대두의 생장 장애 없이 또는 대두의 생장의 영향을 최소화하면서 대두의 유묘기에 발병하는 식물병을 방제하기 위한 종자 처리 방법.Seed treatment for controlling plant diseases affecting soybean seedlings without growing soybeans or minimizing the effects of soybean growth, characterized by treating the soybean seeds with 0.02-50 μM BTH solution for 4-8 hours. Way.
  16. 오이 종자에 2~50 μM BTH 용액을 1초 내지 1시간 동안 처리하는 것을 특징으로 하는 오이의 생장 장애 없이 또는 오이의 생장의 영향을 최소화하면서 오이의 유묘기에 발병하는 식물병을 방제하기 위한 종자 처리 방법.Seeds for controlling plant diseases affecting seedlings of cucumbers without growing disorders or minimizing the effects of cucumber growth, characterized by treating 2-50 μM BTH solutions for 1 second to 1 hour to cucumber seeds. Treatment method.
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WO1995019443A2 (en) * 1994-01-13 1995-07-20 Ciba-Geigy Ag Chemically regulatable and anti-pathogenic dna sequences and uses thereof
US7005562B2 (en) * 1999-12-15 2006-02-28 Syngenta Participations Ag SAR and pathogen-inducible promoter
US20080207448A1 (en) * 2007-02-23 2008-08-28 Vamtech, L.L.C. Coated seeds and methods of making coated seeds
KR20090051847A (en) * 2007-11-20 2009-05-25 한국생명공학연구원 Composition and method for increasing resistance against plant pathogen by comprising bacterial genetic materials, and plant produced by the method

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US7005562B2 (en) * 1999-12-15 2006-02-28 Syngenta Participations Ag SAR and pathogen-inducible promoter
US20080207448A1 (en) * 2007-02-23 2008-08-28 Vamtech, L.L.C. Coated seeds and methods of making coated seeds
KR20090051847A (en) * 2007-11-20 2009-05-25 한국생명공학연구원 Composition and method for increasing resistance against plant pathogen by comprising bacterial genetic materials, and plant produced by the method

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