WO2024080626A1 - Method for preparing bacillus licheniformis jws22003 strain for promoting fermentation of by-products and plant growth and producing auxin, and method for fermenting by-products and method for promoting plant growth by using same - Google Patents

Method for preparing bacillus licheniformis jws22003 strain for promoting fermentation of by-products and plant growth and producing auxin, and method for fermenting by-products and method for promoting plant growth by using same Download PDF

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WO2024080626A1
WO2024080626A1 PCT/KR2023/014634 KR2023014634W WO2024080626A1 WO 2024080626 A1 WO2024080626 A1 WO 2024080626A1 KR 2023014634 W KR2023014634 W KR 2023014634W WO 2024080626 A1 WO2024080626 A1 WO 2024080626A1
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jws22003
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licheniformis
bacillus licheniformis
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소재우
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주식회사 아그로셀
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    • 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
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22—Bacillus
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/10—Bacillus licheniformis

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  • the present invention relates to a Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P), which is characterized by promoting the fermentation of by-products, auxin production, and plant growth, a by-product fermentation method using the same, and a plant growth promotion method.
  • Bacillus licheniformis JWS22003 strain relates to a method of promoting fermentation of by-products using a 100-fold dilution of the Bacillus licheniformis JWS22003 strain, a method of promoting plant growth using a 100-fold dilution, and Bacillus Method for producing auxin by Bacillus licheniformis JWS22003 strain and EMS (ethyl methane sulfonate) to induce chemical mutation in Bacillus licheniformis JWS22003 strain collected from crop by-products and nearby soil. It relates to a method of producing Bacillus licheniformis JWS22003 strain by processing.
  • B. subtilis, B. thuringiensis, and B. licheniformis are attracting attention as organic farming materials of the Bacillus genus.
  • the previously mentioned B. subtilis and B. thuringiensis are used in various industrial fields as agricultural antibiotics, microbial fertilizers, and transgenic genetic materials.
  • suitable analysis technology for culturing and identification of B. licheniformis has not been established, and as a result, research and industrialization of useful strains are relatively insufficient.
  • Bacillus licheniformis (hereinafter referred to as B. licheniformis) is a Gram-negative, mesophilic bacterium commonly found in soil or crop residues. Depending on the strain, it can survive at temperatures as high as 50°C or higher, so there is a low risk of contamination, deterioration, or death when producing or distributing it as agricultural materials or self-manufactured by farmers, so it has recently been attracting attention for agricultural use.
  • microorganisms some may exhibit various useful functions ecologically through interactions with plant pathogens or plants.
  • the principle is, firstly, antibiosis through antibacterial antibiotics against Streptomyces blastmyces, S. griseus, Peicillium nigricans, Bacillus subtilis, etc., secondly, chitinase and ⁇ -1,3-glucanase, known as fungal outer membrane hydrolysis shows. Lytic action, which decomposes the cell walls of plant pathogens by enzymes; third, plant growth-promoting rhizobacteria (PGPR); fourth, promotes the fermentation of plant residues or by-products to produce highly efficient, high-quality fermented compost.
  • PGPR plant growth-promoting rhizobacteria
  • B. licheniformis is known to produce various enzymes to ferment by-products, promote plant growth, and biosynthesize auxin.
  • This technology focuses on the efficiency of agricultural material production and plant growth promotion (PGPR) through the high-temperature culturability of B. licheniformis known so far, explores new B. licheniformis with high vitality, and transforms it into EMS (Ethyl Methane Sulfonate) chemical mutation. The purpose is to develop a new strain and use the by-products of this strain to promote crop growth and fermentation.
  • PGPR agricultural material production and plant growth promotion
  • the purpose of the present invention is to produce Bacillus licheniformis JWS22003 strain, ferment by-products using the same, improve plant growth, and produce auxin.
  • the present invention aims to produce a Bacillus licheniformis JWS22003 strain that has high vigor in plant growth promotion (PGPR) and efficiency in agricultural material production through the possibility of cultivating B. licheniformis at high temperatures.
  • PGPR plant growth promotion
  • the Lactobacillus paracasei strain according to the present invention is a Bacillus licheniformis JWS22003 strain (Accession number: KACC) that promotes fermentation of by-products and plant growth and produces auxin. 92452P).
  • the method for promoting fermentation of by-products according to the present invention is characterized by promoting fermentation of by-products using a 100-fold dilution of Bacillus licheniformis JWS22003 strain (Accession number: KACC 92452P).
  • the method for promoting plant growth according to the present invention is characterized by promoting plant growth by foliar application of a 100-fold dilution of Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P) to crops. .
  • the method for producing the Bacillus licheniformis JWS22003 strain involves chemical mutation in the Bacillus licheniformis JWS22003 (Bacillus licheniformis) strain collected from crop by-products and nearby soil. It is characterized in that it is manufactured by treating EMS (ethyl methane sulfonate), which causes.
  • EMS ethyl methane sulfonate
  • the present invention has the effect of securing Bacillus licheniformis JWS22003 biological resources using EMS (Ethyl Methane Sulfonate) chemical mutation.
  • EMS Ethyl Methane Sulfonate
  • the present invention has the effect of improving compost fermentation through the Bacillus licheniformis JWS22003 strain.
  • the present invention has the effect of improving crop growth by the Bacillus licheniformis JWS22003 strain.
  • the present invention has the effect of producing auxin by the Bacillus licheniformis JWS22003 strain.
  • Figure 1 is a diagram showing in-flight culture of B. licheniformis JWS22003 strain.
  • Figure 2 shows genetic sequence information of B. licheniformis JWS22003 strain.
  • Figure 3 shows the results of genetic affinity analysis of B. licheniformis JWS22003 strain.
  • Figure 4 shows the results of HPLC analysis of auxin production of B. licheniformis JWS22003 strain.
  • Bacillus licheniformis JWS22003 (Accession Number: KACC 92452P) is characterized by promoting the fermentation of by-products and plant growth and producing auxin.
  • Bacillus licheniformis JWS22003 (Accession number: KACC 92452P) is EMS (ethyl), which induces chemical mutations in Bacillus licheniformis JWS22003 (Bacillus licheniformis) strain collected from crop by-products and nearby soil. It is manufactured by processing methane sulfonate).
  • B. licheniformis was isolated based on samples collected from the residue left as a by-product after harvesting rice and the nearby soil. Purely isolated B. licheniformis strains were selected for vigorous in vitro growth on nutritional agar medium. The selected B. licheniformis strain was cultured in NA liquid medium at a culture temperature of 28°C for 48 hours. The selected B. licheniformis strain was dispensed 10 times at 0.9ml each and then treated with 100 ⁇ l/ml EMS for 10 minutes. 100 ⁇ l of the above suspension was plated on NA medium and cultured at a culture temperature of 28°C for 48 hours. Among these, the strain with the most vigorous in-flight growth was selected and named B. licheniformis JWS22003.
  • B. licheniformis JWS22003 was applied to wood by-product compost and agricultural crops (Hwaseong rice, Daewon bean, green skirt lettuce, Chungwang cabbage, Geogeun pepper, and Hongro apple) in the control group, untreated, treatment group 1 diluted 10 times, treatment group 2, diluted 100 times, treatment group 2. It was treated with 500-fold dilution three times.
  • the physicochemical analysis of this material was conducted according to the AOAC (Association of Official Agricultural Chemists) standard analysis method.
  • B. licheniformis JWS22003 For B. licheniformis, single colonies with vigorous in vitro growth were selected on NA medium. The selected single colony strains were selected to induce EMS mutation, and then those that died or had low growth in vitro were culled, and one EMS mutant strain that grew vigorously in vitro on NA medium was finally selected and designated as B. licheniformis JWS22003. Named ( Figure 1).
  • the new B. licheniformis JWS22003 strain was treated for compost fermentation, and all treatments showed better effects than the untreated control.
  • the ammonia nitrogen content ranged from 20.41 mg/L to 78.99 mg/L, and the nitrate nitrogen content ranged from 76.04 mg/L to 182.13 mg/L.
  • the ammonia nitrogen content ranged from 21.15 mg/L to 63.46 mg/L, and the nitrate nitrogen content ranged from 80.28 mg/L to 173.80 mg/L.
  • Treatments 1 and 2 showed three times the effect compared to the control, and in particular, treatment 2 was found to be economical because the dose of B. licheniformis JWS22003 strain was 1/10 that of treatment 1.
  • Table 1 shows the change in nitrogen content after treatment in compost fermentation of B. licheniformis JWS22003 strain.
  • auxin is one of several regulators of plant germination and growth from seeds, and is especially involved in stem elongation. It is a type of growth hormone. Additionally, auxin is produced at the tips of stems and roots where growth is active, and promotes length growth by elongating cell walls. Accordingly, the production of auxin can promote the growth of plants.
  • B. licheniformis JWS22003 strain culture medium was foliar applied to rice (Hwaseong rice) three times, and it was confirmed that plant height, tiller number, and chlorophyll were improved over the entire growth period in all treatments compared to the control group (Table 2).
  • the plant height of 100-drainage treatment group 2 was 62.46cm for 30 days, 93.64cm for 60 days, 106.73cm for 90 days, tillering number was 10.96 plants/week for 30 days, 13.30 plants/week for 60 days, 13.77 plants/week for 90 days, and chlorophyll was 41.82 for 30 days. , 37.73 at 60 days and 35.48 at 90 days, showing that the B. licheniformis JWS22003 strain improved rice growth.
  • Table 2 shows the effect of improving the growth of rice (Hwaseong rice) after foliar application of B. licheniformis JWS22003 strain.
  • B. licheniformis JWS22003 strain culture medium was foliar applied to soybean (Daewon bean) three times, and it was confirmed that the length, number of branches, number of seeds, grain weight, production, and production rate were all improved compared to the control in all treatments (Table 3 ).
  • 100-fold treatment group No. 2 was the best with a length of 74.97 cm, number of branches 82.94 per week, number of seeds 250.60 grains per week, 27.88 g per hundred grains, production volume 259.56 kg/10a, and production rate 128%, resulting in B. licheniformis JWS22003 strain. was found to improve soybean growth.
  • Table 3 shows the effect of improving the growth of soybean (Daewon bean) after foliar fertilization of B. licheniformis JWS22003 strain.
  • B. licheniformis JWS22003 strain culture medium was foliar applied to lettuce (green chives) three times, and it was confirmed that the above-ground fresh weight and dry weight ratio, below-ground root length, below-ground fresh weight, and below-ground dry weight ratio were improved compared to the control in all treatments (Table 4).
  • 100-fold treatment group No. 2 was the best with 41.35 g above-ground live weight, 3.74% of dry matter, 35.76 cm root length below ground, 4.78 g of below-ground live weight, and 3.72% of below-ground dry matter, showing that the B. licheniformis JWS22003 strain improved lettuce growth. It appeared that it did.
  • Table 4 shows the effect of improving the growth of lettuce (green skirt) after foliar application of the B. licheniformis JWS22003 strain.
  • B. licheniformis JWS22003 strain culture medium was foliar applied to Chinese cabbage (Chungwang) three times, and it was confirmed that bulb weight, bulb height, corm, number of leaves, and chlorophyll were improved in all treatments compared to the control (Table 5).
  • 100-fold treatment group No. 2 was the best with 2.95 kg in bulb weight, 27.72 cm in height, 17.60 cm in diameter, 41.95 leaves, and 35.02 chlorophyll, showing that the B. licheniformis JWS22003 strain improves cabbage growth.
  • Table 5 shows the growth improvement effect of Chinese cabbage (Chungwang) after foliar fertilization treatment of B. licheniformis JWS22003 strain.
  • B. licheniformis JWS22003 strain culture medium was applied foliarly to pepper (Geochanghan) three times, and it was confirmed that plant height, number of leaves, leaf area, fresh weight, dry weight, fruit weight, fruit weight, and dry fruit weight were improved in all treatments compared to the control (Table 6 ).
  • plant height was 152.81cm
  • number of leaves was 2,088.34
  • leaf area was 25,592.94cm2/week
  • fresh weight was 489.13g/week
  • dry weight was 105.31g/week
  • fruit tree was 46.17 pieces/week
  • fruit weight was 537.52g/week
  • dry fruit weight was 84.24.
  • the B. licheniformis JWS22003 strain was found to be the best in terms of g/week, improving pepper growth.
  • Table 6 shows the growth improvement effect of pepper (Geochanghan) after foliar application of B. licheniformis JWS22003 strain.
  • B. licheniformis JWS22003 strain culture was applied to apples (Hongro) three times as foliar application, and it was confirmed that shoot volume, jugular, fruit tree, and chlorophyll were improved in all treatments compared to the control fruit (Table 7).
  • the shoot volume was 64.70 cm
  • the crown was 49.17 mm
  • the number of fruits was 36.09 per week
  • the chlorophyll was 35.97, showing that the B. licheniformis JWS22003 strain improved apple growth.
  • Table 7 shows the effect of improving the growth of apples (Hongro) after foliar fertilization of the B. licheniformis JWS22003 strain.
  • the B. licheniformis JWS22003 strain culture medium was 100 times higher. Treatment with was found to be the best, improving the growth of the B. licheniformis JWS22003 strain.
  • HPLC analysis was performed to determine the amount of auxin produced by the B. licheniformis JWS22003 strain, and the results showed that auxin was biosynthesized by the B. licheniformis JWS22003 strain. You can check that it happens.
  • the B. licheniformis JWS22003 strain focuses on the efficiency of agricultural material production and plant growth promotion (PGPR) through the high-temperature culturability of B. licheniformis and has high vitality, so it can be used to ferment by-products of this strain and promote crop growth. .
  • PGPR agricultural material production and plant growth promotion

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Abstract

Disclosed are: a Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P) for promoting fermentation of by-products, auxin production, and plant growth; a method for promoting fermentation of by-products and plant growth by using a 100-fold dilution of the Bacillus licheniformis JWS22003 strain, and a method for producing auxin by the Bacillus licheniformis JWS22003 strain; and a method for preparing a Bacillus licheniformis JWS22003 strain which is prepared by treating a Bacillus licheniformis JWS22003 strain collected from crop by-products and nearby soil with ethyl methane sulfonate (EMS), which causes chemical mutations.

Description

부산물의 발효와 식물의 생육을 촉진하고, 옥신을 생성하는 바실러스 리체니포르미스 JWS22003 균주의 제조방법 및 이를 이용한 부산물 발효방법과 식물 생육 촉진방법Manufacturing method of Bacillus Licheniformis JWS22003 strain that promotes fermentation of by-products and plant growth and produces auxin, and method of fermenting by-products and method of promoting plant growth using the same
본 발명은 부산물의 발효와 옥신 생성 및 식물의 생육을 촉진하는 것을 특징으로 하는 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P), 이를 이용한 부산물 발효방법, 식물 생육 촉진방법에 관한 것으로, 더욱 상세하게는 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주의 100배 희석액을 활용한 부산물의 발효를 촉진하는 방법, 100배 희석액을 활용한 식물의 생육을 촉진하는 방법, 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주에 의한 옥신을 생성하는 방법 및 농작물 부산물과 인근 토양에서 수집한 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis) 균주에 화학적 돌연변이를 유발하는 EMS(ethyl methane sulfonate)를 처리하여 바실러스 리체 니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주를 제조하는 방법에 관한 것이다.The present invention relates to a Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P), which is characterized by promoting the fermentation of by-products, auxin production, and plant growth, a by-product fermentation method using the same, and a plant growth promotion method. More specifically, it relates to a method of promoting fermentation of by-products using a 100-fold dilution of the Bacillus licheniformis JWS22003 strain, a method of promoting plant growth using a 100-fold dilution, and Bacillus Method for producing auxin by Bacillus licheniformis JWS22003 strain and EMS (ethyl methane sulfonate) to induce chemical mutation in Bacillus licheniformis JWS22003 strain collected from crop by-products and nearby soil. It relates to a method of producing Bacillus licheniformis JWS22003 strain by processing.
현재 Bacillus 속 유기농업자재로 B. subtilis, B. thuringiensis, B. licheniformis 등이 주목받고 있다. 앞서 언급한 B. subtilis와 B. thuringiensis는 다른 미생물과 비교해 농업용 항생제, 미생물 비료, 형질전환 유전재료 등으로 다양한 산업분야에서 활용되고 있다. 반면 B. licheniformis는 배양과 동정에 적합한 분석기술이 정립되지 않았고, 이로 인해 유용 균주에 대한 연구와 산업화도 상대적으로 미흡하다.Currently, B. subtilis, B. thuringiensis, and B. licheniformis are attracting attention as organic farming materials of the Bacillus genus. Compared to other microorganisms, the previously mentioned B. subtilis and B. thuringiensis are used in various industrial fields as agricultural antibiotics, microbial fertilizers, and transgenic genetic materials. On the other hand, suitable analysis technology for culturing and identification of B. licheniformis has not been established, and as a result, research and industrialization of useful strains are relatively insufficient.
Bacillus licheniformis(이하 B. licheniformis)는 토양 혹은 농작물의 잔사물에서 흔하게 발견되는 그람음성, 중온성 세균이다. 균주에 따라서 50℃ 혹은 그 이상의 더 높은 온도에서도 생존할 수 있어 농가에서 자가 제조 혹은 농업자재로 생산ㆍ유통시 오염, 변질, 사멸 등의 우려가 낮기 때문에 최근 농업적으로 활용하기 위해 주목 받고 있다.Bacillus licheniformis (hereinafter referred to as B. licheniformis) is a Gram-negative, mesophilic bacterium commonly found in soil or crop residues. Depending on the strain, it can survive at temperatures as high as 50°C or higher, so there is a low risk of contamination, deterioration, or death when producing or distributing it as agricultural materials or self-manufactured by farmers, so it has recently been attracting attention for agricultural use.
여러 가지 미생물중에는 생태학적으로 식물병원균 또는 식물과의 상호작용에 의해 여러 가지 유용한 기능을 나타내는 경우가 있다. 그 원리는 첫째 Streptomyces blastmyces, S. griseus, Peicillium nigricans, Bacillus subtilis 등에 대한 항균성 항생물질을 통한 억제 항생작용(antibiosis), 둘째, 진균 외막 가수분해쇼로 알려진 chitinase, β-1,3-glucanase와 같은 효소에 의한 식물병원균의 세포벽 분해를 시키는 용균작용, 셋째, 식물 성장 촉진세균(plant growth-promoting rhizobacteria, PGPR), 넷째, 식물 잔사물 또는 부산물의 발효를 촉진하여 고효율, 고품질의 발효 퇴비를 생산할 수 있도록 cellulase 및 chitinase 등의 효소 분비력이 우수한 경우 등이 있다. 특히 B. licheniformis는 다양한 효소를 만들어 부산물 발효와 식물 생육 촉진 및 옥신(auxin)을 생합성하는 것으로 알려져 있다.Among various microorganisms, some may exhibit various useful functions ecologically through interactions with plant pathogens or plants. The principle is, firstly, antibiosis through antibacterial antibiotics against Streptomyces blastmyces, S. griseus, Peicillium nigricans, Bacillus subtilis, etc., secondly, chitinase and β-1,3-glucanase, known as fungal outer membrane hydrolysis shows. Lytic action, which decomposes the cell walls of plant pathogens by enzymes; third, plant growth-promoting rhizobacteria (PGPR); fourth, promotes the fermentation of plant residues or by-products to produce highly efficient, high-quality fermented compost. There are cases where the secretion of enzymes such as cellulase and chitinase is excellent. In particular, B. licheniformis is known to produce various enzymes to ferment by-products, promote plant growth, and biosynthesize auxin.
본 기술은 지금까지 알려진 B. licheniformis의 고온 배양 가능성을 통한 농업자재 생산의 효율성과 식물생장촉진(PGPR)에 주목하여 높은 활력을 지닌 새로운 B. licheniformis를 탐색하고 이를 EMS(Ethyl Methane Sulfonate) 화학돌연변이를 이용하여 새로운 균주을 개발하고, 본 균주의 부산물 발효, 농작물 생육 촉진에 활용하는 것을 목적으로 한다.This technology focuses on the efficiency of agricultural material production and plant growth promotion (PGPR) through the high-temperature culturability of B. licheniformis known so far, explores new B. licheniformis with high vitality, and transforms it into EMS (Ethyl Methane Sulfonate) chemical mutation. The purpose is to develop a new strain and use the by-products of this strain to promote crop growth and fermentation.
본 발명은 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주의 제조 및 이를 이용한 부산물 발효와 식물 생육 개선 및 옥신 생성을 목적으로 한다.The purpose of the present invention is to produce Bacillus licheniformis JWS22003 strain, ferment by-products using the same, improve plant growth, and produce auxin.
또한, 본 발명은 B. licheniformis의 고온 배양 가능성을 통한 농업자재 생산의 효율성과 식물생장촉진(PGPR)에 높은 활력을 지닌 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주의 제조를 목적으로 한다.In addition, the present invention aims to produce a Bacillus licheniformis JWS22003 strain that has high vigor in plant growth promotion (PGPR) and efficiency in agricultural material production through the possibility of cultivating B. licheniformis at high temperatures.
상기와 같은 목적을 달성하기 위해 본 발명에 따른 락토바실러스 파라카제이 균주는 부산물의 발효와 식물의 생육을 촉진하며 옥신을 생성하는 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)인 것을 특징으로 한다.In order to achieve the above object, the Lactobacillus paracasei strain according to the present invention is a Bacillus licheniformis JWS22003 strain (Accession number: KACC) that promotes fermentation of by-products and plant growth and produces auxin. 92452P).
또한, 본 발명에 따른 부산물 발효 촉진 방법은 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)의 100배 희석액을 활용하여 부산물의 발효를 촉진하는 것을 특징으로 한다.In addition, the method for promoting fermentation of by-products according to the present invention is characterized by promoting fermentation of by-products using a 100-fold dilution of Bacillus licheniformis JWS22003 strain (Accession number: KACC 92452P).
또한, 본 발명에 따른 식물 생육 촉진방법은 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)의 100배 희석액을 농작물에 엽면시비하여 식물의 생육을 촉진하는 것을 특징으로 한다.In addition, the method for promoting plant growth according to the present invention is characterized by promoting plant growth by foliar application of a 100-fold dilution of Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P) to crops. .
또한, 본 발명에 따른 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P) 제조방법은 농작물 부산물과 인근 토양에서 수집한 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis) 균주에 화학적 돌연변이를 유발하는 EMS(ethyl methane sulfonate)를 처리하여 제조되는 방법인 것을 특징으로 한다.In addition, the method for producing the Bacillus licheniformis JWS22003 strain (Accession number: KACC 92452P) according to the present invention involves chemical mutation in the Bacillus licheniformis JWS22003 (Bacillus licheniformis) strain collected from crop by-products and nearby soil. It is characterized in that it is manufactured by treating EMS (ethyl methane sulfonate), which causes.
본 발명은 EMS(Ethyl Methane Sulfonate) 화학돌연변이를 이용하여 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 생물 자원을 확보할 수 있다는 효과가 있다.The present invention has the effect of securing Bacillus licheniformis JWS22003 biological resources using EMS (Ethyl Methane Sulfonate) chemical mutation.
또한, 본 발명은 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주를 통해 퇴비 발효를 개선할 수 있다는 효과가 있다.In addition, the present invention has the effect of improving compost fermentation through the Bacillus licheniformis JWS22003 strain.
또한, 본 발명은 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주에 의한 농작물 생육을 개선할 수 있다는 효과가 있다.In addition, the present invention has the effect of improving crop growth by the Bacillus licheniformis JWS22003 strain.
또한, 본 발명은 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주에 의해 옥신을 생성할 수 있다는 효과가 있다.In addition, the present invention has the effect of producing auxin by the Bacillus licheniformis JWS22003 strain.
도 1은 B. licheniformis JWS22003 균주의 기내배양을 나타낸 도면. Figure 1 is a diagram showing in-flight culture of B. licheniformis JWS22003 strain.
도 2는 B. licheniformis JWS22003 균주의 유전염기서열정보.Figure 2 shows genetic sequence information of B. licheniformis JWS22003 strain.
도 3은 B. licheniformis JWS22003 균주의 유전적 유연관계분석 결과. Figure 3 shows the results of genetic affinity analysis of B. licheniformis JWS22003 strain.
도 4는 B. licheniformis JWS22003 균주의 옥신 생성량 HPLC 분석결과.Figure 4 shows the results of HPLC analysis of auxin production of B. licheniformis JWS22003 strain.
바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)는 부산물의 발효와 식물의 생육을 촉진하고 옥신을 생성하는 것을 특징으로 한다.Bacillus licheniformis JWS22003 (Accession Number: KACC 92452P) is characterized by promoting the fermentation of by-products and plant growth and producing auxin.
바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P) 제조방법은 농작물 부산물과 인근 토양에서 수집한 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis) 균주에 화학적 돌연변이를 유발하는 EMS(ethyl methane sulfonate)를 처리하여 제조되는 방법이다.The manufacturing method of Bacillus licheniformis JWS22003 (Accession number: KACC 92452P) is EMS (ethyl), which induces chemical mutations in Bacillus licheniformis JWS22003 (Bacillus licheniformis) strain collected from crop by-products and nearby soil. It is manufactured by processing methane sulfonate).
식물의 생육을 촉진하기 위해 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁 번호: KACC 92452P)의 100배 희석액을 농작물에 엽면시비할 경우, 3회에 걸쳐 엽면시비하는 것이 바람직하다.When foliar application of a 100-fold dilution of Bacillus licheniformis JWS22003 strain (Accession Number: KACC 92452P) to crops to promote plant growth, it is desirable to apply foliar application three times.
이하, 도면을 참고하면서 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail with reference to the drawings.
1. 개발 및 분석방법1. Development and analysis method
벼를 수확한 후 부산물로 남겨진 잔사물 및 그 인근 토양에서 채취한 시료를 토대로 B. licheniformis를 분리하였다. 순수 분리된 B. licheniformis는 세균영양배지(Nutrition agar medium)에서 기내 생육이 왕성한 균주를 선발하였다. 선발된 B. licheniformis 균주 NA 액체배지에서 배양온도 28℃에서 48시간 배양하였다. 선발된 B. licheniformis 균주를 0.9ml씩 10반복으로 분주한 후 100㎕/ml의 EMS를 10분간 처리하였다. 상기의 현탁액을 100㎕을 취해 NA 배지에 평판도말한 후 배양온도 28℃에서 48시간 배양하였다. 이 중에서 기내 생육이 가장 왕성한 균주를 선발하여 B. licheniformis JWS22003로 명명하였다.B. licheniformis was isolated based on samples collected from the residue left as a by-product after harvesting rice and the nearby soil. Purely isolated B. licheniformis strains were selected for vigorous in vitro growth on nutritional agar medium. The selected B. licheniformis strain was cultured in NA liquid medium at a culture temperature of 28°C for 48 hours. The selected B. licheniformis strain was dispensed 10 times at 0.9ml each and then treated with 100㎕/ml EMS for 10 minutes. 100㎕ of the above suspension was plated on NA medium and cultured at a culture temperature of 28°C for 48 hours. Among these, the strain with the most vigorous in-flight growth was selected and named B. licheniformis JWS22003.
명명된 B. licheniformis JWS22003를 목재 부산물 퇴비, 농작물(화성벼, 대원콩, 청치마 상추, 춘광 배추, 거창한 고추, 홍로 사과)에 대조구 무처리, 처리구 1번 10배 희석, 처리구 2번 100배 희석, 처리구 3번 500배 희석으로 처리하였다. 본 자료의 이화학성 분석은 AOAC(Association of Official Agricultural Chemists) 표준 분석법에 따라 분석하였다.The named B. licheniformis JWS22003 was applied to wood by-product compost and agricultural crops (Hwaseong rice, Daewon bean, green skirt lettuce, Chungwang cabbage, Geogeun pepper, and Hongro apple) in the control group, untreated, treatment group 1 diluted 10 times, treatment group 2, diluted 100 times, treatment group 2. It was treated with 500-fold dilution three times. The physicochemical analysis of this material was conducted according to the AOAC (Association of Official Agricultural Chemists) standard analysis method.
2. 개발 결과2. Development results
B. licheniformis는 NA 배지에서 기내 생육이 왕성한 단일 콜로니를 선발하였다. 선발된 단일 콜로니 균주는 EMS 돌연변이를 유발시킨 후 이 중 사멸하거나 기내 생육이 저조한 균주를 도태시키고, NA 배지에서 기내 생육이 왕성한 기내 생육이 왕성한 EMS 돌연변이 균주 1종을 최종 선발하여 B. licheniformis JWS22003로 명명하였다(도 1).For B. licheniformis, single colonies with vigorous in vitro growth were selected on NA medium. The selected single colony strains were selected to induce EMS mutation, and then those that died or had low growth in vitro were culled, and one EMS mutant strain that grew vigorously in vitro on NA medium was finally selected and designated as B. licheniformis JWS22003. Named (Figure 1).
선발된 EMS 돌연변이 B. licheniformis JWS22003 균주의 유전자 염기서열을 정렬 및 분석하였다(도 2).The gene sequences of the selected EMS mutant B. licheniformis JWS22003 strain were aligned and analyzed (Figure 2).
이 유전 정보를 토대로 미국 국립생물공학 정보센터(National Center for Biotechnology Information)에 등록된 기존의 바실러스 서브틸리스와 ITS16s rRNA 유전자 염기 서열의 비교 분석 및 유연관계분석(phylogenetic tree)을 통해 지금까지 밝혀진 동종의 바실러스 서브틸리스(B. licheniformis)와 비교해 다르다는 것을 확인하였다(도 3).Based on this genetic information, the homologous species discovered so far through comparative analysis and phylogenetic tree of the existing Bacillus subtilis and ITS16s rRNA gene sequences registered at the U.S. National Center for Biotechnology Information It was confirmed that it was different compared to Bacillus subtilis (B. licheniformis) (Figure 3).
3. 분석 결과3. Analysis results
본 발명에서 새로운 B. licheniformis JWS22003 균주를 퇴비 발효에 처리한 결과 모든 처리구에서 무처리한 대조구보다 좋은 효과를 보였다. 처리구 1번에서 암모니아태질소 함량 20.41mg/L에서 78.99mg/L, 질산태질소 함량 76.04mg/L에서 182.13mg/L로 나타났다. 처리구 2번에서 암모니아태질소 함량 21.15mg/L에서 63.46mg/L, 질산태 질소 함량 80.28mg/L에서 173.80mg/L로 나타났다. 처리구 1, 2번의 두 처리구에서 대조구와 비교해 3배에 이르는 효과를 보였고, 특히 처리구 2번이 처리구 1과 비해 B. licheniformis JWS22003 균주의 투여량이 1/10 수준이기 때문에 경제적인 것으로 나타났다.In the present invention, the new B. licheniformis JWS22003 strain was treated for compost fermentation, and all treatments showed better effects than the untreated control. In treatment group 1, the ammonia nitrogen content ranged from 20.41 mg/L to 78.99 mg/L, and the nitrate nitrogen content ranged from 76.04 mg/L to 182.13 mg/L. In treatment group 2, the ammonia nitrogen content ranged from 21.15 mg/L to 63.46 mg/L, and the nitrate nitrogen content ranged from 80.28 mg/L to 173.80 mg/L. Treatments 1 and 2 showed three times the effect compared to the control, and in particular, treatment 2 was found to be economical because the dose of B. licheniformis JWS22003 strain was 1/10 that of treatment 1.
표 1은 B. licheniformis JWS22003 균주의 퇴비 발효에 처리후 질소 함량 변화를 나타낸 것이다.Table 1 shows the change in nitrogen content after treatment in compost fermentation of B. licheniformis JWS22003 strain.
처리 전 시료 (mg/L)Sample before treatment (mg/L) 처리후 시료 (mg/L)Sample after treatment (mg/L)
NH₄+NH₄+ NO₃-NO₃- NH₄+NH₄+ NO₃-NO₃-
대조구Control 19.2519.25 75.5875.58 44.4844.48 61.8661.86
처리구1 Treatment area 1 20.4120.41 76.0476.04 78.9978.99 182.13182.13
처리구2Treatment area 2 21.1521.15 80.2880.28 63.4663.46 173.80173.80
처리구3Treatment area 3 21.4921.49 74.9974.99 54.4654.46 86.6886.68
B. licheniformis JWS22003 균주의 옥신 생성량을 알아보기 위해 HPLC 분석을 하였다. 그 결과 B. licheniformis JWS22003 균주에 의해 옥신이 생합성되는 것을 확인할 수 있었다(도 4).여기서 옥신은 식물이 씨에서 발아하여 생장하는 데에는 여러 가지 조절물질 중 하나인데, 특히 줄기의 신장에 관여하는 식물생장호르몬의 일종이다. 또한 옥신은 생장이 왕성한 줄기와 뿌리 끝에서 만들어지며 세포벽을 신장시킴으로써 길이 생장을 촉진한다. 이에 따라 옥신이 생성됨으로 인해 식물체의 생장을 촉진할 수 있다.HPLC analysis was performed to determine the amount of auxin produced by the B. licheniformis JWS22003 strain. As a result, it was confirmed that auxin is biosynthesized by the B. licheniformis JWS22003 strain (Figure 4). Here, auxin is one of several regulators of plant germination and growth from seeds, and is especially involved in stem elongation. It is a type of growth hormone. Additionally, auxin is produced at the tips of stems and roots where growth is active, and promotes length growth by elongating cell walls. Accordingly, the production of auxin can promote the growth of plants.
B. licheniformis JWS22003 균주 배양액을 벼(화성벼)에 3회 걸쳐 엽면시비하였는데, 모든 처리구에서 대조구 대비 초장, 분얼수, 엽록소가 전 생육기간에 걸쳐 개선되는 것을 확인하였다(표 2). 특히 100배액 처리구 2번의 초장이 30일 62.46cm, 60일 93.64cm, 90일 106.73cm, 분얼수 30일 10.96개/주, 60일 13.30개/주, 90일 13.77개/주, 엽록소 30일 41.82, 60일 37.73, 90일 35.48로 가장 우수하여 B. licheniformis JWS22003 균주가 벼 생육을 향상시키는 것으로 나타났다.B. licheniformis JWS22003 strain culture medium was foliar applied to rice (Hwaseong rice) three times, and it was confirmed that plant height, tiller number, and chlorophyll were improved over the entire growth period in all treatments compared to the control group (Table 2). In particular, the plant height of 100-drainage treatment group 2 was 62.46cm for 30 days, 93.64cm for 60 days, 106.73cm for 90 days, tillering number was 10.96 plants/week for 30 days, 13.30 plants/week for 60 days, 13.77 plants/week for 90 days, and chlorophyll was 41.82 for 30 days. , 37.73 at 60 days and 35.48 at 90 days, showing that the B. licheniformis JWS22003 strain improved rice growth.
표 2는 B. licheniformis JWS22003 균주의 엽면시비 처리후 벼(화성벼)의 생육 개선 효과를 나타낸 것이다.Table 2 shows the effect of improving the growth of rice (Hwaseong rice) after foliar application of B. licheniformis JWS22003 strain.
초장 (cm)Height (cm) 분얼수 (ea/plant)Tillering water (ea/plant) 엽록소 (SPAD)Chlorophyll (SPAD)
30일30 days 60일60 days 90일90 days 30일30 days 60일60 days 90일90 days 30일30 days 60일60 days 90일90 days
대조구Control 56.5856.58 84.1384.13 102.27102.27 7.457.45 10.9810.98 12.6312.63 34.0234.02 31.3231.32 32.9132.91
처리구1 Treatment area 1 59.5259.52 85.9585.95 100.41100.41 9.789.78 11.5511.55 11.1311.13 39.6439.64 32.5732.57 34.0334.03
처리구2Treatment area 2 62.4662.46 93.6493.64 106.73106.73 10.9610.96 13.3013.30 13.7713.77 41.8241.82 37.7337.73 35.4835.48
처리구3Treatment area 3 59.1159.11 91.9591.95 105.05105.05 10.9110.91 13.0713.07 13.6613.66 41.3941.39 36.3836.38 33.8233.82
B. licheniformis JWS22003 균주 배양액을 콩(대원콩)에 3회 걸쳐 엽면시비하였는데, 모든 처리구에서 대조구 대비 경장, 분지수, 종자수, 백립중, 생산량, 생산률이 모두 개선되는 것을 확인하였다(표 3). 특히 100배액의 처리구 2번이 경장 74.97cm, 분지수 82.94개/주, 종자수 250.60립/주, 백립중 27.88g, 생산량 259.56kg/10a, 생산률 128%로 가장 우수하여 B. licheniformis JWS22003 균주가 콩 생육을 향상시키는 것으로 나타났다.표 3은 B. licheniformis JWS22003 균주의 엽면시비 처리후 콩(대원콩)의 생육 개선 효과를 나타낸 것이다.B. licheniformis JWS22003 strain culture medium was foliar applied to soybean (Daewon bean) three times, and it was confirmed that the length, number of branches, number of seeds, grain weight, production, and production rate were all improved compared to the control in all treatments (Table 3 ). In particular, 100-fold treatment group No. 2 was the best with a length of 74.97 cm, number of branches 82.94 per week, number of seeds 250.60 grains per week, 27.88 g per hundred grains, production volume 259.56 kg/10a, and production rate 128%, resulting in B. licheniformis JWS22003 strain. was found to improve soybean growth. Table 3 shows the effect of improving the growth of soybean (Daewon bean) after foliar fertilization of B. licheniformis JWS22003 strain.
경장
(cm)light suit
(cm) 		분지수
(개/주)basin number
(recast) 		종자수
(립/주)number of seeds
(lip/state) 		백립중
(g)Baekrip Middle School
(g) 		생산량
(kg/10a)output
(kg/10a) 		생산률
(%)production rate
(%)
대조구Control 67.4067.40 74.3674.36 215.52215.52 24.0624.06 203.25203.25 100100
처리구1 Treatment area 1 72.3872.38 75.1875.18 230.40230.40 26.0326.03 228.69228.69 113113
처리구2Treatment area 2 74.9774.97 82.9482.94 250.60250.60 27.8827.88 259.56259.56 128128
처리구3Treatment area 3 72.2472.24 71.8271.82 243.18243.18 25.2225.22 231.71231.71 114114
B. licheniformis JWS22003 균주 배양액을 상추(청치마)에 3회 걸쳐 엽면시비하였는데, 모든 처리구에서 대조구 대비 지상부 생체중과 건물중 비율, 지하부 뿌리 길이, 지하부 생체중, 지하부 건물중 비율이 개선되는 것을 확인하였다(표 4). 특히 100배액의 처리구 2번이 지상부 생체중 41.35g, 건물중 비중 3.74%, 지하부 뿌리길이 35.76cm, 지하부 생체중 4.78g, 지하부 건물중 비중 3.72%로 가장 우수하여 B. licheniformis JWS22003 균주가 상추 생육을 향상시키는 것으로 나타났다.B. licheniformis JWS22003 strain culture medium was foliar applied to lettuce (green chives) three times, and it was confirmed that the above-ground fresh weight and dry weight ratio, below-ground root length, below-ground fresh weight, and below-ground dry weight ratio were improved compared to the control in all treatments (Table 4). In particular, 100-fold treatment group No. 2 was the best with 41.35 g above-ground live weight, 3.74% of dry matter, 35.76 cm root length below ground, 4.78 g of below-ground live weight, and 3.72% of below-ground dry matter, showing that the B. licheniformis JWS22003 strain improved lettuce growth. It appeared that it did.
표 4는 B. licheniformis JWS22003 균주의 엽면시비 처리후 상추(청치마)의 생육 개선 효과를 나타낸 것이다.Table 4 shows the effect of improving the growth of lettuce (green skirt) after foliar application of the B. licheniformis JWS22003 strain.
지상부above ground 지하부underground part
생체중
(g)live weight
(g) 		건물중 비율
(%)Proportion of buildings
(%) 		뿌리 길이
(cm)root length
(cm) 		생체중
(g)live weight
(g) 		건물중 비율
(%)Proportion of buildings
(%)
대조구Control 33.4633.46 3.583.58 30.7330.73 3.393.39 3.243.24
처리구1 Treatment area 1 38.0738.07 3.593.59 31.0031.00 4.694.69 3.613.61
처리구2Treatment area 2 41.3541.35 3.743.74 35.7635.76 4.784.78 3.723.72
처리구3Treatment area 3 37.5037.50 3.613.61 34.4434.44 3.803.80 4.754.75
B. licheniformis JWS22003 균주 배양액을 배추(춘광)에 3회 걸쳐 엽면시비하였는데, 모든 처리구에서 대조구 대비 구중, 구고, 구경, 엽수, 엽록소가 개선되는 것을 확인하였다(표 5). 특히 100배액의 처리구 2번이 구중 2.95kg, 구고 27.72cm, 구경 17.60cm, 엽수 41.95매, 엽록소 35.02로 가장 우수하여 B.licheniformis JWS22003 균주가 배추 생육을 향상시키는 것으로 나타났다.B. licheniformis JWS22003 strain culture medium was foliar applied to Chinese cabbage (Chungwang) three times, and it was confirmed that bulb weight, bulb height, corm, number of leaves, and chlorophyll were improved in all treatments compared to the control (Table 5). In particular, 100-fold treatment group No. 2 was the best with 2.95 kg in bulb weight, 27.72 cm in height, 17.60 cm in diameter, 41.95 leaves, and 35.02 chlorophyll, showing that the B. licheniformis JWS22003 strain improves cabbage growth.
표 5는 B. licheniformis JWS22003 균주의 엽면시비 처리후 배추(춘광)의 생육 개선 효과를 나타낸 것이다.Table 5 shows the growth improvement effect of Chinese cabbage (Chungwang) after foliar fertilization treatment of B. licheniformis JWS22003 strain.
구중(kg)Weight (kg) 구고(cm)Spherical height (cm) 구경(cm)Diameter (cm) 엽수(매)Leaf tree (hawk) 엽록소(SPAD)Chlorophyll (SPAD)
대조구Control 2.262.26 24.8624.86 16.5116.51 33.7033.70 32.2932.29
처리구1 Treatment area 1 2.762.76 27.6627.66 17.2517.25 35.1135.11 33.1733.17
처리구2Treatment area 2 2.952.95 27.7227.72 17.6017.60 41.9541.95 35.0235.02
처리구3Treatment area 3 2.792.79 26.3126.31 17.1217.12 39.0439.04 34.3634.36
B. licheniformis JWS22003 균주 배양액을 고추(거창한)에 3회 걸쳐 엽면시비하였는데, 모든 처리구에서 대조구 대비 초장, 엽수, 엽면적, 생체중, 건물중, 과수, 과중, 건과중이 개선되는 것을 확인하였다(표 6). 특히 100배액의 처리구 2번에서 초장 152.81cm, 엽수 2,088.34매, 엽면적 25,592.94cm²/주, 생체중 489.13g/주, 건물중105.31g/주, 과수 46.17개/주, 과중 537.52g/주, 건과중 84.24g/주로 가장 우수하여 B. licheniformis JWS22003 균주가 고추 생육을 향상시키는 것으로 나타났다.B. licheniformis JWS22003 strain culture medium was applied foliarly to pepper (Geochanghan) three times, and it was confirmed that plant height, number of leaves, leaf area, fresh weight, dry weight, fruit weight, fruit weight, and dry fruit weight were improved in all treatments compared to the control (Table 6 ). In particular, in treatment group 2 with 100 drainage, plant height was 152.81cm, number of leaves was 2,088.34, leaf area was 25,592.94cm²/week, fresh weight was 489.13g/week, dry weight was 105.31g/week, fruit tree was 46.17 pieces/week, fruit weight was 537.52g/week, and dry fruit weight was 84.24. The B. licheniformis JWS22003 strain was found to be the best in terms of g/week, improving pepper growth.
표 6은 B. licheniformis JWS22003 균주의 엽면시비 처리후 고추(거창한)의 생육 개선 효과를 나타낸 것이다.Table 6 shows the growth improvement effect of pepper (Geochanghan) after foliar application of B. licheniformis JWS22003 strain.
초장
(cm)super long
(cm) 		엽수
(매)ground game
(hawk) 		엽면적
(㎠/주)leaf area
(㎠/week) 		생체중
(g/주)live weight
(g/week) 		건물중
(g/주)In the building
(g/week) 		과수
(개/주)fruit tree
(recast) 		과중
(g/주)overload
(g/week) 		건과중
(g/주)Dry fruit heavy
(g/week)
대조구Control 148.45148.45 2033.752033.75 24822.5924822.59 471.32471.32 101.35101.35 40.9440.94 490.07490.07 81.7381.73
처리구1 Treatment area 1 151.24151.24 2074.602074.60 25445.8125445.81 482.34482.34 103.98103.98 43.0143.01 515.65515.65 83.9283.92
처리구2Treatment area 2 152.81152.81 2088.342088.34 25592.9425592.94 489.13489.13 105.31105.31 46.1746.17 537.52537.52 84.2484.24
처리구3Treatment area 3 150.70150.70 2071.782071.78 25310.7025310.70 484.97484.97 103.54103.54 45.2445.24 517.04517.04 82.0782.07
B. licheniformis JWS22003 균주 배양액을 사과(홍로)에 3회 걸쳐 엽면시비하였는데, 모든 처리구에서 대조과와 비교해 신초량, 경정, 과수, 엽록소가 개선되는 것을 확인하였다(표 7). 특히 100배액의 처리구 2번에서 신초량 64.70cm, 경정 49.17mm, 과수 36.09개/주, 엽록소 35.97로 가장 우수하여 B. licheniformis JWS22003 균주가 사과 생육을 향상시키는 것으로 나타났다.B. licheniformis JWS22003 strain culture was applied to apples (Hongro) three times as foliar application, and it was confirmed that shoot volume, jugular, fruit tree, and chlorophyll were improved in all treatments compared to the control fruit (Table 7). In particular, in treatment group 2 at 100 times the amount, the shoot volume was 64.70 cm, the crown was 49.17 mm, the number of fruits was 36.09 per week, and the chlorophyll was 35.97, showing that the B. licheniformis JWS22003 strain improved apple growth.
표 7은 B. licheniformis JWS22003 균주의 엽면시비 처리후 사과(홍로)의 생육 개선 효과를 나타낸 것이다.Table 7 shows the effect of improving the growth of apples (Hongro) after foliar fertilization of the B. licheniformis JWS22003 strain.
신초량(cm)Shincho amount (cm) 경정(mm)Lightweight (mm) 과수(개/주)Fruit trees (units/week) 엽록소(SPAD)Chlorophyll (SPAD)
대조구Control 59.2359.23 42.3042.30 32.8832.88 30.0530.05
처리구1 Treatment area 1 61.6161.61 44.5044.50 34.1534.15 33.1133.11
처리구2Treatment area 2 64.7064.70 49.1749.17 36.0936.09 35.9735.97
처리구3Treatment area 3 63.1363.13 45.9845.98 35.6535.65 33.4333.43
위와 같은 B. licheniformis JWS22003 균주를 목재 부산물 퇴비, 6종의 원예작물에 처리하여 개선효과를 검정한 결과로써, 부산물의 경우, B. licheniformis JWS22003 균주를 퇴비 발효에 처리한 결과, 아무 것도 처리하지 않은 대조구와 비교해 10배액으로 처리하는 것이 생육이 가장 우수했지만, 종합적으로 볼 때, 100배액으로 처리하는 것이 B. licheniformis JWS22003 균주의 투여량이 1/10 수준이기 때문에 경제적이므로 생육이 가장 우수하여 B. licheniformis JWS22003 균주가 생육을 향상시키는 것으로 나타났다.또한, 식물의 경우, B. licheniformis JWS22003 균주 배양액을 농작물에 3회 걸쳐 엽면시비한 결과, 아무 것도 처리하지 않은 대조구와 비교해 B.licheniformis JWS22003 균주 배양액을 100배액으로 처리하는 것이 가장 우수하여 B.licheniformis JWS22003 균주가 생육을 향상시키는 것으로 나타났다.그 외에, B. licheniformis JWS22003 균주의 옥신 생성량을 알아보기 위해 HPLC 분석을 한 결과 B. licheniformis JWS22003 균주에 의해 옥신이 생합성되는 것을 확인할 수 있다.As a result of testing the improvement effect of the above-mentioned B. licheniformis JWS22003 strain on wood by-product compost and 6 types of horticultural crops, in the case of by-products, the B. licheniformis JWS22003 strain was treated with compost fermentation, and as a result, it was not treated with anything. Compared to the control, treatment with 10 times the solution had the best growth, but overall, treatment with 100 times the solution was economical because the dosage of B. licheniformis JWS22003 strain was 1/10, and the growth was the best for B. licheniformis. The JWS22003 strain was found to improve growth. In addition, in the case of plants, as a result of foliar application of the B. licheniformis JWS22003 strain culture medium to crops three times, compared to the untreated control, the B. licheniformis JWS22003 strain culture medium was 100 times higher. Treatment with was found to be the best, improving the growth of the B. licheniformis JWS22003 strain. In addition, HPLC analysis was performed to determine the amount of auxin produced by the B. licheniformis JWS22003 strain, and the results showed that auxin was biosynthesized by the B. licheniformis JWS22003 strain. You can check that it happens.
이에 따라 B. licheniformis JWS22003 균주는 B. licheniformis의 고온 배양 가능성을 통한 농업자재 생산의 효율성과 식물생장촉진(PGPR)에 주목하여 높은 활력을 지녔으므로 본 균주의 부산물 발효, 농작물 생육 촉진에 활용할 수 있다.Accordingly, the B. licheniformis JWS22003 strain focuses on the efficiency of agricultural material production and plant growth promotion (PGPR) through the high-temperature culturability of B. licheniformis and has high vitality, so it can be used to ferment by-products of this strain and promote crop growth. .

Claims (4)

  1. 부산물의 발효와 식물의 생육을 촉진하고 옥신을 생성하는 것을 특징으로 하는 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P).Bacillus licheniformis JWS22003 strain (Accession number: KACC 92452P), which is characterized by promoting fermentation of by-products and plant growth and producing auxin.
  2. 제1항에 따른 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)의 100배 희석액을 활용하여 부산물의 발효를 촉진하는 것을 특징으로 하는 부산물 발효 촉진 방법.A method for promoting fermentation of by-products, characterized in that the fermentation of the by-products is promoted by using a 100-fold dilution of the Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P) according to claim 1.
  3. 제1항에 따른 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)의 100배 희석액을 농작물에 엽면시비하여 식물의 생육을 촉진하는 것을 특징으로 하는 식물 생육 촉진 방법.A method for promoting plant growth, characterized in that foliar application of a 100-fold dilution of the Bacillus licheniformis JWS22003 strain (accession number: KACC 92452P) according to claim 1 to crops to promote plant growth.
  4. 제1항에 따른 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주(기탁번호: KACC 92452P)는 농작물 부산물과 인근 토양에서 수집한 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis) 균주에 화학적 돌연변이를 유발하는 EMS(ethyl methane sulfonate)를 처리하여 제조되는 것을 특징으로 하는 바실러스 리체니포르미스 JWS22003(Bacillus licheniformis JWS22003) 균주 제조방법.The Bacillus licheniformis JWS22003 strain (Accession number: KACC 92452P) according to paragraph 1 is a strain that causes chemical mutations in the Bacillus licheniformis JWS22003 strain collected from crop by-products and nearby soil. A method for producing a Bacillus licheniformis JWS22003 strain, characterized in that it is produced by treatment with EMS (ethyl methane sulfonate).
PCT/KR2023/014634 2022-10-11 2023-09-25 Method for preparing bacillus licheniformis jws22003 strain for promoting fermentation of by-products and plant growth and producing auxin, and method for fermenting by-products and method for promoting plant growth by using same WO2024080626A1 (en)

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