[논문]식물 유용 방선균 2종의 배양 및 포자생성 최적화 조건 탐색

김다란; 곽연식

doi:10.5423/rpd.2023.29.2.174

[국내논문] 식물 유용 방선균 2종의 배양 및 포자생성 최적화 조건 탐색
Optimization of Culture and Sporulation for Two Plant Beneficial Streptomyces Strains 원문보기

Research in plant disease = 식물병연구, v.29 no.2, 2023년, pp.174 - 183

김다란 (경상국립대학교 생명과학연구원) , 곽연식 (경상국립대학교 생명과학연구원)

초록
AI-Helper

관행적 화학농약의 식물병 관리 및 치료의 한계적인 효과는 유익한 세균이나 미생물을 이용한 식물병 제어 방법 개발의 필요성을 나타낸다. 지속 가능한 농업은 특히 다양한 항생물질과 이차 대사 생성물을 생산하는 방선균의 농업적 중요성을 시사하며, Streptomyces globiosporus SP6C4균주와 Streptomyces sp. S8은 강력한 항균 작용을 가지고 있으며, 지속가능한 농업에서 작물 생장을 개선하기 위한 우수한 균주로 인정되고 있다. 본 연구에서는 다양한 질소원과 탄소원을 활용하여 대상 미생물 Streptomyces 두 균주의 생장을 촉진시키는 방법을 조사하였다. L-글루타민과 L-시스테인이 첨가된 조건에서 S. globiosporus SP6C4와 Streptomyces sp. S8 균주의 포자 생성 능력이 증가하였으며, 각 균주의 생장이 촉진되었다. 이러한 결과는 유용 미생물의 대량배양 기술의 범위 확장과 농업미생물의 실용화에 기여할 것으로 생각된다.

Abstract ▼ AI-Helper

The limited effectiveness of current plant disease management treatments necessitates the development of new methods for controlling diseases using beneficial microbes. Demanding sustainable agriculture is increasingly highlighted as a biocontrol approach, particularly Streptomyces species known to produce a variety of antibiotic compounds and secondary metabolites. The Streptomyces globisporus SP6C4 strain and Streptomyces sp. S8 have been reported as potent antifungal agents and are gaining attention for improving crop growth in sustainable agriculture. In this study, we investigated the use of Streptomyces species formulations to enhance bacterial growth with nitrogen sources. Specifically, the addition of L-glutamic acid and L-cysteine resulted in earlier sporulation and bacterial growth in Streptomyces strains, respectively. This approach could expand the range of fermentation techniques in agriculture and be useful for controlling plant growth-promoting bacteria.

주제어

표/그림 (9)

그림 Fig. 1. Plant growth-promoting rhizobacteria activity of Streptomyces sp. S8 and S. globisporus SP6C4. The spore stock solution (10 μl) was added to each enzyme assay medium that contained an 8 mm diameter filter paper disk. (A) Chitinase enzyme production (left), Protease enzyme production (right). (B) Siderophore production, phosphate-solubilization activity, nitrogenase enzyme production, and indole- 3-acetic acid production. The upper panel in each figure showed S8 and the lower panel presented SP6C4 strain.
표 Table 1. Plant growth-promoting related enzyme assay of Streptomyces sp. S8 and S. globisporus SP6C4
그림 Fig. 2. Variation with pH in bacterial growth. The spore stock (10 μl) of the suspension was added to each well of a 96-well plate. The plates were incubated, and the optical density was measured with orbital shaking using a Synergy H1 Hybrid Multi-Mode microplate reader for a period of 24 hr at a temperature of 28ºC. (A) S8 strain. (B) SP6C4 strain. The left panel showed bacterial growth in each pH condition. The right panel presented the maximum growth speed of the strain.
그림 Fig. 3. The emergence of phenotypic in colonies of Streptomyces sp. S8 and S. globisporus SP6C4. Five different media, ISP2, ISP4, ISP6, ISP7, and MS (from left to right) were tested. The plates were incubated at 28ºC for 72 hr, and observations were made at three different time points: (A) 24 hr after incubation, (B) 48 hr after incubation, and (C) 72 hr after incubation. The upper panel showed S8 and the lower panel presented SP6C4 strain.
그림 Fig. 4. Biolog phenotype array for nitrogen utilization by PM3B plate. The spore stock was inoculated onto PM3B plates. A redox dye mix A was added to the plate, and the plate was further incubated at 37ºC. Numbers (1-12) and alphabet (A-H) represented well position and each position contained different nutrient sources.
그림 Fig. 5. Effect of differences in nitrogen sources to bacterial growth. The nitrogen stock solution was added to basal media, followed by the addition of bacterial spore stock standardized to an optical density of 0.2 at 600 nm. The mixture was then incubated with shaking at 28ºC for a duration of 50 hr (x-axis).
그림 Fig. 6. Spore chain formation with different carbon and nitrogen sources by Streptomyces sp. S8. Six different types of nitrogen sources and three types of carbon sources were added to basal media at a final concentration of 0.2%. The sporulation was visualized under a 1,000× microscope (scale bars=10 μm). (A) L-asparagine. (B) L-cysteine. (C) L-glutamic acid. (D) L-proline. (E) L-valine. (F) Peptone. (G) Starch. (H) Mannitol. (I) Yeast extract.
그림 Fig. 7. Spore chain formation with different carbon and nitrogen sources by Streptomyces globisporus SP6C4. Basal media was supplemented with six different nitrogen sources and three different carbon sources, each at a final concentration of 0.2%. The sporulation was observed under a 1,000× microscope (scale bars=10 μm). (A) L-asparagine. (B) L-cysteine. (C) L-glutamic acid. (D) L-proline. (E) L-valine. (F) Peptone. (G) Starch. (H) Mannitol. (I) Yeast extract.
표 Table 2. Validation of spore formation ability under microscopy as affected by nitrogen and carbon sources

참고문헌 (43)

Andleeb, S., Shafique, I., Naseer, A., Abbasi, W. A., Ejaz, S., Liaqat, I. et？al. 2022. Molecular characterization of plant growth-promoting？vermi-bacteria associated with Eisenia fetida gastrointestinal？tract. PLoS ONE 17: e0269946.
Bentley, S. D., Chater, K. F., Cerdeno-Tarraga, A.-M., Challis, G. L.,？Thomson, N. R., James, K. D. et al. 2002. Complete genome？sequence of the model actinomycete Streptomyces coelicolor？A3(2).？Nature 417: 141-147.

상세보기
Berkelmann, D., Schneider, D., Hennings, N., Meryandini, A. and？Daniel, R. 2020. Soil bacterial community structures in relation？to different oil palm management practices.？Sci. Data？7: 421.
Bintarti, A. F., Sulesky-Grieb, A., Stopnisek, N. and Shade, A. 2022.？Endophytic microbiome variation among single plant seeds.？Phytobiomes J. 6: 45-55.

상세보기
Bokhari, A., Essack, M., Lafi, F. F., Andres-Barrao, C., Jalal, R., Alamoudi, S. et al. 2019. Bioprospecting desert plant Bacillus endophytic strains for their potential to enhance plant stress tolerance. Sci. Rep. 9: 18154.

상세보기
Cha, J.-Y., Han, S., Hong, H.-J., Cho, H., Kim, D., Kwon, Y., et al. 2016.？Microbial and biochemical basis of a Fusarium wilt-suppressive？soil.？ISME J.？10: 119-129.

상세보기
Chen, S., Chen, X. and Xu, J. 2016. Impacts of climate change on？agriculture: evidence from China. J. Environ. Econ. Manage. 76:？105-124.

상세보기
Costa, E., Teixido, N., Usall, J., Atares, E. and Vinas, I. 2002. The effect？of nitrogen and carbon sources on growth of the biocontrol？agent Pantoea agglomerans strain CPA-2.？Lett. Appl. Microbiol.？35:117-120.

상세보기
Deshpande, B. S., Ambedkar, S. S. and Shewale, J. G. 1988. Biologically active secondary metabolites from Streptomyces.？Enzyme？Microb. Technol.？10: 455-473.

상세보기
Ducray, H. A. G., Globa, L., Pustovyy, O., Morrison, E., Vodyanoy, V.？and Sorokulova, I. 2019. Yeast fermentate prebiotic improves？intestinal barrier integrity during heat stress by modulation of？the gut microbiota in rats.？J. Appl. Microbiol.？127: 1192-1206.

상세보기
Du, J., Li, Y., Ur-Rehman, S., Mukhtar, I., Yin, Z., Dong, H. et al. 2021.？Synergistically promoting plant health by harnessing synthetic？microbial communities and prebiotics. iScience 24: 102918.

상세보기
Fidan, O. and Zhan, J. 2019. Discovery and engineering of an endophytic Pseudomonas strain from Taxus chinensis for efficient？production of zeaxanthin diglucoside. J. Biol. Eng. 13: 66.

상세보기
Harir, M., Bendif, H., Bellahcene, M., Fortas, Z. and Pogni, R. 2018.？Streptomyces secondary metabolites.？In: Basic Biology and Applications of Actinobacteria, ed. by？S. Enany, pp. 99-122. IntechOpen, London, UK.
Hannula, S. E., Zhu, F., Heinen, R. and Bezemer, T. M. 2019. Foliarfeeding insects acquire microbiomes from the soil rather than？the host plant. Nat. Commun. 10: 1254.

상세보기
Herrera-Quiterio, A., Toledo-Hernandez, E., Aguirre-Noyola, J. L.,？Romero, Y., Ramos, J., Palemon-Alberto, F. et al. 2020. Antagonic？and plant growth-promoting effects of bacteria isolated from？mine tailings at El Fraile, Mexico.？Rev. Argent. Microbiol.？52: 231-239.
Jagermeyr, J., Robock, A., Elliott, J., Muller, C., Xia, L., Khabarov, N., et？al. 2020. A regional nuclear conflict would compromise global？food security. Proc. Natl. Acad. Sci. U. S. A. 117: 7071-7081.

상세보기
Jeon, C.-W., Kim, D.-R. and Kwak, Y.-S. 2019. Valinomycin, produced？by Streptomyces sp. S8, a key antifungal metabolite in large？patch disease suppressiveness. World J. Microbiol. Biotechnol. 35: 128.

상세보기
Jung, Y.-J., Joung, Y. and Ahn, T.-S. 2011. Characterization of Actinomyces isolated from freshwater sponges in lake Baikal.？Korean J.？Microbiol.？47: 130-136.
Kalaiyarasi, M., Ahmad, P. and Vijayaraghavan, P. 2020. Enhanced？production antibiotics using green gram husk medium by？Streptomyces sp. SD1 using response surface methodology.？J.？King Saud Univ. Sci.？32: 2134-2141.
Kim, D.-R., Cho, G., Jeon, C.-W., Weller, D. M., Thomashow, L. S., Paulitz, T. C., et al. 2019a. A mutualistic interaction between Streptomyces bacteria, strawberry plants and pollinating bees. Nat. Commun. 10: 4802.

상세보기
Kim, D.-R., Jeon, C.-W., Cho, G., Thomashow, L. S., Weller, D. M., Paik,？M.-J. et al. 2021. Glutamic acid reshapes the plant microbiota to？protect plants against pathogens. Microbiome 9: 244.

상세보기
Kim, D.-R., Jeon, C.-W., Shin, J.-H., Weller, D. M., Thomashow, L. and？Kwak, Y. S. 2019b. Function and distribution of a lantipeptide in？strawberry Fusarium wilt disease-suppressive soils.？Mol. Plant-Microbe Interact.？32: 306-312.

상세보기
Kim, M.-J., Chae, D.-H., Cho, G., Kim, D.-R. and Kwak, Y.-S. 2019c.？Characterization of antibacterial strains against kiwifruit bacterial canker pathogen.？Plant Pathol. J.？35: 473-485.
Lau, J. A., Lennon, J. T. and Heath, K. D. 2017. Trees harness the power of microbes to survive climate change. Proc. Natl. Acad. Sci. U.？S. A. 114: 11009-11011.

상세보기
Lee, J. H., Lee, Y. S. and Kim, Y. C. 2021. Effect of temperature and？culture media composition on sporulation, mycelial growth,？and antifungal activity of Isaria javanica pf185. Res. Plant Dis. 27:？99-106.
Lee, N., Hwang, S., Kim, J., Cho, S., Palsson, B. and Cho, B.-K. 2020.？Mini review: genome mining approaches for the identification？of secondary metabolite biosynthetic gene clusters in Streptomyces.？Comput. Struct. Biotechnol. J. 18: 1548-1556.

상세보기
Lindow, S. E. and Brandl, M. T. 2003. Microbiology of the phyllosphere.？Appl. Environ. Microbiol.？69: 1875-1883.

상세보기
Lu, T., Ke, M., Lavoie, M., Jin, Y., Fan, X., Zhang, Z., et al. 2018. Rhizosphere microorganisms can influence the timing of plant flowering. Microbiome 6: 231.

상세보기
Mostafa, S. A. 1979. Activity of L-asparaginase in cells of Streptomyces karnatakensis.？Zentralbl. Bakteriol. Natuwiss.？134: 343-351.
Oviedo-Pereira, D. G., Lopez-Meyer, M., Evangelista-Lozano, S.,？Sarmiento-Lopez, L. G., Sepulveda-Jimenez, G. and Rodriguez-Monroy, M. 2022. Enhanced specialized metabolite, trichome？density, and biosynthetic gene expression in Stevia rebaudiana？(Bertoni) Bertoni plants inoculated with endophytic bacteria？Enterobacter hormaechei. PeerJ 10: e13675.

상세보기
Padilla, G., Hindle, Z., Callis, R., Corner, A., Ludovice, M., Liras, P. et al.？1991. The relationship between primary and secondary metabolism in Streptomycetes. In: Genetics and Product Formation in？Streptomyces, eds. by S. Baumberg, H. Krugel and D. Noack, pp.？35-45. Federation of European Microbiological Societies Symposium Series, Vol. 55. Springer, Boston, MA.
Park, J.-K., Kim, J., Lee, C.-W., Song, J., Seo, S.-I., Bong, K.-M. et al.？2019. Mass cultivation and characterization of multifunctional？Bacillus velezensis GH1-13.？Korean J. Org. Agric.？27: 65-76.
Park, J.-K., Seo, S.-I., Han, G. H., Kim, K.-M., Kim, D.-H., Song, J. et al.？2018. Development of practical media and fermentative technique for mass cultivation from agricultural and livestock microorganism.？Trends Agric. Life Sci.？56: 23-33.

상세보기
Patel, D. and Parmar, P. 2013. Isolation and screening of phosphate？solubilizing bacteria from sunflower rhizosphere. Glob. J. Biosci.？Biotechnol. 2: 438-441.
Redford, A. J., Bowers, R. M., Knight, R., Linhart, Y. and Fierer, N. 2010.？The ecology of the phyllosphere: geographic and phylogenetic？variability in the distribution of bacteria on tree leaves.？Environ.？Microbiol.？12: 2885-2893.
Reese, A. T., Pereira, F. C., Schintlmeister, A., Berry, D., Wagner, M.,？Hale, L. P. et al. 2018. Microbial nitrogen limitation in the mammalian large intestine.？Nat. Microbiol.？3: 1441-1450.

상세보기
Rico, A. and Preston, G. M. 2008. Pseudomonas syringae pv. tomato？DC3000 uses constitutive and apoplast-induced nutrient assimilation pathways to catabolize nutrients that are abundant？in the tomato apoplast.？Mol. Plant-Microbe Interact.？21: 269-282.

상세보기
Ripa, E. A., Nikkon, K., Zaman, S. and Khondkar, P. 2009. Optimal？conditions for antimicrobial metabolites production from a？new Streptomyces sp. RUPA-08PR isolated from Bangladeshi？soil.？Mycobiology？37: 211-214.
Rodriguez, P. A., Rothballer, M., Chowdhury, S. P., Nussbaumer, T.,？Gutjahr, C. and Falter-Braun, P. 2019.？Systems biology of plant-microbiome interactions.？Mol. Plant 12: 804-821.

상세보기
Voelker, F. and Altaba, S. 2001. Nitrogen source governs the patterns of growth and pristinamycin production in 'Streptomyces？pristinaespiralis'.？Microbiology？147: 2447-2459.
Xiong, C., Singh, B. K., He, J.-Z., Han, Y.-L., Li, P.-P., Wan, L.-H., et al.？2021. Plant developmental stage drives the differentiation in？ecological role of the maize microbiome. Microbiome 9: 171.

상세보기
Yague, P., Lopez-Garcia, M. T., Rioseras, B., Sanchez, J. and Manteca,？A. 2012. New insights on the development of Streptomyces and？their relationships with secondary metabolite production.？Curr.？Trends Microbial.？8: 65-73.
Xu, Y., Ge, Y., Song, J. and Rensing, C. 2019. Assembly of root-associated microbial community of typical rice cultivars in different？soil types. Biol. Fertil. Soils 56: 249-260.

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증