ارزیابی تأثیر منابع کربن و نیتروژن، سطوح رطوبت بر بهینه‌سازی تولید جیبرلیک‌اسید توسط قارچFusarium fujikuroi در سیستم‌های کشت جامد و مایع

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه ولی عصر (عج)، رفسنجان، ایران

2 مرکز تحقیقات پسته، موسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، رفسنجان، ایران

چکیده

قارچ  Fusarium fujikuroi به عنوان یکی از منابع اصلی تولید هورمون جیبرلیک اسید زیست فعال شناخته می‌شود. پژوهش حاضر به بررسی تأثیر منابع نیتروژن آلی و کربن معدنی در سطح رطوبت متعدد بر تولید جیبرلیک‌اسید توسط جدایه 134 این قارچ، جداسازی شده از استان خراسان‌شمالی، می‌پردازد. فرآیند بهینه‌سازی تولید جیبرلیک‌اسید در دو سیستم کشت جامد و مایع و طی مراحل غربالگری متوالی انجام شد. ارزیابی کیفی متابولیت با روش‌ طیف‌سنجی صورت گرفت. در کشت جامد، چهار بستر آلی (کنجاله سویا، تفاله مرکبات، سبوس گندم و سبوس برنج) بررسی شد که کنجاله سویا با میانگین تولید 16/66 میلی‌گرم بر گرم، به طور معنی‌داری به دلیل محتوای پروتئین بالا و قابلیت تجزیه مناسب، بهترین عملکرد را داشت. سه بستر دیگر از نظر آماری تفاوت معنی‌داری نداشتند. همچنین، تأثیر سطوح مختلف رطوبت اولیه (20، 10،15 و 5 میلی‌لیتر) نشان داد که تیمار ۱۰ میلی‌لیتر آب بهینه‌ترین سطح بود. در کشت مایع، از میان چهار منبع کربن (گلیسرول، گلوکز، مانیتول و ملاس)، مانیتول مؤثرترین بود و پس از آن به ترتیب ملاس و گلیسرول و گلوکز قرار گرفتند. این مطالعه با شناسایی بهترین جدایه (134)، بستر ارزان (کنجاله سویا)، منبع کربن (مانیتول) و شرایط رطوبتی بهینه (۱۰ میلی‌لیتر)، راهکارهای عملی و مقرون به صرفه‌ای را برای تولید صنعتی جیبرلیک‌اسید ارائه می‌دهد. افزایش بازده سویه برتر با به‌کارگیری روش‌های ژنتیکی، توسعه فرآیند تولید در مقیاس نیمه‌صنعتی از طریق بهینه‌سازی همزمان فاکتورهای کلیدی و استفاده از بیورآکتور و ارزیابی اثربخشی محصول نهایی روی گیاهان هدف و تضمین ایمنی زیستی سویه صنعتی‌شده از راهکار­های پیشنهادی برای پژوهش ­های آتی می­ باشد. 

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation of the effect of carbon, nitrogen sources, and moisture levels on optimizing gibberellic acid production by Fusarium fujikuroi in solid and submerged culture systems

نویسندگان [English]

  • Fatemeh Rahimi 1
  • Ebrahim Sedaghati 1
  • Hossein Alaei 1
  • Rosa Dargahi 2
1 Department of Plant Protection, Agriculture Faculty,Vali-e-asr University, Rafsanjan, Iran
2 Pistachio Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rafsanjan, Iran
چکیده [English]

Fusarium fujikuroi is recognized as a primary natural source of bioactive gibberellic acid hormone production. The present study investigates the effects of organic nitrogen sources and inorganic carbon at different moisture levels on the production of gibberellic acid by F. fujikuroi (isolate 134), isolated from North Khorasan province, Iran. The optimization of gibberellic acid production was carried out in two culture systems, solid and submerged, and through successive screening steps. Qualitative evaluation of the metabolite was carried out using spectroscopic methods. In solid culture, four organic substrates were investigated, with soybean meal yielding an average production of 16.66 mg/g, significantly outperforming the others due to its high protein content and good degradability. The other three substrates (citrus pulp, rice bran, and wheat bran) were not significantly different. Also, the effect of initial moisture content (5, 10, 15, and 20 mL) showed that 10 mL of water was the most optimal level. In submerged culture, among the four carbon sources, including glycerol, glucose, mannitol, and molasses, mannitol was the most effective, followed by molasses, glycerol, and glucose, respectively. This study provides practical and cost-effective solutions for the industrial production of gibberellic acid by identifying the best isolate (134), inexpensive substrate (soybean meal), carbon source (mannitol), and optimal moisture conditions (10 mL). Increasing the yield of the superior strain by using genetic methods, developing the production process on a semi-industrial scale through simultaneous optimization of key factors and the use of a bioreactor, evaluating the effectiveness of the final product on target plants, and ensuring the biosafety of the industrialized strain are among the proposed solutions for future research.

کلیدواژه‌ها [English]

  • Agricultural Waste Recycling
  • Bakanae
  • Secondary Metabolites
  • Solid and Submerged Fermentation
  • Spectrophotometer
Adeyemo AA, Otunola GA, 2017. Gibberellic acid production: a review on the impact of nutritional substrates. Critical Reviews in Microbiology 43(4): 460–478.
Barnett YA, Barnett CR, 2008. Aging Methods and Protocols. Springer Science & Business 11(3): 100–113.
Bashyal BM, 2018. Etiology of an emerging disease: Bakanae of rice. Indian Phytopathology 71(4): 485–494.
Berríos J, Illanes A, Aroca G, 2006. Spectrophotometric method for determining gibberellic acid in fermentation broths. Biotechnology Letters 1(9):  26:67–70. https://doi.org/10.1023/B:BILE.0000009463.98203.8b
Castro-Camba R, Sánchez C, Vidal N, Vielba JM, 2022. Plant development and crop yield: The role of gibberellins. Plants 11(19): 26–50.
Crous PW, Verkley GJM, Groenewald JZ, 2019. Fungal Biodiversity Westerdijk Laboratory Manual Series 1. 2nd edition, Westerdijk Fungal Biodiversity Institute, Utrecht Press. 349 pp.
Crous PW, Lombard L, Sandoval-Denis M, Seifert KA, Schroers HJ, et al., 2021. Fusarium: more than a node or a foot-shaped basal cell. Studies in Mycology 98: 100–116. https://doi.org/10.1016/j.simyco.2021.100116.
da Silva LRI, de Andrade CJ, de Oliveira D, 2021. Solid-state fermentation in brewer’s spent grains by Fusarium fujikuroi for gibberellic acid production. Biointerface Research in Applied Chemistry 11(5): 13042–13052.
de Oliveira J, Rodrigues C, Vandenberghe LP, Câmara MC, Libardi N, et al., 2017. Gibberellic acid production by different fermentation systems using citric pulp as substrate/support. Biological Medical Research International 17(1): 56–73.
Hernández Rodríguez A, Díaz Pacheco A, Martínez Tolibia SE, Meléndez Xicohtencatl Y, Granados Balbuena SY, et al., 2024. Bioprocess of gibberellic acid by Fusarium fujikuroi: The challenge of regulation, raw materials, and product yields. Journal of Fungi 10(6): 418–425.
Karakoc SB, and Aksoez NILUFER, 2004. Optimization of carbon-nitrogen ratio for production of gibberellic acid by Pseudomonas sp. Polish Journal of Microbiology 53(2): 117–120.
Kumar A, Sharma S, Mehta P, 2021. Advances in submerged fermentation for gibberellic acid production by Fusarium fujikuroi: Process optimization and scale-up. Biotechnology Reports 32: 659–687.
Lee JH, Kim BK, Kim JH, 2011. Production of gibberellin by Fusarium fujikuroi using rice bran as a carbon source. Journal of Microbiology & Biotechnology 21(4): 309–313.
Murray JD, 2020. Microbial biosynthesis of plant hormones and their roles in sustainable agriculture. Frontiers in Plant Science 11: 580–590.
N’Guessan KJY, Adahi B, Konan-Waidhet AB, Masayoshi S, Assidjo NE. 2023. Assessment of climate change impact on water requirement and rice productivity. Rice Science 30(4): 276–293.
Omojasola PF, Adejoro DO, 2018. Gibberellic acid production by Fusarium moniliforme and Aspergillus niger using submerged fermentation of banana peel. Notulae Scientia Biologicae 10(1): 60–67.
Oyedeji S, Patel N, Krishnamurthy R, Fatoba PO, 2024. Agricultural wastes to value-added products: economic and environmental perspectives for waste conversion. In: Bayer T,  Kircher M (eds). Biowaste to Value-added Products; Economics and Technologies. Springer. pp. 215–248.
Pandey A, Soccol CR, Larroche C, 2008. Current developments in solid-state fermentation. Springer Science & Business  4(12): 26–37.
Pandya JB, Patani AN, Raval VH, Rajput KN, Panchal RR, 2023. Understanding the Fermentation Potentiality for Gibberellic Acid (GA3) Production Using Fungi. Current Microbiology 80(12): 385–400.
Patil RH, Patil MP, Maheshwari VL, 2016. Bioactive secondary metabolites from endophytic fungi: a review of biotechnological production and their potential applications. Studies in Natural Products Chemistry 49: 189–205.
Rezvi HUA, Tahjib‐Ul‐Arif M, Azim MA, Tumpa TA, Tipu MMH, et al.,2023. Rice and food security: Climate change implications and the prospects for nutritional security. Food & Energy Security 12(1): 430–447.
Ríos-Iribe EY, Flores-Cotera LB, Chávira MMG, González-Alatorre Gy, Escamilla-Silva EM, 2011. Inductive effect produced by a mixture of carbon sources in the production of gibberellic acid by Gibberella fujikuroiWorld Journal of Microbiology & Biotechnology 27(6): 1499–1505.
Rodrigues C, Vandenberghe  LP, de Oliveira J, 2012. New perspectives on gibberellic acid production: a review. Critical Reviews in Biotechnology 32(3): 263–273.
Rodrigues C, Vandenberghe LP, Goyzueta LD, Soccol CR, 2016. Production, extraction, and purification of gibberellic acid by solid-state fermentation using citric pulp and soy husk. BAOJ Chemistry 2(2): 14–22.
Rojas JA, García LC, Báez S, 2013. Effect of citrus peels on gibberellin production in fermentation processes. Citrus Research 34(2): 1–6.
Singh R, Kumar A, Dwivedi A, 2021. Improved gibberellic acid production via solid-state fermentation using agro-industrial residues by Fusarium fujikuroi: Process optimization and molecular studies. Bioresource Technology Reports 2(9): 20–33. https://doi.org/10.1016/j.biteb.2021.100698 
Süntar I, Çetinkaya S, Haydaroğlu ÜS, 2021. Bioproduction process of natural products and biopharmaceuticals: biotechnological aspects. Biotechnology Advances 50: 68–77. 
Takahashi N, Phinney BO, MacMillan J. 2012. Gibberellins. 2th edition. Springer Science & Business Media. 524 pp.
Tomasini A, Fajardo C, Barrios González J, 1997. Gibberellic acid production using different solid-state fermentation systems. World Journal of Microbiol Biotechnology 13: 203–206. 
Wang HN, Ke X, Zhou JP, Liu ZQ, Zheng YG, 2022. Recent advances in metabolic regulation and bioengineering of gibberellic acid biosynthesis in Fusarium fujikuroiWorld Journal of Microbiology & Biotechnology 38(8): 1–16.
Wang H, Ke X, Jia R, Huang L, Liu Z, et al., 2023. Gibberellic acid overproduction in Fusarium fujikuroi using regulatory modification and transcription analysis. Applied Microbiology & Biotechnology 107(9): 13–84.
Zhan L, Chen L, Hou Y, Zeng Y, Ji Z, 2024. Bakanae disease resistance in rice: current status and future considerations. Agronomy 14: 1507–1515. htps. doi. org/10.3390/agronomy14071507.
Zhang J, Cheng Y, Zhang H, 2012. Impact of rice bran on the production of gibberellins by Fusarium fujikuroi in submerged fermentation. Bioresource Technology 114: 408–413.
Zhou H, Wu Q, 2011. Optimization of fermentation conditions for gibberellin production by Fusarium fujikuroi in solid-state fermentation. Journal of Biological Engineering 5(1): 1–10.