بررسی توانایی تولید هورمون‌های محرک رشد جیبرلین و اندول اسید استیک در باکتری‌های اندوفیت سودوموناس پوتیدا در سویا
محورهای موضوعی : میکروب شناسی گیاهیفایقه اطمینانی 1 , ادیبه اطمینانی 2 , شعله درویشی 3
1 - کارشناس ارشد، باشگاه پژوهشگران جوان و نخبگان، دانشگاه آزاد اسلامی، واحد سنندج، سنندج
2 - دانشجوی دکتری، گروه زراعت، دانشگاه آزاد اسلامی، واحد سنندج، سنندج
3 - دانشیار، گروه علوم و صنایع غذایی، دانشگاه آزاد اسلامی، واحد سنندج، سنندج
کلید واژه: سویا, سودوموناس پوتیدا, باکتری اندوفیت,
چکیده مقاله :
سابقه و هدف: باکتری‌های اندوفیت بدون ایجاد علایم مشخص قادر به زندگی در بخش‌های داخلی گیاه میزبان هستند و در بسیاری از موارد به عنوان باکتری‌های محرک رشد گزارش شده‌اند. این مطالعه با هدف بررسی توانایی تولید هورمون‌های محرک رشد در باکتری‌های اندوفیت سویا انجام شد. مواد و روش ها: به منظور جداسازی باکتری اندوفیت از بخش‌های مختلف سویا رقم TMS ، نمونه‌برداری از مزرعه کشاورزی دانشگاه آزاد اسلامی واحد سنندج انجام شد. پس از استخراج DNA ژنومی، به منظور تکثیر ژن 16S rDNA از روش PCR استفاده گردید. برای شناسایی باکتری جداسازی شده، محصول PCR تعیین توالی و ترادف بازی آن BLAST گردید. سویه‌ها از نظر تولید هورمون اندول اسید استیک و جیبرلین به صورت آماری در سه تکرار و در قالب طرح کاملاً تصادفی مورد بررسی قرار گرفتند. یافته‌ها: باکتری‌های جداسازی شده قادر به تولید هورمون اندول اسید استیک در حضور تریپتوفان در مقادیر 7.2 تا 15.14 میکروگرم در میلی‌لیتر و در غیاب تریپتوفان در محدوده 3.2 تا 12.14 میکروگرم در میلی‌لیتر بودند. باکتری‌ها توانایی تولید هورمون جیبرلین در مقادیر 0.11 تا 0.20 میکروگرم در میلی‌لیتر را داشتند. نتایج تعیین توالی نشان داد که باکتری جداسازی شده متعلق به سودوموناس پوتیدا است که با سویه تیپ شباهت 99 درصدی دارد. نتیجه‌گیری: این مطالعه اولین گزارش جداسازی باکتری اندوفیت سودوموناس پوتیدا از سویارقم TMS است. باکتری‌های اندوفیت جدا شده می‌توانند در افزایش رشد گیاه به کار روند.
Background & Objectives: Endophytic bacteria live inside tissues of their plant host without causing visible symptoms, and in more cases are reported as plant growth promoting bacteria. This study was conducted to determine plant growth promoting affects of endophytic bacteria associated with Soybean (cultivar TMS). Materials & Methods: In order to isolate endophytic bacteria from various parts of soybean (cultivar TMS), samples were collected from the farm of Islamic Azad University of Sanandaj. After genomic DNA extraction, 16S rDNA gene was amplified using PCR. Then, the PCR product was sequenced by BLAST. Strains were surveyed for IAA, and GA production ability was carried out using a randomized complete design in three replications. Results: The isolated bacteria were able to produce IAA in various amount 3.2 -12.14 µg/ml without tryptophan and in the presence of it, 7.2 -15.14 µg/ml. GA producing abilities were also 0.11-0.20 µg/ml for these isolates. Based on the 16S rDNA sequence, the isolated bacteria was belonged to Pseudomonas putida with 99% similarity. Conclusion: This study is the first report of isolation of P. putida from Soybean (TMS cultivar). The endophytic bacteria isolated in this study can be used to promote plant growth.
1. Schulz B, Boyle C. What are endophytes. In: Schulz B, Boyle C, Sieber TN, editors. Microbial Root Endophytes. Springer-Verlag, Berlin; 2006: 1–13.
2. Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. Bacterial endophytes in agricultural crops. Can J Microbiol. 1997; 43: 895-914.
3. Spaepen S, Vanderleyden J, Remans R. Indole-3-acetic acid in microbial and microorganism plant signaling. FEMS Microbiol Rev. 2007; 31: 425-448.
4. Ali SZ, Sandhya V, Grover M, Kishore N, Rao LV, Venkateswarlu B. Pseudomonas sp. strain AKM-P6 enhances tolerance of sorghum seedlings to elevated temperatures. Biol Fert Soils. 2009; 46: 45-55.
5. Reinhold-Hurek B, Hurek T. Living inside plants: bacterial endophytes. Curr Opin Plant Biol. 2011; 14: 435-443.
6. Martin DN, Proebsting WM, Hedden P. Mendels dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins. Proc. Natl Acad Sci USA. 1997; 94: 8907-8911.
7. Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev. 2003; 67: 491-502.
8. Kobayashi DY, Palumbo JD. Bacterial endophytes and their effects on plants and uses in |agriculture. In: Bacon CW, White JF, editors. Microbial endophytes. Marcel Dekker, New York; 2000: 199-233.
9. Wilcox JR. Soybean: Improvement, productions and uses. Madison, Wisconsin, USA; 1987.
10. Jasim B, Joseph AA, John CJ, Mathew J, Radhakrishnan EK. Isolation and characterization of plant growth promoting endophytic bacteria from the rhizome of Zingiber officinale. Biotechnol J. 2014; 3(4): 197-204.
11. Schaad NW. Initial identification of common genra. In: Schaad NW, Jones JB, Chun W, editors. Laboratory Guide for Identification of Plant Pathogenic Bacteria. The American Phytopathological Society; 2001: 1-15.
12. Sambrook J, Russell, DW. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 2001.
13. Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991; 173: 697-703.
14. Rahman A, Sitepu IR, Tang SY, Hashidoko Y. Salkowski’s reagent test as a primary screening index for functionalities of Rhizobacteria isolated from wild dipterocarp saplings growing naturally on medium-strongly acidic tropical peat soil. Biosci Biotechnol Biochem. 2010; 74: 2202-2208.
15. Hidayati U, Chaniago IA, Munif A, Andreas Santosa S, Andreas Santosa S. Potency of plant growth promoting endophytic bacteria from rubber plants (Hevea brasiliensis Mull. Arg.). J Agron. 2014; 13: 147-152.
16. UmaMaheswari T, Anbukkarasi K, Hemalatha T, Chendrayan K. Studies on phytohormone producing ability of indigenous endophytic bacteria isolated from tropical legume crops. Int J Current Microbiol Appl Sci. 2013; 2: 127-136.
17. Panchal H, Ingle S. Isolation and characterization of endophytes from the root of medicinal plant Chlorophytum borivilianum (Safed musli). J Adv Dev Res. 2011; 2: 205-209.
18. Pirttila AM. Endophytic bacteria in tree shoot tissues and their effects on host. In: Pirttila AM, Frank AC, editors. Endophytes of forest trees. Springer Science and Business; 2011: 139-172.
19. Ozaktan H, Caklr B, Gul A, Yolageldi L, Akkopru A, Fakhraei D, Akbaba M. Isolation and evaluation of endophytic bacteria against Fusarium oxysporum f. sp. cucumerinum infecting cucumber plants. Austin J Plant Biol. 2015; 1: 1003-1009.
20. Khakipour N, Khavazi K, Mojallali H, Pazira E, Asadirahmani H. Production of auxin hormone by fluorescent Pseudomonads. Am Eurasian J Agric Environ Sci. 2008; 4: 687-692.
21. Asghar HN, Zahir ZA, Arshad M, Khaliq A. Relationship between in invitro production of auxins by rhizobacteria and their growth-promoting activities in Brassica juncea L. Biol Fertil Soils. 2002; 35: 231-237.
22. Shokri D, Emtiazi G. Purification and optimization of auxin (Indole-3-Acetic Acid) hormone in Rhizobium Bacterium. Iran Biol J. 2012; 25(2): 194-204. [In Persian]
23. Khan AL, Waqas M, Kang SM, Al-Harrasi A, Hussain J, Al-Rawahi A, Al-Khiziri S, Ullah I, Ali L, Young Jung H, Lee IJ. Bacterial Endophyte Sphingomonas sp. LK11 Produces Gibberellins and IAA and Promotes Tomato Plant Growth. J Microbiol. 2014; 52(8): 689-695.
24. Kang SM, Khan AL, Waqas M, You YH, Kim JH, Kim JG, Hamayun M, Lee IJ. Plant growth promoting rhizobacteria reduce adverse effects of salinity and osmotic stress by regulating phytohormones and antioxidants in Cucumis sativus. J Plant Interact. 2014; 9(1): 673-682.
25. Joo GJ, Kim YM, Lee IJ, Song KS, Rhee IK. Growth promotion of red pepper plug seedlings and the production of gibberellins by Bacillus cereus, Bacillus macroides and Bacillus pumilus. Biotechnol Lett. 2004; 26: 191-487.
26. Oteino N, Lally RD, Kiwanuka S, Lioyd A, Ryan D, Germaine KJ, Dowling DN. Plant growth promotion induced by phosphate solubilizing endophytic Pseudomonas isolates. Front Microbiol. 2015; 6: 1-9.
27. Etesami H, Alikhani HA. The quantitative and qualitative assessment of auxin hormone production ability of same of the Iranian soils indigenous Rhizobial strains. J Water Soil. 2011; 25(1): 61-69. [In Persian]
28. Drigo B, Kowalchuk GA, Van Veen JA. Climate change goes underground: effects of elevated atmospheric CO2 on microbial community structure and activities in the rhizosphere. Biol Fertil Soils. 2008; 44: 667-679.
29. Kuklinsky-Sobral J, Araujo WL, Mendes R, Geraldi IO, Pizzirani-Kleiner AA, Azevedo JL. Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion. Environ Microbiol. 2004; 6: 1244-1251.
_||_1. Schulz B, Boyle C. What are endophytes. In: Schulz B, Boyle C, Sieber TN, editors. Microbial Root Endophytes. Springer-Verlag, Berlin; 2006: 1–13.
2. Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. Bacterial endophytes in agricultural crops. Can J Microbiol. 1997; 43: 895-914.
3. Spaepen S, Vanderleyden J, Remans R. Indole-3-acetic acid in microbial and microorganism plant signaling. FEMS Microbiol Rev. 2007; 31: 425-448.
4. Ali SZ, Sandhya V, Grover M, Kishore N, Rao LV, Venkateswarlu B. Pseudomonas sp. strain AKM-P6 enhances tolerance of sorghum seedlings to elevated temperatures. Biol Fert Soils. 2009; 46: 45-55.
5. Reinhold-Hurek B, Hurek T. Living inside plants: bacterial endophytes. Curr Opin Plant Biol. 2011; 14: 435-443.
6. Martin DN, Proebsting WM, Hedden P. Mendels dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins. Proc. Natl Acad Sci USA. 1997; 94: 8907-8911.
7. Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev. 2003; 67: 491-502.
8. Kobayashi DY, Palumbo JD. Bacterial endophytes and their effects on plants and uses in |agriculture. In: Bacon CW, White JF, editors. Microbial endophytes. Marcel Dekker, New York; 2000: 199-233.
9. Wilcox JR. Soybean: Improvement, productions and uses. Madison, Wisconsin, USA; 1987.
10. Jasim B, Joseph AA, John CJ, Mathew J, Radhakrishnan EK. Isolation and characterization of plant growth promoting endophytic bacteria from the rhizome of Zingiber officinale. Biotechnol J. 2014; 3(4): 197-204.
11. Schaad NW. Initial identification of common genra. In: Schaad NW, Jones JB, Chun W, editors. Laboratory Guide for Identification of Plant Pathogenic Bacteria. The American Phytopathological Society; 2001: 1-15.
12. Sambrook J, Russell, DW. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 2001.
13. Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991; 173: 697-703.
14. Rahman A, Sitepu IR, Tang SY, Hashidoko Y. Salkowski’s reagent test as a primary screening index for functionalities of Rhizobacteria isolated from wild dipterocarp saplings growing naturally on medium-strongly acidic tropical peat soil. Biosci Biotechnol Biochem. 2010; 74: 2202-2208.
15. Hidayati U, Chaniago IA, Munif A, Andreas Santosa S, Andreas Santosa S. Potency of plant growth promoting endophytic bacteria from rubber plants (Hevea brasiliensis Mull. Arg.). J Agron. 2014; 13: 147-152.
16. UmaMaheswari T, Anbukkarasi K, Hemalatha T, Chendrayan K. Studies on phytohormone producing ability of indigenous endophytic bacteria isolated from tropical legume crops. Int J Current Microbiol Appl Sci. 2013; 2: 127-136.
17. Panchal H, Ingle S. Isolation and characterization of endophytes from the root of medicinal plant Chlorophytum borivilianum (Safed musli). J Adv Dev Res. 2011; 2: 205-209.
18. Pirttila AM. Endophytic bacteria in tree shoot tissues and their effects on host. In: Pirttila AM, Frank AC, editors. Endophytes of forest trees. Springer Science and Business; 2011: 139-172.
19. Ozaktan H, Caklr B, Gul A, Yolageldi L, Akkopru A, Fakhraei D, Akbaba M. Isolation and evaluation of endophytic bacteria against Fusarium oxysporum f. sp. cucumerinum infecting cucumber plants. Austin J Plant Biol. 2015; 1: 1003-1009.
20. Khakipour N, Khavazi K, Mojallali H, Pazira E, Asadirahmani H. Production of auxin hormone by fluorescent Pseudomonads. Am Eurasian J Agric Environ Sci. 2008; 4: 687-692.
21. Asghar HN, Zahir ZA, Arshad M, Khaliq A. Relationship between in invitro production of auxins by rhizobacteria and their growth-promoting activities in Brassica juncea L. Biol Fertil Soils. 2002; 35: 231-237.
22. Shokri D, Emtiazi G. Purification and optimization of auxin (Indole-3-Acetic Acid) hormone in Rhizobium Bacterium. Iran Biol J. 2012; 25(2): 194-204. [In Persian]
23. Khan AL, Waqas M, Kang SM, Al-Harrasi A, Hussain J, Al-Rawahi A, Al-Khiziri S, Ullah I, Ali L, Young Jung H, Lee IJ. Bacterial Endophyte Sphingomonas sp. LK11 Produces Gibberellins and IAA and Promotes Tomato Plant Growth. J Microbiol. 2014; 52(8): 689-695.
24. Kang SM, Khan AL, Waqas M, You YH, Kim JH, Kim JG, Hamayun M, Lee IJ. Plant growth promoting rhizobacteria reduce adverse effects of salinity and osmotic stress by regulating phytohormones and antioxidants in Cucumis sativus. J Plant Interact. 2014; 9(1): 673-682.
25. Joo GJ, Kim YM, Lee IJ, Song KS, Rhee IK. Growth promotion of red pepper plug seedlings and the production of gibberellins by Bacillus cereus, Bacillus macroides and Bacillus pumilus. Biotechnol Lett. 2004; 26: 191-487.
26. Oteino N, Lally RD, Kiwanuka S, Lioyd A, Ryan D, Germaine KJ, Dowling DN. Plant growth promotion induced by phosphate solubilizing endophytic Pseudomonas isolates. Front Microbiol. 2015; 6: 1-9.
27. Etesami H, Alikhani HA. The quantitative and qualitative assessment of auxin hormone production ability of same of the Iranian soils indigenous Rhizobial strains. J Water Soil. 2011; 25(1): 61-69. [In Persian]
28. Drigo B, Kowalchuk GA, Van Veen JA. Climate change goes underground: effects of elevated atmospheric CO2 on microbial community structure and activities in the rhizosphere. Biol Fertil Soils. 2008; 44: 667-679.
29. Kuklinsky-Sobral J, Araujo WL, Mendes R, Geraldi IO, Pizzirani-Kleiner AA, Azevedo JL. Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion. Environ Microbiol. 2004; 6: 1244-1251.