باکتری‌های همراه بذر چغندرقند: شناسایی و اثر آن‌ها بر جوانه‌زنی بذر

نوع مقاله : کامل علمی - پژوهشی

نویسندگان

1 گروه گیاهپزشکی دانشگاه بوعلی سینا همدان

2 همدان گروه گیاهپزشکی دانشگاه بوعلی سینا

چکیده

بذر چغندرقند دارای اهمیت ویژه‌ای است زیرا به‌عنوان انتقال‌دهنده باکتری‌های مفید و مضر نقش ایفا می‌کند. جهت شناسایی و تعیین اثر باکتری­های همراه بذر چغندرقند بر جوانه­زنی، بذر بوجاری­شده و ضد­عفونی نشده از بخش تحقیقات چغندرقند مرکز تحقیقات کشاورزی استان همدان تهیه گردید. درمجموع تعداد 80 استرین اندوفیت و اپی­فیت از بذر چغندرقند جداسازی شد. بر پایه الگوی پروتئینی به روش لاملی، تعداد سه الکتروتیپ نماینده انتخاب‌شده و خصوصیات بیوشیمیایی و مولکولی آن‌ها بررسی گردید. بر اساس نتایج آزمون‌های بیوشیمیایی و تعیین توالی ژن 16S rRNA استرین­های نماینده، گونه‌های Stenotrophomonas rhizophila و Pseudomonas geniculata به‌عنوان گونه‌های اپی­فیت و گونه Stenotrophomonas maltophilia به‌عنوان تنها گونه اندوفیت بذر چغندرقند شناسایی شدند. جهت انجام آزمون جوانه‌زنی بذر، سه گروه متفاوت ازلحاظ تأثیر استرین­ها بر جوانه‌زنی بذور چغندرقند ایجاد گردید. با تلقیح پنج استرین نماینده از هر گروه در قالب طرح کاملاً تصادفی با چهار تکرار و هر تکرار شامل یک پتری دیش با 10 عدد بذر، آزمون جوانه‌زنی بذر انجام شد. بر اساس نتایج، بذور تلقیح شده با استرین­های نماینده S. maltophilia، بیشترین میزان درصد جوانه­زنی (90 درصد)، سرعت جوانه‌زنی (0/0174 بر ساعت)، وزن‌تر ساقه­چه (100 میلی­گرم) و ریشه‌چه (126/2 میلی‌گرم)، وزن خشک ساقه­چه (50 میلی‌گرم) و ریشه‌چه (43/7 میلی‌گرم)، طول ساقه­چه (6/5 سانتی‌متر) و ریشه‌چه (8 سانتی‌متر) و کم‌ترین میزان یکنواختی بذر (138/7 ساعت) را نسبت به شاهد دارا بودند. بنابراین می‌توان پیشنهاد کرد که به‌منظور بهبود شاخص‌های جوانه‌زنی و رشد گیاهچه تلقیح بذور چغندرقند با باکتری S. maltophilia صورت گیرد.

کلیدواژه‌ها


عنوان مقاله [English]

Sugar beet seed-associated bacteria: their identification and effects on seed germination

نویسندگان [English]

  • Milad Aeini 1
  • Gholam Khodakaramian 2
چکیده [English]

Sugar beet seed plays an important role in the transmission of beneficial or detrimental bacteria. To identify and evaluate the effects of sugar beet seed-associated bacteria on seed germination, cleaned but not disinfected seeds were provided from Sugar Beet Research Department at Hamadan Agricultural Research Center. A total of 80 endophyte and epiphyte bacterial strains were isolated from the seeds. Based on the protein pattern and according to Laemmli method, three representative electrotypes were selected and their biochemical and molecular characteristics were evaluated. Based on the biochemical test results and sequencing of the 16S rRNA gene of the representative strains Stenotrophomonas rhizophila and Pseudomonas geniculata were identifiedas epiphytic and Stenotrophomonas maltophilia as endophytic species, respectively. To perform seed germination test, three different groups were generated based on the strain effect on seed germination. Seed germination test was conducted based on the inoculation of five representative strains from each group in completely randomized design with four replications. Each replication included a Petri dish containing 10 seeds. Results showed that seeds inoculated by S. maltophilia strain had the highest germination percentage (90), germination rate (0.0174 per hour), shoot fresh weight (100 mg), radicle fresh weight (126.2 mg), shoot dry weight (50 mg), radicle dry weight (43.7 mg), shoot length (6.5 cm), radicle length (8 cm), and the lowest seed uniformity compared with control. In order to improve germination indices and seedling growth, it is recommended to inoculate sugar beet seed with S. maltophilia bacterium.

کلیدواژه‌ها [English]

  • Endophyte
  • epiphyte
  • Seed germination
  • sugar beet
Banerjee M, Yesmin L. Sulfur-oxidizing plant growth promoting rhizobacteria for enhanced canola performance. US Patent. 2002; 07491535.
Bashan Y, Holguin G, De-Bashan L. Azospirillum-plant relationships: physiological, molecular, agricultural, and environmental advances. Can J Microbiol. 2004; 50: 521-577.
Berg G, Egamberdieva D, Lugtenberg B, Hagemann M. Symbiotic plant–microbe interactions: stress protection, plant growth promotion, and biocontrol by Stenotrophomonas. In Seckbach, J. and Grube, M. Symbiosis and Stress. Cellular Origin, Life in Extreme Habitats and Astrobiology. 2010; 4: 445–460.
Bergey DH, Holt JG, Noel RK. Bergey’s Manual of Systematic Bacteriology (Ninth edition). Baltimore, MD. Williams & Wilkins. 1994; 1: 1935-2045.
Compant S, Duffy B, Nowak J, Clement C, Barka EA. Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Appl Environ Microbiol. 2005; 71: 4951–4959.
De Freitas JR, Banerjee MR, Germida JJ. Phosphate-solubilizing rhizobacteria enhance the growth and yield but not phosphorus uptake of canola (Brassica napus L.). Biol Fertl Soils. 1997; 24:358–364.
Denton M, Kerr KG. Microbiological and clinical aspects of infection associated with Stenotrophomonas maltophilia. Clin Microbiol Rev. 1998; 11: 57–80.
Fahy PC, Persley GJ. Plant Bacterial Disease: A Diagnostic Guide. Academic Press. Sydney, Australia. 1983; 393p.
Grondeau C, Samson R, Sands DDC. A review of thermotherapy to free plant materials from pathogens, especially seeds from bacteria. Crit Rev Plant Sci. 2011; 13: 57–75.
Habazar T, Yanti Y, Ritonga C. Formulation of indigenous rhizobacterial isolates from healthy soybean’s root, which ability to promote growth and yield of soybean.Int J Sci Engg Tech. 2014; 4: 2088-5334.
Halmer P. Methods to improve seed performance. In: Benech-Arnold RL, Sanchez RA (eds) Seed Physiology, Applications to Agriculture. Food Product Press, New York. 2003.
Hayward AC, Fegan NM, Stirling GR. Stenotrophomonas and Lysobacter: ubiquitous plant-associated gamma-proteobacteria of developing significance in applied microbiology. J Appl Microbiol. 2010; 108: 756–770.
Holland MA, Polacco JC. PPFMs and other covert contaminants: is there more to plants physiology than just plants. Annu Rev Plant Physiol Plant Mol Biol. 1994; 45: 197–209.
Hardoim PR, Hardoim CCP, van Overbeek LS, van Elsas JD. Dynamics of seed-borne rice endophytes on early plant growth stages. PLoS ONE. 2012; 7: e30438. 
Jagadish DR. Evaluation of different methods of application of Pseudomonas B-25 strain for biological control of early blight of tomato caused by Alternaria solani Mill. M. Sc. (Agri.) Thesis, Univ. of Agri. Sci. Dharwad (India) 2006.
Johnston-Monje D, Raizada MN. Conservation and diversity of seed associated endophytes in Zeaacross boundaries of evolution, ethnography and ecology. PLoS ONE. 2011;6: e20396.
Kaga H, Mano H, Tanaka F, Watanabe A, Kaneko S, Morisaki H. Rice seeds as sources of endophytic bacteria. Microbes Environ. 2009; 24: 154–162.
Kremer RJ. Identity and properties of bacteria inhabiting seeds of selected broadleaf weed species. Microb Ecol. 1987; 14: 29-37
Laemmli UK. Cleavage of structural protein during the assembly of the head of bacteriophage.Nature. 1970; 227: 680- 685.
Li J, Kremer RJ. Growth response of weed and crop seedlings to deleterious rhizobacteria. Biol Control. 2006; 39: 58–65.
Lucero ME, Unc A, Cooke P, Dowd S, Sun S. Endophyte microbiome diversity in micropropagated Atriplex canescens and Atriplex torreyi var griffithsii. PLoS One. 2011 Mar 17; 6(3):e17693.
 Nezarat S, Gholami A. The effect of plant growth promoting rhizobacteria (PGPR) on germination, seedling growth and yield of maize. Pak J Biol Sci. 2009; 12(1):26-32.
Paradis E, Goyer C, Hodge NC, Houge R, Robert ES, Beaulieu C. Fatty acid and protein profiles of Streptomyces scabies strains isolated in eastern Canada. IntJSysBacteriol. 1994; 44: 561- 564.
Park MKim CYang JLee HShin WKim SSa T.Isolation and characterization of diazotrophic growth promoting bacteria from rhizosphere of agricultural crops of Korea. Microbiol Res. 2005;160: 127–133.
Rastogi G, Sbodio A, Tech JJ, Suslow TV, Coaker GL, Leveau JHJ. Leaf microbiota in an agroecosystem: spatiotemporal variation in bacterial community composition on field grown lettuce. ISME Journal. 2012; 6: 1812–1822.
Rudgers JA, Afkhami ME, Rua MA, Davitt AJ, Hammer S, Huguet VM. A fungus among us: broad patterns of endophyte distribution in the grasses. Ecology. 2009; 90: 1531–1539.
Schaad NW, Jones JB, Chun W. Laboratory guide for identification of plant pathogenic bacteria. American Phtopathology Society Minnesota. 2001; 373pp.
Schippers B, Bakker AW, Bakker AHM. Interactions of deleterious and beneficial rhizosphere microorganism and the effect of cropping practices. Ann Rev Phytopathol. 1987; 25: 339-358.
Soltani A, Zeinali E, Galeshi S, Latifi N. Genetic variation for and interrelationships among seed vigor traits in wheat from the Caspian Sea Coast of Iran. Seed Sci and Technol. 2001; 29: 653-662.
Suckstorff I, Berg G. Evidence for dose-dependent effects on plant growth by Stenotrophomonas strains from different origins. J Appl Microbiol. 2003; 95: 656–663.
Vauterin L, Vantomme R, Pot B, Hoste B, Swings J, Kersters K. Taxonomic analysis of Xhantomonascampestris pv. begonidae and X. campestris pv. pelargonii by means of phythopathological, phenotypic, protein electrophoretic and DNA hybridization methods. Syst Appl Bacteriol. 1990; 13: 166–167.
Watson L, Dallwitz MJ. The Families of Flowering Plants: Descriptions, Illustrations, Identification, and Information Retrieval. Version: 15th October 1998.
Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S Ribosomal DNA Amplification for Phylogenetic Study. J Bacteriol. 1991; 173: 697-703.
William GE, Asher MJC. Selection of rhizobacteria for the control of Pythium ultimum and aphanomyces cochioides on sugar beet seedling. Crop prot. 1996; 15: 479-486.
Wolf A, Fritze A, Hagemann M, Gabriele B. Stenotrophomonas rhizophila sp. nov. a novel plant-associated bacterium with antifungal properties. Int J Syst Evol Microbiol. 2002; 52: 1937–1944.
Zaidi SFA. Inoculation with Bradyrhizobium japonicum and fluorescent Pseudomonas to control Rhizoctonia solani in soybean [Glycine max L. Merr]. Ann Agr Res. 2003; 24: 151-153.