تاثیر باکتری اندوفیت .Enterobacter sp جداشده از ریشه ریحان روی تحریک رشد و مهار بیماری شانکر باکتریایی گیاهچه گوجه فرنگی

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانشجوی کارشناسی ارشد ، دانشکده کشاورزی، دانشگاه ایلام، ایلام، ایران

2 استادیار گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه ایلام . ایلام .ایران

3 دانشیار گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه ایلام، ایلام، ایران

4 استادیار گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه ایلام، ایلام، ایران

چکیده

شانکر باکتریایی گوجه‌فرنگی با عامل(Cmm) Clavibacter michiganensis subsp. michiganensis از بیماری‌های محدود کننده کشت گوجه‌فرنگی است. در این پژوهش 16 باکتری اندوفیت جدا شده از ریحان از نظر خاصیت ضد باکتریایی در آزمایشگاه علیه عامل شانکر باکتریایی و تحریک رشد و افزایش فنل کل گیاهچه‌های گوجه‌فرنگی آزمایش شدند. در مرحله بعد بهترین باکتری جهت مهار شانکر باکتریایی روی گیاهچه‌های گوجه‌فرنگی در شرایط گلخانه آزمایش گردید. همچنین تغییرات فیزیولوژیکی مرتبط با فنل کل،کاتالاز و پراکسیداز در گیاهچه‌ها مورد بررسی قرار گرفتند. نهایتا شناسایی مولکولی بهترین جدایه باکتری توسط آغازگرهای RNA ریبوزومی 16S صورت گرفت. هیچکدام از باکتری-های اندوفیت خاصیت ضد باکتریایی علیه شانکر باکتریایی در شرایط آزمایشگاه نشان ندادند، اما جدایه ReA1 بیشترین ویژگی‌های تحریک رشد شامل وزن‌تر و خشک، ارتفاع ساقه و افزایش فنل کل را در مقایسه با سایر باکتری‌ها نشان داد. با فرض اثر احتمالی جدایه ReA1 در مکانیسم القای مقاومت در گوجه فرنگی، این جدایه علیه باکتری Cmm استفاده شد. نتایج نشان داد که این باکتری سبب کاهش پنجاه درصدی بیماری و افزایش فاکتورهای رشدی در گوجه فرنگی می‌شود. همچنین آنزیم های آنتی‌اکسیدانی شامل پراکسیداز،کاتالاز و میزان فنل کل در گیاهان تیمار شده با جدایه ReA1 در مقایسه با شاهد افزایش نشان دادند. براساس نتایج حاصل از توالی یابی جدایه ReA1 به عنوان باکتری sp. Enterobacter تشخیص داده شد. ریحان گیاه غیرمیزبان برای Cmm می‌باشد و باکتری‌های اندوفیت ریحان اثر مستقیم ضد باکتریایی علیه Cmm نداشتند. در عین حال sp. Enterobacter جدایه ReA1 قادر به مهار زیستی شانکر باکتریایی گوجه‌فرنگی با مکانیسم احتمالی القاء مقاومت بود.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

The effect of endophytic bacterium Enterobacter sp. isolated from basil on growth stimulation and control of tomato seedling bacterial canker disease

نویسندگان [English]

  • T. Salimi 1
  • M. R. Alymanesh 2
  • A. Fazeli 3
  • M. Bagnazari 4
1 Master student, Faculty of agriculture, Ilam University, Ilam, Iran
2 Assistant Professor, Department of Plant Protection, Faculty of agriculture, Ilam University, Ilam, Iran
3 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of agriculture, Ilam University, Ilam, Iran
4 Assistant Professor, Department of Horticulture, Faculty of agriculture, Ilam University, Ilam, Iran
چکیده [English]

Background and Objectives

Bacterial canker of tomato caused by Clavibacter michiganensis subsp. michiganensis, Cmm, is one of the limiting diseases of tomato cultivation. Because of the chemical control problems, biological control methods are used to control this disease in the recent investigation. Basil (Ocimum basilicum L.) is an annual herbaceous plant from the Lamiaceae family, and various endophytic bacteria have been reported from this plant. Also, many studies have shown that endophytic bacteria have the ability to stimulate growth and control many plant diseases. Therefore, in recent research, it was tried for the first time to test endophytic bacteria isolated from basil plant against tomato bacterial canker.

Materials and Methods

In this research, 16 endophytic bacteria isolated from basil. They were tested for their antibacterial properties against tomato bacterial canker in laboratory conditions using chloroform vapor method. Also, the effect of these bacteria on increasing total phenol and growth factors, including fresh weight, dry weight and seedling height were investigated. In the next step, the best bacteria in terms of growth factors were used to control bacterial canker, on tomato seedlings, under greenhouse conditions. Next, the physiological changes related to total phenol, catalase and peroxidase were investigated. Finally, molecular identification was done by PCR using universal 16SrRNA gene-specific primers ,27f and 1492r, DNA sequencing and BlastN in NCBI Genebank.

Results

None of the endophytic bacteria showed antibacterial properties against bacterial canker under laboratory conditions. ReA1 isolate showed the highest growth stimulation and total phenol increasing compared to other bacteria therefore, it was used against tomato bacterial canker and reduced the disease by nearly fifty percent. Also, the growth factors were increased compared to the infected and healthy controls after inoculation with ReA1. In addition, the amount of peroxidase, catalase and total phenol increased in plants treated with ReA1 isolate compared to the control. So that the amount of total phenol, peroxidase and catalase, respectively from the highest to the lowest, was related to the application of ReA1 and Cmm, ReA1 alone, Cmm alone and the control without bacteria. ReA1 isolate was detected as Enterobacter sp. by DNA sequencing results.

Discussion

Recent research showed that Enterobacter sp. strain ReA1 isolated from a plant belonging to the Lamiaceae family (basil) was tested on tomato plants from the Solanaceae family, which was able to reduce tomato bacterial canker disease without direct antibacterial effect. The main possible effects of this bacterium are related to the phenomenon of induced resistance in the plant and the increase of defensive and antimicrobial compounds in the plant. This control effect on the different plant species belonging to Solanaceae, the possibility that this bacterium could be used as a biocontrol agent on many kinds of plants and against a wide range of diseases. It also stimulated the growth of the plant, which makes possible the simultaneous use of this bacterium as a fertilizer and biocontrol agent.

کلیدواژه‌ها [English]

  • Keywords: Clavibacter michiganensis subsp. michiganensis
  • Biological Control
  • Total phenol
  • Induction of Resistance
Abdelshafy Mohamad, O. A., Ma, J.-B., Liu, Y.-H., Zhang, D., Hua, S., Bhute, S., Hedlund, B. P., Li, W.-J., & Li, L. (2020). Beneficial endophytic bacterial populations associated with medicinal plant Thymus vulgaris alleviate salt stress and confer resistance to Fusarium oxysporum. Frontiers in plant science, 11, 47. https://doi.org/10.3389/fpls.2020.00047.
Aebi, H. (1984). Catalase in vitro. In Methods in enzymology (Vol. 105, pp. 121-126).
Afzal, I., Shinwari, Z. K., Sikandar, S., & Shahzad, S. (2019). Plant beneficial endophytic bacteria: Mechanisms, diversity, host range and genetic determinants. Microbiological research, 221, 36-49. https://doi: 10.1016/j.micres.2019.02.001.
Alaylar, B. (2022). Isolation and characterization of culturable endophytic plant growth-promoting Bacillus species from Mentha longifolia L. Turkish Journal of Agriculture and Forestry, 46(1), 73-82. https://doi: 10.3906/tar-2109-24.
Ali, B., Hafeez, A., Javed, M. A., Afridi, M. S., Abbasi, H. A., Qayyum, A., Batool, T., Ullah, A., Marc, R. A., & Al Jaouni, S. K. (2022). Role of endophytic bacteria in salinity stress amelioration by physiological and molecular mechanisms of defense: A comprehensive review. South African Journal of Botany, 151, 33-46. https://doi.org/10.1016/j.sajb.2022.09.036.
Al-Maawali, S. S., Al-Sadi, A. M., Ali Khalifa Alsheriqi, S., Nasser Al-Sabahi, J., & Velazhahan, R. (2021). The potential of antagonistic yeasts and bacteria from tomato phyllosphere and fructoplane in the control of Alternaria fruit rot of tomato. All Life, 14(1), 34-48. https://doi: 10.1080/26895293.2020.1858975.
Ansari, M., Taghavi, S. M., Hamzehzarghani, H., Valenzuela, M., Siri, M. I., & Osdaghi, E. (2019). Multiple introductions of tomato pathogen Clavibacter michiganensis subsp. michiganensis into Iran as revealed by a global-scale phylogeographic analysis. Applied and Environmental Microbiology, 85(24), e02098-19. https://doi.org/10.1128/AEM.02098-19.
Bacon, C. W., & Hinton, D. M. (2006). Bacterial endophytes: the endophytic niche, its occupants, and its utility. In Plant-associated bacteria (pp. 155-194). Springer, Dordrecht. 10.1007/978-1-4020-4538-7_5.
Blee, K. A., Choi, J. W., O'Connell, A. P., Schuch, W., Lewis, N. G., & Bolwell, G. P. (2003). A lignin-specific peroxidase in tobacco whose antisense suppression leads to vascular tissue modification. Phytochemistry, 64(1), 163-176. https://doi: 10.1016/s0031-9422(03)00212-7.
Boudyach, E. H., Fatmi, M., Akhayat, O., Benizri, E., & Aoumar, A. A. B. (2001). Selection of antagonistic bacteria of Clavibacter michiganensis subsp. michiganensis and evaluation of their efficiency against bacterial canker of tomato. Biocontrol Science and Technology11(1), 141-149. https://doi.org/10.1080/09583150020029817.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry72(1-2), 248-254. https://doi.org/10.1016/0003-2697(76)90527-3.
Dasa, I., Pandaa, M. K., Rathb, C. C., & Tayungc, K. (2017). Bioactivities of bacterial endophytes isolated from leaf tissues of Hyptis suaveolens against some clinically significant pathogens. Journal of Applied Pharmaceutical Science, 7(8), 131-136. https://doi:10.7324/JAPS.2017.70818.
Egamberdieva, D., Wirth, S., Behrendt, U., Ahmad, P., & Berg, G. (2017). Antimicrobial activity of medicinal plants correlates with the proportion of antagonistic endophytes. Frontiers in microbiology, 8, 199. https://doi: 10.3389/fmicb.2017.00199.
Egert, M., & Tevini, M. (2002). Influence of drought on some physiological parameters symptomatic for oxidative stress in leaves of chives (Allium schoenoprasum). Environmental and Experimental Botany, 48(1), 43-49. https://doi: 10.1016/S0098-8472(02)00008-4.
Eichenlaub, R., & Gartemann, K.-H. (2011). The Clavibacter michiganensis subspecies: molecular investigation of gram-positive bacterial plant pathogens. Annual Review of Phytopathology, 49(1). https://doi: 10.1146/annurev-phyto-072910-095258.
Frank, J.A., Reich, C.I., Sharma, S., Weisbaum, J.S., Wilson, B.A. and Olsen, G.J. (2008). Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes. Applied and environmental microbiology74(8), pp.2461- 2470. https://doi: 10.1128/AEM.02272-07.
Gartemann, K.-H., Abt, B., Bekel, T., Burger, A., Engemann, J., Flügel, M., Gaigalat, L., Goesmann, A., Gräfen, I., & Kalinowski, J. r. (2008). The genome sequence of the tomato-pathogenic actinomycete Clavibacter michiganensis subsp. michiganensis NCPPB382 reveals a large island involved in pathogenicity. Journal of Bacteriology, 190(6), 2138-2149. https:// doi: 10.1128/JB.01595-07.
Gill, S. S., & Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant physiology and biochemistry, 48(12), 909-930. https:// doi: 10.1016/j.plaphy.2010.08.016.
Hallmann, J., Quadt-Hallmann, A., Mahaffee, W., & Kloepper, J. (1997). Bacterial endophytes in agricultural crops. Canadian journal of microbiology, 43(10), 895-914. https://doi: 10.1139/m97-13.
Hernández-Pacheco, C. E., del Carmen Orozco-Mosqueda, M., Flores, A., Valencia-Cantero, E., & Santoyo, G. (2021). Tissue-specific diversity of bacterial endophytes in Mexican husk tomato plants (Physalis ixocarpa Brot. ex Horm.), and screening for their multiple plant growth-promoting activities. Current Research in Microbial Sciences, 2, 100028. https://doi: 10.1016/j.crmicr.2021.100028.
Herzog, V., & Fahimi, H. D. (1973). A new sensitive colorimetric assay for peroxidase using 3, 3′-diaminobenzidine as hydrogen donor. Analytical biochemistry, 55(2), 554-562. https:// doi: 10.1016/0003-2697(73)90144-9.
Hassanein, M. A. F., & Al-Amari, A. (2021). Endophytic Bacteria as Apotential Agent for Control of Tomato Wilt Caused by Fusarium oxysporum f. sp LycopersiciAnnals of the Romanian Society for Cell Biology, 3119-3132. 10.3389/fmicb.2021.731764. eCollection 2021.
Jacob, J., Krishnan, G. V., Thankappan, D., & Amma, D. K. B. N. S. (2020). Endophytic bacterial strains induced systemic resistance in agriculturally important crop plants. In Microbial Endophytes (pp. 75-105). Woodhead Publishing. https://doi.org/10.1016/B978-0-12-819654-0.00004-1.
Jang, H., Kim, S. T., & Sang, M. K. (2022). Suppressive Effect of Bioactive Extracts of Bacillus sp. H8-1 and Bacillus sp. K203 on Tomato Wilt Caused by Clavibacter michiganensis subsp. michiganensis. Microorganisms, 10(2), 403. https:// doi: 10.3390/microorganisms10020403.
Knapp, S., & Peralta, I. E. (2016). The tomato (Solanum lycopersicum L., Solanaceae) and its botanical relatives. The tomato genome, 7-21. 10.1007/978-3-662-53389-5_2.
Kawaguchi, A., & Tanina, K. (2014). Genetic groups of Clavibacter michiganensis subsp. michiganensis identified by DNA fingerprinting and the effects of inoculation methods on disease development. European journal of plant pathology, 140(3), 399-406. https://doi: 10.1007/s10658-014-0475-9.
Lanna-Filho, R., Souza, R. M., Magalhães, M. M., Villela, L., Zanotto, E., Ribeiro-Júnior, P. M., & Resende, M. L. (2013). Induced defense responses in tomato against bacterial spot by proteins synthesized by endophytic bacteria. Tropical Plant Pathology38, 295-302.https://doi.org/10.1590/S1982-56762013005000011.
Ma, Y., Rajkumar, M., Zhang, C., & Freitas, H. (2016). Beneficial role of bacterial endophytes in heavy metal phytoremediation. Journal of environmental management, 174, 14-25.‏ https://doi.org/10.1016/j.jenvman.2016.02.047.
Madhurama, G., Sonam, D., Urmil, P. G., & Ravindra, N. K. (2014). Diversity and biopotential of endophytic actinomycetes from three medicinal plants in India. African Journal of Microbiology Research, 8(2), 184-191. https://doi: 10.5897/AJMR2012.2452.
Mandal, S. (2010). Induction of phenolics, lignin and key defense enzymes in eggplant (Solanum melongena L.) roots in response to elicitors. African Journal of Biotechnology, 9(47), 8038-8047. https://doi: 10.5897/AJB10.984.
Mazarei, M., & Orumchi, S. (1993). Investigation of bacterial canker of tomato in West Azarbaidjan. In Proceedings of the 11th Plant Protection Congress of Iran 28 Aug.-2 Sep. 1993 Rasht.
Meng, X. J., Medison, R. G., Cao, S., Wang, L. Q., Cheng, S., Tan, L. T., ... & Zhou, Y. (2023). Isolation, identification, and biocontrol mechanisms of endophytic Burkholderia vietnamiensis C12 from Ficus tikoua Bur against Rhizoctonia solaniBiological Control, 178, 105132. https://doi.org/10.1016/j.biocontrol.2022.105132.
M'piga, P., Belanger, R., Paulitz, T., & Benhamou, N. (1997). Increased resistance to Fusarium oxysporumf. sp. radicis-lycopersiciin tomato plants treated with the endophytic bacterium Pseudomonas fluorescensstrain 63-28. Physiological and molecular plant pathology, 50(5), 301-320. https://doi: 10.1006/pmpp.1997.0088.
Nawed, A., & Chandra, R. (2015). Endophytic bacteria: optimizaton of isolation procedure from various medicinal plants and their preliminary characterization. Asian Journal of Pharmaceutical and Clinical Research, 8(4), 233-238.
Omomowo, O. I., & Babalola, O. O. (2019). Bacterial and fungal endophytes: tiny giants with immense beneficial potential for plant growth and sustainable agricultural productivity. Microorganisms, 7(11), 481. https:// doi: 10.3390/microorganisms7110481.
Ordookhani, K., Sharafzadeh, S., & Zare, M. (2011). Influence of PGPR on growth, essential oil and nutrients uptake of sweet basil. Advances in Environmental Biology, 5(4), 672-677.
Panigrahi, S., Mohanty, S., & Rath, C. (2020). Characterization of endophytic bacteria Enterobacter cloacae MG00145 isolated from Ocimum sanctum with Indole Acetic Acid (IAA) production and plant growth promoting capabilities against selected crops. South African Journal of Botany, 134, 17-26. https://doi.org/10.1016/j.sajb.2019.09.017.
Parida, A. K., & Das, A. B. (2005). Salt tolerance and salinity effects on plants: a review. Ecotoxicology and environmental safety, 60(3), 324-349. https:// doi: 10.1016/j.ecoenv.2004.06.010.
Pieterse, C. M., Zamioudis, C., Berendsen, R. L., Weller, D. M., Van Wees, S., & Bakker, P. A. (2014). Induced systemic resistance by beneficial microbes. Annual review of phytopathology, 52, 347-375. https:// doi: 10.1146/annurev-phyto-082712-102340. Epub 2014 Jun 2.
Ryan, A. D., Kinkel, L. L., & Schottel, J. L. (2004). Effect of pathogen isolate, potato cultivar, and antagonist strain on potato scab severity and biological control. Biocontrol Science and Technology, 14(3), 301-311. https://doi/abs/10.1080/09583150410001665187.
Sahu, P. K., Singh, S., Gupta, A. R., Gupta, A., Singh, U. B., Manzar, N., Bhowmik, A., Singh, H. V., & Saxena, A. K. (2020). Endophytic bacilli from medicinal-aromatic perennial Holy basil (Ocimum tenuiflorum L.) modulate plant growth promotion and induced systemic resistance against Rhizoctonia solani in rice (Oryza sativa L.). Biological control, 150, 104353. https://doi.org/10.1016/j.biocontrol.2020.104353.
Safara, S., Harighi, B., Bahramnejad, B. and Ahmadi, S. (2022). Antibacterial activity of endophytic bacteria against sugar beet root rot agent by volatile organic compound production and induction of systemic resistance. Frontiers in Microbiology, 13. https:// doi: 10.3389/fmicb.2022.921762.
Sánchez-Pérez, B. N., Zenteno-Rojas, A., Rincón-Molina, C. I., Ruíz-Valdiviezo, V. M., Gutiérrez-Miceli, F. A., Vences-Guzmán, M. A., Villalobos-Maldonado, J. J., & Rincón-Rosales, R. (2020). Rhizosphere and endophytic bacteria associated to Ocimum basilicum L. with decaclorobiphenyl removal potential. Water, Air, & Soil Pollution, 231(3), 1-15. https://doi.org/10.1007/s11270-020-04481-6.
Shahrajabian, M. H., Sun, W., & Cheng, Q. (2020). Chemical components and pharmacological benefits of Basil (Ocimum basilicum): A review. International Journal of Food Properties23(1), 1961-1970. https://doi.org/10.1080/10942912.2020.1828456.
Seevers, P., Daly, J., & Catedral, F. (1971). The role of peroxidase isozymes in resistance to wheat stem rust disease. Plant physiology, 48(3), 353-360. https://doi.org/10.1104/pp.48.3.353.
Sen, Y., van der Wolf, J., Visser, R. G., & van Heusden, S. (2015). Bacterial canker of tomato: current knowledge of detection, management, resistance, and interactions. Plant Disease, 99(1), 4-13. https:// doi: 10.1094/PDIS-05-14-0499-FE.
Sharma, M., Sood, G., & Chauhan, A. (2021). Bioprospecting beneficial endophytic bacterial communities associated with Rosmarinus officinalis for sustaining plant health and productivity. World Journal of Microbiology and Biotechnology, 37(8), 1-17. https://doi.org/10.1007/s11274-021-03101-7.
Shternshis, M., Beljaev, A., Shpatova, T., Bokova, J., & Duzhak, A. (2002). Field testing of Bacticide®, Phytoverm® and Chitanase for control of the raspberry midge blight in Siberia. Biological Control, 47(6), 697-706. https://doi.org/10.1023/A:1020574914831.
Simova-Stoilova, L., Demirevska, K., Petrova, T., Tsenov, N., & Feller, U. (2008). Antioxidative protection in wheat varieties under severe recoverable drought at seedling stage. Plant, Soil and Environment, 54(12), 529-36.
Yanti, Y. (2019). Involvement of Jasmonic Acid in the Induced Systemic Resistance of Tomato against Ralstonia syzigii subsp. indonesiensis by Indigenous Endophyte Bacteria. In IOP Conference Series: Earth and Environmental Science (Vol. 347, No. 1, p. 012024). IOP Publishing. 10.1088/1755-1315/347/1/012024.
Yarte, M. E., Gismondi, M. I., Llorente, B. E., & Larraburu, E. E. (2022). Isolation of endophytic bacteria from the medicinal, forestal and ornamental tree Handroanthus impetiginosus. Environmental Technology, 43(8), 1129-1139. https://doi.org/10.1080/09593330.2020.1818833.
Wang, H., Liu, R., You, M. P., Barbetti, M. J., & Chen, Y. (2021). Pathogen biocontrol using plant growth-promoting bacteria (PGPR): Role of bacterial diversity. Microorganisms, 9(9), 1988. https:// doi: 10.3390/microorganisms9091988.
Wu, W., Chen, W., Liu, S., Wu, J., Zhu, Y., Qin, L., & Zhu, B. (2021). Beneficial relationships between endophytic bacteria and medicinal plants. Frontiers in plant science, 12, 646146. https://doi: 10.3389/fpls.2021.646146.
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