بررسی بیان ژن‌های کیتیناز و گلوکاناز در گندم تیمار شده با اسیدسالیسیلیک و باکتری Pseudomonas flourescens در برهمکنش با قارچ عامل سوختگی برگ گندم

نوع مقاله: علمی پژوهشی

نویسندگان

1 دانشجوی دکتری گروه بیماری‌شناسی گیاهی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

2 استادیار گروه زیست فراورده‌های گیاهی، پژوهشکده زیست‌فناوری کشاورزی، پژوهشگاه ملی مهندسی ژنتیک و زیست‌فناوری، تهران، ایران

3 استاد گروه بیماری شناسی گیاهی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

4 دانشیار گروه بیماری شناسی گیاهی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران.

چکیده

القاء مقاومت به بیمارگرها از جمله راه‌کارهای مقابله با تنش‌های زیستی در گیاهان می‌باشد. آنزیم‌های کیتیناز و بتا 1و3 گلوکوناز از جمله پروتئین-های دفاعی قابل‌القاء هستند که در پاسخ به‎عوامل بیماریزا توسط گیاه، تولید می‌شوند. در این تحقیق، در ابتدا تاثیر تیمار اسیدسالیسیلیک و باکتری Pseudomonas flourescens در کاهش علایم بیماری سوختگی برگ گندم مورد بررسی قرار گرفت. سپس تغییرات بیان ژن‌های کیتیناز و بتا 1و3 گلوکوناز در نمونه‌های گیاهی تیمار شده و کنترل در روزهای 0، 4، 14 و 24 سیکل بیماری به‎روش RT-PCR نیمه کمی بررسی شدند. نتایج نشان‌دهندۀ افزایش بیان هر دو ژن در روز چهارم پس از تلقیح بیمارگر بود و کاربرد اسیدسالیسیک و باکتری بیوکنترل به‎صورت جداگانه توانست بیان آن‌ها را افزایش دهد، ولی کاربرد همزمان هر دو تیمار، کاهش معنی‌داری از نظر آماری ایجاد نکرد. بر اساس نتایج، افزایش بیان آنزیم‌های کیتیناز و بتا-1و3 گلوکوناز با کاربرد اسیدسالیسیک و باکتری بیوکنترل P. flourescens می‌تواند در القاء مقاومت میزبانی در مقابله با بیماری سوختگی برگ گندم نقش داشته باشد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Evaluation of chitinase and β-1,3glucanase expression profile in Pseudomonas flourescens and salicylic acid treated wheat against Septoria tritici bloch (STB)

نویسندگان [English]

  • Elham Zamani 1
  • Forogh Sanjarian 2
  • Ebrahim Mohammadi-goltapeh 3
  • Naser Safaie 4
1 Ph.D. student, Department of Plant Pathology, Tarbiat Modares University, Faculty of Agriculture, Tehran, Iran
2 Assistant Professor, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
3 Professor, Department of Plant Pathology, Tarbiat Modares University, Faculty of Agriculture, Tehran, Iran
4 Associate Professor, Department of Plant Pathology, Tarbiat Modares University, Faculty of Agriculture, Tehran, Iran
چکیده [English]

Induced resistance is one the ways by which plants cope with the biotic stresses. Pathogenesis-related proteins (PRs), including chitinase and β-1,3-glucanase are synthesized by plant in response to pathogenic infection. In present work, we have carried out inoculation experiment in wheat to investigate the effect of salicylic acid (SA) and Pseudomonas flourescens on Septoria tritici Bloch (STB) symptoms. Subsequently, the expression profiles of chitinase and β-1,3-glucanase were analyzed with semi-quantitative RT-PCR at 0, 4, 14 and 24 days after inoculation in both treated and control plants. Results showed that the expression of both chitinase and β-1,3-glucanase genes increased in interaction with Z. tritici and the application of SA and P. flourescens. On the other hand, the application together had no significant effect on gene expression. Briefly, it can be concluded that increment of chitinase and β-1,3-glucanase expression level with the application of SA and P. flourescens would play role induction of plant defense against STB.

کلیدواژه‌ها [English]

  • “Salicylic acid
  • ” Pseudomonas flourescens
  • “Chitinase
  • “ Glucanase
  • “Septoria tritici bloch
Ahangar L, Babaeezad V, Ranjbar G, Najafizarini H, Biabani A (2015) Expression profile of defense-related genes in susceptible and resistant wheat cultivars in response to powdery mildew infection. J. Mod. Genet. 10: 33-46.

Appu M, Muthukrishnan S (2014) Foliar application of salicylic acid stimulates flowering and induce defense related proteins in finger millet plants. Universal J. Plant Sci. 2: 14-18.

Bockus WW, Bowden R, Hunger R, Murray T, Smiley, R (2010) Compendium of wheat diseases and pests, American Phytopathological Society (APS Press).

Bol J, Linthorst H, Cornelissen B (1990) Plant pathogenesis-related proteins induced by virus infection. Annu. Rev. Phytopathol. 28: 113-138.

Brown J, Kema G, Forrer HR, Verstappen E, Arraiano L, Brading P, Foster E, Fried P, Jenny E (2001) Resistance of wheat cultivars and breeding lines to septoria tritici blotch caused by isolates of Mycosphaerella graminicola in field trials. Plant Pathol. 50: 325-338.

Chartrain L, Brading P, Makepeace J, Brown J (2004) Sources of resistance to septoria tritici blotch and implications for wheat breeding. Plant Pathol. 53: 454-460.

Dean R, Van Kan JA, Pretorius ZA, Hammond‐Kosack KE, Di Pietro A, Spanu PD, Rudd JJ, Dickman M, Kahmann R, Ellis J (2012) The Top 10 fungal pathogens in molecular plant pathology. Mol. Plant Pathol. 13: 414-430.

Eyal Z (1987) The Septoria diseases of wheat: concepts and methods of disease management, CIMMYT.

Gholamnejad J, Mohammadi-goltapeh E, Sanjarian F, Safaie N, Razavi Kh (2014). The evaluation of salicylic acid effect on septoria diseases by Mycospharella graminicola.  J. Res. Plant Dis. 2: 37-48.

Goodwin SB, M’barek SB, Dhillon B, Wittenberg AH, Crane CF, Hane JK, Foster AJ, Van der Lee T, Grimwood J, Aerts A (2011) Finished genome of the fungal wheat pathogen Mycosphaerella graminicola reveals dispensome structure, chromosome plasticity, and stealth pathogenesis. PLoS Genet. doi: 10.1371/journal.pgen.1002070

Gorjanović S (2009) A review: biological and technological functions of barley seed Pathogenesis‐related proteins (PRs). J. Inst. Brew. 115: 334-360.

Gozzo F (2003) Systemic acquired resistance in crop protection: from nature to a chemical approach. J. Agri. Food Chem. 51: 4487-4503.

Guo JR, Verreet JA (2008). Formation and germination of Septoria tritici secondary conidia as affected by environmental factors. Phytopathology 156: 635–637.

Habibi M, Mirakhorli N, Shiran B, Mardi M (2013) Study of resistance related gene expression pattern to Septoria tritici Blotch (STB) in wheat (Triticum aestivum). J. Mod. Genet. 8:149-158.

Hajimashaalah Bazaz S, Razavi M, Ghasemi A (2014) Evaluation of Pseudomonas fluorescens for biological control of wheat crown and root rot disease (Fusarium culmorum). Biocontrol. Plant. Protec. 1: 1-16

Hammerschmidt R (1995) Induced resistance to disease in plants. Springer Science and Business Media.

Ji C, Kuc J (1994) Purification and characterization of an acidic beta-1, 3-glucanase from cucumber and its relationship to systemic disease resistance induced by Colletotrichum lagenarium and tobacco necrosis virus. Mol. Plant-Microbe Interact. 8: 899-905.

Jongedijk E, Tigelaar H, Van Roekel JS, Bres-Vloemans SA, Dekker I, van den Elzen PJ, Cornelissen BJ, Melchers LS (1995) Synergistic activity of chitinases and β-1,3glucanases enhances fungal resistance in transgenic tomato plants. Euphytica 85: 173-180.

Kachroo A, Lapchyk L, Fukushige H, Hildebrand D, Klessig D, Kachroo P (2003) Plastidial fatty acid signaling modulates salicylic acid–and jasmonic acid–mediated defense pathways in the Arabidopsis ssi2 mutant.  Plant Cell 15: 2952-2965.

Kang Z, Buchenauer H (2002) Studies on the infection process of Fusarium culmorum in wheat spikes: degradation of host cell wall components and localization of trichothecene toxins in infected tissue. In: Mycotoxins in Plant Disease. Springer, pp 653-660.

Kema G, Annone JG, Sayoud R, Van Silfhout CH, Van Ginkel M, De Bree J (1996) Genetic variation for virulence and resistance in the wheat-Mycosphaerella graminicola pathosystem I. Interactions between pathogen isolates and host cultivars. Phytopathology 86: 200-212.

Kemp G, Botha A-M, Kloppers F, Pretorius Z. (1999) Disease development and β-1, 3-glucanase expression following leaf rust infection in resistant and susceptible near-isogenic wheat seedlings. Physiol. Mol. Plant Pathol. 55: 45-52.

Kini KR, Vasanthi N, Shetty HS (2000) Induction of β-1, 3-glucanase in seedlings of pearl millet in response to infection by Sclerospora graminicola. Eur. J. Plant Pathol. 106: 267-274.

Kloek AP, Verbsky ML, Sharma SB, Schoelz JE, Vogel J, Klessig DF, Kunkel BN (2001) Resistance to Pseudomonas syringae conferred by an Arabidopsis thaliana coronatine‐insensitive (coi1) mutation occurs through two distinct mechanisms. Plant J. 26: 509-522.

Kumar V, Kumar A, Verma V, Gond S, Kharwar R (2007) Induction of defense enzymes in Pseudomonas fluorescens treated chickpea roots against Macrophomina phaseolina. Indian Phytopathol. 60: 289-295.

Leah R, Tommerup H, Svendsen I, Mundy J (1991) Biochemical and molecular characterization of three barley seed proteins with antifungal properties. J. Biol. Chem. 266: 1564-1573.

Leubner-Metzger G, Meins Jr F (1999) Functions and Regulation of Plant β-(PR-2). In: Datta SK, Muthukrishnan S (eds) Pathogenesis-related proteins in plant, CRC Press LLC, Boca Raton, Florida, pp, 49-76.

Li YZ, Zheng XH, Tang HI, Zhu JW,  Yang JM (2003) Increase of ß-1, 3-glucanase and chitinase activities in cotton callus cells treated by salicylic acid and toxin of Verticillium dahliae. Acta Bot. Sinica 45: 802-808.

Ma BC, Tang WL, Ma LY, Li LL, Zhang LB, Zhu SJ, Zhuang C,  Irving D (2009) The role of chitinase gene expression in the defense of harvested banana against anthracnose disease. J. Am. Soc. Hortic Sci. 134: 379-386.

Manikandan R, Raguchander T (2015) Pseudomonas fluorescens (Pf1) mediated chitinolytic activity in tomato plants against Fusarium oxysporum f. sp. lycopersici. Afr. J. Microbiol. Res. 9: 1331-1337.

Mauch F, Mauch-Mani B, Boller T (1988) Antifungal hydrolases in pea tissue II. Inhibition of fungal growth by combinations of chitinase and β-1, 3-glucanase. Plant Physiol. 88: 936-942.

Mehrabi R (2006). Signaling pathways involved in pathogenicity and development of the fungal wheat pathogen Mycosphaerella graminicola, Dissertation, University of Wageningen.

Niki T, Mitsuhara I, Seo S, Ohtsubo N, Ohashi Y (1998) Antagonistic effect of salicylic acid and jasmonic acid on the expression of pathogenesis-related (PR) protein genes in wounded mature tobacco leaves. Plant Cell Physiol. 39: 500-507.

Pieterse C M, van Loon LC (1999) Salicylic acid-independent plant defence pathways. Trends Plant Sci. 4: 52-58.

Ponomarenko A, Goodwin SB,  Kema GH (2011) Septoria tritici blotch (STB) of wheat. Plant Health Instructor Index. doi:10.1094/PHI-I-2011-0407-01

Punja ZK (2001) Genetic engineering of plants to enhance resistance to fungal pathogens-a review of progress and future prospects. Can. J. Plant Pathol. 23: 216-235.

Punja ZK, Zhang YY. (1993) Plant chitinases and their roles in resistance to fungal diseases. J. Nematol. 25: 526-540.

Saikia R, Singh BP, Kumar R, Arora DK (2005) Detection of pathogenesis related proteins-chitinase and-1, 3-glucanase in induced chickpea. Curr. Sci. 89: 659-663.

Sharma N, Sharma K, Gaur R,  Gupta V. (2011) Role of chitinase in plant defense. Asian J. Biochem. 6: 29-37.

Sharifi Tehrani A, Farzaneh M, Afshari F, Behboudi K, Kellenbergeer S, Pechy Tarr M, Keel CH, Mascher F (2011) A study of the effect of Pseudomonas fluorescens CHA0mcherry on the degree of root colonization in some wheat varieties and an induction of systemic resistance against leaf rust disease. Iran. J. Plant Protec. Sci. 42: 85-94.

Sharma N, Sharma KP, Guar RK, Gupa VK (2011) Role of Chitinase in Plant Defense Asian. J. Biochem. 6: 29-37

Simmons CR (1994) The physiology and molecular biology of plant 1, 3-β-D-glucanases and 1, 3; 1, 4-β-D-glucanases. Crit. Rev. Plant Sci. 13: 325-387.

Spoel SH, Koornneef A, Claessens SM, Korzelius JP, Van Pelt J A, Mueller MJ, Buchala A J, Métraux J-P, Brown R, Kazan K (2003) NPR1 modulates cross-talk between salicylate-and jasmonate-dependent defense pathways through a novel function in the cytosol. Plant Cell 15: 760-770.

Sun J, Peng M, Wang Y, Zhao P, Xia Q (2011) Isolation and characterization of antagonistic bacteria against Fusarium wilt and induction of defense related enzymes in banana. Afri. J. Microbiol. Res. 5: 509-515.

Van der Does D, Leon-Reyes A, Koornneef A, Van Verk MC, Rodenburg N, Pauwels L, Goossens A, Körbes AP, Memelink J, Ritsema T (2013) Salicylic acid suppresses jasmonic acid signaling downstream of SCFCOI1-JAZ by targeting GCC promoter motifs via transcription factor ORA59. Plant Cell 25:744-761.

Van Loon L, Van Strien E (1999) The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol. Molecu. Plant Pathol. 55: 85-97.

Yoshikawa M, Yamaoka N, Takeuchi Y (1993) Elicitors: their significance and primary modes of action in the induction of plant defense reactions. Plant Cell Physiol. 34: 1163-1173.