متابولومیکس مقایسه ای گیاهچه های دو لاین تقریباً آیزوژنیک مقاوم و حساس گندم نسبت به بیماری زنگ برگی گندم

نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 هیات علمی

2 بخش گیاهپزشگی، دانشکده کشاورزی،دانشگاه شیراز،شیراز، ایران

3 گروه بیوتکنولوژی دارویی، دانشکده داروسازی،دانشگاه علوم پزشکی شیراز، شیراز، ایران.

چکیده

شناسایی مسیرهای دفاعی و متابولیت‌های مرتبط با مقاومت به عنوان زیست‌نشانگر برای غربال‌گری یا تشخیص سطوح مقاومت ارقام گندم به زنگ برگی گندم (Puccinia triticina) ابزاری مفید جهت افزایش سرعت، دقت و کاهش هزینه غربالگری مقاومت است. استخراج پروفیل متابولیتی بخش قطبی لاین‌های تقریباً آیزوژنیک گندم حساس (Thatcher Lr22b) و مقاوم (Thatcher Lr25) به زنگ برگی گندم در مرحله دو برگچه‌ای 24 ساعت پس از مایه‌زنی با بیمارگر، پودر تالک و آب مقطر استریل شده در سه تکرار در مخلوط متانول-آب انجام شد. تجزیه واریانس برای انتخاب متابولیت های موثر نشان داد که فراوانی 26 متابولیت ‌بین فاکتورهای آزمایشی شامل لاین-ها، مایه‌زنی شده‌ها و برهمکنش لاین‌ها و مایه‌زنی شده‌ها اختلاف معنی‌داری داشتند. این متابولیت‌ها برای گروه‌بندی مشاهدات از نظر تشابهات و تفاوت‌های متابولیکی در فضای متابولوم بررسی شده تحت تجزیه ممیزی کانونی و تجزیه خوشه‌ای سلسله مراتب قرار گرفتند. نهایتاً 16 متابولیت مرتبط با مقاومت و 21 متابولیت مرتبط با بیماریزایی شناسایی شد که اکثر متابولیت‌های مرتبط با مقاومت هم در مقاومت القایی و هم در مقاومت ساختاری نقش داشتند که در مسیرهای سنتز دیواره سلولی، تولید ترکیبات فنولی و آلکالوئیدی، تجمع اسید آمینه‌ها، تولید پروتئین‌های مرتبط با مقاومت، به عنوان پیام رسان و فعالیت‌های ضد میکروبی در طول دفاع نقش داشتند. فعال‌تر شدن تعدادی از مسیرهای مهم در اثر حمله قارچ به گندم بحث می‌شود.

کلیدواژه‌ها


عنوان مقاله [English]

Comparative metabolomics of seedlings of two susceptible and resistant wheat near isogenic lines to wheat leaf rust disease

نویسندگان [English]

  • Habiballah Hamzehzarghani 1
  • Zahra Amjadi 2
  • Yunes Ghasemi 3
1 Faculty (Associate Professor)
2 Department of Plant Protection, School of Agriculture, Shiraz University, shiraz, Iran
3 Department of pharmaceutical biotechnology, Faculty of pharmacy, Shiraz University of medical sciences, Shiraz, Iran.
چکیده [English]

Identifying resistance related metabolites as biomarker for screening wheat germplasms resistant to leaf rust (Puccinia triticina) is a useful approach to increase the speed, accuracy and cost effectiveness of screening disease resistance. The relatively polar metabolites were extracted in a mixture of methanol-water from near isogenic line (NILs) of wheat susceptible (Thatcher Lr22b) or resistant (Thatcher Lr25) to leaf rust in the two leaf stage, 24 h after inoculation with the pathogen, talcum powder or sterilized distilled water, in three replications. Metabolites were analyzed using a GC/MS. ANOVA showed that abundance of 26 metabolites had significant differences among experimental factors including near isogenic lines (NILs), inoculation (I) and NILs ×I interaction. Abundance of 26 significant metabolites was subjected to canonical discriminant analysis and hierarchical cluster analysis to group observations of metabolite profiles based on similarities and differences in the investigated metabolic space. Finally 13 resistance related metabolites (RR) and 13 Pathogenesis related metabolites (PR) were identified that the majority of RR metabolites involved in induced resistance and constitutive resistance. Significant metabolites included metabolites with potential signaling and/or antimicrobial activity as well as molecules used as precursors for cell wall reinforcement, phenolic and alkaloids compounds, and accumulation of amino acids and proteins associated with resistance. Up-regulation of the number pathways upon infection of wheat to leaf rust fungus is discussed.
Key words: metabolic profiling, Wheat leaf rust, screening resistance, gas chromatography/mass spectrometry.

کلیدواژه‌ها [English]

  • Wheat leaf rust
  • screening resistance
  • gas chromatography/mass spectrometry
  • metabolic profiling
Abbasi M., Ershad D. and Hedjaroude G. A. 2005. Taxonomy of Puccinia recondita s. lat. causing brown rust on grasses. in Iran. Iranian Journal of Plant Pathology 41: 245-256.
Arner R. J., Prabhu K. S., Thompson J. T., Hildenbrandt G. R., Liken A. D. and Reddy C. C. 2001. Myo-Inositol oxygenase: molecular cloning and expression of a unique enzyme that oxidizes myo-inositol and D-chiro-inositol. Biochemical Journal 360(2): 313-320.
Beckles D. M., Roessner U. 2012. Plant metabolomics: Applications and opportunities for agricultural biotechnology. Inc: 67-81.
Benkeblia N., Shinano T. and Osaki M. 2007. Metabolite profiling and assessment of metabolome compartmentation of soybean leaves using non-aqueous fractionation and GC-MS analysis. Metabolomics 3: 297–305.
Bolton M. D., Kolmer J. A. and Garvin D. F. 2008a. Wheat leaf rust caused by Puccinia triticina. Molecular Plant Pathology 9: 563-575.
Bolton M. D., James A. K., Xu W. W. and Garvin, D. F. 2008b. Lr34-Mediated Leaf Rust Resistance in Wheat: Transcript Profiling Reveals a High Energetic Demand Supported by Transient Recruitment of Multiple Metabolic Pathways. Molecular Plant-Microbe Intraction 21: 1515–1526.
Chong J., Soufan O., Li C., Caraus I., Li, S.,  Bourque G.,  Wishart,D.S.,  Xia J. 2018. MetaboAnalyst 4.0: towards more transparent and integrative metabolomics analysis. Nuc. Acids Res, gky310: 1-9.
Cloutier S., McCallum B. D., Loutre C., Banks T. W., Wicker T., Feuillet C., Keller B. and Jordan M. C. 2007. Leaf rust resistance gene Lr1, isolated from bread wheat (Triticum aestivum L.) is a member of the large psr567 gene family. Plant Molecular Biology 65:93-106.
Driscoll C. J., Anderson L. M. 1967. Cytogenetic studies of Transec a wheat-rye translocation line. Canadian Journal of Genetics and Cytology 9: 375-380.
Driscoll C. J., Bielig L. M. 1968. Mapping of the Transec wheat-rye translocation. Canadian Journal of Genetics and Cytology 10: 421-425.
Driscoll C. J., Jensen N. F. 1964. Characteristics of leaf rust resistance transferred from rye to wheat. Crop Science 4: 372-374
Ershad D. 2009. Fungi of iran. 3th edition. Iranian Research Institute of Plant Protection. Tehran. Iran. 531pp.
Farmer E. E., Weber H. and Vollenweider S. 1998. Fatty acid signaling in Arabidopsis. Planta 206: 167-174.
Fiehn O., Kopka J., Trethewey R. N. and Willimitzer L. 2000. Identification of uncommon plant metabolites based on calculation of elemental compositions using gas chromatography and quadrupole mass spectrometry. Analytical Chemistry 72: 3573-3580.
Gogoi R., Singh D. V. and Srivastava K. D. 2001. Phenols as a biochemical basis of resistance in wheat against karnal bunt. Plant Pathology 50: 470-476.
Hamzehzarghani H., Kushalappa A. C., Dion Y., Rioux S., Comeau A., Yaylaya V., Marshall W. D. and Mather D. E. 2005. Metabolic profiling and factor analysis to discriminate quantitative resistance in wheat cultivars against fusarium head blight. Physiological and Molecular Plant Pathology 66: 119–133.
Hamzehzarghani H. 2007. Metabolic profiling and multivariate analysis to phenotype cultivars of wheat varying in resistance to fusarium head blight. Ph.D. thesis In Department of plant science McGill University Montreal, Quebec, Canada. pp 182.
Johnson D. E. 1998. Applied multivariate methods for data analysts. Duxbury Press, New York, 576 pp.
Lea P. J., Sodek L., Parry M. A. J., Shewry P. R. and Halford N. G. 2006. Asparagine in plant. Annals of Applied Biology 150: 1–26.
Li W. L., Faris J. D., Chittoor J. M., Leach J. E., Hulbert S. H., and Liu D. J. 1999. Genomic mapping of defense response genes in wheat. Theoretical and Applied Genetics 98: 226-33.
Loewus F. A., Kelly S. and Neufeld E. F. 1962. Metabolism of myo-inositol in plant: convertion to pectin, hemicelluloses, d-xylose, and sugar acids. Biochemistry 48: 421-425.
Lopez-Gresa M. P., Maltese F., Belles J. M., Conejero V., Kim H. K. and Verpoorte R. 2010. Metabolic response of tomato leaves upon different plant-pathogen intractions. Phytochemical Analysis 21: 89-94.
Mahdavi V., Ghanati, F., Ghassempour A. 2016. Integrated pathway-based and network-based analysis of GC-MS rice metabolomics data under diazinon stress to infer affected biological pathways. Anal. Biochem 494: 31-36.
Mert-T.rk F. 2006. Saponins versus plant fungal pathogens. Journal of Cell and Molecular Biology 5: 13-17.
Mitchell H. J., Ayliffe M. A., Rashid K. Y. and Pryor A. J. 2006. A rust-inducible gene from flax (fis1) is involved in proline catabolism. Planta 223: 213–222.
Morkunas, I., & Ratajczak, L. 2014. The role of sugar signaling in plant defense responses against fungal pathogens. Acta Physiologiae Plantarum 36(7):1607-1619
Pe´rez-Garcı´a A., Pereira S., Pissarra J., Garcı´a Gutie´rrez A., Cazorla F. M., Salema R., de Vicente A. and Ca´novas F.M. 1998. Cytosolic localization in tomato mesophyll cells of a novel glutamine synthetase induced in response to bacterial infection or phosphinothricin treatment. Planta 206: 426–434.
Roelfs A. P., Singh R. P. and Saari E. E. 1992. Rust Diseases of Wheat: Concepts and Methods of Disease Management. Mexico, DF: Cimmyt.
Roessner U., Wagner C., Kopka J., Trethewey R. N. and Willmitzer L. 2000 Simultaneous analysis of metabolites in potato tuber by gas chromatography-mass spectrometry. Plant Journal 23: 131–142.
Rozan P., Kuo Y. H. and Lambein F. 2001. Nonprotein amino acids in edible lentil and garden pea seedlings. Amino Acids 20: 319–324.
Scarpari L. M., Meinhardt L. W., Mazzafera P., Pomella A. W. V., Schiavinato M. A., Cascardo J. C. M., Pereira G. A. G. 2005. Biochemical changes during the development of witches’ broom: the most important disease of cocoa in Brazil caused by Crinipellis perniciosa. Journal of Experimental Botany 56: 865–877.
Sumner L. W., Mendes P. and Dixon R. A. 2003. Plant metabolomics: large-scale phytochemistry in the functional genomics era. Phytochemical 62: 817-836.
Yendo A. C. A. and Costa F. D. 2010. Production of Plant Bioactive Triterpenoid Saponins: Elicitation Strategies and Target Genes to Improve Yields. Molecular Biotechnology 46:94–104.
Szabados L. and Savoure A. 2009. Proline: a multifunctional amino acid. Cell 15: 89-97.
Tang Y. J., Martin H. G., Myers S., Rodriguez S., Baidoo E. E. and Keasling J. D. 2009. Advances in analysis of microbial metabolic fluxes via 13C isotopic labeling. Mass spectrometry reviews 28(2): 362-375.
Trouvelot S., Héloir M. C., Poinssot B., Gauthier A., Paris F., GUILLIER C. ... and Adrian M. 2014. Carbohydrates in plant immunity and plant protection: roles and potential application as foliar sprays. Frontiers in plant science 5, 592.
Zhu X., Tang T. and Galili G. 2000. The catabolic function of the a-aminoadipic acid pathway in plants is associated with unidirectional activity of lysine–oxoglutarate reductase, but not saccharopine dehydrogenase. Biochemical Journal 315: 215-220.