الگوی پروتئینی برگ ارقام متحمل و حساس کلزا تحت تنش خشکی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه بهنژادی و بیوتکنولوژی گیاهی، دانشکده کشاورزی، دانشگاه تبریز،

2 دانشگاه تبریز دانشکده کشاورزی گروه بیوتکنولوژی

چکیده

هدف: تنش خشکی یکی از عمده‌ترین محدودیت‌ها در تولید محصولات زراعی است که اثر نامطلوبی بر کمیت و کیفیت آن‌ها دارد. کلزا همانند بسیاری از گیاهان زراعی از تنش ناشی از کمبود آب متاثر می‌شود. در سطح سلولی گیاهان از طریق سنتز پروتئین‌های خاص به تنش خشکی پاسخ می‌دهند. از این رو پژوهشی با هدف مطالعه مکانیسم پاسخ کلزا به تنش خشکی و تعیین پروتئین‌های دخیل در تحمل تنش انجام شد.
مواد و روش‌ها: الگوی پروتئینی برگ ارقام Hyola308 و  Sarigol بترتیب به عنوان ارقام حساس و متحمل با استفاده از تکنیک پروتئومیک فاقد ژل/فاقد (شاتگان پروتئومیکس) برچسب تحت سطوح مختلف تنش خشکی (6/. و 2/1 مگا پاسکال (MPa) به همراه شاهد) مورد بررسی قرار گرفت. جهت اعتبارسنجی تغییر محتوای پروتئین‌های شناسایی شده از وسترن‌بلات استفاده شد.
نتایج: در کل 56 پروتئین شامل 16 پروتئین به صورت اختصاصی برای Hyola308، 16 پروتئین به صورت اختصاصی برای Sarigol و 12 پروتئین مشترک بین هر دو شناسایی شد. در Sarigol فراوانی اکثر پروتئین‌های درگیر در متابولیسم پروتئین، فتوسنتر و انرژی در مواجهه با سطوح مختلف تنش خشکی کاهش نشان داد، در مقابل در Hyola308 افزایش در فراوانی پروتئین‌های درگیر در متابولیسم انرژی، فتوسنتز و دفاع آنتی‌اکسیدان به صورت اختصاصی مشاهده شد.
نتیجه‌گیری: استنباط می‌شود که افزایش فراوانی این پروتئین‌ها در برگ‌های Hyola308 قسمتی از مکانیسم تحمل این رقم در مواجهه با تنش می‌باشد و تقلیل کارایی چرخه کلوین و کاهش تولید قند و انرژی در Sarigol، کاهش رشد این رقم را تحت تنش خشکی می‌تواند توجیه کند.

کلیدواژه‌ها


عنوان مقاله [English]

Leaf protein pattern of tolerant and susceptible canola cultivars under drought stress

نویسندگان [English]

  • Bita Kazemi Oskuei 1
  • Ali Bandehagh 2
1 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
2 Breeding for Abiotic Stresses Lab., Dept. of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz 5166616471, Iran.
چکیده [English]

Objective
Drought stress is the main restriction factor in crop production that has an adverse effect on crop quantity and quality. Canola, like many crops, is affected by stress due to water deficit. At the cellular level, plants respond to drought stress by synthesizing specific proteins. Therefore, a research with the aim of studying the response mechanism of canola to drought stress and determining proteins involved in mediating stress tolerance was carried out.
 
Materials and methods
In order to comprehend a mechanism of canola plant response to drought stress, the protein profiles of the drought-tolerant Hyola308 and drought-sensitive Sarigol leaf under different drought stress conditions based on a gel-free/label-free proteomic technique were investigated. To validate the content variation of proteins identified in the proteomic analysis, Western blot analysis was used.
 
Results
A total of 56 proteins were identified in Sarigol and Hyola308, 16 proteins were specific to Hyola308 and 16 proteins were specific to Sarigol, respectively. Of the identified proteins, 12 proteins were commonly detected between Sarigol and Hyola308. In Sarigol under different drought stress conditions, the abundance of proteins related to protein metabolism, photosynthesis and energy metabolism decreased; whereas, in Hyola308, an enhancement in proteins abundance involved in photosynthesis, energy metabolism and antioxidant defense was observed.
 
Conclusions
It is inferred that enhancement of these protein abundance in Hyola308 leaf may be a part of tolerance mechanism of this cultivar exposed to stress and decrease in the Kelvin cycle efficiency and production of sugar and energy in Sarigol may justify growth reduction of this cultivar.

کلیدواژه‌ها [English]

  • Abiotic stress
  • canola
  • Immuno-blot
  • proteomic
Adam Z, Rudella A, van Wijk KJ (2006) Recent advances in the study of Clp, FtsH and other proteases located in chloroplasts. Curr Opin Plant Biol 9, 234-240.
Alvim FC, Carolino SMB, Cascardo JCM et al. (2001) Enhanced accumulation of BiP in transgenic plants confers tolerance to water stress. Plant Physiol 126, 10420-11054.
Anderson JV, Davis DG (2004) Abiotic stress alters transcript profiles and activity of glutathione S-transferase, glutathione peroxidase, and glutathione reductase in Euphorbia Esula. Physiol Plant 120, 421-433.
Blum A (1989) Breeding methodes for drought resistance. In: Plant Under Stress. Jones HG, Flowers TJ, Jones MB (eds). Combridge University Press, UK, pp. 195-215.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248-254.
Bybordi A, Tabatabaei SJ, Ahmade A (2010) Effect of drought on the growth and peroxidase and IAA oxidase activities in canola. J Food Agri Environ 8, 109-112.
Cazale AC, Clement M, Chiarenza S et al. (2009) Altered expression of cytosolic/nuclear HSC70-1 molecular chaperone affects development and abiotic stress tolerance in Arabidopsis thaliana. J Exp Bot 60, 2653-2664.
Caruso G, Cavaliere C, Foglia P et al. (2009) Analysis of drought responsive proteins in wheat (Triticum durum) by 2D-PAGE and MALDI-TOF mass spectrometry. Plant Sci 177, 570-576.
Castillejo MA, Maldonado AM, Ogueta S, Jorrín JV (2008) Proteomic analysis of responses to drought stress in sunflower (Helianthus annuus) leaves by 2DE gel electrophoresis and mass spectrometry. Open Proteomics J 1, 59-71. 
Chugh A, Khurana P (2002) Gene expression during drought stress recent advances. Curr Sci 83, 6-25.
Degenkolbe T, Do PT, Kopka J et al. (2013) Identification of drought tolerance markers in a diverse population of rice cultivars by expression and metabolite profiling. PLoS One 8, 63637.
De la Torre-González A, Navarro-León E, Blasco B, Ruiz JM (2020) Nitrogen and photorespiration pathways, salt stress genotypic tolerance effects in tomato plants (Solanum lycopersicum L.). Acta Physiol Plant 42, 2.
Ding N, Wang A, Zhang X et al. (2017) Identification and analysis of glutathione S-transferase gene family in sweet potato reveal divergent GST-mediated networks in aboveground and underground tissues in response to abiotic stresses. BMC Plant Biol 17, 225.
Dolatabadi N, Toorchi M, Valizadeh M, Bandehagh A (2018) The proteomic analysis of leaf in Rapeseed (Brassica napus L) under salt stress. J Agric Biotech 9, 51-64.
Dubey H, Grover A (2001) Current initiatives in proteomics research: The plant perspective. Curr Sci 80, 262-269.
Ford KL, Cassin A, Bacic A (2011) Quantitative proteomic analysis of wheat cultivars with differing drought stress tolerance. Front Plant Sci 2, 44.
Ghaffari M, Toorchi M, Valizadeh M, Komatsu S (2013) Differential response of root proteome to drought stress in drought sensitive and tolerant sunflower inbred lines. Funct Plant Biol 40, 609-617. 
Ghaffari A, Gharechahi J, Nakhoda B, Salekdeh GH (2014) Physiology and proteome responses of two contrasting rice mutants and their wild type parent under salt stress conditions at the vegetative stage. J Plant Physiol 171, 31-44.
Gong P, Zhang J, Li H et al. (2010) Transcriptional profiles of drought-responsive genes in modulating transcription signal transduction, and biochemical pathways in tomato. J Exp Bot 61, 3563-3575.
Gupta SC, Sharma A, Mishra M et al. (2010) Heat shock proteins in toxicology:How close and How far? Life Sci 86, 377-384.
Hashiguchi A, Ahsan N, Komatsu S (2010) Proteomics application of crops in the context of climatic changes. Food Res Int 43, 1803-1813.
Hosseini Salkdeh GH, Nasrabadi D (2011) Proteomic analysis of root and leaf in rice under salinity stress. J Crop Biotechnol 1, 1-11. 
Ingle RA, Schmidt UG, Farrant JM et al. (2007) Proteomic analysis of leaf proteins during dehydration of the resurrection plant Xerophyta viscosa. Plant Cell Environ 30, 435-446. 
Irar S, Brini F, Goday A et al (2010) Proteomic analysis of wheat embryos with 2-DE and liquid-phase chromatography (ProteomeLab PF-2D)--a wider perspective of the proteome. J Proteomics 73, 1707-1721. 
Janska H, Piechota J, Kwasniak M (2010) ATP-dependent proteases in biogenesis and maintenance of plant mitochondria. Biochim Biophys Acta 1797, 1071-1075.
Juliann G, Kiang G, Tsokos C (1998) Heat shock protein 70 Kda: Molecular Biology, Biochemistry, and Physiology. Pharmacol Ther 80, 183-201.
Jungkunz I, Link K, Vogel F et al. (2011) AtHSP70-15-deficient Arabidopsis plants are characterized by reduced growth, a constitutive cytosolic protein response and enhanced resistance to TuMV. Plant J 66, 983-95.
Kakaei M, Zabarjadi, A Mostafaei A (2010) Comparison of genetic and morpho-physiological distance via SDS-PAGE marker in some rapeseed genotypes. J Agric Biotech 1, 79-94.
Kappachery S, Baniekal Hiremath G, Yu JW, Park SW (2015) Effect of over-and under-expression of glyceraldehyde3-phosphate dehydrogenase on tolerance of plants to water-deficit stress. Plant Cell Tissue Organ Cult 121, 97-107.
Karlin S, Brocchieri L (1998) Heat shock protein 70 family:multiple sequence comparisons, function, and evolution. J Mol Evol 47, 565-577.
Kato Y, Sakamoto W (2009) Protein quality control in chloroplasts: a current model of D1 protein degradation in the photosystem II repair cycle. J Biochem 146, 463-469.
Kausar R, Arshad M, Shahzad A, Komatsu S (2013) Proteomics analysis of sensitive and tolerant barley genotypes under drought stress. J Amino Acids 44, 345-359.
Kazemi Oskuei B, Bandehagh A, Sarikhani MR, Komatsu S (2018) Protein profiles underlying the effect of plant growth-promoting rhizobacteria on canola under osmotic stress. J Plant Growth Regul 37, 560-574.
Ke Y, Han G, He H, Li J (2009) Differential regulation of proteins and phosphoproteins in rice under drought stress. Biochem Biophys Res Commun 379, 133-138.
Khalili M, Naghavi MR (2016) Evaluation of responsive common proteins in tolerant and sensitive spring wheat cultivars under drought stress. J Crop Biotechnol 16, 31-44.
Khalili M, Naghavi M (2017a) Evaluation of spring canola cultivars in terms of some morphological and physiological traits under drought stress and proteome analysis of the most tolerant and susceptible ones. J Agric Biotech, 9, 59-82.
Khalili M, Naghavi MR (2017b) Assessment of changes in protein expression pattern in tolerant and sensitive cultivars of rapeseed under salt stress. Iranian J Field Crop Sci 48, 721-735.
Kim SR, An G (2013) Rice chloroplast-localized heat shock protein 70, OsHsp70CP1, is essential for chloroplast development under high-temperature conditions. J Plant Physiol 170, 854-863.
Kissoudis C, Kalloniati C, Flemetakis E et al. (2015) Stress-inducible GmGSTU4 shapes transgenic tobacco plants metabolome towards increased salinity tolerance. Acta Physiol Plant 37, 128. 
Klein WL, Luo Y (2010) Heat shock treatment reduces beta amyloid toxicity in vivo by diminishing oligomers. Neurobiol Aging 31, 1055-1058.
Komatsu S, Han C, Nanjo Y et al. (2013) Label-free quantitative proteomic analysis of abscisic acid effect in early-stage soybean under flooding. J Proteome Res 12, 4769-4784.
Kozuleva M, Goss T, Twachtmann M et al. (2016) Ferredoxin:NADP(H) Oxidoreductase abundance and location influences redox poise and stress tolerance. Plant physiol 172, 1480-1493. 
Krugman T, Chagué V, Peleg Z et al. (2010) Multilevel regulation and signalling processes associated with adaptation to terminal drought in wild emmer wheat. Funct Integr Genomics 10, 167-186.
Kumar S, Trivedi PK (2018) Glutathione S-Transferases: role in combating abiotic stresses including arsenic detoxification in plants. Front Plant Sci 9, 751. 
Lee JH, Schoeffl F (1996) An Hsp70 antisense gene affects the expression of HSP70/HSC70, the regulation of HSF, and the acquisition of thermotolerance in transgenic Arabidopsis thaliana. Mol Gen Genet 252, 11-19.
Lintala M, Lehtimäki N, Benz JP et al. (2012) Depletion of leaf-type ferredoxin-NADP(+) oxidoreductase results in the permanent induction of photoprotective mechanisms in Arabidopsis chloroplasts. Plant J 70, 809-817.
Michel BE, Kaufmann MR (1973) The osmotic potential of polyethylene glycol 6000. Plant Physiol 51, 914-916.
Mohammadi PP, Moieni A, Komatsu S (2012) Comparative proteome analysis of drought-sensitive and drought-tolerant rapeseed roots and their hybrid F1 line under drought stress. J Amino Acids 43, 2137-2152.
Muller B, Pantin F, Génard M et al. (2011) Water deficits uncouple growth from photosynthesis, increase C content, and modify the relationships between C and growth in sink organs. J Exp Biol 62, 1715-1729.
Mundree SG, Farrant JM (2000) Some physiological and molecular insights into the mechanisms of desiccation tolerance in the resurrection plant Xerophyta viscosa Baker. In: Plant tolerance to abiotic stresses in agriculture: role of geneticengineering. Cherry JH, Locy RD, Rychter A (eds). KluwerAcademic Publishers, Dordrecht. pp. 201-222.
Nanjo Y, Skultety L, Uváčková L et al. (2012) Mass spectrometry-based analysis of proteomic changes in the root tips of flooded soybean seedlings. J Proteome Res 11, 372-385.
Palatnik JF, Tognetti VB, Poli HO et al. (2003) Transgenic tobacco plants expressing antisense ferredoxin-NADP(H) reductase transcripts display increased susceptibility to photo-oxidative damage. Plant J 35, 332-341.
Pawar VV, Lokhande PK (2015) Effect of osmotic stress on osmolyte accumulation and ammonia assimilating enzymes in chickpea cultivars. Indian J Plant Physiol 20, 276-280.
Penrose DM, Glick BR (2003) Methods for isolating and characterizing ACC deaminase containing plant growth promoting rhizobacteria. J Plant Physiol 118, 10-18.
Plucken H, Müller B, Grohmann D et al. (2002) The HCF136 protein is essential for assembly of the photosystem II reaction center in Arabidopsis thaliana. FEBS Lett 532, 85-90.
Rodriguez RE, Lodeyro A, Poli HO et al. (2007) Transgenic tobacco plants overexpressing chloroplastic ferredoxinNADP(H) reductase display normal rates of photosynthesis and increased tolerance to oxidative stress. Plant Physiol 143, 639-649.
Sakamoto W (2006) Protein degradation machineries in plastids. Annu Rev Plant Biol 57, 599-621.
Schroda M, Vallon O, Wollman FA, Beck CF (1999) A chloroplast-targeted heat shock protein 70 (HSP70) contributes to the photoprotection and repair of photosystem II during and after photoinhibition. Plant Cell 11, 1165-1178.
Singh NB, Singh D, Singh A (2015) Biological seed proming mitigates the effects of water stress in sunflower seedlings. Physiol Mol Biol Plants 21, 207-214.
Su PH, Li HM (2008) Arabidopsis stromal 70-kD heat shock proteins are essential for plant development and important for thermotolerance of germinating seeds. Plant Physiol 146, 1231-1241.
Sung DY, Guy CL (2003) Physiological and molecular assessment of altered expression of Hsc70‐1 in Arabidopsis. Evidence for pleiotropic consequences. Plant Physiol 132, 979-987. 
Takahashi S, Murata N (2008) How do environmental stresses accelerate photo inhibition? Trends Plant Sci 13, 178-182.
Tamoi M, Miyazaki T, Fukamizo T, Shigeoka S (2005) The Calvin cycle in cyanobacteria is regulated by CP12 via the NAD(H)/NADP(H) ratio under light/dark conditions. Plant J 42, 504-513.
Tran DQ, Konishi A, Morokuma M et al. (2020) NaCl-stimulated ATP synthesis in mitochondria of a halophyte Mesembryanthemum crystallinum L. Plant Prod Sci 23, 129-135.
Usadel B, Obayashi T, Mutwil M et al. (2009) Co-expression tools for plant biology: opportunities for hypothesis generation and caveats. Plant Cell Environ 32, 1633-1651.
Vincent D, Lapierre C, Pollet B et al. (2005) Water deficits affect caffeate O-methyltransferase, lignification, and related enzymes in maize leaves. A proteomic investigation. Plant Physiol 137, 949-960.
Von Ballmoos C, Dimroth P (2007) Two distinct proton binding sites in the ATP synthase family. Biochemistry 46, 11800-11809.
Wang W, Vinocur B, Shoseyov O, Altman A (2004) Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 9, 244-252. 
Wassmann R, Jagadish SVK, Sumfleth K et al. (2009) Regional vulnerability of climate change impacts on Asian rice production and scope for adaptation. Adv Agron 3, 91-133.
Xiong X, Chang L, Khalid M et al. (2018) Alleviation of Drought Stress by Nitrogen Application in Brassica campestris ssp. Chinensis L. Agronomy 8, 66.
Xu Ch, Huang B (2010) Comparative analysis of drought responsive proteins in Kentucky bluegrass cultivars contrasting in drought tolerance. Crop Sci 50, 2543-2552.
Xu C, Sibicky T, Huang B (2010) Protein profile analysis of salt responsive proteins in leaves and roots in two cultivars of creeping bent grass differing in salinity tolerance. Plant Cell Rep 29, 595-615.
Yang Y, Han C, Liu Q et al. (2008) Effect of drought and low light on growth and enzymatic antioxidant system of Picea asperata seedlings. Acta Physiol Plant 30, 433-440.
Ye J, Wang S, Zhang F et al. (2013) Proteomic analysis of leaves of different wheat genotypes subjected to PEG6000 stress and rewatering. Plant Omics 6, 286-294.
Yin X, Komatsu S (2016) Nuclear proteomics reveals the role of protein synthesis and chromatin structure in root tip of soybean during the initial stage of flooding stress. J Proteome Res 15, 2283-2298.
Yu A1, Li P, Tang T et al. (2015) Roles of Hsp70s in stress responses of microorganisms, plants, and animals. Biomed Res Int 2015, 510319.
Yue G, Hu X, He Y et al. (2010) Identification and characterization of two members of the FtsH gene family in maize (Zea mays L.). Mol Biol Rep 37, 855-863.
Zaltsman A, Ori N, Adam Z (2005) Two types of FtsH protease subunits are required for chloroplast biogenesis and photosystem II repair in Arabidopsis. Plant Cell 17, 2782-2790.
Zhang H, Whitelegge JP, Cramer WA (2001) Ferredoxin:NADP+ oxidoreductase is a subunit of the chloroplast cytochrome b6f complex. J Biol Chem 276, 38159-38165.