بررسی بیان افتراقی پروتئین های محافظ در گیاه آرابیدوپسیس تحت تنش شوری با روش iTRAQ-2DLC-MS/MS و آنالیز شبکه ژنی مربوطه

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری اصلاح نباتات، دانشگاه علوم کشاورزی و منابع طبیعی ساری

2 استادیار و عضو هیئت علمی دانشگاه علوم کشاورزی و منابع طبیعی ساری.

چکیده

شناسایی و کمیتسنجی پروتئینها و ژنهای بیان شده در شرایط تنش شوری میتواند منجر به درک هرچه بهتر مکانیسمهای پاسخی شود. بدین منظور در این تحقیق از روش پروتئومیکس مقایسهای iTRAQ جهت بررسی تغییرات پروتئینی در شرایط تنش شوری و کمیتسنجی آنها استفاده شد. سپس جهت بررسیهای بیشتر پیرامون عملکرد ژنهای شناسایی شده از اطلاعات بیان ژن و همبیانی در پایگاهداده و آنالیز توالی‌های مربوطه استفاده شد. با توجه به نتایج به‎دست آمده از iTRAQ-2DLC-MS/MS مجموعهای از پروتئینها از جمله P5CS1، KIN1، KIN2، ERD10، ERD14 و COR47 شناسایی شد. نتایج بیان ژنهای مربوطه نشان داد که بیان این ژنها تنها به تنش شوری محدود نمیشود و در پاسخ به سایر تنشهای اسمزی نیز دخالت دارند. شبکهی ژنی پروتئینهای شناسایی شده با نرمافزار String بررسی گردید. بر اساس نتایج به‎دست آمده در طی بیان این ژنها ترکیبات محافظ از جمله مواد محلول سازگار، پروتئین‌های دهایدرین و غیره در سلول تولید میشود که حضور آنها منجر به القای مکانیسمهای مقاومت و تحمل در گیاه در برابر تنشهای اسمزی از جمله شوری می شود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Investigation of differential protein expression of protective proteins under salinity stress in Arabidopsis by iTRAQ-2DLC-MS/MS and analysis of gene expression and related gene network

نویسندگان [English]

  • Mohammad Amin Baghery 1
  • Hamid Najafi Zarini 2
1 Ph.D Student of Plant Breeding, Sari Agriculture Sciences and Natural Resources University (SANRU), Sari, Iran
2 Assistant Professor of Plant Biotechnology, Sari Agriculture Sciences and Natural Resources University (SANRU), Sari, Iran.
چکیده [English]

Identification and quantification of proteins and genes that expressed in the salinity stress conditions could lead to better understanding of the response mechanisms. So, in this study iTRAQ comparative proteomic method was used to investigate the protein changes and quantification of them under salinity stress. Then, more studies about the function of identified genes were done using gene expression and co-expression data obtained from databases and analysis of related sequences. According to the results of iTRAQ-2DLC-MS/MS a set of proteins including P5CS1, KIN1, KIN2, ERD10, ERD14 and COR47 were identified. Results showed that expressions of the related genes were not just restricted to salinity stress. They were also involved in other osmotic stresses. Gene network based on identified proteins was evaluated by String software. Based on the results, during the expression of these genes, protective compounds such as compatible solutes, dehydrin proteins and etc. were produced in the cells. Presence of the compounds led to induction of resistance and tolerance mechanisms in plants against osmotic stress such as salinity.

کلیدواژه‌ها [English]

  • Arabidopsis
  • iTRAQ Comparative proteomics
  • Protective compounds
  • Gene network
  • Salinity

مراجع

Abrahám E, Rigó G, Székely G, Nagy R, Koncz C, Szabados L (2003) Light-dependent induction of proline biosynthesis by abscisic acid and salt stress is inhibited by brassinosteroid in Arabidopsis. Plant Mol. Biol. 51(3): 363-372.
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 55: 373-399.
Ashraf M, Foolad M (2007) Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ. Exp. Bot. 59(2): 206-216.
Bray EA (1993) Molecular responses to water deficit. Plant Physiol. 103(4): 1035-1040.
Brini F, Hanin M, Lumbreras V, Amara I, Khoudi H, Hassairi A, Pages M, Masmoudi K (2007) Overexpression of wheat dehydrin DHN-5 enhances tolerance to salt and osmotic stress in Arabidopsis thaliana. Plant Cell Rep. 26(11): 2017-2026.
Chong PK, Gan CS, Pham TK, Wright PC (2006) Isobaric tags for relative and absolute quantitation (iTRAQ) reproducibility: implication of multiple injections. J. Proteome Res. 5(5): 1232-1240.
Dure Iii L, Crouch M, Harada J, Ho T-HD, Mundy J, Quatrano R, Thomas T, Sung Z (1989) Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol. Biol. 12(5): 475-486.
Fabro G, Kovács I, Pavet V, Szabados L, Alvarez ME (2004) Proline accumulation and atp5cs2 gene activation are induced by plant-pathogen incompatible interactions in Arabidopsis. Mol. Plant Microbe Interact. 17(4): 343-350.
Hanin M, Brini F, Ebel C, Toda Y, Takeda S, Masmoudi K (2011) Plant dehydrins and stress tolerance: versatile proteins for complex mechanisms. Plant Signal Behav. 6(10): 1503-1509.
Hare P, Cress W, Van Staden J (2002) Disruptive effects of exogenous proline on chloroplast and mitochondrial ultrastructure in Arabidopsis leaves. S. Afr. J. Bot. 68(3): 393-396.
Heuer B (2003) Influence of exogenous application of proline and glycinebetaine on growth of salt-stressed tomato plants. Plant Sci. 165(4): 693-699.
Hoque MA, Okuma E, Banu MNA, Nakamura Y, Shimoishi Y, Murata Y (2007) Exogenous proline mitigates the detrimental effects of salt stress more than exogenous betaine by increasing antioxidant enzyme activities. J. Plant Physiol. 164(5): 553-561.
Hsieh EJ, Cheng MC, Lin TP (2013) Functional characterization of an abiotic stress-inducible transcription factor aterf53 in Arabidopsis thaliana. Plant Mol. Biol. 82(3): 223-237.
Hu C, Delauney AJ, Verma D (1992) A bifunctional enzyme (Delta 1-Pyrroline-5-Carboxylate Synthetase) catalyzes the first two steps in proline biosynthesis in plants. Proc. Natl. Acad. Sci. U.S.A. 89(19): 9354-9358.
Ingram J, Bartels D (1996) The molecular basis of dehydration tolerance in plants. Annu. Rev. Plant Biol. 47(1): 377-403.
Jain M, Mathur G, Koul S, Sarin N (2001) Ameliorative effects of proline on salt stress-induced lipid peroxidation in cell lines of groundnut (Arachis hypogaea L.). Plant Cell Rep. 20(5): 463-468.
Jung Y, Park J, Choi Y, Yang JG, Kim D, Kim BG, Roh K, Lee DH, Auh CK, Lee S (2010) Expression analysis of proline metabolism‐related genes from halophyte Arabis stelleri under osmotic stress conditions. J. Integr. Plant Biol. 52(10): 891-903.
Kaya C, Tuna AL, Ashraf M, Altunlu H (2007) Improved salt tolerance of melon (Cucumis melo L.) by the addition of proline and potassium nitrate. Environ. Exp. Bot. 60(3): 397-403.
Khedr AH, Abbas M, Wahid AA, Quick WP, Abogadallah GM (2003) Proline induces the expression of salt‐stress‐responsive proteins and may improve the adaptation of Pancratium maritimum L. to salt‐stress. J. Exp. Bot. 54(392): 2553-2562.
Kim K, Jang YJ, Lee SM, Oh BT, Chae JC, Lee KJ (2014) Alleviation of salt stress by enterobacter sp. EJ01 in tomato and Arabidopsis is accompanied by up-regulation of conserved salinity responsive factors in plants. Mol. Cells 37(2): 109-117.
Knight H, Veale EL, Warren GJ, Knight MR (1999) The sfr6 mutation in Arabidopsis suppresses low-temperature induction of genes dependent on the CRT/DRE sequence motif. Plant Cell 11(5): 875-886.
Konstantinova T, Parvanova D, Atanassov A, Djilianov D (2002) Freezing tolerant tobacco, transformed to accumulate osmoprotectants. Plant Sci. 163(1): 157-164.
Larsson C, Widell S, Kjellbom P (1987) Preparation of high-purity plasma-membranes. Meth. Enzymol. 148: 558-568.
Li J, Han Y, Zhao Q, Li C, Xie Q, Chong K, Xu Y (2013) The E3 ligase AtRDUF1 positively regulates salt stress responses in Arabidopsis thaliana. PLoS ONE 8(8): e71078.
Lin CC, Kao CH (2001) Cell wall peroxidase against ferulic acid, lignin, and NaCl-reduced root growth of rice seedlings. J. Plant Physiol. 158(5): 667-671.
Maggio A, Miyazaki S, Veronese P, Fujita T, Ibeas JI, Damsz B, Narasimhan ML, Hasegawa PM, Joly RJ, Bressan RA (2002) Does proline accumulation play an active role in stress‐induced growth reduction? Plant J. 31(6): 699-712.
Mani S, Van De Cotte B, Van Montagu M, Verbruggen N (2002) Altered levels of proline dehydrogenase cause hypersensitivity to proline and its analogs in Arabidopsis. Plant Physiol. 128(1): 73-83.
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu. Rev. Plant Biol. 59: 651-681.
Nakashima K, Satoh R, Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K (1998) A gene encoding proline dehydrogenase is not only induced by proline and hypoosmolarity, but is also developmentally regulated in the reproductive organs of Arabidopsis. Plant Physiol. 118(4): 1233-1241.
Nanjo T, Fujita M, Seki M, Kato T, Tabata S, Shinozaki K (2003) Toxicity of free proline revealed in an Arabidopsis t-DNA-tagged mutant deficient in proline dehydrogenase. Plant Cell Physiol. 44(5): 541-548.
Parre E, Ghars MA, Leprince AS, Thiery L, Lefebvre D, Bordenave M, Richard L, Mazars C, Abdelly C, Savouré A (2007) Calcium signaling via phospholipase C is essential for proline accumulation upon ionic but not nonionic hyperosmotic stresses in Arabidopsis. Plant Physiol. 144(1): 503-512.
Roosens NH, Thu TT, Iskandar HM, Jacobs M (1998) Isolation of the ornithine-δ-aminotransferase cDNA and effect of salt stress on its expression in Arabidopsis thaliana. Plant Physiol. 117(1): 263-271.
Roychoudhury A, Roy C, Sengupta DN (2007) Transgenic tobacco plants overexpressing the heterologous lea gene Rab16A from rice during high salt and water deficit display enhanced tolerance to salinity stress. Plant Cell Rep. 26(10): 1839-1859.
Savouré A, Hua XJ, Bertauche N, Van Montagu M, Verbruggen N (1997) Abscisic acid-independent and abscisic acid-dependent regulation of proline biosynthesis following cold and osmotic stresses in Arabidopsis thaliana. Mol. Gen. Genet. 254(1): 104-109.
Savouré A, Jaoua S, Hua XJ, Ardiles W, Van Montagu M, Verbruggen N (1995) Isolation, characterization, and chromosomal location of a gene encoding the δ 1-pyrroline-5-carboxylate synthetase in Arabidopsis thaliana. FEBS Lett. 372(1): 13-19.
Skopelitis DS, Paranychianakis NV, Paschalidis KA, Pliakonis ED, Delis ID, Yakoumakis DI, Kouvarakis A, Papadakis AK, Stephanou EG, Roubelakis-Angelakis KA (2006) Abiotic stress generates ROS that signal expression of anionic glutamate dehydrogenases to form glutamate for proline synthesis in tobacco and grapevine. Plant Cell 18(10): 2767-2781.
Strizhov N, Abraham E, Ökrész L, Blickling S, Zilberstein A, Schell J, Koncz C, Szabados L (1997) Differential expression of two P5CS genes controlling proline accumulation during salt-stress requires ABA and is regulated by ABA1, ABI1 and AXR2 in Arabidopsis. Plant J. 12(3): 557-569.
Székely G, Ábrahám E, Cséplő Á, Rigó G, Zsigmond L, Csiszár J, Ayaydin F, Strizhov N, Jásik J, Schmelzer E (2008) Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. Plant J. 53(1): 11-28.
Trovato M, Mattioli R, Costantino P (2008) Multiple roles of proline in plant stress tolerance and development. Rendiconti Lincei 19(4): 325-346.
Verbruggen N, Villarroel R, Van Montagu M (1993) Osmoregulation of a pyrroline-5-carboxylate reductase gene in Arabidopsis thaliana. Plant Physiol. 103(3): 771-781.
Wang H, Datla R, Georges F, Loewen M, Cutler AJ (1995) Promoters from kin1 and cor6.6, two homologous Arabidopsis thaliana genes: transcriptional regulation and gene expression induced by low temperature, ABA, osmoticum and dehydration. Plant Mol. Biol. 28(4): 605-617.
Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. Plant Cell 6(2): 251-264.
Yeo A, Caporn S, Flowers T (1985) The effect of salinity upon photosynthesis in rice (Oryza sativa L.): gas exchange by individual leaves in relation to their salt content. J. Exp. Bot. 36(8): 1240-1248.
Yoshiba Y, Kiyosue T, Katagiri T, Ueda H, Mizoguchi T, Yamaguchi‐Shinozaki K, Wada K, Harada Y, Shinozaki K (1995) Correlation between the induction of a gene for δ1‐pyrroline‐5‐carboxylate synthetase and the accumulation of proline in Arabidopsis thaliana under osmotic stress. Plant J. 7(5): 751-760.
Yoshiba Y, Kiyosue T, Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K (1997) Regulation of levels of proline as an osmolyte in plants under water stress. Plant Cell Physiol. 38(10): 1095-1102.
Zhu JK (2001) Plant salt tolerance. Trends Plant Sci. 6(2): 66-71.

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