با همکاری مشترک دانشگاه پیام نور و انجمن بیوتکنولوژی جمهوری اسلامی ایران

نوع مقاله : علمی پژوهشی

نویسندگان

1 کارشناس‌ارشد اصلاح‌نباتات، پژوهشکده ژنتیک و زیست‌فناوری کشاورزی طبرستان، دانشگاه علوم کشاورزی و منابع طبیعی ساری.

2 استاد پژوهشکده ژنتیک و زیست‌فناوری کشاورزی طبرستان، دانشگاه علوم کشاورزی و منابع طبیعی ساری.

3 استادیار گروه بیوتکنولوژی کشاورزی، دانشکده مهندسی انرژی و فناوری‌های نوین، دانشگاه شهید بهشتی، تهران.

چکیده

شوری خاک با ایجاد تنش اسمزی و بر هم زدن تعادل یونی تولیدات گیاهی را محدود می­کند و منجر به خسارت در سطح مولکولی و نهایتاً مرگ سلول می­گردد. در این تحقیق آنالیز بیان ژن مبتنی بر تکنیک cDNA-AFLP جهت مقایسه پروفایل بیانی ناشی از تنش شوری KCl در سه سطح 0 (شاهد)، 200 و 400 میلی مولار در گیاه آلوروپوس لیتورالیس که نزدیک­ترین خانواده به غلات می­باشد، انجام شد. از میان 34 عدد EST  جداسازی شده، 27 عدد EST با طول میانگین 280 جفت باز بدست آمد. در این میان 80% از رونوشت­ها با پروتئین­ها و توالی­های اسید نوکلئیک شناسایی شده در ارگانیسم­های دیگر، شباهت نشان دادند. از طرفی دیگر، 6 رونوشت به عنوان ژن­های جدید در نظر گرفته شدند. در نهایت 27 عدد EST  در بانک ژن به ثبت رسید که مهمترین آنها شامل گروه­های پروتئینی سینتاکسین، آدنوزیل متیونین دکربوکسیلاز و پروتئین­های ریبوزومی می‌باشند. گروه­بندی EST­ها بر مبنای نوع پاسخ به تنش­ها، آنالیز عملکردی و خصوصیت یابی ژن­های پاسخگو در ریشه گیاه آلوروپوس لیتورالیس به تنش­ مذکور را در مطالعات آتی روی این گیاه که از اعضای خانواده غلات می‌باشد، تسهیل خواهد کرد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification and Isolation of Transcripts in Response to KCl from Aeluropus littoralis

نویسندگان [English]

  • Seyyedeh farzaneh Fatemi ardestani 1
  • Ghorbanali Nematzadeh 2
  • Hossein Askari 3
  • Hamidreza Hashemi 1

1 M.Sc. of Plant Breeding, Genetic and Agricultural Biotechnology Institute of Tabarestan, University of Agriculture and Natural Resources of Sari, Sari, Iran,

2 Professor of Biotechnology, Genetic and Agricultural Biotechnology Institute of Tabarestan, University of Agriculture and Natural Resources of Sari, Sari, Iran,

3 Assistant Professor, Department of Biotechnology, Faculty of New Technologies and Energy Engineering, Shahid Beheshti University, Tehran, Iran

چکیده [English]

Soil salinity limits crop production by creating osmotic stress and disruption of ion homeostasis, leads to damage at the molecular level and finally cell death. In this study, gene expression analysis based on cDNA-AFLP technique was used to compare the expression profiles of KCl stress at three levels: 0 (control), 200mM and 400mM, in Aeluropus littoralis which is the closest family to cereal. Among 34 isolated ESTs, 27 ESTs were obtained with the average length of 280 bp. The nucleotide sequences were compared with those in the GenBank database. Approximately 80% of the ESTs show homology to nucleotide or amino acid sequences in the GenBank database and 6 ESTs show no significant similarity in the GenBank database which considered as novel genes. Finally, 27 ESTs were recorded in NCBI database which are included potassium transporter, ribosomal protein, NADH dehydrogenase and golgin. The result of this research is very important to understand molecular basis and resistance mechanisms of drought stress for breeding and genetic engineering to improve crop resistance against stress and the production of resistant plants. EST classification based on responses to stress, will facilitate performance analysis, characterization of responsive genes in plant roots of Aeluropus littoralis to stress in future studies on this herb that is a member of the Poaceae family.

کلیدواژه‌ها [English]

  • Aeluropus littoralis
  • cDNA-AFLP
  • Gene expression
  • Salinity
Akhani H, Ghorbanli M (1993) A contribution to the halophytic vegetation and flora of Iran. In Towards the rational use of high salinity tolerant plants. Springer Netherlands. 1: 35-44.
Bachem CWB, Van der hoven RS, De Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF (1996) Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development. The Plant J. 9(5): 745-753.
Batanouny KH (1994) Halophytes and halophytic plant communities in the Arab Region. In Halophytes as a resource for livestock and for rehabilitation of degraded lands (pp. 139-163). Springer Netherlands.
Bauer D, Warthoe P, Rohde M, Struss M (1994) PCR methods and applications manual supplement, pp S805-809. Cold Spring Harbor University Press, Cold Spring Harbor, NY.
Breyne P, Zabeau M (2001) Genome-wide expression analysis of plant cell cycle modulated genes, Curr. Opin. Plant Biol. 4: 136–142.
Craciun AR, Courbot M, Bourgis F, salis P, saumitou-laprade P, Verbruggen N (2006) Comparative cDNA-AFLP analysis of cd-tolerante and sensitive genotypes derived from crosses between the cd hyperaccumulator Arabidopsis halleri and Arabidopsis lyrata ssp. Petraea. J. Exp. Bot. 2967-2983.
Diachenko LB, Ledesma J, Chenchik AA, Siebert PD (1996) Combining the technique of RNA fingerprinting and differential display to obtain differentially expressed mRNA. Biochem. Biophys Res Com 219: 824–828.
Evans SJ, Datson NA, Kabbaj M, Thompson RC, Vreugdenhil E, De Kloet ER, Akil H (2002) Evaluation of Affymetrix Gene Chip sensitivity in rat hippocampal tissue using SAGE analysis. Eur. J. Neurosci. 16(3): 409-413.
Fatemi F, Nematzadeh G, Askari H, Hashemi H (2013) Differential gene expression of Aeluropus littoralis in response to NaCl. Annals of Biological Research. 4(11): 110-115.
Fukumura R, Takahashi H, Saito T, Tsutsumi Y, Fujimori A, Sato S, Abe M (2003) A sensitive transcriptome analysis method that can detect unknown transcripts. Nucleic Acids Res. 31(16): e94-e94.
Grattan SR, Grieve CM (1992) Mineral element acquisition and growth response of plants grown in saline environments. Agr. Ecosyst. Environ. 38.4: 275-300.
Gulzar S, E Khan (2001) Seed germination of a halophytic grass Aeluropus logopoides. Ann. Botany. 87(3): 319-324.
Guo YQ, Tian ZY, Qin GY, Yan DL, Zhang J, Zhou WZ, Qin P (2009) Gene expression of halophyte Kosteletzkya virginica seedlings under salt stress at early stage. Genetica, 137(2): 189-199.
Hasegawa PM, Bressan RA, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu. Rev. Plant Physiol. Plant Mol. Biol. 51: 463–499.
Ivanova NB, Belyavsky AV (1995) Identification of differentially expressed genes by restriction endonuclease-based gene expression fingerprinting. Nucl. Acids Res. 23: 2954–2958.
Jayaraman A, Puranik S, Rai NK, Vidapu S, Sahu P P, Lata C, Prasad M (2008) cDNA-AFLP analysis reveals differential gene expression in response to salt stress in foxtail millet (Setaria italica L.). Mol. Biotechnol. 40(3) P: 241-251.
Kawasaki S, Borchert C, Deyholos M, Wang H, Brazille S, Kawai K, Bohnert HJ (2001) Gene expression profiles during the initial phase of salt stress in rice. The Plant Cell Online, 13(4): 889-905.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol. Plant. 15: 473-97.
Roy M, Wu R (2002) Overexpression of adenosylmethionine decarboxylase gene in rice increases polyamine level and enhance sodium chloride-stress tolerance. Plant Sci. 163(5): 987-992.
Sanderfoot AA, Assaad FF, Raikhel NV (2000) The Arabidopsis genome: An abundance of soluble N-ethylmaleimide-sensitive factor adaptor protein receptors. Plant Physiol. 124: 1558–1569.
Santos C (1998) Study of some mechanisms of osmotic regulation and the expression of glutamine synthetase in Helianthus annus L. cells exposed to salt stress: selection of salt tolerant cells. PhD thesis, University of Aveiro, Portugal (English summery).
Santos CL, Vieira A, Campos H, Azevedo, Caldeira G (2001) In situe and in vitro senescence induced by KCl stress: nutritional imbalance, lipid peroxidation and antioxidant metabolism. J. Exp. Bot. vol. 52, No. 355, pp: 351-360.
Shabala S, Cuin TA (2007) Potassium transport and plant salt tolerance. Physiologia Plantarum, 133(4): 651-669.
Singh KB, Foley RC, Oñate-Sánchez L (2002). Transcription factors in plant defense and stress responses. Curr. Opin. Plant Biol., 5(5): 430-436.
Torbati-nejad NM, Maghsud-Lorad H, Gharebash AM (2001) Determination of feed value of two spescies of Aeluropus logopoides and Aeluropus littoralis in sheep. J. Agric. Sci. Natur. Resour; 7(2): 31-45.
Umezawa T, K Mizuno, Fujimura T (2002) Discrimination of genes expressed in response to the ionic or osmotic effect of salt stress in soybean with cDNA-AFLP. Plant, Cell Environ 25: 1617–1625.
Wang Z, Li P, Fredricksen M, Gong Z, Kim CS, Zhang C, Bohnert HJ, Zhu JK, Bressan RA, Hasegawa PM, Zhao Y, Zhang H (2004) Expressed sequence tags from Thellungiella halophila, a new model to study plant salt-tolerance. Plant Sci. 166: 609–614.
Wi SJ, Kim WT and Park KY (2006) Overexpression of carnation S-adenosylmethionine decarboxylase gene generates a broad-spectrum tolerance to abiotic stresses in transgenic tobacco plants. Plant Cell Rep., 25(10): 1111-1121.
Zhao S, Ooi SL, Pardee AB (1995) New primer strategy improves precision of differential display. Bio.Techniques 18: 842–850.
Zhu JK (2001) Plant salt tolerance. Trends Plant Sci. 6: 66-71.