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

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری بیوتکنولوژی کشارزی، گروه زراعت و اصلاح نباتات، دانشکده پردیس کشاورزی و منابع طبیعی کرج، دانشگاه تهران، کرج، ایران.

2 استادیار رشته اصلاح نباتات مولکولی، پژوهشکده ژنتیک و زیست فناوری کشاورزی طبرستان، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران.

3 دانشیار رشته بیوتکنولوژی گیاهی، گروه زراعت و اصلاح نباتات، دانشکده پردیس کشاورزی و منابع طبیعی کرج، دانشگاه تهران، کرج، ایران.

4 استادیار اصلاح نباتات مولکولی، پژوهشکده ژنتیک و زیست فناوری کشاورزی طبرستان، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران.

چکیده

در این پژوهش نوع و فراوانی عناصر تنظیمی در نواحی پروموتری خانواده ژنی DVL در گیاه آلوروپوس لیتورالیس مورد مطالعه قرار گرفت. همچنین بیان ژن AlDVL8 به‌عنوان یکی از اعضای این خانواده تحت تیمار تنش شوری، هورمون‌های ‌اسید‌سالیسیلیک، ‌اسید‌‌جیبرلیک و سیتوکینین اندازه‌گیری شد. نتایج بررسی پروموتری حاکی از آن بود که این خانواده ژنی عناصر تنظیمی متفاوتی برای پاسخ به انواع تنش‌ها و هورمون‌ها دارا می‌باشد. برخی از این عناصر تنظیمی در ناحیه پروموتری تمامی ژن‌ها موجودند که احتمالاً بیانگر نقش عمومی DVLها است. همچنین، گروهی از عناصر تنظیمی تنها در ناحیه پروموتری بعضی از ژن‌ها موجودند که می‌توانند مرتبط با فعالیت اختصاصی آن‌ها باشند. تیمارهای اعمال شده به غیر از سیتوکینین سبب افزایش بیان ژن در ‌اندام‌هوایی در زمان‌های 3، 12 و 24 ساعت و کاهش بیان در زمان 6 ساعت پس از اعمال تنش شد. تیمار سیتوکینین در همه زمان‌ها بیان ژن را در ‌اندام هوایی افزایش داد. در بافت ریشه تقریباً روند معکوس بیان ژن دیده شد، به صورتی که در زمان 6 ساعت افزایش بیان ژن و در زمان 12 و 24 ساعت کاهش بیان در تمامی تیمارها مشاهده شد. نتایج این تحقیق نشان داد که بیان ژن AlDVL8 در ‌اندام هوایی و زمینی با تیمارهای آزمایشی القا شد و بیان آن در دو بافت به‌صورت معکوس بود. با توجه به تغییرات هورمون‌ها طی تنش و القای بیان این خانواده ژنی و وجود عناصر پاسخ‌‌دهنده به تنش در نواحی پروموتری این ژن‌ها، این خانواده ژنی می‌تواند به‌عنوان کاندیدی برای تحمل به تنش‌ها پیشنهاد گردد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification and evaluation of relation between regulatory elements of the promoter region of DLV gene family and gene expression in Aeluropus littoralis

نویسندگان [English]

  • Maryam Chalekaei 1
  • Ali Dehestani 2
  • Ali reza Abbasi 3
  • Seyyed Hamidreza Hashemi-petroudi 4

1 Ph.D. Candidate of Agricultural Biotechnology, Department of Agronomy and Plant Breeding, Agricultural College, University of Tehran, Karaj, Iran.

2 Assistant Professor of Molecular Plant Breeding, Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

3 Associate Professor, Department of Agronomy and Plant Breeding, Agricultural College, University of Tehran, Karaj, Iran.

4 Assistant professor, Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

چکیده [English]

In this study, the type and frequency of regulatory elements in the promoter regions of DVL gene family in Aeluropus littoralis were studied. Relative expression of AlDVL8 gene as a member of this family was also measured under the salinity stress, salicylic acid, gibberellic acid and cytokinin. The results of the promoter study indicated that, this gene family has different regulatory elements for responding to stresses and hormones. Some of these regulatory elements are present in the promoter region of all genes, possibly indicating the general role of DVLs. Some others are present only in the promoter region of some genes that may be related to their specific activity. Treatments other than cytokinin increased gene expression in the shoot at 3, 12, and 24 hours and decreased expression at 6 hours. Cytokinin treatment at all times increased gene expression. In the root, almost the reverse trend of gene expression was observed, so that at 6 hours, increased gene expression was observed in all treatments, and at 12 and 24 hours, decreased expression was observed in all treatments. The results of this study showed that the expression of AlDVL8 gene in shoot and root organs was induced by experimental treatments and its expression was inverse in these two different tissues. Due to the changes in hormones during stress, expression induction of this gene family, and the presence of stress-responsive elements in the promoter regions of these genes, this gene family can be suggested as a candidate for stress tolerance.

کلیدواژه‌ها [English]

  • Gene expression
  • Gene family
  • Halophyte plant
  • Regulatory elements
  • Salt stress
 
 
Abe H, Urao T, Ito T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant. Cell. 15(1):63-78.
Abe H, Yamaguchi-Shinozaki K, Urao T, Iwasaki T, Hosokawa D, Shinozaki K (1997) Role of Arabidopsis MYC and MYB homologs in drought-and abscisic acid-regulated gene expression. Plant. Cell. 9(10): 1859-1868.
Baker SS, Wilhelm KS, Thomashow MF (1994) The 5′-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold, drought-and ABA-regulated gene expression. Plant. Mol. Biol. 24(5):701-713.
Bari R, Jones JD (2009) Role of plant hormones in plant defence responses. Plant. Mol. Biol. 69(4):473-488.
Boter M, Ruíz-Rivero O, Abdeen A, Prat S (2004) Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis. Genes. Dev. 18(13):1577-1591.
Brown RL, Kazan K, McGrath KC, Maclean DJ, Manners JM (2003) A role for the GCC-box in jasmonate-mediated activation of the PDF1. 2 gene of Arabidopsis. Plant Physiol. 132(2):1020-1032.
Busk PK, Pagès M (1998) Regulation of abscisic acid-induced transcription. Plant. Mol. biol. 37(3):425-435.
Chow C-N, Zheng H-Q, Wu N-Y, et al. (2016) PlantPAN 2.0: an update of plant promoter analysis navigator for reconstructing transcriptional regulatory networks in plants. Nucleic. Acids. Res. 44(D1):D1154-D1160.
Dabrowska G, Mierek-Adamska A, Goc A (2012) Plant metallothineins: putative functions identified by promoter analysis in silico. Acta. Biol. Cracov. Bot. 54(2).
Du H, Zhang L, Liu L (2009) Biochemical and molecular characterization of plant MYB transcription factor family. Biochemistry. (Moscow.). 74(1):1-11.
Dubouzet JG, Sakuma Y, Ito Y, Kasuga M, Dubouzet EG, Miura S, Yamaguchi‐Shinozaki K (2003) OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought‐, high‐salt‐and cold‐responsive gene expression. Plant. J. 33(4), 751-763.
Dunn MA, White AJ, Vural S, Hughes MA (1998) Identification of promoter elements in a low-temperature-responsive gene (blt4. 9) from barley (Hordeum vulgare L.). Plant molecular biology 38(4):551-564.
Finkelstein RR, Lynch TJ (2000) The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor. Plant. Cell. 12(4):599-609.
Guo P, Yoshimura A, Ishikawa N, Yamaguchi T, Guo Y, Tsukaya H (2015) Comparative analysis of the RTFL peptide family on the control of plant organogenesis. J. Plant. Res. 128(3): 497-510.
Hashemi SH, Ghorbani M (2021) RT-qPCR analysis of some members of DEVIL gene family in Aeluropus littoralis (Gouan) Parl. under salinity stress. Environ. Stresses. Crop. Sci. 13(4):1245-1258.
Hashemi SH, Nematzadeh G, Ahmadian G, Yamchi A, Kuhlmann M (2016) Identification and validation of Aeluropus littoralis reference genes for Quantitative Real-Time PCR Normalization. J. Biol. Res. (Thessalon.). 23(1):1-13.
Hashemi SH, Nematzade G, Kuhlmann M (2019) Identification and analysis of a DEVIL paralog gene cluster in Aeluropus littoralis by a comparative genomic approach. Crop Biotech. 9(25): 75- 87.
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic. Acids. Res. 27(1): 297-300.
Hudson ME, Quail PH (2003) Identification of promoter motifs involved in the network of phytochrome A-regulated gene expression by combined analysis of genomic sequence and microarray data. Plant. Physiol. 133(4):1605-1616.
Ibraheem O, Botha CE, Bradley G (2010) In silico analysis of cis-acting regulatory elements in 5′ regulatory regions of sucrose transporter gene families in rice (Oryza sativa Japonica) and Arabidopsis thaliana. Comput Biol. Chem. 34(5-6):268-283.
Kim SY, Chung HJ, Thomas TL (1997) Isolation of a novel class of bZIP transcription factors that interact with ABA‐responsive and embryo‐specification elements in the Dc3 promoter using a modified yeast one‐hybrid system. PlantJ 11(6):1237-1251.
Larue CT, Wen J, Walker JC (2010) Interactions between a NAC-domain transcription factor and the putative small protein encoding DVL/ROT gene family. Plant. Mol. Biol. Rep. 28(1), 162.
Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic. Acids. Res. 30(1): 325-327.
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. methods. 25(4):402-408.
Mena M, Cejudo FJ, Isabel-Lamoneda I, Carbonero P (2002) A role for the DOF transcription factor BPBF in the regulation of gibberellin-responsive genes in barley aleurone. Plant. Physiol. 130(1):111-119 .
Mohanty B, Krishnan S, Swarup S, Bajic VB (2005) Detection and preliminary analysis of motifs in promoters of anaerobically induced genes of different plant species. Ann. Bot. 96(4):669-681.
Mongkolsiriwatana C, Pongtongkam P, Peyachoknagul S (2009) In silico promoter analysis of photoperiod-responsive genes identified by DNA microarray in rice (Oryza sativa L.). J Nat Sci 43:164-77.
Narita NN, Moore S, Horiguchi G, et al. (2004) Overexpression of a novel small peptide ROTUNDIFOLIA4 decreases cell proliferation and alters leaf shape in Arabidopsis thaliana. Plant. J. 38(4):699-713.
Park HC, Kim ML, Kang YH, et al. (2004) Pathogen-and NaCl-induced expression of the SCaM-4 promoter is mediated in part by a GT-1 box that interacts with a GT-1-like transcription factor. Plant. physiol. 135(4):2150-2161.
Rahnama H, Haghighat V (2014) Cloning and functional analysis of inducible promoter Rd29A in transgenic tobacco plants. Cell. Mol. Res. 26(4):480-490.
Redman J, Whitcraft J, Johnson C, Arias J (2002) Abiotic and biotic stress differentially stimulate as-1 element activity in Arabidopsis. Plant. Cell. Rep. 21(2):180-185.
Ross EJ, Stone JM, Elowsky CG, Arredondo-Peter R, Klucas RV, Sarath G (2004) Activation of the Oryza sativa non-symbiotic haemoglobin-2 promoter by the cytokinin-regulated transcription factor, ARR1. J. Exp. Bot. 55(403):1721-1731.
Ryu H, Cho Y-G (2015) Plant hormones in salt stress tolerance. J. Plant. Biol. 58(3):147-155
Sakai H, Aoyama T, Oka A (2000) Arabidopsis ARR1 and ARR2 response regulators operate as transcriptional activators. Plant. J. 24(6):703-711.
Simpson SD, Nakashima K, Narusaka Y, Seki M, Shinozaki K, Yamaguchi‐Shinozaki K (2003) Two different novel cis‐acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark‐induced senescence. Plant. J. 33(2):259-270.
Sudan J, Sharma D, Mustafiz A, Kumari S (2018) Signaling Peptides: Hidden Molecular Messengers of Abiotic Stress Perception and Response in Plants.  In: Zargar SM,  Zargar MY (eds) Abiotic Stress-Mediated Sensing and Signaling in Plants: An Omics Perspective, 1st Ed. Springer, Singapore, pp 95-125
Sutoh K, Yamauchi D (2003) Two cis‐acting elements necessary and sufficient for gibberellin‐upregulated proteinase expression in rice seeds. Plant. J. 34(5):635-645.
Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K (1993) An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant. Cell. 5(11):1529-1539.
Valdivia ER, Hertweck KL, Cho SK, Walker JC (2013) DVL/RTFL. In: Kastin AJ (ed) Handbook of Biologically Active Peptides, 2nd Ed. Academic Press, Waltham, pp 15-19.
Van Helden J (2003) Regulatory sequence analysis tools. Nucleic. Acids. Res. 31(13):3593-3596.
Wen J, Lease KA, Walker JC (2004) DVL, a novel class of small polypeptides: overexpression alters Arabidopsis development. Plant. J. 37(5): 668-677.
Xie T, Zeng L, Chen X, Rong H, Wu J, Batley J, Jiang J, Wang Y (2020) Genome-Wide Analysis of the Lateral Organ Boundaries Domain Gene Family in Brassica Napus. Genes. 11(3): 280.
Xue GP (2002) Characterisation of the DNA‐binding profile of barley HvCBF1 using an enzymatic method for rapid, quantitative and high‐throughput analysis of the DNA‐binding activity. Nucleic. Acids. Res. 30(15): e77-e77.
Yu D, Chen C, Chen Z (2001) Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant. Cell. 13(7):1527-1540.
Zhang J, Jia W, Yang J, Ismail AM (2006) Role of ABA in integrating plant responses to drought and salt stresses. Field. Crops. Res. 97(1):111-119.
Zhang ZL, Xie Z, Zou X, Casaretto J, Ho THD, Shen QJ (2004) A rice WRKY gene encodes a transcriptional repressor of the gibberellin signaling pathway in aleurone cells. Plant. Physiol. 134(4): 1500-1513.
Zhou M-L, Ma JT, Pang JF, Zhang ZL, Tang YX, Wu YM (2010) Regulation of plant stress response by dehydration responsive element binding (DREB) transcription factors. Afr. J. Biotechnol. 9(54):9255-9269.
Zouari N, Saad RB, Legavre T, et al. (2007) Identification and sequencing of ESTs from the halophyte grass Aeluropus littoralis. Gene. 404(1-2):61-69.