بررسی بیان miRNAهای کنترل کننده فاکتورهای رونویسی مرتبط با مسیرهای سیگنالینگ اکسین، جیبرلین و اسید آبسزیک، تحت شرایط تنش خشکی در گندم (Triticum aestivum L.)

نوع مقاله: علمی پژوهشی

نویسندگان

1 کارشناس‌ارشد، بخش ژنومیکس، پژوهشکده بیوتکنولوژی کشاورزی ایران، کرج،

2 استادیار، دانشکده کشاورزی، دانشگاه زنجان، زنجان.

3 دانشجوی دکتری، بخش ژنومیکس، پژوهشکده بیوتکنولوژی کشاورزی ایران، کرج.

چکیده

رشد و متابولیسم گیاه تحت تأثیر انواع محرک‌های زنده و غیرزنده‌ از جمله تنش‌های محیطی قرار می‌گیرد که گیاه از طریق هورمون‌ها به آن‌ها پاسخ می‌دهد. miRNAها، گروهی از RNAهای کوچک غیر کدکننده هستند که برخی از آن‌ها در سیگنالینگ هورمون‌های گیاهی نقش دارند. در این مطالعه با استفاده از تکنیک qRT-PCR، الگوی بیان miR159a,b، miR160، miR167a,b و miR171a، که به‌ترتیب در تنظیم بیان فاکتورهای رونویسی MYB، ARF، ARF، SCL نقش دارند، در دو رقم حساس و متحمل به تنش خشکی در گندم مورد بررسی قرار گرفتند. بررسی میزان شباهت نوکلئوتیدی نشان داد که بیشترین شباهت در هر یک از این خانواده‌ها در ناحیه تولید کننده miRNA بالغ می‌باشد. آنالیزهای qRT-PCR نشان داد تحت شرایط تنش خشکی  miR159a,bدر رقم حساس و miR160 و miR167a,b در رقم متحمل افزایش بیان معنی‌داری داشتند. تحت این شرایط در miR171a تغییر بیان معنی‌داری در هر دو رقم مشاهده نشد. احتمالاً افزایش بیان miR159a,b در رقم حساس منجر به کاهش پاسخ ژن‌های MYB درگیر در تنش خشکی خواهد شد. افزایش بیان miR160 و miR167a,b در رقم متحمل، باعث تنظیم اثر متقابل مسیرهای سیگنالینگ اکسین و آبسیزیک اسید و احتمالاً کمک به تحمل تنش، در رقم متحمل به تنش خشکی می‌شوند. همچنین با عدم تغییر بیان miR171a در هر دو رقم، احتمالاً تحت این شرایط بیان ژن‌های SCL از طریق سایر مکانیسم‌های گیاهی تنظیم می‌شوند.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Expression Analysis Of miRNAs That Regulate Transcription factors related to Auxin, Gibberellin And ABA Signaling Pathways, Under Water Stress In Wheat (Triticum aestivum L.)

نویسندگان [English]

  • Mahdieh Safarzadeh 1
  • Reza Fotovat 2
  • Mohammadreza Azimi 2
  • Ehsan Mohsenifard 3
  • Behnam Bakhshi 3
1 M.Sc. Department of Genomics, Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran.
2 Assistant Professors, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.
3 Ph.D. Student, Department of Genomics, Agricultural Biotechnology Research Institute of Iran, Karaj, Iran.
چکیده [English]

Growth and metabolism of plants are affected by a variety of stimuli, including biotic and abiotic environmental stresses that could leads to responses of the plant through hormone regulation. miRNAs, are a group of Non-coding RNAs that some of them are involved in signaling of plant hormones. In this study, the expression patterns of miR159a,b, miR160, miR167a,b and miR171a have been studied in both drought susceptible and drought tolerant varieties in wheat using qRT-PCR. miR159a,b, miR160, miR167a,b and miR171a could play important roles in MYB, ARF, ARF, and SCL, transcription factors regulation, respectively. High conservation among the studied miRNA families was observed in the mature miRNA producer regions by multiplex alignment of pre-miRNAs. Results of qRT-PCR analysis indicated that expressions of miR160 and miR167a,b in tolerant Variety and miR159a,b in susceptible Variety are increased significantly. However, no significant changes in expression were observed for miR171a in both tolerant and sensitive varieties. Presumably, up-regulation miR159a,b in susceptible variety could be resulted to reduction in the expression of  MYB genes involved in drought response. On the other hand, up-regulation of miR160 and miR167a,b in tolerant variety, may lead to regulation of auxin and abscisic acid pathways interaction and probably these miRNAs could contribute in stress tolerance in tolerant variety. In addition, no significant change in miR171a expression demonstrated that expression of SCL could be regulated through other mechanisms in plant.

کلیدواژه‌ها [English]

  • miRNA
  • Hormone signaling
  • Wheat
  • Drought stress
Achard P, Herr A, Baulcombe DC, Harberd NP (2004) Modulation of floral development by a gibberellin-regulated microRNA. Science Signalling. 131(14):3357.

Arenas-Huertero C, Pérez B, Rabanal F, Blanco-Melo D, De la Rosa C, Estrada-Navarrete G, Sanchez F, Covarrubias AA, Reyes JL (2009) Conserved and novel miRNAs in the legume Phaseolus vulgaris in response to stress. Plant Molecular Biology 70(4):385-401.

Barrera-Figueroa BE, Gao L, Diop NN, Wu Z, Ehlers JD, Roberts PA, Close TJ, Zhu JK, Liu R (2011) Identification and comparative analysis of drought-associated microRNAs in two cowpea genotypes. BMC Plant Biology. 11(1):127.

Barrera-Figueroa BE, Gao L, Wu Z, Zhou X, Zhu J, Jin H, Liu R, and Zhu J-K (2012) High throughput sequencing reveals novel and abiotic stress-regulated microRNAs in the inflorescences of rice. BMC Plant Biology 12(1):132.

Bartel DP, Chen CZ (2004) Micromanagers of gene expression: the potentially widespread influence of metazoan microRNAs. Nature Reviews Genetics 5(5):396-400.

Blanca E Barrera-Figueroa LG, Ndeye N Diop, Zhigang Wu, Jeffrey D Ehlers, Philip A Roberts,Timothy J Close, Jian-Kang Zhu, Renyi Liu (2011) Identification and comparative analysis ofdrought-associated microRNAs in two cowpea genotypes. BMC Plant Biology 11:127.

Bray EA, Bailey-Serres J, and Weretilnyk E (2000) Responses to abiotic stresses. Biochemistry and Molecular Biology of Plants:1158-1203.

Chaves M, Oliveira M (2004) Mechanisms underlying plant resilience to water deficits: prospects for water-saving agriculture. Journal of Experimental Botany 55(407): 2365-2384.

Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR (2005) Real-time quantification of microRNAs by stem–loop RT–PCR. Nucleic Acids Research 33(20):e179-e179.

Ding Y, Tao Y, Zhu C (2013) Emerging roles of microRNAs in the mediation of drought stress response in plants. Journal of Experimental Botany 64(11): 3077-3086.

Ehya F, Monavarfeshani A, Fard EM, Farsad LK, Nekouei MK, Mardi M, Salekdeh GH (2013) Phytoplasma-Responsive microRNAs Modulate Hormonal, Nutritional, and Stress Signalling Pathways in Mexican Lime Trees. PloS one 8(6):e66372.

Eldem V, Okay S, Unver T (2013) Plant microRNAs: new players in functional genomics. Turk. J. Agric. For. 37:1-21.

Faghani E, Khavari-Nejad RA, Salekdeh GH, Najafi F (2012) Evaluation of Cuticular Wax Deposition, Stomata and Carbohydrate of Wheat Leaves for Screening Drought Tolerance. Advances in Environmental Biology. 6(13):4035-4040.

Gielen H, Remans T, Vangronsveld J, Cuypers A (2012) MicroRNAs in Metal Stress: Specific Roles or Secondary Responses? International Journal of Molecular Sciences. 13(12): 15826-15847.

Gill BS, Appels R, Botha-Oberholster AM, Buell CR, Bennetzen JL, Chalhoub B, Chumley F, Dvořák J, Iwanaga M, Keller B (2004) A workshop report on wheat genome sequencing international genome research on wheat consortium. Genetics. 168(2):1087-1096.

Golldack D, Li C, Mohan H, Probst N (2013) Gibberellins and abscisic acid signal crosstalk: living and developing under unfavorable conditions. Plant Cell Reports:1-10.

Gray WM (2004) Hormonal regulation of plant growth and development. PLoS Biology 2(9):e311.

Guo A-Y, Zhu Q-H, Gu X, Ge S, Yang J, Luo J (2008) Genome-wide identification and evolutionary analysis of the plant specific SBP-box transcription factor family. Gene 418(1): 1-8.

Hagen G, Guilfoyle T (2002) Auxin-responsive gene expression: genes, promoters and regulatory factors. Plant Molecular Biology 49(3-4): 373-385.

Huq E (2006) Degradation of negative regulators: a common theme in hormone and light signaling networks? Trends in Plant Science 11(1):4-7.

Jain M, Nijhawan A, Tyagi AK, Khurana JP (2006) Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochem. Biophys. Res. Commun. 345(2):646-651.

Jones-Rhoades MW, Bartel DP, Bartel B (2006) MicroRNAs and their regulatory roles in plants. Annu. Rev. Plant Biol. 57:19-53.

Kantar M, Unver T, Budak H (2010) Regulation of barley miRNAs upon dehydration stress correlated with target gene expression. Functional & integrative genomics 10(4):493-507.

Li B, Qin Y, Duan H, Yin W, Xia X (2011a)Genome-wide characterization of new and drought stress responsive microRNAs in Populus euphratica. Journal of Experimental Botany 62(11):3765-3779.

Li H, Dong Y, Yin H, Wang N, Yang J, Liu X, Wang Y, Wu J, Li X (2011b) Characterization of the stress associated microRNAs in Glycine max by deep sequencing. BMC Plant Biology 11(1):170.

Liu H-H, Tian X, Li Y-J, Wu C-A, and Zheng C-C (2008a) Microarray-based analysis of stress-regulated microRNAs in Arabidopsis thaliana. RNA 14(5): 836-843.

Liu HH, Tian X, Li YJ, Wu CA, Zheng CC (2008b) Microarray-based analysis of stress-regulated microRNAs in Arabidopsis thaliana. RNA 14(5):836-843.

Liu PP, Montgomery TA, Fahlgren N, Kasschau KD, Nonogaki H, Carrington JC (2007) Repression of AUXIN RESPONSE FACTOR10 by microRNA160 is critical for seed germination and post‐germination stages. The Plant Journal 52(1):133-146.

Liu Q, Chen Y-Q (2009) Insights into the mechanism of plant development: interactions of miRNAs pathway with phytohormone response. Biochemical and Biophysical Research Communications. 384(1):1-5.

Liu Q, Zhang Y-C, Wang C-Y, Luo Y-C, Huang Q-J, Chen S-Y, Zhou H, Qu L-H, Chen Y-Q (2009) Expression analysis of phytohormone-regulated microRNAs in rice, implying their regulation roles in plant hormone signaling. FEBS Letters 583(4):723-728.

Lu C, Fedoroff N (2000) A mutation in the Arabidopsis HYL1 gene encoding a dsRNA binding protein affects responses to abscisic acid, auxin, and cytokinin. The Plant Cell Online 12(12): 2351-2365.

Lu W, Li J, Liu F, Gu J, Guo C, Xu L, Zhang H, Xiao K (2011) Expression pattern of wheat miRNAs under salinity stress and prediction of salt-inducible miRNAs targets. Frontiers of Agriculture in China:1-10.

Marin E, Jouannet V, Herz A, Lokerse AS, Weijers D, Vaucheret H, Nussaume L, Crespi MD, Maizel A (2010) miR390, Arabidopsis TAS3 tasiRNAs, and their AUXIN RESPONSE FACTOR targets define an autoregulatory network quantitatively regulating lateral root growth. The Plant Cell Online. 22(4): 1104-1117.

Paolacci AR, Tanzarella OA, Porceddu E, Ciaffi M (2009) Identification and validation of reference genes for quantitative RT-PCR normalization in wheat. BMC Molecular Biology 10(1):11.

Phillips JR, Dalmay T, Bartels D (2007) The role of small RNAs in abiotic stress. FEBS Letters 581(19):3592-3597.

Reyes JL, Chua NH (2007) ABA induction of miR159 controls transcript levels of two MYB factors during Arabidopsis seed germination. The Plant Journal. 49(4):592-606.

Sánchez C, Vielba JM, Ferro E, Covelo G, Solé A, Abarca D, De Mier BS, Díaz-Sala C (2007) Two SCARECROW-LIKE genes are induced in response to exogenous auxin in rooting-competent cuttings of distantly related forest species. Tree Physiology. 27(10):1459-1470.

Shukla LI, Chinnusamy V, Sunkar R (2008) The role of microRNAs and other endogenous small RNAs in plant stress responses. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms. 1779(11): 743-748.

Sunkar R, Zhou X, Zheng Y, Zhang W, Zhu JK (2008) Identification of novel and candidate miRNAs in rice by high throughput sequencing. BMC Plant Biology. 8(1):25.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution. 28(10):2731-2739.

Tang Z, Zhang L, Xu C, Yuan S, Zhang F, Zheng Y, Zhao C (2012) Uncovering Small RNA-Mediated Responses to Cold Stress in a Wheat Thermosensitive Genic Male-Sterile Line by Deep Sequencing. Plant Physiology. 159(2): 721-738.

Tuteja N (2007) Abscisic acid and abiotic stress signaling. Plant signaling & behavior 2(3):135-138.

Wang L, Hua D, He J, Duan Y, Chen Z, Hong X, Gong Z (2011) Auxin Response Factor2 (ARF2) and its regulated homeodomain gene HB33 mediate abscisic acid response in Arabidopsis. PLoS Genetics 7(7):e1002172.

Wilkinson S, Davies WJ (2002) ABA‐based chemical signalling: the co‐ordination of responses to stress in plants. Plant, Cell & Environment. 25(2):195-210.

Xin M, Wang Y, Yao Y, Xie C, Peng H, Ni Z, Sun Q (2010) Diverse set of microRNAs are responsive to powdery mildew infection and heat stress in wheat (Triticum aestivum L.). BMC Plant Biology. 10(1):123.

Xiong L, Schumaker KS, Zhu J-K (2002) Cell signaling during cold, drought, and salt stress. The Plant Cell Online 14 (suppl 1): S165-S183.

Yeqin M. Kong AAE, Beibei Chen, Xing Wang Deng (2010) Differential Expression of microRNAs in Maize Inbred and Hybrid Lines during Salt and Drought Stress. American Journal of Plant Sciences 69-76

Yuanyuan Ren LC, Yiyun Zhang, Xiangyang Kang, Zhiyi Zhang, Yanwei Wang (2012) Identification of novel and conserved Populus tomentosa microRNA as components of a response to water stress. Funct Integr Genomics. 12: 327–339.

Zhang B, Pan X, Cobb GP, Anderson TA (2006) Plant microRNA: a small regulatory molecule with big impact. Developmental Biology. 289(1):3-16.

Zhang Z-L, Ogawa M, Fleet CM, Zentella R, Hu J, Heo J-O, Lim J, Kamiya Y, Yamaguchi S, Sun T-p (2011) Scarecrow-like 3 promotes gibberellin signaling by antagonizing master growth repressor DELLA in Arabidopsis. Proceedings of the National Academy of Sciences. 108(5):2160-2165.

Zhou L, Liu Y, Liu Z, Kong D, Duan M, Luo L (2010) Genome-wide identification and analysis of drought-responsive microRNAs in Oryza sativa. Journal of Experimental Botany. 61(15):4157-4168.

Zhu J-K (2001) Cell signaling under salt, water and cold stresses. Current Opinion in Plant Biology 4(5): 401-406.