Identification and investigation of microRNAs associated with drought stress in barley

Document Type : Research Paper

Authors

1 Ph.D.Candidate, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.

2 Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.

3 Associate Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Shiraz University, Shiraz, Iran.

Abstract

Study of stress tolerance in barley can help for a better understanding of the genetic basis of drought stress tolerance mechanisms and lead to improve the genetic properties associated with drought tolerance through modern molecular genetic techniques. To this end, microRNAs associated with drought stress in barley leaf and root ESTs were analyzed in Nimruz and spantaneum barley genotypes. Bioinformatics databases were mined for relevant EST libraries and bioinformatics services were used for pre-processing and identify genes with different expressions among libraries. The expression profile of candidate genes was studied, by using Real time-PCR in a factorial-split plot design, including Nimruz as tolerant and Spontaneum as a drought sensitive in pots with three replications. Sampling time was also considered at 0, 24 and 72 hours after drought stress as sub factor. Results of this study led to identification of three highly-expressed miRNAs (ath-miR414, os-miR2102-5p and osa-miR414). The expression analysis showed that miR414 and miR2102 expression was significantly (P< 0.05) increased in both genotypes in response to drought stress. After 72h in Nimruz and Spontaneum, the expression of miR414 reached 2.61 and 2-fold and the expression of miR2102 was 2.4 and 2.8-fold of that of control (Spontaneum at control condition at 0 times), respectively.

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Allen E, Xie Z, Gustafson AM, Carrington JC (2005) microRNA-directed phasing during trans-acting siRNA biogenesis in plants. Cell, 121(2): 207-221.
Audic S, Claverie J-M (1997) The significance of digital gene expression profiles. Genome research 7 (10): 986-995
Bakhshi B, Bihamta M, Mohseni Fard A, Tohid Far M (2014) Comprehensive review of the expression of miRNAs in response to abiotic stresses. The first international congress and thirteenth Iranian Congress of Genetics, Tehran, Iran's Genetics Society.
Barati M, Majidi MM, Safari M, Mirlohi A, Zeinalinejad KH (2017) Evaluation of Drought Tolerance Indices and Physiological Traits in Cultivated and Wild Barley. Journal of Crop Production and Processing, 7(2):1-18.
Covarrubias AA, Reyes JL (2010) Post‐transcriptional gene regulation of salinity and drought responses by plant microRNAs. Plant, Cell & Environ. 33(4): 481-489.
Ding Y, Tao Y, Zhu C (2013) Emerging roles of microRNAs in the mediation of drought stress response in plants. J. exp. botany, 64(11): 3077-3086.
FAO (2018) Food and Agriculture Organization of the United Nations.
Guleria P, Yadav SK (2011) Identification of miR414 and expression analysis of conserved miRNAs from Stevia rebaudiana. Genomics Proteomics Bioinf. 9(6): 211-217.
Hackenberg M, Gustafson P, Langridge P, Shi BJ (2015) Differential expression of micro RNA s and other small RNA s in barley between water and drought conditions. Plant Biotechnol. J. 13(1): 2-13.
Hsieh CH, Chen WM, Hsieh YS, Fan YC, Yang PE, Kang ST, Liao CT (2018) A Novel Multi-Gene Detection Platform for the Analysis of miRNA Expression. Scientific reports, 8(1), p.10684.
Jaiswal S, Iquebal MA, Arora V, Sheoran S, Sharma P, Angadi UB, Dahiya V, Singh R, Tiwari R, Singh GP, Rai A (2019) Development of species specific putative miRNA and its target prediction tool in wheat (Triticum aestivum L.). Scientific reports, 9(1), p.3790.
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. Funct. Integr. Genomics. 10(4): 493-507.
Khraiwesh B, Zhu JK, Zhu J (2012) Role of miRNAs and siRNAs in biotic and abiotic stress responses of plants. Bioch. Bioph. Acta (BBA)-Gene Regul. Mechan. 1819(2):137-148.
Kim JC, Lee SH, Cheong YH, Yoo CM, Lee SI, Chun HJ, Yun DJ, Hong JC, Lee SY, Lim CO, Cho MJ (2001) A novel cold‐inducible zinc finger protein from soybean, SCOF‐1, enhances cold tolerance in transgenic plants. The Plant J. 25(3): 247-259.
Kong Y, Elling AA, Chen B, Deng X (2010) Differential expression of microRNAs in maize inbred and hybrid lines during salt and drought stress. Am. J. Plant Sci. 1(02): 69-76.
Kuhkan Sh, Lak M, Yousefi A, Lak I, Fallahi H, Barati A, Akbari H, Rostami H, Jahanbin A, Kekha Gh (2011) Nimrooz, A New Barley Cultivar for Sistan and Warm Areas of Southern Provinces of Iran. Seed and Plant Improv J. 4(1): 727-728
Lim LP, Glasner ME, Yekta S, Burge CB, Bartel DP (2003) Vertebrate microRNA genes. Sci. 299(5612): 1540-1540.
Liu HH, Tian X, Li YJ, Wu CA, Zheng CC (2008) Microarray-based analysis of stress-regulated microRNAs in Arabidopsis thaliana. Rna, 14(5): 836-843.
Masoudi-Nejad A, Tonomura K, Kawashima S, Moriya Y, Suzuki M, Itoh M, Kanehisa M, Endo T, Goto S (2006) EGassembler: online bioinformatics service for large-scale processing, clustering and assembling ESTs and genomic DNA fragments. Nucleic Acids Res. 34: 459-462.
Mohseni Fard EM, Bakhshi B, Keshavarznia R, Nikpay N, Shahbazi M, Salekdeh GH (2017) Drought responsive microRNAs in two barley cultivars differing in their level of sensitivity to drought stress. Plant Physiol. Biochem. 118:121-129.
Numnark S, Mhuantong W, Ingsriswang S, Wichadakul D (2012) C-mii: a tool for plant miRNA and target identification. BMC genomics. 13(7): 120-137.
Ozhuner E, Eldem V, Ipek A, Okay S, Sakcali S, Zhang B, Boke H, Unver T (2013) Boron stress responsive microRNAs and their targets in barley. PloS one, 8(3): e59543.
Sharma D, Tiwari M, Lakhwani D, Tripathi RD, Trivedi PK (2015) Differential expression of microRNAs by arsenate and arsenite stress in natural accessions of rice. Metallomics, 7(1): 174-187.
Soleimani A, Valizadeh M, Darvishzadeh R, Aharizad S, Alipour H (2017) Evaluation of Yield and Yield Component of Spring Barely Genotypes under Late Season Drought Stress. Journal of Crop Breeding. 9(23): 105-106
Varkonyi-Gasic E, Wu R, Wood M, Walton EF, Hellens RP (2007) Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant methods, 3(1): 1-12.
Wang Y, Makedon FS, Ford JC, Pearlman J (2004) HykGene: a hybrid approach for selecting marker genes for phenotype classification using microarray gene expression data. Bioinformatics, 21(8): 1530-1537.
Xie Z, Allen E, Fahlgren N, Calamar A, Givan SA, Carrington JC (2005) Expression of Arabidopsis MIRNA genes. Plant physiol. 138(4): 2145-2154.
Yin ZJ, Shen FF (2010) Identification and characterization of conserved microRNAs and their target genes in wheat (Triticum aestivum). Genet. Mol. Res. 9(2):1186-1196.
Zhang BH, Pan XP, Wang Q, Cobb GP, Anderson TA (2006) Computational identification of microRNAs and their targets.        Comput. Biol. Chem. 30: 395-407.
Zhao B, Liang R, Ge L, Li W, Xiao H, Lin H, Ruan K, Jin Y (2007) Identification of drought-induced microRNAs in rice. Biochem. Biophys. Res. Commun. 4(2): 585-590.