Gene Expression Patterns of some Transcription Factors (MYB and WRKY) under Salt Stress in the seedling stage of Alfalfa using qPCR

Shoushi Dezfuli, Ahmad Ali Shoushi Dezfuli1; Kalantar Ahmadi, Ahmad

doi:10.30473/cb.2018.5110

Gene Expression Patterns of some Transcription Factors (MYB and WRKY) under Salt Stress in the seedling stage of Alfalfa using qPCR

Document Type : Research Paper

Authors

Assistant Professor Seed and Plant Improvement Institute Department, Safiabad Agricultural and Natural Resource Research Center, AREEO, Dezful, Iran.

10.30473/cb.2018.5110

Abstract

Salinity is one the major problems for production and increasing the area under cultivation around the world and Iran. Understanding of defense mechanisms and genes involved could improve tolerance to different stresses in crops by using some methods such as genetic manipulation. Regulation in the gene transcription phase is one the most methods to control stress in plants. Transcription factors thought binding with transcription elements in DNA promoters regulate genes expression which plays a key role in tolerance to salinity stress in plants. An experiment was conducted to evaluate four genes expression of transcription factors of MYB (MYB14 and MYB112) and WRKY (WRKY53 and WRKY70) in leaf and root tissue of Yazdi genotype (tolerant genotype to salinity) and Diabloverde (sensitive genotype to salinity) under salinity stress. The selection of these genes was based on the statistical analysis of the microarray data that was related to a study on the effect of salinity stress on Medicago truncatula. Short-term salinity stress caused a significant variation in the expression of these genes in leaf and root tissues of Yazdi and Diabloverde genotypes. Real-Time PCR analysis revealed that higher expression of transcription factors (MYB112 and MYB14) associated with more tolerance to salinity stress. This finding could be assisted plant breeders to apply these transcriptional factors to choose tolerant genotypes to salinity in alfalfa.

Keywords

20.1001.1.22520783.1397.8.23.3.9

Main Subjects

Biotic and Abiotic stress

References

Bartels D, Sunkars R (2005) Drought and salt tolerance in plants. CRC. Crit. Rev. Plant. Sci. 24: 23–58.

Chen L, Song Y, Li SH, Zhang L, Zou SH, Yu D (2011) The role of WRKY transcription factors in plant abiotic stresses. Biochim. Biophys. Acta. 9(2): 1-8.

Chen Y, Chen Z, Kang J, Kang D, Gu H, Qin G (2013) AtMYB14 regulates cold tolerance in Arabidopsis. Plant. Mol. Biol. Report. 31: 87–97.

Chinnusamy V, Jagendorf A, Zhu JK (2005) Understanding and Improving Salt Tolerance in Plants. Crop. Sci. 45: 437-448.

Cook DR (1999) Medicago truncatula, a model in the making! Curr. Opin. Plant. Biol. 2: 301–304.

Du H, Zhang L, Liu L, Tang XF, Yang WJ, Wu YM, Huang YB, Tang YX (2009) Biochemical and Molecular Characterization of Plant MYB Transcription Factor Family. Biochemistry (Moscow). 74: 1-11.

Dai X, Xu Y, Ma Q, Xu W, Wang T, Xue Y, Chong K (2007) Overexpression of an R1R2R3 MYB gene OsMYB3R-2, increases tolerance to freezing, drought, salt stress in transgenic Arabidopsis. Plant Physiol 143: 739-1751.

Duan D, Fischer S, Merz P, Bogs J, Riemann M, Nick P (2016) An ancestral allele of grapevine transcription factor MYB14 promotes plant defence. J. Exp. Bot. 67(6): 1795-1804.

Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L (2010) MYB transcription factors in Arabidopsis. Trends Plant Sci. 15: 573-581.

Heidari Sharif Abad H (2001) Plants and salinity. Research Institute of Forests and Rangelands press. Tehran. (In Persian).

Holl J, Vannozzi A, Czemmel S, D’Onofrio C, Walker AR, Rausch T, Lucchin M, Boss PK, Dry IB, Bogs J (2013) The R2R3-MYB Transcription Factors MYB14 and MYB15 Regulate Stilbene Biosynthesis in Vitis vinifera. Plant. Cell. 25: 4135–4149.

Jiang Y, Deyholos MK, (2006) Comprehensive transcriptional prowling of NaCl-stressed Arabidopsis roots reveals novel classes of responsive genes. BMC. Plant. Biol. 6: 1471–2229.

Jiang Y, Deyholos MK, (2009) Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses. Plant. Mol. Biol. 69: 91-105.

Jianga C, Gua J, Chopraa S, Gua X, Peterson T (2004) Ordered origin of the typical two- and three- repeat Myb genes. Gene. 326: 13–229.

Kafi M, Borzouei A, Salehi M (2010) Physiology of environmental stresses in plants. Ferdowsi University Press, Mashhad. (In Persian).

Kawaura K, Mochida K, Ogihara Y (2008) Genome-wide analysis for identification of salt-responsive genes in common wheat. Funct. Integr. Genomics. 8: 277–86.

Kim, IH, Nguven NH, Jeong CY, Nguyen NT, Hong S, Lee H (2013). Loss of the R2R3 MYB AtMyb73 causes hyper-induction of the SOS1 and SOS3 genes in response to high salinity in Arabidopsis. J. Plant Physiol. 170: 1461-1465.

Li D, Zhang Y, Hu X, Shen X, Ma L, Su Z, Wang T, Dong J (2011) Transcriptional profiling of Medicago truncatula under salt stress identified novel CBF transcription factor MtCBF4 that plays an important role in abiotic stress response. BMC. Plant. Biol. 11: 109.

Li D, Su Z, Dong J, Wang T (2009) An expression database for roots of the model legume Medicago truncatula under salt stress. BMC. Genomics. 10: 517.

Li J, Besseau S, Toronen P, Sipari N, Kollist H, Holm L, Palva ET (2013) Defense-related transcription factors WRKY70 and WRKY54 modulate osmotic stress tolerance by regulating stomatal aperture in Arabidopsis. New. Phytol. 200: 457–472.

Lin-Wa K, Bolitho K, Grafton K, Kortstee A, Karunairetnam S (2010) An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae. BMC. Plant. Biol. 10: 3-17.

Lotkowska ME, Tohge T, Fernie AR, Xue GP, Balazadeh S, Mueller-Roeber B (2015) The Arabidopsis Transcription Factor MYB112 Promotes Anthocyanin Formation during Salinity and under High Light Stress. Plant. Physiol. (169): 1862–1880.

Mokhtari F, Rafiei F, Shabani L, Shiran B (2017) Differential expression pattern of transcription factors across annual Medicago genotypes in response to salinity stress. Biol plantarum. 61 (2): 227-234.

Motahari M, Namaki Shoushtari A, Klantari M (2005) Study of the effects of salinity stress on nitrogen fixation system of Agrobacterium tumefaciens in two cultivars of alfalfa. Master thesis of Shahid Bahonar University of Kerman. (In Persian).

Mott IW, Wang RRC (2007) Comparative transcriptome analysis of salt-tolerant wheat germplasm lines using wheat genome arrays. Plant. Sci. 173: 327–339.

Niu CF, Wei W, Zhou QY, Tian AG, Hao YJ (2012) Wheat WRKY genes TaWRKY2 and TaWRKY19 regulate abiotic stress tolerance in transgenic Arabidopsis plants. Plant Cell Environ 35:1156-1170.

Noble CL, Halloran GM, West DW (1984) Identification and selection for salt tolerance in lucerne. Aust. J. Agri. Res. 35: 239-252.

Paz-Ares J, Ghosal D, Wienand U, Peterson P, Saedler H, (1987) The regulatory c1 locus of Zea mays encodes a protein with homology to myb oncogene products and with structural similarities to transcriptional activators. EMBO. J. 6: 3553-3558.

Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT–PCR. Nucleic. Acids. Res. 29(9): 44-45.

Postnikova OA, Shao J, Nemchinov LG (2013). Analysis of the Alfalfa Root Transcriptome in Response to Salinity Stress. Plant Cell Physiology 54(7): 1041–1055.

Rahaei M, Gomarian M, Alizadeh H, Malboubi M, Naghavi MR (2011) Analysis of expression of transcription factors under long-term salinity stress conditions in two susceptible wheat genotypes using Norton Blot reverse method. Iran. J. Crop. Sci. 13(3): 580-595.

Ramsay NA, Glover BJ (2005) MYB– bHLH–WD40 protein complex and the evolution of cellular diversity. Trends. Plant. Sci. 10: 63-70.

Riechmann JL, Heard J, Martin G, Reuber L, Jiang C, Keddie J, Adam L, Pineda O, Ratcliffe OJ, Samaha RR (2000) Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes. Science. 290: 2105–2110.

Rushton PJ, Somssich IE, Ringler P, Shen QJ (2010) WRKY transcription factors. Trends. Plant. Science. 15(5): 247-258.

Scasta JD, Trostle CL, Foster MA (2012) Evaluating Alfalfa (Medicago sativa L.) Cultivars for Salt Tolerance Using Laboratory, Greenhouse and Field Methods. J. Agric. Sci. 4(9): 90-10.

Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat. Protoc. 3: 1101-1108.

Seo JS, Sohn HB, Noh K, Jung C, An JH, Donovan CM, Somers DA, Kim DI, Jeong SC, Kim C, Kim HM, Lee S, Choi YD, Moon TW, Kim CH, Cheong J (2012). Expression of the Arabidopsis AtMYB44 gene confers drought/salt-stress tolerance in transgenic soybean. Mol. Breeding 29: 601-608

Shoushi Dezfuli AA, Mohammady Dehcheshmeh S, Rafiei F, Shiran B (2016) Evaluation of Salinity Tolerance of Alfalfa Genotypes during Germination Stage Using Multivariate Analysis. 6(3): 51-56.

Shoushi Dezfuli AA, Mohammady Dehcheshmeh S, Rafiei F, Shiran B (2017a) Variation among Iranian alfalfa genotypes for absolute growth rates and salt stress tolerance indices. Environ. Conserv. J. 18(1): 27-39.

Shoushi Dezfuli AA, Paknegad AR, Asareh A, Zarifinia N (2017b) Evaluation of Alfalfa Echotype's Tolerance to Salinity by Using some Morphological and Chemical Characteristic. Crop. Physiol. J. 35: 105-120. (In Persian).

Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr. Opin. Plant. Biol. 4:447–456.

Ulker B, Shahid Mukhtar M, Somssich IE (2007) The WRKY70 transcription factor of Arabidopsis influences both the plant senescence and defense signaling pathways. Planta. 226: 125–137.

Wang H, Miyazaki S, Kawai K, Deyholos M, Galbraith DW, Bohnert HJ (2003) Temporal progression of gene expression responses to salt shock in maize roots. Plant. Mol. Biol. 52: 873–891.

Wang YJ, Zhang ZG, He XJ, Zhou HL, Wen YX, Dai JX, Zhang JS, Chen SY (2003) A rice transcription factor OsbHLH1 is involved in cold stress response. Theor. Appl. Genet. 107: 1402–1409.

Yarnia M., Heydari H, Hashemi Dezfuli A, Rahimzadeh Khouei F, Galavand A (2001) Evaluation of alfalfa (medicage sativa) lines to salinity. Iran. J. Crop. Sci. 3(2): 12-26.

Yamaguchi-Shinazaki K, Shinozaki K, (2005) Organization of cis-acting regulatory elements in osmotic and cold-stress-responsive promoters. Trends. Plant. Sci. 10: 88–94.

Yang A, Xiaoyan Dai X, Zhang WH (2012) A R2R3-type MYB gene, OsMYB2, is involved in salt, cold, and dehydration tolerance in rice. J Exp Bot 63(7):2541–2556.

Young ND, Cannon SB, Sato S, Kim DJ, Cook DR, Town CD, Roe BA, Tabata S (2005) Sequencing the gene spaces of Medicago truncatula and Lotus japonicus. Plant. Physiol. 137: 1174–1181.