Abstract
MYB transcription factors are one of the most important mediators for the survival of plants under multiple stress responses. In the present study, EaMYB18, encoding a single R3 repeat MYB DNA binding domain was isolated from stress-tolerant wild relative species of sugarcane Erianthus arundinaceus. In silico analysis of 948 bp coding mRNA sequence of EaMYB18 exhibited the presence of four exons and three introns. Further, the EaMYB18 gene was transformed in tobacco and its stable inheritance was confirmed through antibiotic resistance screening, PCR amplification and Southern hybridization blotting. Results of the estimation of MDA, proline, total chlorophyll and antioxidant activities of EaMYB18 transgenic tobacco lines exhibited least oxidative damage under drought and cold stress over the untransformed ones, the over-expression of EaMYB18 has improved drought and cold stress tolerance ability in tobacco. The comparative physiological and biochemical analysis of transgenic tobacco plants overexpressing SoMYB18, SsMYB18 and EaMYB18, revealed that the EaMYB18 and SsMYB18 transgenic plants demonstrated effective tolerance to drought and cold stresses, while SoMYB18 showed improved tolerance to salt stress alone. Amongst these three genes, EaMYB18 displayed the highest potential for drought and cold stress tolerances as compared to SoMYB18 and SsMYB18 genes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agarwal PK, Shukla PS, Gupta K, Jha B (2013) Bioengineering for salinity tolerance in plants: state of the art. Mol Biotechnol 54:102–123. https://doi.org/10.1007/s12033-012-9538-3
Ambawat S, Sharma P, Yadav NR, Yadav RC (2013) MYB transcription factor genes as regulators for plant responses: an overview. Physiol Mol Biol Plants 19:307–321. https://doi.org/10.1007/s12298-013-0179-1
Augustine SM, Ashwin NJ, Syamaladevi DP, Appunu C, Chakravarthi M, Ravichandran V, Tuteja N, Subramonian N (2014) Overexpression of EaDREB2 and pyramiding of EaDREB2 with the pea DNA helicase gene (PDH45) enhance drought and salinity tolerance in sugarcane (Saccharum spp. hybrid). Plant Cell Rep 34:247–263. https://doi.org/10.1007/s00299-014-1704-6
Baldoni E, Genga A, Medici A, Coraggio I, Locatelli F (2013) The OsMyb4 gene family: stress response and transcriptional auto-regulation mechanisms. Biol Plant 57:691–700. https://doi.org/10.1007/s10535-013-0331-3
Baranowskij N, Frohberg C, Prat S, Willmitzer L (1994) A novel DNA binding protein with homology to Myb oncoproteins containing only one repeat can function as a transcriptional activator. EMBO J 13:5383–5392
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Chen T, Li W, Hu X, Guo J, Liu A, Zhang B (2015) A cotton MYB transcription factor, GbMYB5, is positively involved in plant adaptive response to drought stress. Plant Cell Physiol 56:917–929. https://doi.org/10.1093/pcp/pcv019
Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L (2010) MYB transcription factors in Arabidopsis. Trends Plant Sci 15:573–581. https://doi.org/10.1016/j.tplants.2010.06.005
El-kereamy A, Bi YM, Ranathunge K, Beatty PH, Good AG, Rothstein SJ (2012) The rice R2R3-MYB transcription factor OsMYB55 Is involved in the tolerance to high temperature and modulates amino acid metabolism. PLoS ONE. https://doi.org/10.1371/journal.pone.0052030
Ganesan G, Sankararamasubramanian HM, Harikrishnan M, Ganpudi A, Ashwin G, Parida A (2012) A MYB transcription factor from the grey mangrove is induced by stress and confers NaCl tolerance in tobacco. J Exp Bot 63:4549–4561. https://doi.org/10.1093/jxb/ers135
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930. https://doi.org/10.1016/j.plaphy.2010.08.016
Hare PD, Cress WA, Van SJ (1998) Dissecting the roles of osmolyte accumulation during stress. Plant Cell Environ 21:535–553. https://doi.org/10.1046/j.1365-3040.1998.00309.x
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for hybridization revealed the expected. Science 227:1229–1231. https://doi.org/10.1126/science.227.4691.1229
Kale PB, Mirajkar SJ, Shingote PR (2017) A practical manual for basic techniques in molecular biology. Lambert Academic Publishing, Germany (ISBN 978-3-330-03194-4)
Li J, Bin-Luan YS, Liu Z (2015) Overexpression of SpWRKY1 promotes resistance to Phytophthora nicotianae and tolerance to salt and drought stress in transgenic tobacco. Physiol. Plant. 155:248–266. https://doi.org/10.1111/ppl.12315
Li C, Ng CKY, Fan LM (2014a) MYB transcription factors, active players in abiotic stress signaling. Environ Exp Bot. https://doi.org/10.1016/j.envexpbot.2014.06.014
Li HL, Guo D, Peng SQ (2014b) Molecular and functional characterization of the JcMYB1, encoding a putative R2R3-MYB transcription factor in Jatropha curcas. Plant Growth Regul 75:45–53. https://doi.org/10.1007/s10725-014-9930-z
Liu H, Zhou X, Dong N, Liu X, Zhang H, Zhang Z (2011) Expression of a wheat MYB gene in transgenic tobacco enhances resistance to Ralstonia solanacearum, and to drought and salt stresses. Funct Integr Genomics 11:431–443. https://doi.org/10.1007/s10142-011-0228-1
Ma Q, Dai X, Xu Y, Guo J, Liu Y, Chen N, Xiao J, Zhang D, Xu Z, Zhang X, Chong K (2009) Enhanced tolerance to chilling stress in OsMYB3R-2 transgenic rice is mediated by alteration in cell cycle and ectopic expression of stress genes. Plant Physiol 150:244–256. https://doi.org/10.1104/pp.108.133454
Mahajan M, Yadav SK (2014) Overexpression of a tea flavanone 3-hydroxylase gene confers tolerance to salt stress and Alternaria solani in transgenic tobacco. Plant Mol Biol 85:551–573. https://doi.org/10.1007/s11103-014-0203-z
Mao X, Jia D, Li A, Zhang H, Tian S, Zhang X, Jia J, Jing R (2011) Transgenic expression of TaMYB2A confers enhanced tolerance to multiple abiotic stresses in Arabidopsis. Funct Integr Genomics 11:445–465. https://doi.org/10.1007/s10142-011-0218-3
Movahedi A, Zhang J, Gao P, Yang Y, Wang L, Yin T, Kadkhodaei S, Ebrahimi M, Zhuge Q (2014) Expression of the chickpea CarNAC3 gene enhances salinity and drought tolerance in transgenic poplars. Plant Cell Tissue Organ Cult 120:141–154. https://doi.org/10.1007/s11240-014-0588-z
Muley AB, Shingote PR, Patil AP, Dalvi SG, Suprasanna P (2019) Gamma radiation degradation of chitosan for application in growth promotion and induction of stress tolerance in potato (Solanum tuberosum L.). Carbohydr Polym 210:289–301. https://doi.org/10.1016/j.carbpol.2019.01.056
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Planta 15(3):473–497
Pagariya MC, Devarumath RM, Kawar PG (2012) Biochemical characterization and identification of differentially expressed candidate genes in salt stressed sugarcane. Plant Sci 184:1–13. https://doi.org/10.1016/j.plantsci.2011.12.002
Piperidis G, Christopher MJ, Carroll BJ, Berding N, D’Hont A (2000) Molecular contribution to selection of intergeneric hybrids between sugarcane and the wild species Erianthus arundinaceus. Genome 43:1033–1037. https://doi.org/10.1139/g00-059
Prabu G, Kawar PG, Pagariya MC, Prasad DT (2011) Identification of water deficit stress upregulated genes in sugarcane. Plant Mol Biol Report 29:291–304. https://doi.org/10.1007/s11105-010-0230-0
Rahaie M, Xue GP, Naghavi MR, Alizadeh H, Schenk PM (2010) A MYB gene from wheat (Triticum aestivum L.) is up-regulated during salt and drought stresses and differentially regulated between salt-tolerant and sensitive genotypes. Plant Cell Rep 29:835–844. https://doi.org/10.1007/s00299-010-0868-y
Ramegowda V, Senthil-Kumar M, Nataraja KN, Reddy MK, Mysore KS, Udayakumar M (2012) Expression of a finger millet transcription factor, ECNAC1, in tobacco confers abiotic stress-tolerance. PLoS ONE. https://doi.org/10.1371/journal.pone.0040397
RoyChoudhury A, Roy C, Sengupta DN (2007) Transgenic tobacco plants overexpressing the heterologous lea gene Rab16A from rice during high salt and water deficit display enhanced tolerance to salinity stress. Plant Cell Rep 26:1839–1859. https://doi.org/10.1007/s00299-007-0371-2
Sairam RK, Srivastava GC, Agarwal S, Meena RC (2005) Differences in antioxidant activity in response to salinity stress in tolerant and susceptible wheat genotypes. Biol Plant 49:85–91. https://doi.org/10.1007/s10535-005-5091-2
Seo PJ, Xiang F, Qiao M, Park JY, Lee YN, Kim SG, Lee YH, Park WJ, Park CM (2009) The MYB96 transcription factor mediates abscisic acid signaling during drought stress response in Arabidopsis. Plant Physiol 151:275–289. https://doi.org/10.1104/pp.109.144220
Shingote PR, Kawar PG, Pagariya MC, Kuhikar RS, Thorat AS, Babu KH (2015) SoMYB18, a sugarcane MYB transcription factor improves salt and dehydration tolerance in tobacco. Acta Physiol Plant 37:217. https://doi.org/10.1007/s11738-015-1961-1
Shingote PR, Kawar PG, Pagariya MC, Rathod PR, Kharte SB (2016) Ectopic Expression of SsMYB18, a novel MYB transcription factor from Saccharum spontaneum augments salt and cold tolerance in tobacco. Sugar tech. https://doi.org/10.1007/s12355-016-0466-6
Wang RK, Cao ZH, Hao YJ (2014) Over-expression of a R2R3 MYB gene MdSIMYB1 increases tolerance to multiple stresses in transgenic tobacco and apples. Physiol Plant 150:76–87. https://doi.org/10.1111/ppl.12069
Xiong H, Li J, Liu P, Duan J, Zhao Y, Guo X, Li Y, Zhang H, Ali J, Li Z (2014) Overexpression of OsMYB48-1, a novel MYB-related transcription factor, enhances drought and salinity tolerance in rice. PLoS ONE 9:1–13. https://doi.org/10.1371/journal.pone.0092913
Yang A, Dai X, Zhang WH (2012) A R2R3-type MYB gene, OsMYB2, is involved in salt, cold, and dehydration tolerance in rice. J Exp Bot 63:2541–2556. https://doi.org/10.1093/jxb/err431
Zhang X, Ju HW, Chung MS, Huang P, Ahn SJ, Kim CS (2011) The R-R-type MYB-like transcription factor, AtMYBL, is involved in promoting leaf senescence and modulates an abiotic stress response in Arabidopsis. Plant Cell Physiol 52:138–148. https://doi.org/10.1093/pcp/pcq180
Zhang D, Jiang S, Pan J, Kong X, Zhou Y, Liu Y, Li D (2014a) The over-expression of a maize mitogen-activated protein kinase gene (ZmMPK5) confers salt stress tolerance and induces defence responses in tobacco. Plant Biol 16:558–570. https://doi.org/10.1111/plb.12084
Zhang L, Liu G, Zhao G, Xia C, Jia J, Liu X, Kong X (2014b) Characterization of a wheat R2R3-MYB transcription factor gene, TaMYB19, involved in enhanced abiotic stresses in Arabidopsis. Plant Cell Physiol 55:1802–1812. https://doi.org/10.1093/pcp/pcu109
Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273. https://doi.org/10.1146/annurev.arplant.53.091401.143329
Acknowledgements
The authors appreciatively recognize the Director General, Vasantdada Sugar Institute, Pune, India for financial and research support, and Dr. S. Anandhan, Directorate of Onion and Garlic Research, Rajgurunagar, India and Dr. Som Dutt, Central Potato Research Institute, Shimla, India for their precious suggestions during this research work and for critically proof reading the manuscript. The financial grant in terms of CREST fellowship to PGK by DBT, New Delhi is also thankfully accredited.
Author information
Authors and Affiliations
Contributions
The overall concept was conceived by PGK and PRS, Experiments were designed by PRS, PGK, KHB and MCP, Experiments were performed by PRS and ABM.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Shingote, P.R., Kawar, P.G., Pagariya, M.C. et al. Isolation and functional validation of stress tolerant EaMYB18 gene and its comparative physio-biochemical analysis with transgenic tobacco plants overexpressing SoMYB18 and SsMYB18. 3 Biotech 10, 225 (2020). https://doi.org/10.1007/s13205-020-02197-2
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-020-02197-2