The miR156/miR529/miR535 superfamily, showing extremely high sequence identity, has been well documented to modulate growth and development. However, their roles in abiotic stress responses are rarely reported. Here, in this study, we reported the negative regulatory function of OsmiR535 in cold stress responses. The induced expression of OsmiR535 by cold stress was identified through semi-quantitative RT-PCR and quantitative real-time PCR analyses. By comparing the phenotype of the wild type and OsmiR535 overexpression lines, we showed that OsmiR535 overexpression repressed the early seedling growth under cold stress. Our studies further revealed that OsmiR535 overexpression aggravated the cold-induced cell death, affected the ROS accumulation and scavenging, and influenced the osmotic regulation under cold stress. In addition, OsmiR535 overexpression altered the expression of the OsCBF1, OsCBF2, and OsCBF3 genes, the core components of the CBF-mediated cold signaling, and the cold stress–responsive marker genes downstream of the CBF signaling pathway. Expectedly, the transcriptional levels of three SPL genes OsSPL14/11/4, which were predicted to be OsmiR535 targets, were downregulated in the OsmiR535 overexpression lines. Taken together, results in this study suggest that OsmiR535 negatively regulates rice cold stress responses possibly by targeting the SPL target genes and mediating the CBF-mediated cold signaling pathway.
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Arshad M, Feyissa BA, Amyot L, Aung B, Hannoufa A (2017a) MicroRNA156 improves drought stress tolerance in alfalfa (Medicago sativa) by silencing SPL13. Plant Sci 258:122–136. https://doi.org/10.1016/j.plantsci.2017.01.018
Arshad M, Gruber MY, Wall K, Hannoufa A (2017b) An insight into microRNA156 role in salinity stress responses of alfalfa. Front Plant Sci 8:356. https://doi.org/10.3389/fpls.2017.00356
Arshad M, Gruber MY, Hannoufa A (2018) Transcriptome analysis of microRNA156 overexpression alfalfa roots under drought stress. Sci Rep 8(1):9363. https://doi.org/10.1038/s41598-018-27088-8
Chen T, Zhang B (2016) Measurements of proline and malondialdehyde content and antioxidant enzyme activities in leaves of drought stressed cotton. Bio Protoc 6(17):e1913. https://doi.org/10.21769/BioProtoc.1913
Cui LG, Shan JX, Shi M, Gao JP, Lin HX (2014) The miR156-SPL9-DFR pathway coordinates the relationship between development and abiotic stress tolerance in plants. Plant J 80(6):1108–1117. https://doi.org/10.1111/tpj.12712
Cui N, Sun XL, Sun MZ, Jia BW, Duanmu HZ, Lv DK, Duan X, Zhu YM (2015) Overexpression of OsmiR156k leads to reduced tolerance to cold stress in rice (Oryza Sativa). Mol Breeding 35(11):214. https://doi.org/10.1007/s11032-015-0402-6
Dong CH, Pei HX (2014) Over-expression of miR397 improves plant tolerance to cold stress in Arabidopsis thaliana. J Plant Biol 57(4):209–217. https://doi.org/10.1007/s12374-013-0490-y
Gou J, Debnath S, Sun L, Flanagan A, Tang Y, Jiang Q, Wen J, Wang ZY (2018) From model to crop: functional characterization of SPL8 in M. truncatula led to genetic improvement of biomass yield and abiotic stress tolerance in alfalfa. Plant Biotechnol J 16(4):951–962. https://doi.org/10.1111/pbi.12841
Hou H, Jia H, Yan Q, Wang X (2018) Overexpression of a SBP-box gene (VpSBP16) from Chinese wild vitis species in Arabidopsis improves salinity and drought stress tolerance. Int J Mol Sci 19(4). https://doi.org/10.3390/ijms19040940
Iwakawa HO, Tomari Y (2015) The functions of microRNAs: mRNA decay and translational repression. Trends Cell Biol 25(11):651–665. https://doi.org/10.1016/j.tcb.2015.07.011
Jiao Y, Wang Y, Xue D, Wang J, Yan M, Liu G, Dong G, Zeng D, Lu Z, Zhu X, Qian Q, Li J (2010) Regulation of OsSPL14 by OsmiR156 defines ideal plant architecture in rice. Nat Genet 42(6):541–544. https://doi.org/10.1038/ng.591
Kaur N, Sharma I, Kirat K, Pati PK (2016) Detection of reactive oxygen species in Oryza sativa L. (rice). Bio Protoc 6(24):e2061. https://doi.org/10.21769/BioProtoc.2061
Lu Z, Yu H, Xiong G, Wang J, Jiao Y, Liu G, Jing Y, Meng X, Hu X, Qian Q, Fu X, Wang Y, Li J (2013) Genome-wide binding analysis of the transcription activator ideal plant architecture1 reveals a complex network regulating rice plant architecture. Plant Cell 25(10):3743–3759. https://doi.org/10.1105/tpc.113.113639
Luo L, Li W, Miura K, Ashikari M, Kyozuka J (2012) Control of tiller growth of rice by OsSPL14 and strigolactones, which work in two independent pathways. Plant Cell Physiol 53(10):1793–1801. https://doi.org/10.1093/pcp/pcs122
Lv DK, Bai X, Li Y, Ding XD, Ge Y, Cai H, Ji W, Wu N, Zhu YM (2010) Profiling of cold-stress-responsive miRNAs in rice by microarrays. Gene 459(1–2):39–47. https://doi.org/10.1016/j.gene.2010.03.011
Ma C, Burd S, Lers A (2015) miR408 is involved in abiotic stress responses in Arabidopsis. Plant J 84(1):169–187. https://doi.org/10.1111/tpj.12999
Matthews C, Arshad M, Hannoufa A (2018) Alfalfa response to heat stress is modulated by microRNA156. Physiol Plant. https://doi.org/10.1111/ppl.12787
Megha S, Basu U, Kav NNV (2018) Regulation of low temperature stress in plants by microRNAs. Plant Cell Environ 41(1):1–15. https://doi.org/10.1111/pce.12956
Miao C, Wang Z, Zhang L, Yao J, Hua K, Liu X, Shi H, Zhu J-K (2019) The grain yield modulator miR156 regulates seed dormancy through the gibberellin pathway in rice. Nat Commun 10:3822. https://doi.org/10.1038/s41467-019-11830-5
Miura K, Ikeda M, Matsubara A, Song XJ, Ito M, Asano K, Matsuoka M, Kitano H, Ashikari M (2010) OsSPL14 promotes panicle branching and higher grain productivity in rice. Nat Genet 42(6):545–549. https://doi.org/10.1038/ng.592
Ning K, Chen S, Huang H, Jiang J, Yuan H, Li H (2017) Molecular characterization and expression analysis of the SPL gene family with BpSPL9 transgenic lines found to confer tolerance to abiotic stress in Betula platyphylla Suk. Plant Cell Tissue Organ Cult 130(3):469–481. https://doi.org/10.1007/s11240-017-1226-3
Nour-Eldin HH, Hansen BG, Norholm MH, Jensen JK, Halkier BA (2006) Advancing uracil-excision based cloning towards an ideal technique for cloning PCR fragments. Nucleic Acids Res 34(18):e122. https://doi.org/10.1093/nar/gkl635
Nv P, Pa V, Vemanna RS, Ms S, Makarla U (2017) Quantification of membrane damage/cell death using Evan’s Blue staining technique. Bio Protoc 7(16):e2519. https://doi.org/10.21769/BioProtoc.2519
Peever TL, Higgins VJ (1989) Electrolyte leakage, lipoxygenase, and lipid peroxidation induced in tomato leaf tissue by specific and nonspecific elicitors from Cladosporium fulvum. Plant Physiol 90(3):867–875
Shi Y, Ding Y, Yang S (2018) Molecular regulation of CBF signaling in cold acclimation. Trends Plant Sci 23(7):623–637. https://doi.org/10.1016/j.tplants.2018.04.002
Song JB, Gao S, Wang Y, Li BW, Zhang YL, Yang ZM (2016) miR394 and its target gene LCR are involved in cold stress response in Arabidopsis. Plant Gene 5:56–64. https://doi.org/10.1016/j.plgene.2015.12.001
Stief A, Altmann S, Hoffmann K, Pant BD, Scheible WR, Baurle I (2014) Arabidopsis miR156 regulates tolerance to recurring environmental stress through SPL transcription factors. Plant Cell 26(4):1792–1807. https://doi.org/10.1105/tpc.114.123851
Sun MZ, Yang JK, Cai XX, Shen Y, Cui N, Zhu YM, Jia BW, Sun XL (2018) The opposite roles of OsmiR408 in cold and drought stress responses in Oryza sativa. Mol Breeding 38(10):120–112. https://doi.org/10.1007/s11032-018-0877-z
Sun M, Shen Y, Li H, Yang J, Cai X, Zheng G, Zhu Y, Jia B, Sun X (2019) The multiple roles of OsmiR535 in modulating plant height, panicle branching and grain shape. Plant Sci 283:60–69. https://doi.org/10.1016/j.plantsci.2019.02.002
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. Mol Biol Evol 28(10):2731–2739. https://doi.org/10.1093/molbev/msr121
Thiebaut F, Rojas CA, Almeida KL, Grativol C, Domiciano GC, Lamb CR, Engler Jde A, Hemerly AS, Ferreira PC (2012) Regulation of miR319 during cold stress in sugarcane. Plant Cell Environ 35(3):502–512. https://doi.org/10.1111/j.1365-3040.2011.02430.x
Wang B, Wang H (2017) IPA1: a new “green revolution” gene? Mol Plant 10(6):779–781. https://doi.org/10.1016/j.molp.2017.04.011
Wang L, Zhang Q (2017) Boosting rice yield by fine-tuning SPL gene expression. Trends Plant Sci 22(8):643–646. https://doi.org/10.1016/j.tplants.2017.06.004
Wang ST, Sun XL, Hoshino Y, Yu Y, Jia B, Sun ZW, Sun MZ, Duan XB, Zhu YM (2014) MicroRNA319 positively regulates cold tolerance by targeting OsPCF6 and OsTCP21 in rice (Oryza sativa L.). PLoS One 9(3):e91357. https://doi.org/10.1371/journal.pone.0091357
Wang L, Sun S, Jin J, Fu D, Yang X, Weng X, Xu C, Li X, Xiao J, Zhang Q (2015) Coordinated regulation of vegetative and reproductive branching in rice. Proc Natl Acad Sci U S A 112(50):15504–15509. https://doi.org/10.1073/pnas.1521949112
Xie G, Kato H, Imai R (2012a) Biochemical identification of the OsMKK6-OsMPK3 signalling pathway for chilling stress tolerance in rice. Biochem J 443(1):95–102. https://doi.org/10.1042/BJ20111792
Xie K, Shen J, Hou X, Yao J, Li X, Xiao J, Xiong L (2012b) Gradual increase of miR156 regulates temporal expression changes of numerous genes during leaf development in rice. Plant Physiol 158(3):1382–1394. https://doi.org/10.1104/pp.111.190488
Yang CH, Li DY, Mao DH, Liu X, Ji CJ, Li XB, Zhao XF, Cheng ZK, Chen CY, Zhu LH (2013) Overexpression of microRNA319 impacts leaf morphogenesis and leads to enhanced cold tolerance in rice (Oryza sativa L.). Plant Cell Environ 36(12):2207–2218. https://doi.org/10.1111/pce.12130
Yue E, Li C, Li Y, Liu Z, Xu JH (2017a) MiR529a modulates panicle architecture through regulating SQUAMOSA PROMOTER BINDING-LIKE genes in rice (Oryza sativa). Plant Mol Biol 94(4–5):469–480. https://doi.org/10.1007/s11103-017-0618-4
Yue E, Liu Z, Li C, Li Y, Liu Q, Xu JH (2017b) Overexpression of miR529a confers enhanced resistance to oxidative stress in rice (Oryza sativa L.). Plant Cell Rep 36(7):1171–1182. https://doi.org/10.1007/s00299-017-2146-8
Zhang Z, Huang R (2013) Analysis of malondialdehyde, chlorophyll, proline, soluble sugar, and glutathione content in Arabidopsis seedling. Bio Protoc 3(14):e817. https://doi.org/10.21769/BioProtoc.817
Zhang Q, Chen Q, Wang S, Hong Y, Wang Z (2014) Rice and cold stress: methods for its evaluation and summary of cold tolerance-related quantitative trait loci. Rice (N Y) 7(1):24. https://doi.org/10.1186/s12284-014-0024-3
Zhang SD, Ling LZ, Zhang QF, Xu JD, Cheng L (2015) Evolutionary comparison of two combinatorial regulators of SBP-box genes, miR156 and miR529, in plants. PLoS One 10(4):e0124621. https://doi.org/10.1371/journal.pone.0124621
Zhang XN, Wang W, Wang M, Zhang HY, Liu JH (2016) The miR396b of Poncirus trifoliata functions in cold tolerance by regulating ACC oxidase gene expression and modulating ethylene-polyamine homeostasis. Plant Cell Physiol 57(9):1865–1878. https://doi.org/10.1093/pcp/pcw108
Zhang ZY, Li JH, Li F, Liu HH, Yang WS, Chong K, Xu YY (2017) OsMAPK3 phosphorylates OsbHLH002/OsICE1 and inhibits its ubiquitination to activate OsTPP1 and enhances rice chilling tolerance. Dev Cell 43(6):731–743. https://doi.org/10.1016/j.devcel.2017.11.016
Zheng Y, Jagadeeswaran G, Gowdu K, Wang N, Li S, Ming R, Sunkar R (2013) Genome-wide analysis of microRNAs in sacred lotus, Nelumbo nucifera (Gaertn). Trop Plant Biol 6(2–3):117–130. https://doi.org/10.1007/s12042-013-9127-z
We would like to thank the lab members and friends who are not listed in the authorship for their work in data collection.
This work was supported by the Major Special Projects for New Varieties of Genetically Modified Organisms (grant number 2018ZX0800956B-003); the National Natural Science Foundation of China (grant number 31671596); the Startup Foundation of Heilongjiang Bayi Agricultural University (grant number XYB201903); the Heilongjiang Bayi Agricultural University Support Program for San Heng San Zong (grant number ZRCQC201902); and the Graduate Student Scientific Research Innovation Projects of Heilongjiang Bayi Agricultural University (grant number YJSCX2019-Y06).
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Sun, M., Shen, Y., Yang, J. et al. miR535 negatively regulates cold tolerance in rice. Mol Breeding 40, 14 (2020). https://doi.org/10.1007/s11032-019-1094-0
- Cold tolerance
- CBF signaling
- SPL family genes