Abstract
The miR171 is a conserved microRNA (miRNA) family and has been shown to participate in plant growth and development. However, the precise function of miR171 in Pinus densata remains largely unclear. Mature miR171 sequence comparison reveals high similarity between Arabidopsis thaliana and P. densata and the pre-miR171 could fold into a characteristic stem-loop hairpin structure. Genes encoding GRAS (GAI-RGA-SCR) family transcription factors and actin binding protein were identified as targets of pde-miR171 using a modified RNA ligase mediated 5’ rapid amplification of cDNA ends (RLM-RACE). Furthermore, the interaction between pde-miR171 and Arabidopsis SCL6 (SCARECROW-LIKE6) was further validated through transient co-expression of both genes in Nicotiana benthamiana leaves. Next, results of real-time quantitative PCR demonstrated that the expression of pde-miR171 was significantly up-regulated in miR171-overexpressing plants than in wild-type plants, which was inversely correlated with the expression of Arabidopsis SCL6 genes. In addition, overexpression of pde-miR171 in Arabidopsis induced larger leaves and earlier flowering under long-day conditions compared with the wild type. The findings presented here suggest that miR171 derived from a P. densata precursor together with its target gene SCL6 may play important roles in the regulation of primary root growth, leaf shape, and flowering time in plants.
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Abbreviations
- GRAS :
-
GAI-RGA-SCR
- mRNA:
-
messenger RNA
- miRNA:
-
microRNA
- RLM-RACE:
-
RNA ligase mediated 5’ rapid amplification of cDNA ends
- SCL6 :
-
SCARECROW-LIKE6
References
Asha, S., Nisha, J., Soniya, E.V.: In silico characterization and phylogenetic analysis of two evolutionarily conserved miRNAs (miR166 and miR171) from black pepper (Piper nigrum L.). — Plant mol. Biol. Rep. 31: 707–718, 2013.
Axtell, M.J., Bowman, J.L.: Evolution of plant microRNAs and their targets. — Trends Plant Sci. 13: 343–349, 2008.
Bartel, D.: MicroRNAs: genomics, biogenesis, mechanism, and function. — Cell 116: 281–297, 2004.
Bechtold, N., Pelletier, G.: In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration methods. — Mol. Biol. 82: 259–266, 1998.
Cuperus, J.T., Fahlgren, N., Carrington, J.C.: Evolution and functional diversification of miRNA genes. — Plant Cell 23: 431–442, 2011.
Curaba, J., Talbot, M., Li, Z., Helliwell, C.: Over-expression of microRNA171 affects phase transitions and floral meristem determinancy in barley. — BMC Plant Biol. 13: 6, 2013.
Fan, T., Li, X., Yang, W., Xia, K., Ouyang, J., Zhang, M.: Rice osa-miR171c mediates phase change from vegetative to reproductive development and shoot apical meristem maintenance by repressing four OsHAM transcription factors. — PLoS ONE 10: e0125833, 2015.
Guo, H.S., Xie, Q., Fei, J.F., Chua, N.H.: MicroRNA directs mRNA cleavage of the transcription factor NAC1 to downregulate auxin signals for Arabidopsis lateral root development. — Plant Cell 17: 1376–1386, 2005.
Hofferek, V., Mendrinna, A., Gaude, N., Krajinski, F., Devers, E.A.: MiR171h restricts root symbioses and shows like its target NSP2 a complex transcriptional regulation in Medicago truncatula. — BMC Plant Biol. 14: 199, 2014.
Huang, W., Peng, S., Xian, Z., Lin, D., Hu, G., Yang, L., Ren, M., Li, Z.: Overexpression of a tomato miR171 target gene SlGRAS24 impacts multiple agronomical traits via regulating gibberellin and auxin homeostasis. — Plant Biotechnol. J. 15: 472–488, 2017.
Li, W., Wang, T., Zhang, Y., Li, Y.: Overexpression of soybean miR172c confers tolerance to water deficit and salt stress, but increases ABA sensitivity in transgenic Arabidopsis thaliana. — J. exp. Bot. 67: 175–194, 2016.
Li, W.F., Zhang, S.G., Han, S.Y., Wu, T., Zhang, J.H., Qi, L.W.: The post-transcriptional regulation of LaSCL6 by miR171 during maintenance of embryogenic potential in Larix kaempferi (Lamb.) Carr.. — Tree Genet. Genomes 10: 223–229, 2014.
Livak, K.J., Schmittgen, T.D.: Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. — Methods 25: 402–408, 2001.
Ma, Z.X., Hu, X.P., Cai, W.J., Huang, W.H., Zhou, X., Luo, Q., Yang, H.Q.: Arabidopsis miR171-targeted scarecrow-like proteins bind to GT cis-elements and mediate gibberellinregulated chlorophyll biosynthesis under light conditions. — PLoS Genet. 10: e1004519, 2014.
McCurdy, D.W., Kovar, D.R., Staiger, C.J.: Actin and actinbinding proteins in higher plants. — Protoplasma 215: 89–104, 2001.
Qiu, Z.B., Li, X.J., Zhao, Y.Y., Zhang, M.M., Wan, Y.L., Cao, D.C., Lu, S.F., Lin, J.X.: Genome-wide analysis reveals dynamic changes in microRNAs expression during vascular cambium development in Chinese fir (Cunninghamia lanceolata). — J. exp. Bot. 66: 3041–3054, 2015.
Qiu, Z.B., Yuan, M.M., Hai, B.Z., Wang, L., Zhang, L.: Characterization and expression analysis of conserved miRNAs and their targets in Pinus densata. — Biol. Plant. 60: 427–434, 2016.
Ronemus, M., Vaughn, M.W., Martienssen, R.A.: MicroRNAtargeted and small interfering RNA-mediated mRNA degradation is regulated by Argonaute, Dicer, and RNAdependent RNA polymerase in Arabidopsis. — Plant Cell 18: 1559–1574, 2006.
Shivaraj, S.M., Dhakate, P., Mayee, P., Negi, M.S., Singh, A.: Natural genetic variation in MIR172 isolated from Brassica species. — Biol. Plant. 58: 627–640, 2014.
Sunkar, R., Kapoor, A., Zhu, J. K.: Post-transcriptional induction of two Cu/Zn superoxide dismutase genes in Arabidopsisis mediated by down-regulation of miR398 and important for oxidative stress tolerance. — Plant Cell 18: 2051–2065, 2006.
Voinnet, O.: Origin, biogenesis, and activity of plant microRNAs. — Cell 136: 669–687, 2009.
Wan, L.C., Zhang, H.Y., Lu, S.F., Zhang, L., Qiu, Z.B., Zhao, Y.Y., Zeng, Q.Y., Lin, J.X.: Transcriptome-wide identification and characterization of miRNAs from Pinus densata. — BMC Genomics 13: 132, 2012.
Wang, B., Mao, J.F., Gao, J., Zhao, W., Wang, X.R.: Colonization of the Tibetan Plateau by the homoploid hybrid pine Pinus densata. — Mol. Ecol. 18: 3796–3811, 2011.
Wang, L., Mai, Y.X., Zhang, Y.C., Luo, Q., Yang, H.Q.: MicroRNA171c-targeted SCL6-II, SCL6-III, and SCL6-IV genes regulate shoot branching in Arabidopsis. — Mol. Plant 3: 794–806, 2010.
Zhang, Y.: miRU: an automated plant miRNA target prediction server. — Nucl. Acids Res. 33: W701–W704 2005.
Zheng, Y., Li, Y.F., Sunkar, R., Zhang, W.: SeqTar: an effective method for identifying microRNA guided cleavage sites from degradome of polyadenylated transcripts in plants. — Nucl. Acids Res. 40: e28, 2012.
Zuker, M.: Mfold web server for nucleic acid folding and hybridization prediction. — Nucl. Acids Res. 13: 3406–3415, 2003.
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Acknowledgments: This work was supported by the National Natural Science Foundation of China (31500499 and U1404304), the Program for Science and Technology Innovation Talents in Universities of Henan Province (16HASTIT019), the Innovative Research Team (in Science and Technology) in University of Henan Province (No. 15IRTSTHN020) and the Key Science and Technology Project of Henan Province (162102310442).
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Hai, B.Z., Qiu, Z.B., He, Y.Y. et al. Characterization and primary functional analysis of Pinus densata miR171. Biol Plant 62, 318–324 (2018). https://doi.org/10.1007/s10535-018-0774-7
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DOI: https://doi.org/10.1007/s10535-018-0774-7