Abstract
Gibberellin 2-oxidases (GA2oxs) irreversibly convert bioactive gibberellins (GAs) and their immediate precursors into inactive GAs via 2-β hydroxylation and so regulate gibberellin content in plants. However, to the best of our knowledge, little has been known about the GA2oxs and its function in cool season turfgrass Poa pratensis. In this study, rapid amplification of cDNA end (RACE) was employed to isolate PpGA2ox from P. pratensis. The open reading frame of PpGA2ox was 1 047 bp in length, corresponding to 348 amino acids. PpGA2ox was localized in both nucleus and cytoplasm. The expression of PpGA2ox could be up-regulated by 10 μM gibberellic acid, 5 μM methyl jasmonate, or 10 μM indole-3-acetic acid. In addition, its native promoter could drive GUS expression in both leaf apex and shoot apical region. Moreover, overexpression of PpGA2ox in Arabidopsis led to GA-deficiency leading to dwarf phenotype, delayed flowering time, and increased chlorophyll content. Our study suggests that PpGA2ox could be a candidate gene for breeding new cultivars of P. pratensis.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- ABA:
-
abscisic acid
- FLC :
-
FLOWERING LOCUS C
- FT :
-
FLOWERING LOCUS T
- GA:
-
gibberellin
- GA2oxs:
-
GA2-oxidases
- GA3 :
-
gibberellic acid
- GUS:
-
β-glucuronidase
- IAA:
-
indole-3-acetic acid
- MeJA:
-
methyl jasmonate
- MS:
-
Murashige and Skoog
- PAC:
-
paclobutrazol
- qPCR:
-
quantitative polymerase chain reactions
- SOC1 :
-
SUPPRESSOR OF OVEREXPRESSION OF CO1
- WT:
-
wild type
References
El-Sharkawy I., El Kayal, W., Prasath, D., Fernandez, H., Bouzayen, M., Svircev, A.M., Jayasankar, S.: Identification and genetic characterization of a gibberellin 2-oxidase gene that controls tree stature and reproductive growth in plum. - J. exp. Bot. 63: 1225–1239, 2012.
Frigerio, M., Alabadi, D., Perez-Gomez, J., Garcia-Carcel, L., Phillips, A.L., Hedden, P., Blazquez, M.A.: Transcriptional regulation of gibberellin metabolism genes by auxin signaling in Arabidopsis. - Plant Physiol. 142: 553–563, 2006.
Gallego-Giraldo, L., Ubeda-Tomas, S., Gisbert, C., Garcia- Martinez, J.L., Moritz, T., Lopez-Diaz, I.: Gibberellin homeostasis in tobacco is regulated by gibberellin metabolism genes with different gibberellin sensitivity. - Plant Cell Physiol. 49: 679–690, 2008.
Gargul, J.M., Mibus, H., Serek, M.: Constitutive overexpression of Nicotiana GA(2)ox leads to compact phenotypes and delayed flowering in Kalanchoe blossfeldiana and Petunia hybrida. - Plant Cell Tissue Organ Cult. 115: 407–418, 2013.
Hedden, P., Phillips, A.L.: Gibberellin metabolism: new insights revealed by the genes. - Trends Plant Sci. 5: 523–530, 2000.
Hedden, P., Thomas, S.G.: Gibberellin biosynthesis and its regulation. - Biochem. J. 444: 11–25, 2012.
Hepworth, S.R., Valverde, F., Ravenscroft, D., Mouradov, A., Coupland, G.: Antagonistic regulation of flowering-time gene SOC1 by CONSTANS and FLC via separate promoter motifs. - EMBO J. 21: 4327–4337, 2002.
Hisamatsu, T., King, R.W.: The nature of floral signals in Arabidopsis. II. Roles for FLOWERING LOCUS T (FT) and gibberellin. - J. exp. Bot. 59:3821–3829, 2008.
Hoagland, D.R., Arnon, D.I.: The water-culture method for growing plants without soil. - Calif. Agr. Exp. Stn. Circ. 347: 357–359, 1950.
Jia, N., Liu, X.M., Gao, H.B.: A DNA2 homolog is required for DNA damage repair, cell cycle regulation, and meristem maintenance in plants. - Plant Physiol. 171: 318–333, 2016.
Kim, S.Y., Park, B.S., Kwon, S.J., Kim, J., Lim, M.H., Park, Y.D., Kim, D.Y., Suh, S.C., Jin, Y.M., Ahn, J.H., Lee, Y.H.: rapa L.: ssp. pekinensis). - Plant Cell Rep. 26: 327–336, 2007.
Lee, D.H., Lee, I.C., Kim, K.J., Kim, D.S., Na, H.J., Lee, I.J., Kang, S.M., Jeon, H.W., Le, P.Y., Ko, J.H.: Expression of gibberellin 2-oxidase 4 from Arabidopsis under the control of a senescence-associated promoter results in a dominant semi-dwarf plant with normal flowering. - J. Plant Biol. 57: 106–116, 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.
Olszewski, N., Sun, T.P., Gubler, F.: Gibberellin signaling: biosynthesis, catabolism, and response pathways. - Plant Cell 14 (Suppl): S61–S80, 2002.
Petersen, T.N., Brunak, S., Von Heijne, G., Nielsen, H.: Signal P 4.0: discriminating signal peptides from transmembrane regions. - Nat. Methods 8: 785–786, 2011.
Puyang, X.H., An, M.Y., Han, L.B., Zhang, X.Z.: Protective effect of spermidine on salt stress induced oxidative damage in two Kentucky bluegrass (Poa pratensis L.) cultivars. - Ecotox. Environ. Safety 117: 96–106, 2015.
Sparkes, I.A., Runions, J., Kearns, A., Hawes, C.: Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. - Natur. Protocols 1: 2019–2025, 2006.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S.: MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. - Mol. Biol. E Vol. 28: 2731–2739, 2011.
Teng, K., Chang, Z.H., Li, X., Sun., X.B., Liang, X.H., Xu, L.X., Chao, Y.H., Han, L.B.: Functional and RNAsequencing analysis revealed expression of a novel staygreen gene from Zoysia japonica (ZjSGR) caused chlorophyll degradation and accelerated senescence in Arabidopsis. - Front. Plant Sci. 7: 1894, 2016.
Ueguchi-Tanaka, M., Ashikari, M., Nakajima, M., Itoh, H., Katoh, E., Kobayashi, M., Chow, T.Y., Hsing, Y.I.C., Kitano, H., Yamaguchi, I., Matsuoka, M.: Gibberellin insensitive dwarf1 encodes a soluble receptor for gibberellin. - Nature 437: 693–698, 2005.
Wuddineh, W.A., Mazarei, M., Zhang, J.Y., Poovaiah, C.R., Mann, D.G.J., Ziebell, A., Sykes, R.W., Davis, M.F., Udvardi, M.K., Stewart, C.N.: Identification and overexpression of gibberellin 2-oxidase (GA2ox) in switchgrass (Panicum virgatum L.) for improved plant architecture and reduced biomass recalcitrance. - Plant Biotechnol. J. 13: 636–647, 2015.
Xiao, Z., Fu, R.P., Li, J.Y., Fan, Z.Q., Yin, H.F.: Overexpression of the gibberellin 2-oxidase gene from Camellia lipoensis induces dwarfism and smaller flowers in Nicotiana tabacum. - Plant mol. Biol. Rep. 34: 182–191, 2016.
Yamaguchi, S.: Gibberellin metabolism and its regulation. - Annu. Rev. Plant Biol. 59: 225–251, 2008.
Yan, J.D., Liao, X.Y., He, R.Q., Zhong, M., Feng, P.P., Li, X.M., Tang, D.Y., Liu, X.M., Zhao, X.Y.: Ectopic expression of GA 2-oxidase 6 from rapeseed (Brassica napus L.) causes dwarfism, late flowering and enhanced chlorophyll accumulation in Arabidopsis thaliana. - Plant Physiol. Biochem. 111: 10–19, 2017.
Yin, S., Zhou, S., Kong, X., Han, Y., Wang, W.: Altered gibberellin content affects growth and development in trasgenic tobacco lines overexpressing a wheat gene encoding F-box protein. - Biol. Plant. 61: 349–358, 2017.
Zhou, Y.C., Underhill, S.J.R.: Breadfruit (Artocarpus altilis) gibberellin 2-oxidase genes in stem elongation and abiotic stress response. - Plant Physiol. Biochem. 98: 81–88, 2016.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Acknowledgments: We are very grateful to Dr. Y.-W. Jiang from Department of Agronomy, Purdue University and Miss Z.-S. Sun from School of Foreign Languages, Beihang University for critically reviewing the manuscript. This research was supported by the China Postdoctoral Science Foundation (2017M620677), the Beijing Postdoctoral Research Foundation (No.2017ZZ-087), the Postdoctoral Fund of the Beijing Academy of Agriculture and Forestry Sciences, the National Natural Science Foundation of China (No. 31672477), and the Innovative Project of Beijing Academy of Agriculture and Forestry Sciences (KJCX20151202).
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Tan, PH., Zhang, L., Yin, SX. et al. Heterologous expression of a novel Poa pratensis gibberellin 2-oxidase gene, PpGA2ox, caused dwarfism, late flowering, and increased chlorophyll accumulation in Arabidopsis. Biol Plant 62, 462–470 (2018). https://doi.org/10.1007/s10535-018-0788-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10535-018-0788-1