Abstract
Brassinosteroids (BRs) are a class of plant-specific steroid hormones and play an essential role in plant growth and development. The DWARF4 (DWF4) gene encodes a C-22 hydroxylase, which is a rate-limiting enzyme in BR biosynthesis pathway. Here, the DWF4 (ZmDWF4) in maize (Zea mays L.), the ortholog of Arabidopsis DWF4 (CYP90B1), was transformed into elite inbred Q319 and driven maize ubiquitin promoter. In transgenic lines, the level of the endogenous BR precursor canosterone (CS) was significantly enhanced. ZmDWF4 overexpression greatly improved grain yield per ear from 28.3 to 33.5% in transgenic lines compared with that in non-transgenic Q319 plants. Moreover, ZmDWF4 overexpression significantly enhanced heterosis of combinations by increasing both seed number and seed weight up to 20.4% in transgenic hybrids. Further analysis revealed that ZmDWF4 overexpression significantly improved several agronomic traits and leaf photosynthetic ability in transgenic lines. Finally, RNA-seq analysis suggested that overexpressing ZmDWF4 improved cell growth, cell division, and nutrient assimilation in transgenic kernels. Altogether, our results demonstrate that manipulating BR level by overexpressing ZmDWF4 effectively improves maize agronomic traits, and thus, provides a useful biotechnological strategy for the genetic improvement and further commercially favorable maize breeding.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Best NB, Hartwig T, Budka J, Fujioka S, Johal G, Schulz B, Dilkes BP (2016) nana plant2 encodes a maize ortholog of the Arabidopsis brassinosteroid biosynthesis gene DWARF1, identifying developmental interactions between brassinosteroids and gibberellins. Plant Physiol 171:2633–2647. https://doi.org/10.1104/pp.16.00399
Choe S, Dilkes BP, Fujioka S, Takatsuto S, Sakurai A, Feldmann KA (1998) The DWF4 gene of Arabidopsis encodes a cytochrome P450 that mediates multiple 22α-hydroxylation steps in brassinosteroid biosynthesis. Plant Cell 10:231–243. https://doi.org/10.1105/tpc.10.2.231
Choe S, Fujioka S, Noguchi T, Takatsuto S, Yoshida S, Feldmann KA (2001) Overexpression of DWARF4 in the brassinosteroid biosynthetic pathway results in increased vegetative growth and seed yield in Arabidopsis. Plant J 26:573–582. https://doi.org/10.1046/j.1365-313x.2001.01055.x
Choudhary SP, Yu J-Q, Yamaguchi-Shinozaki K, Shinozaki K, Tran L-SP (2012) Benefits of brassinosteroid crosstalk. Trends Plant Sci 17:594–605. https://doi.org/10.1016/j.tplants.2012.05.012
Ding H, Qin C, Luo X, Li L, Chen Z, Liu H, Gao J, Lin H, Shen Y, Zhao M, Lübberstedt T, Zhang Z, Pan G (2014) Heterosis in early maize ear inflorescence development: a genome-wide transcription qnalysis for two maize inbred lines and their hybrid. Int J Mol Sci 15:13892–13915. https://doi.org/10.3390/ijms150813892
Feng Z, Wu C, Wang C, Roh J, Zhang L, Chen J, Zhang S, Zhang H, Yang C, Hu J, You X, Liu X, Yang X, Guo X, Zhang X, Wu F, Terzaghi W, Kim S-K, Jiang L, Wan J (2016) SLG controls grain size and leaf angle by modulating brassinosteroid homeostasis in rice. J Exp Bot 67:4241–4253. https://doi.org/10.1093/jxb/erw204
Fridman Y, Savaldi-Goldstein S (2013) Brassinosteroids in growth control: how, when and where. Plant Sci 209:24–31. https://doi.org/10.1016/j.plantsci.2013.04.002
Fujioka S, Noguchi T, Yokota T, Takatsuto S, Yoshida S (1998) Brassinosteroids in Arabidopsis thaliana. Phytochemistry 48:595–599. https://doi.org/10.1016/S0031-9422(98)00065-X
Fujita S, Ohnishi T, Watanabe B, Yokota T, Takatsuto S, Fujioka S, Yoshida S, Sakata K, Mizutani M (2006) Arabidopsis CYP90B1 catalyses the early C-22 hydroxylation of C27, C28 and C29 sterols. Plant J 45:765–774. https://doi.org/10.1111/j.1365-313X.2005.02639.x
Guo H, Li L, Aluru M, Aluru S, Yin Y (2013) Mechanisms and networks for brassinosteroid regulated gene expression. Curr Opin Plant Biol 16:545–553. https://doi.org/10.1016/j.pbi.2013.08.002
Hannah LC, Futch B, Bing J, Shaw JR, Boehlein S, Stewart JD, Beiriger R, Georgelis N, Greene T (2012) A shrunken-2 transgene increases maize yield by acting in maternal tissues to increase the frequency of seed development. Plant Cell 24:2352–2363. https://doi.org/10.1105/tpc.112.100602
Hartwig T, Chuck GS, Fujioka S, Klempien A, Weizbauer R, Potluri DPV, Choe S, Johal GS, Schulz B (2011) Brassinosteroid control of sex determination in maize. Proc Natl Acad Sci U S A 108:19814–19819. https://doi.org/10.1073/pnas.1108359108
Ishida Y, Hiei Y, Komari T (2007) Agrobacterium-mediated transformation of maize. Nat Protoc 2:1614–1621. https://doi.org/10.1038/nprot.2007.241
Kir G, Ye H, Nelissen H, Neelakandan AK, Kusnandar AS, Luo A, Inzé D, Sylvester AW, Yin Y, Becraft PW (2015) RNA interference knockdown of BRASSINOSTEROID INSENSITIVE1 in maize reveals novel functions for brassinosteroid signaling in controlling plant architecture. Plant Physiol 169:826–839. https://doi.org/10.1104/pp.15.00367
Koh S, Lee S-C, Kim M-K, Koh JH, Lee S, An G, Choe S, Kim S-R (2007) T-DNA tagged knockout mutation of rice OsGSK1, an orthologue of Arabidopsis BIN2, with enhanced tolerance to various abiotic stresses. Plant Mol Biol 65:453–466. https://doi.org/10.1007/s11103-007-9213-4
Li J, Nagpal P, Vitart V, McMorris TC, Chory J (1996) A role for brassinosteroids in light-dependent development of Arabidopsis. Science 272:398–401. https://doi.org/10.1126/science.272.5260.398
Li B, Liu H, Zhang Y, Kang T, Zhang L, Tong J, Xiao L, Zhang H (2013) Constitutive expression of cell wall invertase genes increases grain yield and starch content in maize. Plant Biotechnol J 11:1080–1091. https://doi.org/10.1111/pbi.12102
Li W, Zhao X, Zhang X (2015) Genome-wide analysis and expression patterns of the YUCCA genes in maize. J Genet Genomics 42:707–710. https://doi.org/10.1016/j.jgg.2015.06.010
Li X-J, Guo X, Zhou Y-H, Shi K, Zhou J, Yu J-Q, Xia X-J (2016) Overexpression of a brassinosteroid biosynthetic gene Dwarf enhances photosynthetic capacity through activation of Calvin cycle enzymes in tomato. BMC Plant Biol 16:33. https://doi.org/10.1186/s12870-016-0715-6
Liu T, Zhang J, Wang M, Wang Z, Li G, Qu L, Wang G (2007) Expression and functional analysis of ZmDWF4, an ortholog of Arabidopsis DWF4 from maize (Zea mays L.). Plant Cell Rep 26:2091–2099. https://doi.org/10.1007/s00299-007-0418-4
Liu X, Feng ZM, Zhou CL, Ren YK, Mou CL, Wu T, Yang CY, Liu SJ, Jiang L, Wan JM (2016) Brassinosteroid (BR) biosynthetic gene lhdd10 controls late heading and plant height in rice (Oryza sativa L.). Plant Cell Rep 35:357–368. https://doi.org/10.1007/s00299-015-1889-3
Makiko C et al (2003) A semidwarf phenotype of barley uzu results from a nucleotide substitution in the dene encoding a putative brassinosteroid receptor. Plant Physiol 133:1209–1219. https://doi.org/10.1104/pp.103.026195
Ming L, Yuehua X, Xianbi L, Xiaofeng L, Wei D, Demou L, Lei H, Mingyu H, Yi L, Yan P (2007) GhDET2, a steroid 5α-reductase, plays an important role in cotton fiber cell initiation and elongation. Plant J 51:419–430. https://doi.org/10.1111/j.1365-313X.2007.03144.x
Ren RC, Wang LL, Zhang L, Zhao YJ, Wu JW, Wei YM, Zhang XS, Zhao XY (2020) DEK43 is a P-type pentatricopeptide repeat (PPR) protein responsible for the Cis-splicing of nad4 in maize mitochondria. J Integr Plant Biol 62:299–313. https://doi.org/10.1111/jipb.12843
Shimada S, Komatsu T, Yamagami A, Nakazawa M, Matsui M, Kawaide H, Natsume M, Osada H, Asami T, Nakano T (2015) Formation and dissociation of the BSS1 protein complex regulates plant development via brassinosteroid signaling. Plant Cell 27:375–390. https://doi.org/10.1105/tpc.114.131508
Si J, Sun Y, Wang L, Qin Y, Wang C, Wang X (2016) Functional analyses of Populus euphratica brassinosteroid biosynthesis enzyme genes DWF4 (PeDWF4) and CPD (PeCPD) in the regulation of growth and development of Arabidopsis thaliana. J Biosci 41:727–742. https://doi.org/10.1007/s12038-016-9635-8
Singh AP, Savaldi-Goldstein S (2015) Growth control: brassinosteroid activity gets context. J Exp Bot 66:1123–1132. https://doi.org/10.1093/jxb/erv026
Singh A, Breja P, Khurana JP, Khurana P (2016) Wheat Brassinosteroid-Insensitive1 (TaBRI1) interacts with members of TaSERK gene family and cause early flowering and seed yield enhancement in Arabidopsis. PLoS One 11:e0153273. https://doi.org/10.1371/journal.pone.0153273
Sun X, Cahill J, Van Hautegem T, Feys K, Whipple C, Novák O, Delbare S, Versteele C, Demuynck K, De Block J, Storme V, Claeys H, Van Lijsebettens M, Coussens G, Ljung K, De Vliegher A, Muszynski M, Inzé D, Nelissen H (2017) Altered expression of maize PLASTOCHRON1 enhances biomass and seed yield by extending cell division duration. Nat Commun 8:14752. https://doi.org/10.1038/ncomms14752
Szekeres M, Németh K, Koncz-Kálmán Z, Mathur J, Kauschmann A, Altmann T, Rédei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85:171–182. https://doi.org/10.1016/S0092-8674(00)81094-6
Tian Y, Fan M, Qin Z, Lv H, Wang M, Zhang Z, Zhou W, Zhao N, Li X, Han C, Ding Z, Wang W, Wang Z-Y, Bai M-Y (2018) Hydrogen peroxide positively regulates brassinosteroid signaling through oxidation of the BRASSINAZOLE-RESISTANT1 transcription factor. Nat Commun 9:1063–1063. https://doi.org/10.1038/s41467-018-03463-x
Tomoaki S, Yoichi M, Toshiyuki O, Hidehiko S, Shozo F, Miyako U-T, Masaharu M, Kanzo S, Suguru T, Shigeo Y, Hiroshi T, Hidemi K, Makoto M (2005) Erect leaves caused by brassinosteroid deficiency increase biomass production and grain yield in rice. Nat Biotechnol 24:105–109. https://doi.org/10.1038/nbt1173
Tong H, Xiao Y, Liu D, Gao S, Liu L, Yin Y, Jin Y, Qian Q, Chu C (2012) DWARF AND LOW-TILLERING acts as a direct downstream target of a GSK3/SHAGGY-like kinase to mediate brassinosteroid responses in rice. Plant Cell 24:2562–2577. https://doi.org/10.1105/tpc.112.097394
Tong H, Liu L, Jin Y, Du L, Yin Y, Qian Q, Zhu L, Chu C (2014) Brassinosteroid regulates cell elongation by modulating gibberellin metabolism in rice. Plant Cell 26:4376–4393. https://doi.org/10.1105/tpc.114.132092
Vogler F, Schmalzl C, Englhart M, Bircheneder M, Sprunck S (2014) Brassinosteroids promote Arabidopsis pollen germination and growth. Plant Reprod 27:153–167. https://doi.org/10.1007/s00497-014-0247-x
Wang X, Zhang J, Yuan M, Ehrhardt DW, Wang Z, Mao T (2012) Arabidopsis MICROTUBULE DESTABILIZING PROTEIN40 is involved in brassinosteroid regulation of hypocotyl elongation. Plant Cell 24:4012–4025. https://doi.org/10.1105/tpc.112.103838
Wei Z, Li J (2016) Brassinosteroids regulate root growth, development, and symbiosis. Mol Plant 9:86–100. https://doi.org/10.1016/j.molp.2015.12.003
Wu C-y, Trieu A, Radhakrishnan P, Kwok SF, Harris S, Zhang K, Wang J, Wan J, Zhai H, Takatsuto S, Matsumoto S, Fujioka S, Feldmann KA, Pennell RI (2008) Brassinosteroids regulate grain filling in rice. Plant Cell 20:2130–2145. https://doi.org/10.1105/tpc.107.055087
Wu J, Lawit SJ, Weers B, Sun J, Mongar N, Van Hemert J, Melo R, Meng X, Rupe M, Clapp J, Haug Collet K, Trecker L, Roesler K, Peddicord L, Thomas J, Hunt J, Zhou W, Hou Z, Wimmer M, Jantes J, Mo H, Liu L, Wang Y, Walker C, Danilevskaya O, Lafitte RH, Schussler JR, Shen B, Habben JE (2019) Overexpression of zmm28 increases maize grain yield in the field. Proc Natl Acad Sci U S A 116:23850–23858. https://doi.org/10.1073/pnas.1902593116
Xia XJ, Zhou YH, Shi K, Zhou J, Foyer CH, Yu JQ (2015) Interplay between reactive oxygen species and hormones in the control of plant development and stress tolerance. J Exp Bot 66:2839–2856
Xie G, Li Z, Ran Q, Wang H, Zhang J (2018) Over-expression of mutated ZmDA1 or ZmDAR1 gene improves maize kernel yield by enhancing starch synthesis. Plant Biotechnol J 16:234–244. https://doi.org/10.1111/pbi.12763
Xin P, Li B, Yan J, Chu J (2018) Pursuing extreme sensitivity for determination of endogenous brassinosteroids through direct fishing from plant matrices and eliminating most interferences with boronate affinity magnetic nanoparticles. Anal Bioanal Chem 410:1363–1374. https://doi.org/10.1007/s00216-017-0777-9
Zhang C, Xu Y, Guo S, Zhu J, Huan Q, Liu H, Wang L, Luo G, Wang X, Chong K (2012) Dynamics of brassinosteroid response modulated by negative regulator LIC in rice. PLoS Genet 8:e1002686. https://doi.org/10.1371/journal.pgen.1002686
Zhang C, Bai M-Y, Chong K (2014a) Brassinosteroid-mediated regulation of agronomic traits in rice. Plant Cell Rep 33:683–696. https://doi.org/10.1007/s00299-014-1578-7
Zhang WY, Xu YC, Li WL, Yang L, Yue X, Zhang XS, Zhao XY (2014b) Transcriptional analyses of natural leaf senescence in maize. PLoS One 9:e115617. https://doi.org/10.1371/journal.pone.0115617
Zhao XY, Hong P, Wu JY, Chen XB, Ye XG, Pan YY, Wang J, Zhang XS (2016) The tae-miR408-mediated control of TaTOC1 genes transcription is required for the regulation of heading time in wheat. Plant Physiol 170:1578–1594. https://doi.org/10.1104/pp.15.01216
Funding
This research was supported by the Major Project of China on New varieties of GMO Cultivation (2016ZX08003-003), the National Natural Science Foundation of China (91735301 and 91535109), Taishan Scholars Project (ts201712024), Funds of Shandong “Double Tops” Program (SYL2017YSTD03), and the project (dxkt201707) from State Key Laboratory of Crop Biology.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(DOCX 3904 kb)
Rights and permissions
About this article
Cite this article
Liu, N., Zhao, Y.J., Wu, J.W. et al. Overexpression of ZmDWF4 improves major agronomic traits and enhances yield in maize. Mol Breeding 40, 71 (2020). https://doi.org/10.1007/s11032-020-01152-6
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11032-020-01152-6