DNA Demethylation Induces Tree Peony Flowering with a Low Deformity Rate Compared to Gibberellin by Inducing PsFT Expression under Forcing Culture Conditions
Abstract
:1. Introduction
2. Results
2.1. Effects of 5-azaC and GA Treatments on the Morphological Changes in Tree Peony
2.2. Effects of 5-azaC and GA Treatments on Flowering Quality and GA Biosynthesis Regulation
2.3. Effects of 5-azaC and GA Treatments on the Expression of Flowering-Pathway Genes
2.4. Effects of 5-azaC and GA Treatments on DNA Demethylation-Induced Flowering in Tree Peony
2.5. Transcriptome Data Analysis
2.6. Correlation of Different DEGs and Identification of Potential Regulatory Genes
2.7. Construction of the Potential Network Involved in Tree Peony Flowering Regulation
3. Discussion
4. Conclusions
5. Experimental Procedures
5.1. Plant Materials, Growth Conditions, and Sample Collection
- (1)
- (2)
- 5-azaC treatment group: all buds were supplied with 5-azaC (100 μmol L−1, biological reagent grade, Shanghai Yuanye Bio-Technology Co., Ltd., Shanghai, China) 14, 16, and 18 d after the plants were transferred into the greenhouse.
- (3)
- GA treatment group: all buds were supplied with GA3 (600 mg L−1, analytical grade, Beijing Chemical Reagent Co., Ltd., Beijing, China) at the same time as the 5-azaC treatment group.
- (4)
- A + G treatment group: all buds were supplied with 5-azaC and GA3 at the same concentrations and times described above, and the processing interval was 2 h with 5-azaC as the first treatment.
5.2. Observation of Morphological Changes
5.3. Measurement of Chlorophyll, GA Contents, and Photosynthetic Gas Exchange
5.4. RNA Isolation and RT-qPCR Testing
5.5. Measurement of the DNA Methylation Level
5.6. Data Analysis and cis-Element Prediction
5.7. Transcriptome Sequencing and Bioinformatics Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Xue, Y.Q.; Liu, R.; Xue, J.Q.; Wang, S.L.; Zhang, X.X. Genetic diversity and relatedness analysis of nine wild species of tree peony based on simple sequence repeats markers. Hortic. Plant J. 2021, 7, 579–588. [Google Scholar] [CrossRef]
- Li, J.; Zhang, X.; Zhao, X. Tree Peony of China; Encyclopedia of China Publishing House: Beijing, China, 2011. [Google Scholar]
- Xin, H.; Zhang, Y.X.; Wang, X.T.; Liu, C.Y.; Feng, W.R.; Gai, S.P. Morphological, anatomical and DNA methylation changes of tree peony buds during chilling induced dormancy release. Plant Physiol. Biochem. 2019, 144, 64–72. [Google Scholar] [CrossRef] [PubMed]
- Lange, M.J.P.; Lange, T. Gibberellin biosynthesis and the regulation of plant development. Plant Biol. 2006, 8, 281–290. [Google Scholar] [CrossRef] [PubMed]
- Wu, K.; Xu, H.; Gao, X.H.; Fu, X.D. New insights into gibberellin signaling in regulating plant growth-metabolic coordination. Curr. Opin. Plant Biol. 2021, 63, 102074. [Google Scholar] [CrossRef] [PubMed]
- Hedden, P.; Sponsel, V. A Century of Gibberellin Research. J. Plant Growth Regul. 2015, 34, 740–760. [Google Scholar] [CrossRef] [Green Version]
- Katyayini, N.U.; Rinne, P.L.H.; Tarkowska, D.; Strnad, M.; van der Schoot, C. Dual Role of Gibberellin in Perennial Shoot Branching: Inhibition and Activation. Front. Plant Sci. 2020, 11, 736. [Google Scholar] [CrossRef] [PubMed]
- Auge, G.A.; Penfield, S.; Donohue, K. Pleiotropy in developmental regulation by flowering-pathway genes: Is it an evolutionary constraint? New Phytol. 2019, 224, 55–70. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Andres, F.; Coupland, G. The genetic basis of flowering responses to seasonal cues. Nat. Rev. Genet. 2012, 13, 627–639. [Google Scholar] [CrossRef]
- Samach, A.; Onouchi, H.; Gold, S.E.; Ditta, G.S.; Schwarz-Sommer, Z.; Yanofsky, M.F.; Coupland, G. Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis. Science 2000, 288, 1613–1616. [Google Scholar] [CrossRef] [Green Version]
- Lee, J.; Oh, M.; Park, H.; Lee, I. SOC1 translocated to the nucleus by interaction with AGL24 directly regulates LEAFY. Plant J. 2008, 55, 832–843. [Google Scholar] [CrossRef]
- Wellmer, F.; Riechmann, J.L. Gene networks controlling the initiation of flower development. Trends Genet. 2010, 26, 519–527. [Google Scholar] [CrossRef] [PubMed]
- Conti, L. Hormonal control of the floral transition: Can one catch them all? Dev. Biol. 2017, 430, 288–301. [Google Scholar] [CrossRef] [PubMed]
- Bartels, A.; Han, Q.; Nair, P.; Stacey, L.; Gaynier, H.; Mosley, M.; Huang, Q.Q.; Pearson, J.K.; Hsieh, T.-F.; An, Y.-Q.C.; et al. Dynamic DNA Methylation in Plant Growth and Development. Int. J. Mol. Sci. 2018, 19, 2144. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Huang, H.; Liu, R.E.; Niu, Q.F.; Tang, K.; Zhang, B.; Zhang, H.; Chen, K.S.; Zhu, J.K.; Lang, Z.B. Global increase in DNA methylation during orange fruit development and ripening. Proc. Natl. Acad. Sci. USA 2019, 116, 1430–1436. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Meng, F.R.; Li, Y.C.; Yin, J.; Liu, H.; Chen, X.J.; Ni, Z.F.; Sun, Q.X. Analysis of DNA methylation during the germination of wheat seeds. Biol. Plant. 2012, 56, 269–275. [Google Scholar] [CrossRef]
- Song, Q.X.; Zhang, T.Z.; Stelly, D.M.; Chen, Z.J. Epigenomic and functional analyses reveal roles of epialleles in the loss of photoperiod sensitivity during domestication of allotetraploid cottons. Genome Biol. 2017, 18, 99. [Google Scholar] [CrossRef]
- Yang, H.X.; Chang, F.; You, C.J.; Cui, J.; Zhu, G.F.; Wang, L.; Zheng, Y.; Qi, J.; Ma, H. Whole-genome DNA methylation patterns and complex associations with gene structure and expression during flower development in Arabidopsis. Plant J. 2015, 81, 268–281. [Google Scholar] [CrossRef] [PubMed]
- Kong, X.F.; Hong, Y.C.; Hsu, Y.F.; Huang, H.; Liu, X.; Song, Z.; Zhu, J.K. SIZ1-Mediated SUMOylation of ROS1 Enhances Its Stability and Positively Regulates Active DNA Demethylation in Arabidopsis. Mol. Plant 2020, 13, 1816–1824. [Google Scholar] [CrossRef]
- Law, J.A.; Jacobsen, S.E. Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat. Rev. Genet. 2010, 11, 204–220. [Google Scholar] [CrossRef]
- Burn, J.E.; Bagnall, D.J.; Metzger, J.D.; Dennis, E.S.; Peacock, W.J. DNA methylation, vernalization, and the initiation of flowering. Proc. Natl. Acad. Sci. USA 1993, 90, 287–291. [Google Scholar] [CrossRef] [Green Version]
- Zicola, J.; Liu, L.Y.; Tanzler, P.; Turck, F. Targeted DNA methylation represses two enhancers of FLOWERING LOCUS T in Arabidopsis thaliana. Nat. Plants 2019, 5, 300–307. [Google Scholar] [CrossRef] [PubMed]
- Finnegan, E.J.; Ford, B.; Wallace, X.; Pettolino, F.; Griffin, P.T.; Schmitz, R.J.; Zhang, P.; Barrero, J.M.; Hayden, M.J.; Boden, S.A.; et al. Zebularine treatment is associated with deletion of FT-B1 leading to an increase in spikelet number in bread wheat. Plant Cell Environ. 2018, 41, 1346–1360. [Google Scholar] [CrossRef] [PubMed]
- Zhang, J.Z.; Mei, L.; Liu, R.; Khan, M.R.G.; Hu, C.G. Possible Involvement of Locus-Specific Methylation on Expression Regulation of LEAFY Homologous Gene (CiLFY) during Precocious Trifoliate Orange Phase Change Process. PLoS ONE 2014, 9, e88558. [Google Scholar] [CrossRef] [PubMed]
- Zhou, P.; Zhang, X.; Fatima, M.; Ma, X.; Fang, H.; Yan, H.; Ming, R. DNA methylome and transcriptome landscapes revealed differential characteristics of dioecious flowers in papaya. Hortic. Res. 2020, 7, 81. [Google Scholar] [CrossRef]
- Guan, Y.R.; Xue, J.Q.; Xue, Y.Q.; Yang, R.W.; Wang, S.L.; Zhang, X.X. Effect of exogenous GA3 on flowering quality, endogenous hormones, and hormone- and flowering-associated gene expression in forcing-cultured tree peony (Paeonia suffruticosa). J. Integr. Agric. 2019, 18, 1295–1311. [Google Scholar] [CrossRef]
- Xue, J.Q.; Li, T.T.; Wang, S.L.; Xue, Y.Q.; Hu, F.R.; Zhang, X.X. Elucidation of the mechanism of reflowering in tree peony (Paeonia suffruticosa) ‘Zi Luo Lan’ by defoliation and gibberellic acid application. Plant Physiol. Biochem. 2018, 132, 571–578. [Google Scholar] [CrossRef]
- Kondo, H.; Shiraya, T.; Wada, K.C.; Takeno, K. Induction of flowering by DNA demethylation in Perilla frutescens and Silene armeria: Heritability of 5-azacytidine-induced effects and alteration of the DNA methylation state by photoperiodic conditions. Plant Sci. 2010, 178, 321–326. [Google Scholar] [CrossRef]
- Fleet, C.M.; Sun, T.P. A DELLAcate balance: The role of gibberellin in plant morphogenesis. Curr. Opin. Plant Biol. 2005, 8, 77–85. [Google Scholar] [CrossRef]
- Zhang, Y.X.; Yuan, Y.C.; Liu, Z.J.; Zhang, T.; Li, F.; Liu, C.Y.; Gai, S.P. GA3 is superior to GA4 in promoting bud endodormancy release in tree peony (Paeonia suffruticosa) and their potential working mechanism. BMC Plant Biol. 2021, 21, 323. [Google Scholar]
- Li, H.C.; Torres-Garcia, J.; Latrasse, D.; Benhamed, M.; Schilderink, S.; Zhou, W.K.; Kulikova, O.; Hirt, H.; Bisseling, T. Plant-Specific Histone Deacetylases HDT1/2 Regulate GIBBERELLIN 2-OXIDASE2 Expression to Control Arabidopsis Root Meristem Cell Number. Plant Cell 2017, 29, 2183–2196. [Google Scholar] [CrossRef] [Green Version]
- Spray, C.R.; Kobayashi, M.; Suzuki, Y. The dwarf-1 (dt) mutant of Zea mays blocks three steps in the gibberellin-biosynthetic pathway. Proc. Natl. Acad. Sci. USA 1996, 93, 10515–10518. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bohlenius, H.; Huang, T.; Charbonnel-Campaa, L.; Brunner, A.M.; Jansson, S.; Strauss, S.H.; Nilsson, O. CO/FT regulatory module controls timing of flowering and seasonal growth cessation in trees. Science 2006, 312, 1040–1043. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dong, B.; Deng, Y.; Wang, H.B.; Gao, R.; Stephen, G.K.; Chen, S.M.; Jiang, J.F.; Chen, F.D. Gibberellic Acid Signaling Is Required to Induce Flowering of Chrysanthemums Grown under Both Short and Long Days. Int. J. Mol. Sci. 2017, 18, 1259. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Orbovic, V.; Ravanfar, S.A.; Acanda, Y.; Narvaez, J.; Merritt, B.A.; Levy, A.; Lovatt, C.J. Stress-inducible Arabidopsis thaliana RD29A promoter constitutively drives Citrus sinensis APETALA1 and LEAFY expression and precocious flowering in transgenic Citrus spp. Transgenic Res. 2021, 30, 687–699. [Google Scholar] [CrossRef] [PubMed]
- Ai, Y.J.; Jing, S.L.; Cheng, Z.N.; Song, B.T.; Xie, C.H.; Liu, J.; Zhou, J. DNA methylation affects photoperiodic tuberization in potato (Solanum tuberosum L.) by mediating the expression of genes related to the photoperiod and GA pathways. Hortic. Res. 2021, 8, 181. [Google Scholar] [CrossRef] [PubMed]
- Zhou, S.L.; Li, X.; Liu, Q.; Zhao, Y.; Jiang, W.; Wu, A.Q.; Zhou, D.X. DNA demethylases remodel DNA methylation in rice gametes and zygote and are required for reproduction. Mol. Plant 2021, 14, 1569–1583. [Google Scholar] [CrossRef]
- Zhang, Y.X.; Si, F.H.; Wang, Y.Y.; Liu, C.Y.; Zhang, T.; Yuan, Y.C.; Gai, S.P. Application of 5-azacytidine induces DNA hypomethylation and accelerates dormancy release in buds of tree peony. Plant Physiol. Biochem. 2020, 147, 91–100. [Google Scholar] [CrossRef]
- Li, W.G.; Ma, Y.X.; Zheng, C.K.; Li, G. Variations of Cytosine Methylation Patterns between Staminate and Perfect Flowers within Andromonoecious Taihangia rupestris (Rosaceae) Revealed by Methylation-Sensitive Amplification Polymorphism. J. Plant Growth Regul. 2022, 41, 351–363. [Google Scholar] [CrossRef]
- Liu, Z.Y.; Shi, Y.T.; Xue, Y.Q.; Wang, X.P.; Huang, Z.; Xue, J.Q.; Zhang, X.X. Non-structural carbohydrates coordinate tree peony flowering both as energy substrates and as sugar signaling triggers, with the bracts playing an essential role. Plant Physiol. Biochem. 2021, 159, 80–88. [Google Scholar] [CrossRef]
- Wagner, T.A.; Kohorn, B.D. Wall-associated kinases are expressed throughout plant development and are required for cell expansion. Plant Cell 2001, 13, 303–318. [Google Scholar] [CrossRef] [Green Version]
- Kaye, Y.; Golani, Y.; Singer, Y.; Leshem, Y.; Cohen, G.; Ercetin, M.; Gillaspy, G.; Levine, A. Inositol Polyphosphate 5-Phosphatase7 Regulates the Production of Reactive Oxygen Species and Salt Tolerance in Arabidopsis. Plant Physiol. 2011, 157, 229–241. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Coon, M.J. Cytochrome P450: Nature’s most versatile biological catalyst. Annu. Rev. Pharmacol. Toxicol. 2005, 45, 1–25. [Google Scholar] [CrossRef] [PubMed]
- Chen, M.; Markham, J.E.; Dietrich, C.R.; Jaworski, J.G.; Cahoon, E.B. Sphingolipid long-chain base hydroxylation is important for growth and regulation of sphingolipid content and composition in Arabidopsis. Plant Cell 2008, 20, 1862–1878. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xue, J.Q.; Li, T.; Wang, S.L.; Xue, Y.Q.; Liu, X.W.; Zhang, X.X. Defoliation and gibberellin synergistically induce tree peony flowering with non-structural carbohydrates as intermedia. J. Plant Physiol. 2019, 233, 31–41. [Google Scholar] [CrossRef]
- Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef]
- Lescot, M.; Dehais, P.; Thijs, G.; Marchal, K.; Moreau, Y.; Van de Peer, Y.; Rouze, P.; Rombauts, S. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res. 2002, 30, 325–327. [Google Scholar] [CrossRef]
- Lv, S.; Cheng, S.; Wang, Z.; Li, S.; Jin, X.; Lan, L.; Yang, B.; Yu, K.; Ni, X.; Li, N.; et al. Draft genome of the famous ornamental plant Paeonia suffruticosa. Ecol. Evol. 2020, 10, 4518–4530. [Google Scholar] [CrossRef]
- Tian, T.; Liu, Y.; Yan, H.Y.; You, Q.; Yi, X.; Du, Z.; Xu, W.Y.; Su, Z. agriGO v2.0: A GO analysis toolkit for the agricultural community, 2017 update. Nucleic Acids Res. 2017, 45, W122–W129. [Google Scholar] [CrossRef] [Green Version]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Sun, K.; Xue, Y.; Prijic, Z.; Wang, S.; Markovic, T.; Tian, C.; Wang, Y.; Xue, J.; Zhang, X. DNA Demethylation Induces Tree Peony Flowering with a Low Deformity Rate Compared to Gibberellin by Inducing PsFT Expression under Forcing Culture Conditions. Int. J. Mol. Sci. 2022, 23, 6632. https://doi.org/10.3390/ijms23126632
Sun K, Xue Y, Prijic Z, Wang S, Markovic T, Tian C, Wang Y, Xue J, Zhang X. DNA Demethylation Induces Tree Peony Flowering with a Low Deformity Rate Compared to Gibberellin by Inducing PsFT Expression under Forcing Culture Conditions. International Journal of Molecular Sciences. 2022; 23(12):6632. https://doi.org/10.3390/ijms23126632
Chicago/Turabian StyleSun, Kairong, Yuqian Xue, Zeljana Prijic, Shunli Wang, Tatjana Markovic, Caihuan Tian, Yingying Wang, Jingqi Xue, and Xiuxin Zhang. 2022. "DNA Demethylation Induces Tree Peony Flowering with a Low Deformity Rate Compared to Gibberellin by Inducing PsFT Expression under Forcing Culture Conditions" International Journal of Molecular Sciences 23, no. 12: 6632. https://doi.org/10.3390/ijms23126632