Chinese Journal OF Rice Science >
AP2 Transcription Factor OsERF34 Positively Regulates Seed Dormancy in Rice
Received date: 2025-04-24
Revised date: 2025-06-03
Online published: 2026-03-16
【Objective】 Weakened seed dormancy in rice leads to pre-harvest sprouting (PHS), which severely impacts grain yield and quality, making it crucial to investigate the genetic regulatory mechanisms underlying rice seed dormancy. 【Method】We generated loss-of-function mutants of the AP2/ERF transcription factor OsERF34 in the Zhonghua 11 (ZH11) background using CRISPR/Cas9 technology. 【Result】 The oserf34 mutants exhibited severe pre-harvest sprouting and significantly reduced seed dormancy. Transcriptome analysis revealed that differentially expressed genes (DEGs) between oserf34 and the wild type were significantly enriched in plant hormone signal transduction, ROS metabolism, and glycolytic pathways, suggesting that OsERF34 may maintain seed dormancy by regulating hormone biosynthesis/metabolism, redox homeostasis, and carbohydrate metabolism. Further investigation demonstrated that the oserf34 mutant seeds showed substantially increased levels of auxin (IAA), gibberellins (GA19/GA20), and salicylic acid (SA), along with decreased proline content and elevated hydrogen peroxide (H2O2) and superoxide anion level. Meanwhile, the activities of antioxidant enzymes (SOD, POD, CAT) were upregulated. Additionally, the mutant endosperm exhibited significantly elevated soluble sugar content and α-amylase activity, providing an energy foundation for dormancy release. This study demonstrates that OsERF34 positively regulates rice seed dormancy through its involvement in hormone biosynthesis/metabolism, redox homeostasis regulation, and carbohydrate metabolism. 【Conclusion】 These findings provide novel theoretical insights and genetic resources for deciphering the molecular mechanisms of seed dormancy and germination, as well as for developing PHS-resistant rice varieties.
DU Zhimin, JIA Yinan, DUAN Yingqing, CAO Ni, MA Liuyang, DONG Xinli, XU Hai, JIAO Guiai, TANG Shaoqing, HU Peisong . AP2 Transcription Factor OsERF34 Positively Regulates Seed Dormancy in Rice[J]. Chinese Journal OF Rice Science, 2026 , 40(2) : 196 -209 . DOI: 10.16819/j.1001-7216.2026.250411
| [1] | Muthayya S, Sugimoto J D, Montgomery S, Maberly G F. An overview of global rice production, supply, trade, and consumption[J]. Annals of the New York Academy of Sciences, 2014, 1324: 7-14. |
| [2] | 储零逸. AP2/ERF转录因子OsERF44调控水稻穗发芽的效应及其机制研究[D]. 扬州: 扬州大学, 2021. |
| Chu L Y. Effect and mechanism of AP2/ERF transcription factor OsERF44 on panicle sprouting in rice[D]. Yangzhou: Yangzhou University, 2021. (in Chinese with English abstract) | |
| [3] | 左志梅, 赵凯, 朱海山. 外源ABA对不同品种番茄种子萌发的影响[J]. 长江蔬菜, 2014(18): 35-38. |
| Zuo Z M, Zhao K, Zhu H S. Effects of exogenous abscisic acid on germination of different tomato cultivar seeds[J]. Journal of Changjiang Vegetables, 2014(18): 35-38. (in Chinese with English abstract) | |
| [4] | Nonogaki H, Bassel G W, Bewley J D. Germination: Still a mystery[J]. Plant Science, 2010, 179(6): 574-581. |
| [5] | Bewley J D. Seed germination and dormancy[J]. The Plant Cell, 1997: 1055-1066. |
| [6] | Graeber K, Nakabayashi K, Miatton E, Leubner-Metzger G, Soppe W J J. Molecular mechanisms of seed dormancy[J]. Plant, Cell & Environment, 2012, 35(10): 1769-1786. |
| [7] | Nonogaki H. Seed dormancy and germination emerging mechanisms and new hypotheses[J]. Frontiers in Plant Science, 2014, 5: 233. |
| [8] | Finkelstein R, Reeves W, Ariizumi T, Steber C. Molecular aspects of seed dormancy[J]. Annual Review of Plant Biology, 2008, 59: 387-415. |
| [9] | Sutoh K, Washio K, Imai R, Wada M, Nakai T, Yamauchi D. An N-terminal region of a Myb-like protein is involved in its intracellular localization and activation of a gibberellin-inducible proteinase gene in germinated rice seeds[J]. Bioscience, Biotechnology, and Biochemistry, 2015, 79(5): 747-759. |
| [10] | Finch-Savage W E, Leubner-Metzger G. Seed dormancy and the control of germination[J]. New Phytologist, 2006, 171(3): 501-523. |
| [11] | Del Bel Z, Andrade A, Lindström L, Alvarez D, Vigliocco A, Alemano S. The role of the sunflower seed coat and endosperm in the control of seed dormancy and germination: Phytohormone profile and their interaction with seed tissues[J]. Plant Growth Regulation, 2024, 102(1): 51-64. |
| [12] | 陈兵先, 张琪, 戴彰言, 周旭, 刘军. 水杨酸引发提高低温下水稻种子萌发活力的生理与分子效应[J]. 中国农业科学, 2024, 57(7): 1220-1236. |
| Chen B X, Zhang Q, Dai Z Y, Zhou X, Liu J. Physiological and molecular effects of salicylic acid on rice seed germination at low temperature[J]. Scientia Agricultura Sinica, 2024, 57(7): 1220-1236. (in Chinese with English abstract) | |
| [13] | Nguyen T N, Tuan P A, Ayele B T. Jasmonate regulates seed dormancy in wheat via modulating the balance between gibberellin and abscisic acid[J]. Journal of Experimental Botany, 2022, 73(8): 2434-2453. |
| [14] | Shu K, Zhang H, Wang S, Chen M, Wu Y, Tang S, Liu C, Feng Y, Cao X, Xie Q. ABI4 regulates primary seed dormancy by regulating the biogenesis of abscisic acid and gibberellins in Arabidopsis[J]. PLoS Genetics, 2013, 9(6): e1003577. |
| [15] | Sun M, Shen Y, Chen Y, Wang Y, Cai X, Yang J, Jia B, Dong W, Chen X, Sun X. Osa-miR1320 targets the ERF transcription factor OsERF096 to regulate cold tolerance via JA-mediated signaling[J]. Plant Physiology, 2022, 189(4): 2500-2516. |
| [16] | To A, Valon C, Savino G, Guilleminot J, Devic M, Giraudat J, Parcy F. A network of local and redundant gene regulation governs Arabidopsis seed maturation[J]. The Plant Cell, 2006, 18(7): 1642-1651. |
| [17] | Xie Z, Zhang Z L, Hanzlik S, Cook E, Shen Q J. Salicylic acid inhibits gibberellin-induced alpha-amylase expression and seed germination via a pathway involving an abscisic-acid-inducible WRKY gene[J]. Plant Molecular Biology, 2007, 64(3): 293-303. |
| [18] | Anand A, Kumari A, Thakur M, Koul A. Hydrogen peroxide signaling integrates with phytohormones during the germination of magnetoprimed tomato seeds[J]. Scientific Reports, 2019, 9(1): 8814. |
| [19] | Guha T, Das H, Mukherjee A, Kundu R. Elucidating ROS signaling networks and physiological changes involved in nanoscale zero valent iron primed rice seed germination sensu stricto[J]. Free Radical Biology & Medicine, 2021, 171: 11-25. |
| [20] | Howard T P, Fahy B, Craggs A, Mumford R, Leigh F, Howell P, Greenland A, Smith A M. Barley mutants with low rates of endosperm starch synthesis have low grain dormancy and high susceptibility to preharvest sprouting[J]. The New Phytologist, 2012, 194(1): 158-167. |
| [21] | Li W Y, Chen B X, Chen Z J, Gao Y T, Chen Z, Liu J. Reactive oxygen species generated by NADPH oxidases promote radicle protrusion and root elongation during rice seed germination[J]. International Journal of Molecular Sciences, 2017, 18(1): 110. |
| [22] | 邓本良. 活性氧调控作物种子发芽机理的研究进展[J]. 安徽农业科学, 2020, 48(13): 21-22, 26. |
| Deng B L. Research advances of reactive oxygen species in regulating crop seed germination[J]. Journal of Anhui Agricultural Sciences, 2020, 48(13): 21-22, 26. (in Chinese with English abstract) | |
| [23] | Xiong M, Yu J, Wang J, Gao Q, Huang L, Chen C, Zhang C, Fan X, Zhao D, Liu Q Q, Li Q F. Brassinosteroids regulate rice seed germination through the BZR1-RAmy3D transcriptional module[J]. Plant Physiology, 2022, 189(1): 402-418. |
| [24] | Du L, Xu F, Fang J, Gao S, Tang J, Fang S, Wang H, Tong H, Zhang F, Chu J, Wang G, Chu C. Endosperm sugar accumulation caused by mutation of PHS8/ISA1 leads to pre-harvest sprouting in rice[J]. The Plant Journal, 2018, 95(3): 545-556. |
| [25] | Zhao J, Wang J, Liu J, Zhang P, Kudoyarova G, Liu C J, Zhang K. Spatially distributed cytokinins: Metabolism, signaling, and transport[J]. Plant Communications, 2024, 5(7): 100936. |
| [26] | Faix B, Radchuk V, Nerlich A, Hümmer C, Radchuk R, Neil Emery R J, Keller H, Götz K P, Weschke W, Geigenberger P, Weber H. Barley grains, deficient in cytosolic small subunit of ADP-glucose pyrophosphorylase, reveal coordinate adjustment of C: N metabolism mediated by an overlapping metabolic- hormonal control[J]. The Plant Journal, 2012, 69(6): 1077-1093. |
| [27] | Sugimoto K, Takeuchi Y, Ebana K, Miyao A, Hirochika H, Hara N, Ishiyama K, Kobayashi M, Ban Y, Hattori T, Yano M. Molecular cloning of Sdr4, a regulator involved in seed dormancy and domestication of rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2010, 107(13): 5792-5797. |
| [28] | Zhao B, Zhang H, Chen T, Ding L, Zhang L, Ding X, Zhang J, Qian Q, Xiang Y. Sdr4 dominates pre-harvest sprouting and facilitates adaptation to local climatic condition in Asian cultivated rice[J]. Journal of Integrative Plant Biology, 2022, 64(6): 1246-1263. |
| [29] | Song S, Wang G, Wu H, Fan X, Liang L, Zhao H, Li S, Hu Y, Liu H, Ayaad M, Xing Y. OsMFT2 is involved in the regulation of ABA signaling-mediated seed germination through interacting with OsbZIP23/66/72 in rice[J]. The Plant Journal, 2020, 103(2): 532-546. |
| [30] | Shen J, Zhang L, Wang H, Guo J, Li Y, Tan Y, Shu Q, Qian Q, Yu H, Chen Y, Song S. The phosphatidylethanolamine-binding proteins OsMFT1 and OsMFT2 regulate seed dormancy in rice[J]. The Plant Cell, 2024, 36(9): 3857-3874. |
| [31] | Xu F, Tang J, Wang S, Cheng X, Wang H, Ou S, Gao S, Li B, Qian Y, Gao C, Chu C. Antagonistic control of seed dormancy in rice by two bHLH transcription factors[J]. Nature Genetics, 2022, 54(12): 1972-1982. |
| [32] | Fu F F, Xue H W. Coexpression analysis identifies Rice Starch Regulator1, a rice AP2/EREBP family transcription factor, as a novel rice starch biosynthesis regulator[J]. Plant Physiology, 2010, 154(2): 927-938. |
| [33] | Gu C, Guo Z H, Hao P P, Wang G M, Jin Z M. Multiple regulatory roles of AP2/ERF transcription factor in angiosperm[J]. Botanical Studies, 2017, 58(1): 6. |
| [34] | Ma Z, Hu L, Jiang W. Understanding AP2/ERF transcription factor responses and tolerance to various abiotic stresses in plants: A comprehensive review[J]. International Journal of Molecular Sciences, 2024, 25(2): 893. |
| [35] | Ma Z, Wu T, Huang K, Jin Y M, Li Z, Chen M, Yun S, Zhang H, Yang X, Chen H, Bai H, Du L, Ju S, Guo L, Bian M, Hu L, Du X. A novel AP2/ERF transcription factor, OsRPH1, negatively regulates plant height in rice[J]. Frontiers in Plant Science, 2020, 11: 709. |
| [36] | Yaish M W, El-Kereamy A, Zhu T, Beatty P H, Good A G, Bi Y M, Rothstein S J. The APETALA-2-like transcription factor OsAP2-39 controls key interactions between abscisic acid and gibberellin in rice[J]. PLoS Genetics, 2010, 6(9): e1001098. |
| [37] | Qi W, Sun F, Wang Q, Chen M, Huang Y, Feng Y Q, Luo X, Yang J. Rice ethylene-response AP2/ERF factor OsEATB restricts internode elongation by down-regulating a gibberellin biosynthetic gene[J]. Plant Physiology, 2011, 157(1): 216-228. |
| [38] | Huang K, Lu F, Chen P, Jiao G, Lin H, Zhang J, Zhao S, Cao R, Shao G, Sheng Z, Hu S, Tang S, Hu P, Wei X. A large-scale gene regulatory network for rice endosperm starch biosynthesis and its application in genetic improvement of rice quality[J]. Plant Biotechnology Journal, 2025, 23(7): 2583-2594. |
| [39] | Huijser C, Kortstee A, Pego J, Weisbeek P, Wisman E, Smeekens S. The Arabidopsis sucrose uncoupled-6 gene is identical to ABSCISIC acid insensitive-4: Involvement of abscisic acid in sugar responses[J]. The Plant Journal, 2000, 23(5): 577-585. |
| [40] | Wang C, Wang H, Zhang J, Chen S. A seed-specific AP2-domain transcription factor from soybean plays a certain role in regulation of seed germination[J]. Science in China Series C: Life Sciences, 2008, 51(4): 336-345. |
| [41] | Arenas-Huertero F, Arroyo A, Zhou L, Sheen J, León P. Analysis of Arabidopsis glucose insensitive mutants, gin5 and gin6, reveals a central role of the plant hormone ABA in the regulation of plant vegetative development by sugar[J]. Genes & Development, 2000, 14(16): 2085-2096. |
| [42] | Shu K, Chen Q, Wu Y, Liu R, Zhang H, Wang P, Li Y, Wang S, Tang S, Liu C, Yang W, Cao X, Serino G, Xie Q. ABI4 mediates antagonistic effects of abscisic acid and gibberellins at transcript and protein levels[J]. The Plant Journal, 2016, 85(3): 348-361. |
| [43] | Tuan P A, Kumar R, Rehal P K, Toora P K, Ayele B T. Molecular mechanisms underlying abscisic acid/ gibberellin balance in the control of seed dormancy and germination in cereals[J]. Frontiers in Plant Science, 2018, 9: 668. |
| [44] | 刘建利. 7种激素对蒺藜状苜蓿种子发芽的影响[J]. 干旱地区农业研究, 2010, 28(2): 113-117. |
| Liu J L. Effects of 7 plant growth regulators on seed germination of Medicago truncatula[J]. Agricultural Research in the Arid Areas, 2010, 28(2): 113-117. (in Chinese with English abstract) | |
| [45] | Brady S M, Sarkar S F, Bonetta D, McCourt P. The abscisic acid insensitive 3 (ABI3) gene is modulated by farnesylation and is involved in auxin signaling and lateral root development in Arabidopsis[J]. The Plant Journal, 2003, 34(1): 67-75. |
| [46] | Liu P P, Montgomery T A, Fahlgren N, Kasschau K D, Nonogaki H, Carrington J C. Repression of AUXIN RESPONSE FACTOR10 by microRNA160 is critical for seed germination and post-germination stages[J]. The Plant Journal, 2007, 52(1): 133-146. |
| [47] | Ramaih S, Guedira M, Paulsen G M. Relationship of indoleacetic acid and tryptophan to dormancy and preharvest sprouting of wheat[J]. Functional Plant Biology, 2003, 30(9): 939-945. |
| [48] | Liu X, Zhang H, Zhao Y, Feng Z, Li Q, Yang H Q, Luan S, Li J, He Z H. Auxin controls seed dormancy through stimulation of abscisic acid signaling by inducing ARF-mediated ABI3 activation in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110(38): 15485-15490. |
| [49] | Mallory A C, Bartel D P, Bartel B. microRNA-directed regulation of Arabidopsis AUXIN RESPONSE FACTOR17 is essential for proper development and modulates expression of early auxin response genes[J]. The Plant Cell, 2005, 17(5): 1360-1375. |
| [50] | Wang J W, Wang L J, Mao Y B, Cai W J, Xue H W, Chen X Y. Control of root cap formation by microRNA-targeted auxin response factors in Arabidopsis[J]. The Plant Cell, 2005, 17(8): 2204-2216. |
| [51] | Chapman E J, Estelle M. Mechanism of auxin-regulated gene expression in plants[J]. Annual Review of Genetics, 2009, 43: 265-285. |
| [52] | Ogawa M, Hanada A, Yamauchi Y, Kuwahara A, Kamiya Y, Yamaguchi S. Gibberellin biosynthesis and response during Arabidopsis seed germination[J]. The Plant Cell, 2003, 15(7): 1591-1604. |
| [53] | Kurepa J, Li Y, Smalle J A. Cytokinin signaling stabilizes the response activator ARR1[J]. The Plant Journal, 2014, 78(1): 157-168. |
| [54] | Zhao Z, Andersen S U, Ljung K, Dolezal K, Miotk A, Schultheiss S J, Lohmann J U. Hormonal control of the shoot stem-cell niche[J]. Nature, 2010, 465(7301): 1089-1092. |
| [55] | Nie L, Song S, Yin Q, Zhao T, Liu H, He A, Wang W. Enhancement in seed priming-induced starch degradation of rice seed under chilling stress via GA-mediated α-amylase expression[J]. Rice, 2022, 15(1): 19. |
| [56] | Bailly C, El-Maarouf-Bouteau H, Corbineau F. From intracellular signaling networks to cell death: The dual role of reactive oxygen species in seed physiology[J]. Comptes Rendus Biologies, 2008, 331(10): 806-814. |
| [57] | El-Maarouf-Bouteau H, Bailly C. Oxidative signaling in seed germination and dormancy[J]. Plant Signaling & Behavior, 2008, 3(3): 175-182. |
| [58] | Foyer C H, Noctor G. Oxidant and antioxidant signalling in plants: A re-evaluation of the concept of oxidative stress in a physiological context[J]. Plant, Cell & Environment, 2005, 28(8): 1056-1071. |
| [59] | Gill S S, Tuteja N. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants[J]. Plant Physiology and Biochemistry, 2010, 48(12): 909-930. |
| [60] | Zavaliev R, Dong X. NPR1, a key immune regulator for plant survival under biotic and abiotic stresses[J]. Molecular Cell, 2024, 84(1): 131-141. |
| [61] | Gao W, Liu Y, Huang J, Chen Y, Chen C, Lu L, Zhao H, Men S, Zhang X. MES7 modulates seed germination via regulating salicylic acid content in Arabidopsis[J]. Plants, 2021, 10(5): 903. |
| [62] | Wani A B, Chadar H, Wani A H, Singh S, Upadhyay N. Salicylic acid to decrease plant stress[J]. Environmental Chemistry Letters, 2017, 15(1): 101-123. |
| [63] | Mishra P, Dubey R S. Effect of aluminium on metabolism of starch and sugars in growing rice seedlings[J]. Acta Physiologiae Plantarum, 2008, 30(3): 265-275. |
| [64] | Gómez-Cadenas A, Zentella R, Walker-Simmons M K, Ho T H. Gibberellin/abscisic acid antagonism in barley aleurone cells: Site of action of the protein kinase PKABA1 in relation to gibberellin signaling molecules[J]. The Plant Cell, 2001, 13(3): 667-679. |
| [65] | Wang E, Xu X, Zhang L, Zhang H, Lin L, Wang Q, Li Q, Ge S, Lu B R, Wang W, He Z. Duplication and independent selection of cell-wall invertase genes GIF1 and OsCIN1 during rice evolution and domestication[J]. BMC Evolutionary Biology, 2010, 10: 108. |
| [66] | He Y, Zhu M, Li Z, Jiang S, He Z, Xu S, Chen X, Hu Z, Zhang Z. IPA1 negatively regulates early rice seedling development by interfering with starch metabolism via the GA and WRKY pathways[J]. International Journal of Molecular Sciences, 2021, 22(12): 6605. |
| [67] | Ishibashi Y, Kasa S, Sakamoto M, Aoki N, Kai K, Yuasa T, Hanada A, Yamaguchi S, Iwaya-Inoue M. A role for reactive oxygen species produced by NADPH oxidases in the embryo and aleurone cells in barley seed germination[J]. PLoS One, 2015, 10(11): e0143173. |
/
| 〈 |
|
〉 |