Please wait a minute...
Journal of Integrative Agriculture  2026, Vol. 25 Issue (5): 1961-1970    DOI: 10.1016/j.jia.2025.12.067
Horticulture Advanced Online Publication | Current Issue | Archive | Adv Search |
BrRRG regulates leaf size by controlling cell cycle gene expression in Chinese cabbage

Qianyun Wang1, Rui Yang1, Daling Feng1, Yongcheng Li1, Rui Li1, Mengyang Liu1, Yiguo Hong1, Na Li2#, Wei Ma1#, Jianjun Zhao1#

1 State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Vegetable Germplasm Innovation and Utilization of Hebei/ Collaborative Innovation Center of Vegetable Industry in Hebei, College of Horticulture, Hebei Agricultural University, Baoding 071000, China

School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572025, China

 Highlights 
RETARDED ROOT GROWTH (BrRRG) is the gene responsible for defective leaf and root development in the mini24 mutant.
BrRRG was identified as a key regulator controlling leaf size via E2Fa and cell-cycle genes in Chinese cabbage.
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  大白菜(Brassica rapa subsp. pekinensis)是十字花科芸苔属的一种叶菜类蔬菜。可食用叶片的大小决定其经济价值和营养价值。然而,目前对大白菜叶片发育的认识有限。本研究通过对大白菜叶片发育缺陷突变体mini24的正向遗传分析,我们确定了调控大白菜叶片细胞分裂的BrRRG基因。进一步证明了BrRRG通过调控E2Fa转录因子和细胞周期相关基因的表达来影响白菜的叶片大小。此外,我们还发现BrRRG影响大白菜的生长素和细胞分裂素信号的响应。本研究揭示了BrRRG对地下部(根)和地上部(叶)发育调节的不同机制,BrRRG对大白菜叶片的生长至关重要。

Abstract  

Chinese cabbage (Brassica rapa subsp. pekinensis) is an important leafy vegetable in the Brassica genus of the Brassicaceae family.  The size of its edible leaves is an essential trait that determines its economic and nutritional values.  However, the current understanding of leaf development in Chinese cabbage remains limited.  Here, through forward genetic analysis of the mutant mini24 with defective leaf and root development, we identified the BrRRG gene, which regulates cell division in Chinese cabbage by map-based cloning.  We demonstrated that BrRRG impacts leaf size by regulating the expression of E2Fa transcription factors and cell cycle-related genes in Chinese cabbage.  Furthermore, BrRRG was found to modulate Chinese cabbage’s response to auxin (indole-3-acetic acid, IAA) and cytokinins hormones, revealing a distinct regulatory mechanism by which BrRRG coordinates the development of underground roots and aboveground leaves.  Thus, these results indicate that mutation in BrRRG impairs the growth and development of Chinese cabbage.

Keywords:  Chinese cabbage       BrRRG       cell division       cell cycle gene  
Received: 10 February 2025   Accepted: 29 August 2025 Online: 31 December 2025  
Fund: 

This work was supported by the National Natural Science Foundation of China (32172594, 32330096, and 32222076), and the Innovative Research Group Project of Hebei Natural Science Foundation, China (C2024204246).

About author:  #Correspondence Jianjun Zhao, E-mail: yyzjj@hebau.edu.cn; Wei Ma, E-mail: mawei0720@163.com; Na Li, E-mail: 13521251421@163.com

Cite this article: 

Qianyun Wang, Rui Yang, Daling Feng, Yongcheng Li, Rui Li, Mengyang Liu, Yiguo Hong, Na Li, Wei Ma, Jianjun Zhao. 2026. BrRRG regulates leaf size by controlling cell cycle gene expression in Chinese cabbage. Journal of Integrative Agriculture, 25(5): 1961-1970.

Aboul-Maaty N A F, Oraby H A S. 2019. Extraction of high-quality genomic DNA from different plant orders applying a modified CTAB-based method. Bulletin of the National Research Centre43, 25.

Andriankaja M, Dhondt S, De Bodt S, Vanhaeren H, Coppens F, De Milde L, Mühlenbock P, Skirycz A, Gonzalez N, Beemster G T, Inzé D. 2012. Exit from proliferation during leaf development in Arabidopsis thaliana: a not-so-gradual process. Developmental Cell22, 64–78.

Bogre L, Magyar Z, Lopez-Juez E. 2008. New clues to organ size control in plants. Genome Biology9, 226.

Breuer C, Ishida T, Sugimoto K. 2010. Developmental control of endocycles and cell growth in plants. Current Opinion in Plant Biology13, 654–660.

De Veylder L, Larkin J C, Schnittger A. 2011. Molecular control and function of endoreplication in development and physiology. Trends in Plant Science16, 624–634.

Dewitte W, Scofield S, Alcasabas A A, Maughan S C, Menges M, Braun N, Collins C, Nieuwland J, Prinsen E, Sundaresan V, Murray J A. 2007. Arabidopsis CYCD3 D-type cyclins link cell proliferation and endocycles and are rate-limiting for cytokinin responses. Proceedings of the National Academy of Sciences of the United States of America104, 14537–14542.

Ding Q, Cui B, Li J, Li H, Zhang Y, Lv X, Qiu N, Liu L, Wang F, Gao J. 2018. Ectopic expression of a Brassica rapa AINTEGUMENTA gene (BrANT-1) increases organ size and stomatal density in ArabidopsisScientific Reports8, 10528.

Elliott R C, Betzner A S, Huttner E, Oakes M P, Tucker W Q, Gerentes D, Perez P, Smyth D R. 1996. AINTEGUMENTA, an APETALA2-like gene of Arabidopsis with pleiotropic roles in ovule development and floral organ growth. The Plant Cell8, 155–168.

Fang W, Wang Z, Cui R, Li J, Li Y. 2012. Maternal control of seed size by EOD3/CYP78A6 in Arabidopsis thalianaPlant Journal70, 929–939.

Gonzalez N, Vanhaeren H, Inze D. 2012. Leaf size control: complex coordination of cell division and expansion. Trends in Plant Science17, 332–340.

Gu A X, Zhao J J, Li L M, Wang Y H, Zhao Y J, Hua F, Xu Y C, Shen S X. 2016. Analyses of phenotype and ARGOS and ASY1 expression in a ploidy Chinese cabbage series derived from one haploid. Breeding Science66, 161–168.

Guo X, Liang J, Lin R, Zhang L, Wu J, Wang X. 2021. Series-spatial transcriptome profiling of leafy head reveals the key transition leaves for head formation in Chinese cabbage. Frontiers in Plant Science12, 787826.

Gutierrez C, Ramirez-Parra E, Castellano M M, del Pozo J C. 2002. G(1) to S transition: more than a cell cycle engine switch. Current Opinion in Plant Biology5, 480–486.

He S S, Liu J, Xie Z, O’Neill D, Dotson S. 2004. Arabidopsis E2Fa plays a bimodal role in regulating cell division and cell growth. Plant Molecular Biology56, 171–184.

Herridge R P, Day R C, Macknight R C. 2014. The role of the MCM2-7 helicase complex during Arabidopsis seed development. Plant Molecular Biology86, 69–84.

Hong J K, Oh S W, Kim J H, Lee S B, Lee Y H. 2017. Overexpression of Brassica rapa GROWTH-REGULATING FACTOR genes in Arabidopsis thaliana increases organ growth by enhancing cell proliferation. Journal of Plant Biotechnology44, 271–286.

Hong S Y, Kim O K, Kim S G, Yang M S, Park C M. 2011. Nuclear import and DNA binding of the ZHD5 transcription factor is modulated by a competitive peptide inhibitor in ArabidopsisJournal of Biological Chemistry286, 1659–1668.

Hur Y S, Um J H, Kim S, Kim K, Park H J, Lim J S, Kim W Y, Jun S E, Yoon E K, Lim J, Ohme-Takagi M, Kim D, Park J, Kim G T, Cheon C I. 2015. Arabidopsis thaliana homeobox 12 (ATHB12), a homeodomain-leucine zipper protein, regulates leaf growth by promoting cell expansion and endoreduplication. New Phytologist205, 316–328.

Jeleńska J, Deckert J, Kondorosi E, Legocki A B. 2000. Mitotic B-type cyclins are differentially regulated by phytohormones and during yellow lupine nodule development. Plant Science150, 29–39.

John P C, Mews M, Moore R. 2001. Cyclin/Cdk complexes: their involvement in cell cycle progression and mitotic division. Protoplasma216, 119–142.

Johnson K, Lenhard M. 2011. Genetic control of plant organ growth. New Phytologist191, 319–333.

Kosugi S, Ohashi Y. 2003. Constitutive E2F expression in tobacco plants exhibits altered cell cycle control and morphological change in a cell type-specific manner. Plant Physiology132, 2012–2022.

Kwon S H, Lee B H, Kim E Y, Seo Y S, Lee S, Kim W T, Song J T, Kim J H. 2009. Overexpression of a Brassica rapa NGATHA gene in Arabidopsis thaliana negatively affects cell proliferation during lateral organ and root growth. Plant and Cell Physiology50, 2162–2173.

Lee B H, Ko J H, Lee S, Lee Y, Pak J H, Kim J H. 2009. The Arabidopsis GRF-INTERACTING FACTOR gene family performs an overlapping function in determining organ size as well as multiple developmental properties. Plant Physiology151, 655–668.

Limas J C, Cook J G. 2019. Preparation for DNA replication: the key to a successful S phase. FEBS Letters593, 2853–2867.

Lu D, Wang T, Persson S, Mueller-Roeber B, Schippers J H. 2014. Transcriptional control of ROS homeostasis by KUODA1 regulates cell expansion during leaf development. Nature Communications5, 3767.

Mironov V V, De Veylder L, Van Montagu M, Inze D. 1999. Cyclin-dependent kinases and cell division in plants the nexus. The Plant Cell11, 509–522.

Naouar N, Vandepoele K, Lammens T, Casneuf T, Zeller G, van Hummelen P, Weigel D, Rätsch G, Inzé D, Kuiper M, De Veylder L, Vuylsteke M. 2009. Quantitative RNA expression analysis with Affymetrix Tiling 1.0R arrays identifies new E2F target genes. Plant Journal57, 184–194.

Neelam K, Brown-Guedira G, Huang L. 2013. Development and validation of a breeder-friendly KASPar marker for wheat leaf rust resistance locus Lr21Molecular Breeding31, 233–237.

Ni D A, Sozzani R, Blanchet S, Domenichini S, Reuzeau C, Cella R, Bergounioux C, Raynaud C. 2009. The Arabidopsis MCM2 gene is essential to embryo development and its over-expression alters root meristem function. New Phytologist184, 311–322.

Nisa M, Eekhout T, Bergis C, Pedroza-Garcia J A, He X, Mazubert C, Vercauteren I, Cools T, Brik-Chaouche R, Drouin-Wahbi J, Chmaiss L, Latrasse D, Bergounioux C, Vandepoele K, Benhamed M, De Veylder L, Raynaud C. 2023. Distinctive and complementary roles of E2F transcription factors during plant replication stress responses. Molecular Plant16, 1269–1282.

Omidbakhshfard M A, Fujikura U, Olas J J, Xue G P, Balazadeh S, Mueller-Roeber B. 2018. GROWTH-REGULATING FACTOR 9 negatively regulates Arabidopsis leaf growth by controlling ORG3 and restricting cell proliferation in leaf primordia. PLoS Genetics14, e1007484.

Omidbakhshfard M A, Proost S, Fujikura U, Mueller-Roeber B. 2015. Growth-regulating factors (GRFs): A small transcription factor family with important functions in plant biology. Molecular Plant8, 998–1010.

Ramirez-Parra E, Fründt C, Gutierrez C. 2003. A genome-wide identification of E2F-regulated genes in ArabidopsisPlant Journal33, 801–811.

Ratnayeke N, Baris Y, Chung M, Yeeles J T P, Meyer T. 2023. CDT1 inhibits CMG helicase in early S phase to separate origin licensing from DNA synthesis. Molecular Cell83, 26–42.e13.

Reddy G V, Heisler M G, Ehrhardt D W, Meyerowitz E M. 2004. Real-time lineage analysis reveals oriented cell divisions associated with morphogenesis at the shoot apex of Arabidopsis thalianaDevelopment131, 4225–4237.

Shi J, Zhang Q, Yan X, Zhang D, Zhou Q, Shen Y, Anupol N, Wang X, Bao M, Larkin R M, Luo H, Ning G. 2021. A conservative pathway for coordination of cell wall biosynthesis and cell cycle progression in plants. Plant Journal106, 630–648.

Springer P S, Holding D R, Groover A, Yordan C, Martienssen R A. 2000. The essential Mcm7 protein PROLIFERA is localized to the nucleus of dividing cells during the G(1) phase and is required maternally for early Arabidopsis development. Development127, 1815–1822.

Stortenbeker N, Bemer M. 2019. The SAUR gene family: the plant’s toolbox for adaptation of growth and development. Journal of Experimental Botany1, 17–27.

Sun X, Li X, Lu Y, Wang S, Zhang X, Zhang K, Su X, Liu M, Feng D, Luo S, Gu A, Fu Y, Chen X, Xuan S, Wang Y, Xu D, Chen S, Ma W, Shen S, Cheng F, Zhao J. 2022. Construction of a high-density mutant population of Chinese cabbage facilitates the genetic dissection of agronomic traits. Molecular Plant15, 913–924.

Trimarchi J M, Lees J A. 2002. Sibling rivalry in the E2F family. Nature Reviews Molecular Cell Biology3, 11–20.

Tsukaya H. 2015. Yield increase: GRFs provide the key. Nature Plants, 2, 15210.

Vandepoele K, Vlieghe K, Florquin K, Hennig L, Beemster G T, Gruissem W, Van de Peer Y, Inze D, De Veylder L. 2005. Genome-wide identification of potential plant E2F target genes. Plant Physiology139, 316–328.

Vlieghe K, Boudolf V, Beemster G T, Maes S, Magyar Z, Atanassova A, de Almeida Engler J, De Groodt R, Inze D, De Veylder L. 2005. The DP-E2F-like gene DEL1 controls the endocycle in Arabidopsis thalianaCurrent Biology15, 59–63.

Vlieghe K, Vuylsteke M, Florquin K, Rombauts S, Maes S, Ormenese S, Van Hummelen P, Van de Peer Y, Inze D, De Veylder L. 2003. Microarray analysis of E2Fa-DPa-overexpressing plants uncovers a cross-talking genetic network between DNA replication and nitrogen assimilation. Journal of Cell Science116, 4249–4259.

Wang B, Zhou X, Xu F, Gao J. 2010. Ectopic expression of a Chinese cabbage BrARGOS gene in Arabidopsis increases organ size. Transgenic Research19, 461–472.

Wang F, Zheng T, Wu G, Lang C, Hu Z, Shi J, Jin W, Chen J, Liu R. 2015. Overexpression of miR319a affects the balance between mitosis and endoreduplication in Arabidopsis leaves. Plant Molecular Biology Reporter33, 2006–2013.

Wang G, Kong H, Sun Y, Zhang X, Zhang W, Altman N, DePamphilis C W, Ma H. 2004. Genome-wide analysis of the cyclin family in Arabidopsis and comparative phylogenetic analysis of plant cyclin-like proteins. Plant Physiology135, 1084–1099.

Wang J, Jin D, Deng Z, Zheng L, Guo P, Ji Y, Song Z, Zeng H Y, Kinoshita T, Liao Z, Chen H, Deng X W, Wei N. 2025. The apoplastic pH is a key determinant in the hypocotyl growth response to auxin dosage and light. Nature Plants11, 279–294.

Wang Y, Huang S, Liu Z, Tang X, Feng H. 2018. Changes in endogenous phytohormones regulated by microRNA-target mRNAs contribute to the development of dwarf autotetraploid Chinese cabbage (Brassica rapa L. ssp. pekinensis). Molecular Genetics and Genomics293, 1535–1546.

Xu H, Lantzouni O, Bruggink T, Benjamins R, Lanfermeijer F, Denby K, Schwechheimer C, Bassel G W. 2020. A molecular signal integration network underpinning Arabidopsis seed germination. Current Biology30, 3703–3712.e3704.

Zhang G, Zhai N, Zhu M, Zheng K, Sang Y, Li X, Xu L. 2025. Cell wall remodeling during plant regeneration. Journal of Integrative Plant Biology67, 1060–1076.

Zhou X, Li Q, Chen X, Liu J, Zhang Q, Liu Y, Liu K, Xu J. 2011. The Arabidopsis RETARDED ROOT GROWTH gene encodes a mitochondria-localized protein that is required for cell division in the root meristem. Plant Physiology157, 1793–1804.


[1] Shan Wang, Kailin Shi, Yufan Xiao, Wei Ma, Yiguo Hong, Daling Feng, Jianjun Zhao. The circadian clock shapes diurnal gene expression patterns linked to glucose metabolic processes in Chinese cabbage[J]. >Journal of Integrative Agriculture, 2025, 24(6): 2155-2170.
[2] Xiaochun Wei, Yuanlin Zhang, Yanyan Zhao, Weiwei Chen, Ujjal Kumar Nath, Shuangjuan Yang, Henan Su, Zhiyong Wang, Wenjing Zhang, Baoming Tian, Fang Wei, Yuxiang Yuan, Xiaowei Zhang. Mitotic pollen abnormalities are linked to Ogura cytoplasmic male sterility in Chinese cabbage (Brassica rapa L. ssp. pekinensis)[J]. >Journal of Integrative Agriculture, 2025, 24(3): 1092-1107.
[3] Qi Zeng, Qingguo Sun, Xinru Hou, Lin Chen, Ruixing Zhang, Xue Bai, Xifan Liu, Xiaowu Wang, Lugang Zhang, Baohua Li. Comparative transcriptomic analysis of Chinese cabbage’s defense responses to Alternaria brassicae[J]. >Journal of Integrative Agriculture, 2025, 24(10): 3895-3908.
[4] Lin Chen, Chao Li, Jiahao Zhang, Zongrui Li, Qi Zeng, Qingguo Sun, Xiaowu Wang, Limin Zhao, Lugang Zhang, Baohua Li. Physiological and transcriptome analyses of Chinese cabbage in response to drought stress[J]. >Journal of Integrative Agriculture, 2024, 23(7): 2255-2269.
[5] Liping Song, Xia Li, Liguang Tang, Chuying Yu, Bincai Wang, Changbin Gao, Yanfeng Xie, Xueli Zhang, Junliang Wang, Chufa Lin, Aihua Wang.

Fine mapping and cloning of the sterility gene Bra2Ms in non-heading Chinese cabbage (Brassica rapa ssp. chinensis) [J]. >Journal of Integrative Agriculture, 2024, 23(4): 1195-1204.

[6] ZHANG Rui-xing, ZHANG Ni-nan, WANG Ya-xiu, Khan ABID, MA Shuai, BAI Xue, ZENG Qi, PAN Qi-ming, LI Bao-hua, ZHANG Lu-gang. Blue light induces leaf color change by modulating carotenoid metabolites in orange-head Chinese cabbage (Brassica rapa L. ssp. pekinensis)[J]. >Journal of Integrative Agriculture, 2023, 22(11): 3296-3311.
[7] SU He-nan, YUAN Yu-xiang, YANG Shuang-juan, WEI Xiao-chun, ZHAO Yan-yan, WANG Zhi-yong, QIN Liu-yue, YANG Zhi-yuan, NIU Liu-jing, LI Lin, ZHANG Xiao-wei. Comprehensive analysis of the full-length transcripts and alternative splicing involved in clubroot resistance in Chinese cabbage[J]. >Journal of Integrative Agriculture, 2023, 22(11): 3284-3295.
[8] TANG Zi-kai, SUN Man-yi, LI Jia-ming, SONG Bo-bo, LIU Yue-yuan, TIAN Yi-ke, WANG Cai-hong, WU Jun. Comparative transcriptome analysis provides insights into the mechanism of pear dwarfing[J]. >Journal of Integrative Agriculture, 2022, 21(7): 1952-1967.
[9] XU Yu-chao, HOU Xi-lin, XU Wei-wei, SHEN Lu-lu, Lü Shan-wu, ZHANG Shi-lin, HU Chun-mei. Isolation and characterization of an ERF-B3 gene associated with flower abnormalities in non-heading Chinese cabbage[J]. >Journal of Integrative Agriculture, 2016, 15(3): 528-536.
[10] TIAN Jia, ZENG Bin, LUO Shu-ping, LI Xiu-gen, WU Bin, LI Jiang. Cloning, localization and expression analysis of two fw2.2-like genes in small- and large-fruited pear species[J]. >Journal of Integrative Agriculture, 2016, 15(2): 282-294.
[11] WANG Li, GE Ting-ting, PENG Hai-tao, WANG Cheng, LIU Tong-kun, HOU Xi-lin. Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)[J]. >Journal of Integrative Agriculture, 2013, 12(12): 2149-2156.
No Suggested Reading articles found!