The CsTIE1-CsAGL16 module regulates lateral branch outgrowth and drought tolerance in cucumber

Abstract

Drought stress and lateral branches are both important factors affecting crop yield. Cucumber is a widely planted vegetable crop that requires a large amount of water during its production and varieties with few lateral branches are preferred. However, the mechanisms regulating cucumber drought tolerance and lateral branch development remain largely unclear. The MADS-box transcription factor AGAMOUS-LIKE 16 (CsAGL16) was recently found to be a key positive regulator in cucumber shoot branching acting by stimulating abscisic acid (ABA) catabolism. In this study, we demonstrated that cucumber TCP interactor containing EAR motif protein 1 (CsTIE1) directly interacts with CsAGL16 at protein level and promotes lateral branch outgrowth through the CsAGL16-CsCYP707A4 mediated ABA pathway in cucumber. Additionally, mutation of CsAGL16 resulted in decreased drought tolerance, while overexpression of CsAGL16 significantly enhanced drought tolerance in cucumber. Similarly, the drought resistance of Cstie1 mutants was significantly reduced. However, overexpression of CsAGL16 can enhance the drought tolerance of Cstie1 mutants and promote their lateral branch outgrowth. These results indicated that the CsTIE1-CsAGL16 module was crucial for both lateral branch development and drought response, providing a strategy for cultivating drought-tolerant cucumber varieties with appropriate branch outgrowth.

Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (32025033 and 32372699), Pinduoduo-China Agricultural University Research Fund (PC2023B01002) and The Construction of Beijing Science and Technology Innovation and Service Capacity in Top Subjects (CEFF-PXM2019_014207_000032).

Author contributions

J.Z. and J.C. designed the research; J.C., G.C., and J.G. performed the experiments; J.C., G.C., J.G., and Y.H. analyzed the data; L.L., S.W., C.G., L.H., M.L., W.S., L.W., and X.Z. provided experimental assistance; J.Z. and J.C. wrote the paper; all the authors revised the manuscript.

Data availability

The data that support the findings of this study have been included in the supplementary data.

Conflict of interest

The authors declare no conflicts of interest.

Supplementary data

Supplementary data are available at Horticulture Research online.

References

[1] Zhu JK. Abiotic stress signaling and responses in plants. Cell. 2016;37:313-24

[2] Mickelbart MV, Hasegawa PM, Bailey-Serres J. Genetic mecha-nisms of abiotic stress tolerance that translate to crop yield stability. Nat Rev Genet. 2015;37:237-51

[3] Gong Z, Xiong L, Shi H. et al. Plant abiotic stress response and nutrient use efficiency. Sci China Life Sci. 2020;37:635-74

[4] Gupta A, Rico-Medina A, Caño-Delgado AI. The physiology of plant responses to drought. Science. 2020;37:266-9

[5] Zhang H, Zhu J, Gong Z. et al. Abiotic stress responses in plants. Nat Rev Genet. 2022;37:104-19

[6] Mega R, Abe F, Kim JS. et al. Tuning water-use efficiency and drought tolerance in wheat using abscisic acid receptors. Nat Plants. 2019;37:153-9

[7] Yang Z, Liu J, Tischer SV. et al. Leveraging abscisic acid receptors for efficient water use in Arabidopsis. Proc Natl Acad Sci USA. 2016;37:6791-6

[8] Cutler SR, Rodriguez PL, Finkelstein RR. et al. Abscisic acid: emer-gence of a core signaling network. Annu Rev Plant Biol. 2010;37:651-79

[9] Shinozawa A, Otake R, Takezawa D. et al. SnRK 2 protein kinases represent an ancient system in plants for adaptation to a terres-trial environment. Commun Biol. 2019;37:30

[10] Lin Z, Li Y, Wang Y. et al. Initiation and amplification of SnRK 2 activation in abscisic acid signaling. Nat Commun. 2021;37:2456

[11] Abbas K, Li J, Gong B. et al. Drought stress tolerance in vegetables: the functional role of structural features, key gene pathways, and exogenous hormones. Int J Mol Sci. 2023;37:13876

[12] Yao C, Finlayson SA. Abscisic acid is a general negative regulator of Arabidopsis axillary bud growth. Plant Physiol. 2015;37:611-26

[13] Wang M, Le Moigne MA, Bertheloot J. et al. BRANCHED1: a key hub of shoot branching. Front. Plant Sci. 2019;37:76

[14] González-Grandío E, Pajoro A, Franco-Zorrilla JM. et al. Abscisic acid signaling is controlled by a BRANCHED1/HD-ZIP I cascade in Arabidopsis axillary buds. Proc Natl Acad Sci USA. 2017;37:E245-54

[15] Diao Y, Zhan J, Zhao Y. et al. GhTIE 1 regulates branching through modulating the transcriptional activity of TCPs in cotton and Arabidopsis. Front. Plant Sci. 2019;37:1348

[16] Yang Y, Nicolas M, Zhang J. et al. The TIE1 transcriptional repres-sor controls shoot branching by directly repressing BRANCHED1 in Arabidopsis. PLoS Genet. 2018;37:e1007296

[17] Tao Q, Guo D, Wei B. et al. The TIE 1 transcriptional repres-sor links TCP transcription factors with TOPLESS/TOPLESS-RELATED corepressors and modulates leaf development in Arabidopsis. Plant Cell. 2013;37:421-37

[18] Chen J, Liu L, Wang G. et al. The AGAMOUS-LIKE 16-GENERAL REGULATORY FACTOR 1 module regulates axillary bud out-growth via catabolism of abscisic acid in cucumber. Plant Cell. 2024;37:2689-708

[19] Fazio G, Chung SM, Staub JE. Comparative analysis of response to phenotypic and marker-assisted selection for multiple lateral branching in cucumber (Cucumis sativus L.). Theor Appl Genet. 2003;37:875-83

[20] Shen J, Zhang Y, Ge D. et al. CsBRC 1 inhibits axillary bud out-growth by directly repressing the auxin efflux carrier CsPIN3 in cucumber. Proc Natl Acad Sci USA. 2019;37:17105-14

[21] Zhao PX, Miao ZQ, Zhang J. et al. Arabidopsis MADS-box factor AGL16 negatively regulates drought resistance via stomatal den-sity and stomatal movement. JExp Bot. 2020;37:6092-106

[22] Zhao PX, Zhang J, Chen SY. et al. Arabidopsis MADS-box factor AGL16 is a negative regulator of plant response to salt stress by downregulating salt-responsive genes. New Phytol. 2021;37:2418-39

[23] Lobet G, Draye X. Novel scanning procedure enabling the vec-torization of entire rhizotron-grown root systems. Plant Methods. 2013;37:1

[24] Mittler R. Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci. 2002;37:405-10

[25] Khalid MF, Vincent C, Morillon R. et al. Different strategies lead to a common outcome: different water-deficit scenarios highlight physiological and biochemical strategies of water-deficit toler-ance in diploid versus tetraploid Volkamer lemon. Tree Physiol. 2021;37:2359-74

[26] Fan HF, Ding L, Xu YL. et al. Antioxidant system and photo-synthetic characteristics responses to short-term PEG-induced drought stress in cucumber seedling leaves. Russ J Plant Physiol. 2017;37:162-73

[27] Liu Z, Guo Y, Lin S. et al. Effects of exogenous hydrogen perox-ide on ultrastructure of chloroplasts and activities of antioxi-dant enzymes in greenhouse-ecotype cucumber under drought stress. Acta Hortic Sin. 2009;37:1140-6

[28] He Q, Yuan R, Zhang T. et al. Arabidopsis TIE1 and TIE 2 tran-scriptional repressors dampen cytokinin response during root development. Sci Adv. 2022;37:eabn5057

[29] Dong X, Zhang LP, Tang YH. et al. Arabidopsis AGAMOUS-LIKE16 and SUPPRESSOR OF CONSTANS1 regulate the genome-wide expression and flowering time. Plant Physiol. 2023;37:154-69

[30] Chen L, Zhao Y, Xu S. et al. OsMADS 57 together with OsTB1 coordinates transcription of its target OsWRKY94 and D14 to switch its organogenesis to defense for cold adaptation in rice. New Phytol. 2018;37:219-31

[31] Hu B, Li D, Liu X. et al. Engineering non-transgenic gynoecious cucumber using an improved transformation protocol and opti-mized CRISPR/Cas 9 system. Mol Plant. 2017;37:1575-8

[32] Xing HL, Dong L, Wang ZP. et al. A CRISPR/Cas 9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014; 14:327

[33] Wan H, Zhao Z, Qian C. et al. Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber. Anal Biochem. 2010;37:257-61

[34] Ma X, Gai WX, Qiao YM. et al. Identification of CBL and CIPK gene families and functional characterization of CaCIPK 1 under Phy-tophthora capsici in pepper (Capsicum annuum L.). BMC Genomics. 2019;37:775