CsRAXs negatively regulate leaf size and fruiting ability through auxin glycosylation in cucumber

Jiacai Chen, Liu Liu, Guangxin Chen, Shaoyun Wang, Ye Liu, Zeqin Zhang, Hongfei Li, Liming Wang, Zhaoyang Zhou, Jianyu Zhao, Xiaolan Zhang

PDF
Journal of Integrative Plant Biology ›› 2024, Vol. 66 ›› Issue (5) : 1024-1037. DOI: 10.1111/jipb.13655
Research Article

CsRAXs negatively regulate leaf size and fruiting ability through auxin glycosylation in cucumber

Author information +
History +

Abstract

Leaves are the main photosynthesis organ that directly determines crop yield and biomass. Dissecting the regulatory mechanism of leaf development is crucial for food security and ecosystem turn-over. Here, we identified the novel function of R2R3-MYB transcription factors CsRAXs in regulating cucumber leaf size and fruiting ability. Csrax5 single mutant exhibited enlarged leaf size and stem diameter, and Csrax1/2/5 triple mutant displayed further enlargement phenotype. Overexpression of CsRAX1 or CsRAX5 gave rise to smaller leaf and thinner stem. The fruiting ability of Csrax1/2/5 plants was significantly enhanced, while that of CsRAX5 overexpression lines was greatly weakened. Similarly, cell number and free auxin level were elevated in mutant plants while decreased in overexpression lines. Biochemical data indicated that CsRAX1/5 directly promoted the expression of auxin glucosyltransferase gene CsUGT74E2. Therefore, our data suggested that CsRAXs function as repressors for leaf size development by promoting auxin glycosylation to decrease free auxin level and cell division in cucumber. Our findings provide new gene targets for cucumber breeding with increased leaf size and crop yield.

Keywords

auxin glycosylation / CsRAXs / CsUGT74E2 / fruiting ability / leaf size

Cite this article

Download citation ▾
Jiacai Chen, Liu Liu, Guangxin Chen, Shaoyun Wang, Ye Liu, Zeqin Zhang, Hongfei Li, Liming Wang, Zhaoyang Zhou, Jianyu Zhao, Xiaolan Zhang. CsRAXs negatively regulate leaf size and fruiting ability through auxin glycosylation in cucumber. Journal of Integrative Plant Biology, 2024, 66(5): 1024‒1037 https://doi.org/10.1111/jipb.13655

References

[1]
Andriankaja,M., Dhondt, S., De Bodt,S., Vanhaeren,H., Coppens, F., De Milde,L., Mühlenbock,P., Skirycz, A., Gonzalez,N., Beemster,G.T.S., et al. (2012). Exit from proliferation during leaf development in Arabidopsis thaliana: a not-so-gradual process. Dev. Cell 22: 64–78.
[2]
Avramova,V., Sprangers, K., and Beemster,G.T.S. (2015). The maize leaf: Another perspective on growth regulation. Trends Plant Sci. 20: 787–797.
[3]
Aya,K., Hobo,T., Sato-Izawa,K., Ueguchi-Tanaka,M., Kitano, H., and Matsuoka,M. (2014). A novel AP2-type transcription factor, SMALL ORGAN SIZE1, controls organ size downstream of an auxin signaling pathway. Plant Cell Physiol. 55: 897–912.
[4]
Bancoş,S., Nomura, T., Sato,T., Molnár,G., Bishop, G.J., Koncz,C., Yokota,T., Nagy,F., and Szekeres,M. (2002). Regulation of transcript levels of the Arabidopsis cytochrome p450 genes involved in brassinosteroid biosynthesis. Plant Physiol. 130: 504–513.
[5]
Bartel,B., LeClere, S., Magidin,M., and Zolman,B.K. (2001). Inputs to the active indole-3-acetic acid pool: De novo synthesis, conjugate hydrolysis, and indole-3-butyric acid b-oxidation. J. Plant Growth Regul. 20: 198–216.
[6]
Baute,J., Polyn,S., De Block,J., Blomme, J., Van Lijsebettens,M., and Inzé,D. (2017). F-Box protein FBX92 affects leaf size in Arabidopsis thaliana. Plant Cell Physiol. 58: 962–975.
[7]
Beltramino,M., Ercoli, M.F., Debernardi,J.M., Goldy,C., Rojas,A.M.L., Nota,F., Alvarez, M.E., Vercruyssen,L., Inzé,D., Palatnik, J.F., et al. (2018). Robust increase of leaf size by Arabidopsis thaliana GRF3-like transcription factors under different growth conditions. Sci. Rep. 8: 13447.
[8]
Bernacchi,C.J., Portis, A.R., Nakano,H., von Caemmerer,S., and Long, S.P. (2002). Temperature response of mesophyll conductance. Implications for the determination of Rubisco enzyme kinetics and for limitations to photosynthesis in vivo. Plant Physiol. 130: 1992–1998.
[9]
Braun,N., Wyrzykowska, J., Muller,P., David,K., Couch,D., Perrot-Rechenmann,C., and Fleming,A.J. (2008). Conditional repression of AUXIN BINDING PROTEIN1 reveals that it coordinates cell division and cell expansion during postembryonic shoot development in Arabidopsis and tobacco. Plant Cell 20: 2746–2762.
[10]
Busch,B.L., Schmitz, G., Rossmann,S., Piron,F., Ding,J., Bendahmane,A., and Theres,K. (2011). Shoot branching and leaf dissection in tomato are regulated by homologous gene modules. Plant Cell 23: 3595–3609.
[11]
Cheng,Y., Dai,X., and Zhao,Y. (2007). Auxin synthesized by the YUCCA flavin monooxygenases is essential for embryogenesis and leaf formation in Arabidopsis. Plant Cell 19: 2430–2439.
[12]
Czesnick,H., and Lenhard, M. (2015). Size control in plants—Lessons from leaves and flowers. Cold Spring Harb. Perspect. Biol. 7: a019190.
[13]
del Pozo,J.C., Boniotti, M.B., and Gutierrez,C. (2002a). Arabidopsis E2Fc functions in cell division and is degraded by the ubiquitin-SCFAtSKP2 pathway in response to light. Plant Cell 14: 3057–3071.
[14]
del Pozo,J.C., Dharmasiri, S., Hellmann,H., Walker,L., Gray,W.M., and Estelle,M. (2002b). AXR1-ECR1-dependent conjugation of RUB1 to the Arabidopsis Cullin AtCUL1 is required for auxin response. Plant Cell 14: 421–433.
[15]
Donnelly,P.M., Bonetta, D., Tsukaya,H., Dengler,R.E., and Dengler, N.G. (1999). Cell cycling and cell enlargement in developing leaves of Arabidopsis. Dev. Biol. 215: 407–419.
[16]
Ecker,J.R. (1995). The ethylene signal transduction pathway in plants. Science 268: 667–675.
[17]
Ellis,C.M., Nagpal, P., Young,J.C., Hagen,G., Guilfoyle, T.J., and Reed,J.W. (2005). AUXIN RESPONSE FACTOR1 and AUXIN RESPONSE FACTOR2 regulate senescence and floral organ abscission in Arabidopsis thaliana. Development 132: 4563–4574.
[18]
Eloy,N.B., de Freitas Lima, M., Van Damme,D., Vanhaeren,H., Gonzalez, N., De Milde,L., Hemerly,A.S., Beemster, G.T., Inzé,D., and Ferreira,P.C. (2011). The APC/C subunit 10 plays an essential role in cell proliferation during leaf development. Plant J. 68: 351–363.
[19]
Eloy,N.B., Gonzalez, N., Van Leene,J., Maleux,K., Vanhaeren, H., De Milde,L., Dhondt,S., Vercruysse, L., Witters,E., Mercier,R., et al. (2012). SAMBA, a plant-specific anaphase-promoting complex/cyclosome regulator is involved in early development and A-type cyclin stabilization. Proc. Natl. Acad. Sci. U.S.A. 109: 13853–13858.
[20]
Faralli,M., and Lawson, T. (2020). Natural genetic variation in photosynthesis: An untapped resource to increase crop yield potential? Plant J. 101: 518–528.
[21]
Fujino,K., Matsuda, Y., Ozawa,K., Nishimura,T., Koshiba, T., Fraaije,M.W., and Sekiguchi,H. (2008). NARROW LEAF 7 controls leaf shape mediated by auxin in rice. Mol. Genet. Genomics 279: 499–507.
[22]
Gao,D., Zhang,C., Zhang,S., Hu, B., Wang,S., Zhang,Z., and Huang, S. (2017). Mutation in a novel gene SMALL AND CORDATE LEAF 1 affects leaf morphology in cucumber. J. Integr. Plant Biol. 59: 736–741.
[23]
González,N., and Inzé, D. (2015). Molecular systems governing leaf growth: from genes to networks. J. Exp. Bot. 66: 1045–1054.
[24]
Gonzalez,N., Vanhaeren, H., and Inzé,D. (2012). Leaf size control: Complex coordination of cell division and expansion. Trends Plant Sci. 17: 332–340.
[25]
Gray,W.M., del Pozo, J.C., Walker,L., Hobbie,L., Risseeuw, E., Banks,T., Crosby,W.L., Yang,M., Ma,H., and Estelle, M. (1999). Identification of an SCF ubiquitin-ligase complex required for auxin response in Arabidopsis thaliana. Genes Dev. 13: 1678–1691.
[26]
Hu,B., Li,D., Liu,X., Qi, J., Gao,D., Zhao,S., Huang,S., Sun,J., and Yang, L. (2017). Engineering non-transgenic gynoecious cucumber using an improved transformation protocol and optimized CRISPR/Cas9 system. Mol. Plant 10: 1575–1578.
[27]
Hu,Y., Xie,Q., and Chua,N.H. (2003). The Arabidopsis auxin-inducible gene ARGOS controls lateral organ size. Plant Cell 15: 1951–1961.
[28]
Jeifetz,D., David-Schwartz, R., Borovsky,Y., and Paran,I. (2011). CaBLIND regulates axillary meristem initiation and transition to flowering in pepper. Planta 234: 1227–1236.
[29]
Kalve,S., and De Vos, D. (2014). Beemster GTS: Leaf development: A cellular perspective. Front. Plant Sci. 5: 362.
[30]
Keller,T., Abbott, J., Moritz,T., and Doerner,P. (2006). Arabidopsis REGULATOR OF AXILLARY MERISTEMS1 controls a leaf axil stem cell niche and modulates vegetative development. Plant Cell 18: 598–611.
[31]
Kepinski,S. (2006). Integrating hormone signaling and patterning mechanisms in plant development. Curr. Opin. Plant Biol. 9: 28–34.
[32]
Kim,J.H., and Kende, H. (2004). A transcriptional coactivator, AtGIF1, is involved in regulating leaf growth and morphology in Arabidopsis. Proc. Natl. Acad. Sci. U.S.A. 101: 13374–13379.
[33]
Kim,J.H., and Tsukaya, H. (2015). Regulation of plant growth and development by the GROWTH-REGULATING FACTOR and GRF-INTERACTING FACTOR duo. J. Exp. Bot. 66: 6093–6107.
[34]
Kim,J.H., Choi,D., and Kende,H. (2003). The AtGRF family of putative transcription factors is involved in leaf and cotyledon growth in Arabidopsis. Plant J. 36: 94–104.
[35]
Leakey,A.D., Xu,F., Gillespie,K.M., McGrath,J.M., Ainsworth, E.A., and Ort,D.R. (2009). Genomic basis for stimulated respiration by plants growing under elevated carbon dioxide. Proc. Natl. Acad. Sci. U.S.A. 106: 3597–3602.
[36]
Leyser,O. (2001). Auxin. Curr. Biol. 11: R728.
[37]
Li,Q., Fu,C., Liang,C., Ni, X., Zhao,X., Chen,M., and Ou, L. (2022). Crop lodging and the roles of lignin, cellulose, and hemicellulose in lodging resistance. Agronomy 12: 1795.
[38]
Lincoln,C., Britton, J.H., and Estelle,M. (1990). Growth and development of the axr1 mutants of Arabidopsis. Plant Cell 2: 1071–1080.
[39]
Liu,X., Chen,J., and Zhang,X. (2021). Genetic regulation of shoot architecture in cucumber. Hortic. Res. 8: 143.
[40]
Lucas,W.J., Groover, A., Lichtenberger,R., Furuta,K., Yadav,S.R., Helariutta,Y., He,X.Q., Fukuda, H., Kang,J., Brady,S.M., et al. (2013). The plant vascular system: Evolution, development and functions. J. Integr. Plant Biol. 55: 294–388.
[41]
Ludwig-Müller,J., and Epstein, E. (1993). Indole-3-butyric acid in Arabidopsis thaliana - II. In vivo metabolism. Plant Growth Regul. 13: 189–195.
[42]
Müller,D., Schmitz, G., and Theres,K. (2006). Blind homologous R2R3 Myb genes control the pattern of lateral meristem initiation in Arabidopsis. Plant Cell 18: 586–597.
[43]
Nicotra,A.B., Cosgrove, M.J., Cowling,A., Schlichting,C.D., and Jones, C.S. (2007). Leaf shape linked to photosynthetic rates and temperature optima in South African Pelargonium species. Oecologia 154: 625–635.
[44]
Nicotra,A.B., Leigh,A., Boyce,C.K., Jones, C.S., Niklas,K.J., Royer,D.L., and Tsukaya, H. (2011). The evolution and functional significance of leaf shape in the angiosperms. Funct. Plant Biol. 38: 535–552.
[45]
Okushima,Y., Mitina, I., Quach,H.L., and Theologis,A. (2005). AUXIN RESPONSE FACTOR 2 (ARF2): A pleiotropic developmental regulator. Plant J. 43: 29–46.
[46]
Powell,A.E., and Lenhard, M. (2012). Control of organ size in plants. Curr. Biol. 22: R360–R367.
[47]
Qi,J., Liu,X., Shen,D., Miao, H., Xie,B., Li,X., Zeng,P., Wang,S., Shang, Y., Gu,X., et al. (2013). A genomic variation map provides insights into the genetic basis of cucumber domestication and diversity. Nat. Genet. 45: 1510–1515.
[48]
Rodriguez,R.E., Debernardi, J.M., and Palatnik,J.F. (2014). Morphogenesis of simple leaves: regulation of leaf size and shape. Wiley Interdiscip. Rev. Dev. Biol. 3: 41–57.
[49]
Rojas,C.A., Eloy,N.B., Lima,M.F., Rodrigues, R.L., Franco,L.O., Himanen,K., Beemster, G.T., Hemerly,A.S., and Ferreira,P.C. (2009). Overexpression of the Arabidopsis anaphase promoting complex subunit CDC27a increases growth rate and organ size. Plant Mol. Biol. 71: 307–318.
[50]
Schmitz,G., Tillmann, E., Carriero,F., Fiore,C., Cellini, F., and Theres,K. (2002). The tomato Blind gene encodes a MYB transcription factor that controls the formation of lateral meristems. Proc. Natl. Acad. Sci. U.S.A. 99: 1064–1069.
[51]
Schruff,M.C., Spielman, M., Tiwari,S., Adams,S., Fenby,N., and Scott,R.J. (2006). The AUXIN RESPONSE FACTOR 2 gene of Arabidopsis links auxin signalling, cell division, and the size of seeds and other organs. Development 133: 251–261.
[52]
Tamura,K., Stecher, G., Peterson,D., Filipski,A., and Kumar, S. (2013). MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30: 2725–2729.
[53]
Tax,F.E., and Durbak, A. (2006). Meristems in the movies: Live imaging as a tool for decoding intercellular signaling in shoot apical meristems. Plant Cell 18: 1331–1337.
[54]
Tobeña-Santamaria,R., Bliek,M., Ljung,K., Sandberg,G., Mol, J.N., Souer,E., and Koes,R. (2002). FLOOZY of petunia is a flavin mono-oxygenase-like protein required for the specification of leaf and flower architecture. Genes Dev. 16: 753–763.
[55]
Tognetti,V.B., Van Aken, O., Morreel,K., Vandenbroucke,K., van de Cotte, B., De Clercq,I., Chiwocha,S., Fenske, R., Prinsen,E., Boerjan,W., et al. (2010). Perturbation of indole-3-butyric acid homeostasis by the UDP-glucosyltransferase UGT74E2 modulates Arabidopsis architecture and water stress tolerance. Plant Cell 22: 2660–2679.
[56]
Tsukaya,H. (2005). Leaf shape: Genetic controls and environmental factors. Int. J. Dev. Biol. 49: 547–555.
[57]
Tsukaya,H. (2006). Mechanism of leaf-shape determination. Annu. Rev. Plant Biol. 57: 477–496.
[58]
Vodermaier,H.C. (2004). APC/C and SCF: Controlling each other and the cell cycle. Curr. Biol. 14: R787–R796.
[59]
Wang,Y., Li,B., Du,M., Eneji, A.E., Wang,B., Duan,L., Li,Z., and Tian,X. (2012). Mechanism of phytohormone involvement in feedback regulation of cotton leaf senescence induced by potassium deficiency. J. Exp. Bot. 63: 5887–5901.
[60]
Wang,Z., Li,N., Jiang,S., Gonzalez, N., Huang,X., Wang,Y., Inzé, D., and Li,Y. (2016). SCFSAP controls organ size by targeting PPD proteins for degradation in Arabidopsis thaliana. Nat. Commun. 7: 11192.
[61]
Wu,L., Zhang,D., Xue,M., Qian, J., He,Y., and Wang,S. (2014). Overexpression of the maize GRF10, an endogenous truncated growth-regulating factor protein, leads to reduction in leaf size and plant height. J. Integr. Plant Biol. 56: 1053–1063.
[62]
Xing,H.L., Dong,L., Wang,Z.P., Zhang, H.Y., Han,C.Y., Liu,B., Wang,X.C., and Chen,Q.J. (2014). A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 14: 327.
[63]
Yang,L., Liu,H., Zhao,J., Pan, Y., Cheng,S., Lietzow,C.D., Wen,C., Zhang,X., and Weng, Y. (2018). LITTLELEAF (LL) encodes a WD40 repeat domain-containing protein associated with organ size variation in cucumber. Plant J. 95: 834–847.
[64]
Yin,J., Niu,L., Li,Y., Song, X., Ottosen,C.O., Wu,Z., Jiang,F., and Zhou,R. (2023). The effects of waterlogging stress on plant morphology, leaf physiology and fruit yield in six tomato genotypes at anthesis stage. Veg. Res. 3: 31.
[65]
Yue,B., Xue,W.Y., Luo,L.J., and Xing, Y.Z. (2006). QTL analysis for flag leaf characteristics and their relationships with yield and yield traits in rice. Acta Genet. Sin. 33: 824–832.
[66]
Zhang,B., Ye,W., Ren,D., Tian, P., Peng,Y., Gao,Y., Ruan,B., Wang,L., Zhang, G., Guo,L., et al. (2015). Genetic analysis of flag leaf size and candidate genes determination of a major QTL for flag leaf width in rice. Rice 8: 39.
[67]
Zhao,Y., Christensen, S.K., Fankhauser,C., Cashman,J.R., Cohen,J.D., Weigel,D., and Chory, J. (2001). A role for flavin monooxygenase-like enzymes in auxin biosynthesis. Science 291: 306–309.
[68]
Zhong,R., and Ye, Z.H. (2001). Alteration of auxin polar transport in the Arabidopsis ifl1 mutants. Plant Physiol. 126: 549–563.
[69]
Zhu,X.G., Long,S.P., and Ort,D.R. (2010). Improving photosynthetic efficiency for greater yield. Annu. Rev. Plant Biol. 61: 235–261.
[70]
Zolman,B.K., Martinez, N., Millius,A., Adham,A.R., and Bartel, B. (2008). Identification and characterization of Arabidopsis indole-3-butyric acid response mutants defective in novel peroxisomal enzymes. Genetics 180: 237–251.
[71]
Zolman,B.K., Silva,I.D., and Bartel,B. (2001). The Arabidopsis pxa1 mutant is defective in an ATP-binding cassette transporter-like protein required for peroxisomal fatty acid beta-oxidation. Plant Physiol. 127: 1266–1278.

RIGHTS & PERMISSIONS

2024 2024 Institute of Botany, Chinese Academy of Sciences.
PDF

Accesses

Citations

Detail

Sections
Recommended

/