, Volume 249, Issue 5, pp 1391–1403 | Cite as

Brassinosteroids facilitate xylem differentiation and wood formation in tomato

  • Jinsu Lee
  • Seahee Han
  • Hwa-Yong Lee
  • Bomi Jeong
  • Tae-Young Heo
  • Tae Kyung Hyun
  • Kyunghwan Kim
  • Byoung Il Je
  • Horim Lee
  • Donghwan Shim
  • Soon Ju Park
  • Hojin RyuEmail author
Original Article


Main conclusion

BR signaling pathways facilitate xylem differentiation and wood formation by fine tuning SlBZR1/SlBZR2-mediated gene expression networks involved in plant secondary growth.

Brassinosteroid (BR) signaling and BR crosstalk with diverse signaling cues are involved in the pleiotropic regulation of plant growth and development. Recent studies reported the critical roles of BR biosynthesis and signaling in vascular bundle development and plant secondary growth; however, the molecular bases of these roles are unclear. Here, we performed comparative physiological and anatomical analyses of shoot morphological growth in a cultivated wild-type tomato (Solanum lycopersicum cv. BGA) and a BR biosynthetic mutant [Micro Tom (MT)]. We observed that the canonical BR signaling pathway was essential for xylem differentiation and sequential wood formation by facilitating plant secondary growth. The gradual retardation of xylem development phenotypes during shoot vegetative growth in the BR-deficient MT tomato mutant recovered completely in response to exogenous BR treatment or genetic complementation of the BR biosynthetic DWARF (D) gene. By contrast, overexpression of the tomato Glycogen synthase kinase 3 (SlGSK3) or CRISPR-Cas9 (CR)-mediated knockout of the tomato Brassinosteroid-insensitive 1 (SlBRI1) impaired BR signaling and resulted in severely defective xylem differentiation and secondary growth. Genetic modulation of the transcriptional activity of the tomato Brassinazole-resistant 1/2 (SlBZR1/SlBZR2) confirmed the positive roles of BR signaling pathways for xylem differentiation and secondary growth. Our data indicate that BR signaling pathways directly promote xylem differentiation and wood formation by canonical BR-activated SlBZR1/SlBZR2.





Micro Tom


Brassinazole-resistant 1/2


Brassinosteroid-insensitive 2


Glycogen synthase kinase 3


BRI1-EMS-suppressor 1


Secondary cell wall





This work was carried out with the support of the Basic Science Research Program through the National Research Foundation of Korea (2015R1A4A1041869), Korean Ministry of Science, ICT and Future Planning, and the Next-Generation BioGreen 21 Program (no. PJ01313601), Rural Development Administration, Republic of Korea. SJP was supported by a grant from the National Research Foundation (2017R1A4A1015594) funded by the Korean Ministry of Science, ICT and Future Planning.

Supplementary material

425_2019_3094_MOESM1_ESM.pdf (301.5 mb)
Supplementary material 1 (PDF 308710 kb)
425_2019_3094_MOESM2_ESM.xlsx (12 kb)
Supplementary material 2 (XLSX 11 kb)


  1. Bai M-Y, Shang J-X, Oh E, Fan M, Bai Y, Zentella R, T-P Sun, Wang Z-Y (2012) Brassinosteroid, gibberellin and phytochrome impinge on a common transcription module in Arabidopsis. Nat Cell Biol 14(8):810CrossRefPubMedPubMedCentralGoogle Scholar
  2. Bajwa V, Wang X, Blackburn RK, Goshe MB, Mitra SK, Williams EL, Bishop GJ, Krasnyanski S, Allen G, Huber SC (2013) Identification and functional analysis of tomato BRI1 and BAK1 receptor kinase phosphorylation sites. Plant Physiol 113:221465Google Scholar
  3. Barra-Jiménez A, Ragni L (2017) Secondary development in the stem: when Arabidopsis and trees are closer than it seems. Curr Opin Plant Biol 35:145–151CrossRefPubMedGoogle Scholar
  4. Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov V (2013) Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods 9(1):39CrossRefPubMedPubMedCentralGoogle Scholar
  5. Brackmann K, Qi J, Gebert M, Jouannet V, Schlamp T, Grünwald K, Wallner E-S, Novikova DD, Levitsky VG, Agustí J (2018) Spatial specificity of auxin responses coordinates wood formation. Nat Commun 9(1):875CrossRefPubMedPubMedCentralGoogle Scholar
  6. Choe S, Fujioka S, Noguchi T, Takatsuto S, Yoshida S, Feldmann KA (2001) Overexpression of DWARF4 in the brassinosteroid biosynthetic pathway results in increased vegetative growth and seed yield in Arabidopsis. Plant J 26(6):573–582CrossRefPubMedGoogle Scholar
  7. Eriksson ME, Israelsson M, Olsson O, Moritz T (2000) Increased gibberellin biosynthesis in transgenic trees promotes growth, biomass production and xylem fiber length. Nat Biotechnol 18(7):784CrossRefPubMedGoogle Scholar
  8. Hacham Y, Sela A, Friedlander L, Savaldi-Goldstein S (2012) BRI1 activity in the root meristem involves post-transcriptional regulation of PIN auxin efflux carriers. Plant Signal Behav 7(1):68–70CrossRefPubMedPubMedCentralGoogle Scholar
  9. Han S, Cho H, Noh J, Qi J, Jung H-J, Nam H, Lee S, Hwang D, Greb T, Hwang I (2018) BIL1-mediated MP phosphorylation integrates PXY and cytokinin signalling in secondary growth. Nat Plants 4(8):605CrossRefPubMedGoogle Scholar
  10. Ibañes M, Fàbregas N, Chory J, Caño-Delgado AI (2009) Brassinosteroid signaling and auxin transport are required to establish the periodic pattern of Arabidopsis shoot vascular bundles. Proc Natl Acad Sci USA 106(32):13630–13635CrossRefPubMedGoogle Scholar
  11. Jackson D, Veit B, Hake S (1994) Expression of maize KNOTTED1 related homeobox genes in the shoot apical meristem predicts patterns of morphogenesis in the vegetative shoot. Development 120(2):405–413Google Scholar
  12. Jin YL, Tang RJ, Wang HH, Jiang CM, Bao Y, Yang Y, Liang MX, Sun ZC, Kong FJ, Li B (2017) Overexpression of Populus trichocarpa CYP 85A3 promotes growth and biomass production in transgenic trees. Plant Biotechnol J 15(10):1309–1321CrossRefPubMedPubMedCentralGoogle Scholar
  13. Kim T-W, Wang Z-Y (2010) Brassinosteroid signal transduction from receptor kinases to transcription factors. Annu Rev Plant Biol 61:681–704CrossRefPubMedGoogle Scholar
  14. Kondo Y, Ito T, Nakagami H, Hirakawa Y, Saito M, Tamaki T, Shirasu K, Fukuda H (2014) Plant GSK3 proteins regulate xylem cell differentiation downstream of TDIF–TDR signalling. Nat Commun 5:3504CrossRefPubMedGoogle Scholar
  15. Kubo M, Udagawa M, Nishikubo N, Horiguchi G, Yamaguchi M, Ito J, Mimura T, Fukuda H, Demura T (2005) Transcription switches for protoxylem and metaxylem vessel formation. Genes Dev 19(16):1855–1860CrossRefPubMedPubMedCentralGoogle Scholar
  16. Lanza M, Garcia-Ponce B, Castrillo G, Catarecha P, Sauer M, Rodriguez-Serrano M, Páez-García A, Sánchez-Bermejo E, Mohan T, del Puerto YL (2012) Role of actin cytoskeleton in brassinosteroid signaling and in its integration with the auxin response in plants. Dev Cell 22(6):1275–1285CrossRefPubMedGoogle Scholar
  17. Lee J, Shim D, Moon S, Kim H, Bae W, Kim K, Kim Y-H, Rhee S-K, Hong CP, Hong S-Y (2018) Genome-wide transcriptomic analysis of BR-deficient Micro-Tom reveals correlations between drought stress tolerance and brassinosteroid signaling in tomato. Plant Physiol Biochem 127:553–560CrossRefPubMedGoogle Scholar
  18. Li Q-F, He J-X (2013) Mechanisms of signaling crosstalk between brassinosteroids and gibberellins. Plant Signal Behav 8(7):e24686CrossRefPubMedPubMedCentralGoogle Scholar
  19. Li J, Nam KH (2002) Regulation of brassinosteroid signaling by a GSK3/SHAGGY-like kinase. Science 295(5558):1299–1301PubMedGoogle Scholar
  20. Li XJ, Chen XJ, Guo X, Yin LL, Ahammed GJ, Xu CJ, Chen KS, Liu CC, Xia XJ, Shi K (2016) DWARF overexpression induces alteration in phytohormone homeostasis, development, architecture and carotenoid accumulation in tomato. Plant Biotechnol J 14(3):1021–1033CrossRefPubMedGoogle Scholar
  21. Liu T, Zhang J, Wang M, Wang Z, Li G, Qu L, Wang G (2007) Expression and functional analysis of ZmDWF4, an ortholog of Arabidopsis DWF4 from maize (Zea mays L.). Plant Cell Rep 26(12):2091–2099CrossRefPubMedGoogle Scholar
  22. Martí E, Gisbert C, Bishop GJ, Dixon MS, García-Martínez JL (2006) Genetic and physiological characterization of tomato cv. Micro-Tom. J Exp Bot 57(9):2037–2047CrossRefPubMedGoogle Scholar
  23. Moreno-Piovano GS, Moreno JE, Cabello JV, Arce AL, Otegui ME, Chan RL (2017) A role for LAX2 in regulating xylem development and lateral-vein symmetry in the leaf. Ann Bot 120(4):577–590CrossRefPubMedPubMedCentralGoogle Scholar
  24. Nagata N, Asami T, Yoshida S (2001) Brassinazole, an inhibitor of brassinosteroid biosynthesis, inhibits development of secondary xylem in cress plants (Lepidium sativum). Plant Cell Physiol 42(9):1006–1011CrossRefPubMedGoogle Scholar
  25. Nemhauser JL, Mockler TC, Chory J (2004) Interdependency of brassinosteroid and auxin signaling in Arabidopsis. PLoS Biol 2(9):e258CrossRefPubMedPubMedCentralGoogle Scholar
  26. Nie S, Huang S, Wang S, Cheng D, Liu J, Lv S, Li Q, Wang X (2017) Enhancing brassinosteroid signaling via overexpression of tomato (Solanum lycopersicum) SlBRI1 improves major agronomic traits. Front Plant Sci 8:1386CrossRefPubMedPubMedCentralGoogle Scholar
  27. Nolan T, Chen J, Yin Y (2017) Cross-talk of Brassinosteroid signaling in controlling growth and stress responses. Biochem J 474(16):2641–2661CrossRefPubMedPubMedCentralGoogle Scholar
  28. Ruonala R, Ko D, Helariutta Y (2017) Genetic networks in plant vascular development. Annu Rev Genet 51:335–359CrossRefPubMedGoogle Scholar
  29. Ryu H, Kim K, Cho H, Park J, Choe S, Hwang I (2007) Nucleocytoplasmic shuttling of BZR1 mediated by phosphorylation is essential in Arabidopsis brassinosteroid signaling. Plant Cell 19(9):2749–2762CrossRefPubMedPubMedCentralGoogle Scholar
  30. Ryu H, Cho H, Kim K, Hwang I (2010) Phosphorylation dependent nucleocytoplasmic shuttling of BES1 is a key regulatory event in brassinosteroid signaling. Mol Cells 29(3):283–290CrossRefPubMedGoogle Scholar
  31. Ryu H, Cho H, Bae W, Hwang I (2014) Control of early seedling development by BES1/TPL/HDA19-mediated epigenetic regulation of ABI3. Nat Commun 5:4138CrossRefPubMedGoogle Scholar
  32. Saito M, Kondo Y, Fukuda H (2018) BES1 and BZR1 redundantly promote phloem and xylem differentiation. Plant Cell Physiol 59(3):590–600CrossRefGoogle Scholar
  33. Schuetz M, Smith R, Ellis B (2012) Xylem tissue specification, patterning, and differentiation mechanisms. J Exp Bot 64(1):11–31CrossRefPubMedGoogle Scholar
  34. Shen Y, Li Y, Xu D, Yang C, Li C, Luo K (2018) Molecular cloning and characterization of a brassinosteriod biosynthesis-related gene PtoDWF4 from Populus tomentosa. Tree Physiol 38(9):1424–1436CrossRefPubMedGoogle Scholar
  35. Siaud N, Dray E, Gy I, Gerard E, Takvorian N, Doutriaux MP (2004) Brca2 is involved in meiosis in Arabidopsis thaliana as suggested by its interaction with Dmc1. EMBO J 23(6):1392–1401CrossRefPubMedPubMedCentralGoogle Scholar
  36. Szekeres M, Németh K, Koncz-Kálmán Z, Mathur J, Kauschmann A, Altmann T, Rédei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85(2):171–182CrossRefGoogle Scholar
  37. Tian H, Lv B, Ding T, Bai M, Ding Z (2018) Auxin-BR interaction regulates plant growth and development. Front Plant Sci 8:2256CrossRefPubMedPubMedCentralGoogle Scholar
  38. Unterholzner SJ, Rozhon W, Papacek M, Ciomas J, Lange T, Kugler KG, Mayer KF, Sieberer T, Poppenberger B (2015) Brassinosteroids are master regulators of gibberellin biosynthesis in Arabidopsis. Plant Cell 15:00433Google Scholar
  39. Verbančič J, Lunn JE, Stitt M, Persson S (2017) Carbon supply and the regulation of cell wall synthesis. Mol Plant 11(1):75–94CrossRefGoogle Scholar
  40. Wang Z-Y, Nakano T, Gendron J, He J, Chen M, Vafeados D, Yang Y, Fujioka S, Yoshida S, Asami T (2002) Nuclear-localized BZR1 mediates brassinosteroid-induced growth and feedback suppression of brassinosteroid biosynthesis. Dev Cell 2(4):505–513CrossRefPubMedGoogle Scholar
  41. Wu C-Y, Trieu A, Radhakrishnan P, Kwok SF, Harris S, Zhang K, Wang J, Wan J, Zhai H, Takatsuto S (2008) Brassinosteroids regulate grain filling in rice. Plant Cell 20(8):2130–2145CrossRefPubMedPubMedCentralGoogle Scholar
  42. Yamamoto R, Demura T, Fukuda H (1997) Brassinosteroids induce entry into the final stage of tracheary element differentiation in cultured Zinnia cells. Plant Cell Physiol 38(8):980–983CrossRefPubMedGoogle Scholar
  43. Yang JH, Wang H (2016) Molecular mechanisms for vascular development and secondary cell wall formation. Front Plant Sci 7:356PubMedPubMedCentralGoogle Scholar
  44. Yin Y, Wang Z-Y, Mora-Garcia S, Li J, Yoshida S, Asami T, Chory J (2002) BES1 accumulates in the nucleus in response to brassinosteroids to regulate gene expression and promote stem elongation. Cell 109(2):181–191CrossRefPubMedGoogle Scholar
  45. Zhao Q (2016) Lignification: flexibility, biosynthesis and regulation. Trends Plant Sci 21(8):713–721CrossRefPubMedGoogle Scholar
  46. Zheng L, Ma J, Zhang L, Gao C, Zhang D, Zhao C, Han M (2018) Revealing critical mechanisms of BR-mediated apple nursery tree growth using iTRAQ-based proteomic analysis. J Proteomics 173:139–154CrossRefPubMedGoogle Scholar
  47. Zhou J, Zhong R, Ye Z-H (2014) Arabidopsis NAC domain proteins, VND1 to VND5, are transcriptional regulators of secondary wall biosynthesis in vessels. PLoS ONE 9(8):e105726CrossRefPubMedPubMedCentralGoogle Scholar
  48. Zhu J-Y, Sae-Seaw J, Wang Z-Y (2013) Brassinosteroid signalling. Development 140(8):1615–1620CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Jinsu Lee
    • 1
  • Seahee Han
    • 2
  • Hwa-Yong Lee
    • 1
  • Bomi Jeong
    • 3
  • Tae-Young Heo
    • 3
  • Tae Kyung Hyun
    • 4
  • Kyunghwan Kim
    • 1
  • Byoung Il Je
    • 5
  • Horim Lee
    • 6
  • Donghwan Shim
    • 7
  • Soon Ju Park
    • 8
  • Hojin Ryu
    • 1
    Email author
  1. 1.Department of BiologyChungbuk National UniversityCheongjuRepublic of Korea
  2. 2.National Agrobiodiversity Center, National Academy of Agricultural Science RDAJeonjuRepublic of Korea
  3. 3.Department of Information and StatisticsChungbuk National UniversityCheongjuRepublic of Korea
  4. 4.Department of Industrial Plant Science and TechnologyChungbuk National UniversityCheongjuRepublic of Korea
  5. 5.Department of Horticultural Bioscience, College of Natural Resource and Life SciencePusan National UniversityMiryangRepublic of Korea
  6. 6.Department of BiotechnologyDuksung Women’s UniversitySeoulRepublic of Korea
  7. 7.Department of Forest Bio-ResourcesNational Institute of Forest ScienceSuwonRepublic of Korea
  8. 8.Division of Biological Sciences, Research Institute for Basic ScienceWonkwang UniversityIksanRepublic of Korea

Personalised recommendations