Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Two types of bHLH transcription factor determine the competence of the pericycle for lateral root initiation

Abstract

The molecular basis of the competence of the pericycle cell to initiate lateral root primordium formation is totally unknown. Here, we report that in Arabidopsis, two types of basic helix-loop-helix (bHLH) transcription factors, named PERICYCLE FACTOR TYPE-A (PFA) proteins and PERICYCLE FACTOR TYPE-B (PFB) proteins, govern the competence of pericycle cells to initiate lateral root primordium formation. Overexpression of PFA genes confers hallmark pericycle characteristics, including specific marker gene expression and auxin-induced cell division, and multiple loss-of-function mutations in PFA genes or the repression of PFB target genes results in the loss of this specific pericycle function. PFA and PFB proteins physically interact and are under mutual- and self-regulation, forming a positive feedback loop. This study unveils the transcriptional regulatory system that determines pericycle participation in lateral root initiation.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Fig. 1: PFA proteins confer pericycle characteristics.
Fig. 2: PFA mutants are defective in lateral root formation and auxin-induced cell division in the pericycle.
Fig. 3: Repression of PFB1 target genes disrupts the cell character of the pericycle.
Fig. 4: PFA and PFB proteins specifically interact in yeast and in planta.
Fig. 5: PFA and PFB proteins form a feedback loop.

Data availability

The authors declare that all data supporting the findings of this study are available within the paper and any raw data can be obtained from the corresponding author upon request. Source data are provided with this paper.

References

  1. Motte, H. & Beeckman, T. The evolution of root branching: increasing the level of plasticity. J. Exp. Bot. 70, 785–793 (2019).

    CAS  PubMed  Google Scholar 

  2. Motte, H., Vanneste, S. & Beeckman, T. Molecular and environmental regulation of root development. Annu. Rev. Plant Biol. 70, 465–488 (2019).

    CAS  PubMed  Google Scholar 

  3. Torres-Martínez, H. H., Rodríguez-Alonso, G., Shishkova, S. & Dubrovsky, J. G. Lateral root primordium morphogenesis in angiosperms. Front. Plant Sci. 10, 206 (2019).

    PubMed  PubMed Central  Google Scholar 

  4. Dubrovsky, J. G. et al. Auxin acts as a local morphogenetic trigger to specify lateral root founder cells. Proc. Natl Acad. Sci. USA 105, 8790–8794 (2008).

    CAS  PubMed  Google Scholar 

  5. Moreno-Risueno, M. A. et al. Oscillating gene expression determines competence for periodic Arabidopsis root branching. Science 329, 1306–1311 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Himanen, K. et al. Auxin-mediated cell cycle activation during early lateral root initiation. Plant Cell 14, 2339–2351 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Beeckman, T. & De Smet, I. Pericycle. Curr. Biol. 24, 378–379 (2014).

    Google Scholar 

  8. Torres-Martínez, H. H., Hernández-Herrera, P., Corkidi, G. & Dubrovsky, J. From one cell to many: morphogenetic field of lateral root founder cells in Arabidopsis thaliana is built by gradual recruitment. Proc. Natl Acad. Sci. USA 117, 20943–20949 (2020).

    PubMed  Google Scholar 

  9. Péret, B. et al. Arabidopsis lateral root development: an emerging story. Trends Plant Sci. 14, 399–408 (2009).

    PubMed  Google Scholar 

  10. Benková, E. & Bielach, A. Lateral root organogenesis—from cell to organ. Curr. Opin. Plant Biol. 13, 677–683 (2010).

    PubMed  Google Scholar 

  11. Laplaze, L. et al. GAL4–GFP enhancer trap lines for genetic manipulation of lateral root development in Arabidopsis thaliana. J. Exp. Bot. 56, 2433–2442 (2005).

    CAS  PubMed  Google Scholar 

  12. Dinneny, J. R. et al. Cell identity mediates the response of Arabidopsis roots to abiotic stress. Science 320, 942–945 (2008).

    CAS  PubMed  Google Scholar 

  13. De Smet, I. et al. Receptor-like kinase ACR4 restricts formative cell divisions in the Arabidopsis root. Science 322, 594–597 (2008).

    PubMed  Google Scholar 

  14. Hiratsu, K., Matsui, K., Koyama, T. & Ohme-Takagi, M. Dominant repression of target genes by chimeric repressors that include the EAR motif, a repression domain, in Arabidopsis. Plant J. 34, 733–739 (2003).

    CAS  PubMed  Google Scholar 

  15. Pires, N. & Dolan, L. Origin and diversification of basic-helix-loop-helix proteins in plants. Mol. Biol. Evol. 27, 862–874 (2010).

    CAS  PubMed  Google Scholar 

  16. Carretero-Paulet, L. et al. Genome-wide classification and evolutionary analysis of the bHLH family of transcription factors in Arabidopsis, poplar, rice, moss, and algae. Plant Physiol. 153, 1398–1412 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Zuo, J., Niu, Q. W. & Chua, N. H. An estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. Plant J. 24, 265–273 (2000).

    CAS  PubMed  Google Scholar 

  18. De Rybel, B. et al. A novel Aux/IAA28 signaling cascade activates GATA23-dependent specification of lateral root founder cell identity. Curr. Biol. 20, 1697–1706 (2010).

    PubMed  Google Scholar 

  19. Goh, T. et al. Lateral root initiation requires the sequential induction of transcription factors LBD16 and PUCHI in Arabidopsis thaliana. New Phytol. 224, 749–760 (2019).

    CAS  PubMed  Google Scholar 

  20. Shaner, N. C. et al. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat. Biotechnol. 22, 1567–1572 (2004).

    CAS  PubMed  Google Scholar 

  21. Malamy, J. E. & Benfey, P. N. Organization and cell differentiation in lateral roots of Arabidopsis thaliana. Development 124, 33–44 (1997).

    CAS  PubMed  Google Scholar 

  22. Dubrovsky, J. G., Doerner, P. W., Colón-Carmona, A. & Rost, T. L. Pericycle cell proliferation and lateral root initiation in Arabidopsis. Plant Physiol. 124, 1648–1657 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Colon-Carmona, A., You, R., Haimovitch-Gal, T. & Doerner, P. Spatio-temporal analysis of mitotic activity with a labile cyclin–GUS fusion protein. Plant J. 20, 503–508 (1999).

    CAS  PubMed  Google Scholar 

  24. Amoutzias, G. D., Robertson, D. L., Oliver, S. G. & Bornberg-Bauer, E. Convergent evolution of gene networks by single-gene duplications in higher eukaryotes. EMBO Rep. 5, 274–279 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Gookin, T. E. & Assmann, S. M. Significant reduction of BiFC non-specific assembly facilitates in planta assessment of heterotrimeric G-protein interactors. Plant J. 80, 553–567 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Massari, M. E. & Murre, C. Helix-loop-helix proteins: regulators of transcription in eucaryotic organisms. Mol. Cell. Biol. 20, 429–440 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Lin, Q. et al. GLABRA2 directly suppresses basic helix-loop-helix transcription factor genes with diverse functions in root hair development. Plant Cell 27, 2894–2906 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Zhang, J. et al. Sperm cells are passive cargo of the pollen tube in plant fertilization. Nat. Plants 3, 17079 (2017).

    PubMed  PubMed Central  Google Scholar 

  29. Omelyanchuk, N. A. et al. A detailed expression map of the PIN1 auxin transporter in Arabidopsis thaliana root. BMC Plant Biol. 16, 5 (2016).

    PubMed  PubMed Central  Google Scholar 

  30. Benková, E. et al. Local, efflux-dependent auxin gradients as a common module for plant organ formation. Cell 115, 591–602 (2003).

    PubMed  Google Scholar 

  31. Okushima, Y., Fukaki, H., Onoda, M., Theologis, A. & Tasaka, M. ARF7 and ARF19 regulate lateral root formation via direct activation of LBD/ASL genes in Arabidopsis. Plant Cell 19, 118–130 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Lee, H. W., Kim, N. Y., Lee, D. J. & Kim, J. LBD18/ASL20 regulates lateral root formation in combination with LBD16/ASL18 downstream of ARF7 and ARF19 in Arabidopsis. Plant Physiol. 151, 1377–1389 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Kiryushkin, A. S. et al. Lateral root initiation in the parental root meristem of cucurbits: old players in a new position. Front. Plant Sci. 10, 365 (2019).

    PubMed  PubMed Central  Google Scholar 

  34. Feng, Z., Zhu, J., Du, X. & Cui, X. Effects of three auxin-inducible LBD members on lateral root formation in Arabidopsis thaliana. Planta 236, 1227–1237 (2012).

    CAS  PubMed  Google Scholar 

  35. Ohashi-Ito, K. & Bergmann, D. C. Regulation of the Arabidopsis root vascular initial population by LONESOME HIGHWAY. Development 134, 2959–2968 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Parizot, B. et al. Diarch symmetry of the vascular bundle in Arabidopsis root encompasses the pericycle and is reflected in distich lateral root initiation. Plant Physiol. 146, 140–148 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Atta, R. et al. Pluripotency of Arabidopsis xylem pericycle underlies shoot regeneration from root and hypocotyl explants grown in vitro. Plant J. 57, 626–644 (2009).

    CAS  PubMed  Google Scholar 

  38. Sugimoto, K., Jiao, Y. & Meyerowitz, E. M. Arabidopsis regeneration from multiple tissues occurs via a root development pathway. Dev. Cell 18, 463–471 (2010).

    CAS  PubMed  Google Scholar 

  39. Yin, K. et al. A dual-color marker system for in vivo visualization of cell cycle progression in Arabidopsis. Plant J. 80, 541–552 (2014).

    CAS  PubMed  Google Scholar 

  40. Musielak, T. J., Schenkel, L., Kolb, M., Henschen, A. & Bayer, M. A simple and versatile cell wall staining protocol to study plant reproduction. Plant Reprod. 28, 161–169 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Kurihara, D., Mizuta, Y., Sato, Y. & Higashiyama, T. ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging. Development 142, 4168–4179 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Mitsuda, N. et al. Efficient yeast one-/two-hybrid screening using a library composed only of transcription factors in Arabidopsis thaliana. Plant Cell Physiol. 51, 2145–2151 (2010).

    CAS  PubMed  Google Scholar 

  43. Boulton, S. J. & Jackson, S. P. Saccharomyces cerevisiae Ku70 potentiates illegitimate DNA double-strand break repair and serves as a barrier to error-prone DNA repair pathways. EMBO J. 15, 5093–5103 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Wang, Z. P. et al. Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 16, 144 (2015).

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

Special thanks to F. Tobe, T. Ishizuka and Y. Takiguchi for technical assistance regarding the establishment of the Y2H assay system and screening experiments, Chua N.-H. for the pER8 vector, L. Hausmann for the pLH5000 vector, Q.-J. Chen for the CRISPR vector (pHEE401E), T. Beeckman for GATA23 pro::NLS–GFP, M. Umeda for Cytrap, R. Tsien for tdTomato, S. Assmann for the BiFC vectors, E. Dafa Alla for comments on the manuscripts, and the Arabidopsis Biological Resource Centre for the Arabidopsis seeds. This work was supported by JSPS KAKENHI grant numbers 19H03246 and 19K22430 to T.K., MEXT KAKENHI grant numbers 18H04837 and 20H04886 to T.K., MEXT KAKENHI grant number 25113006 to T.K., N.M. and M.M., and a Grant-in-Aid for JSPS Fellow number 13J01792 to Y.Z.

Author information

Authors and Affiliations

Authors

Contributions

T.K., Y.Z. and N.M. designed the study. Y.Z. performed most of the plant experiments and some of the yeast experiments, and T.K. performed some of the experiments. Y.Z. and T.K. wrote the paper, and N.M. and M.M. commented on the manuscript. N.M. supervised the establishment of the Y2H assay system and the screening experiments. N.M., T.Y., Y.H. and M.M. made the transcription factor library. N.M., Y.O. and M.O.-T. analysed the data obtained from the Y2H assay.

Corresponding author

Correspondence to Tatsuo Kakimoto.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Peer review information Nature Plants thanks Tom Beeckman, Joseph Dubrovsky and Ben Scheres for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Effects of PFAs on the XPP marker expression of J0121.

a-f, Overexpression of PFA2 (b), PFA3(d), and PFA4 (f) caused ectopic expression of the xylem pole-pericycle marker GFP in the J0121 line. J0121 plants carrying PFA2, PFA3 or PFA4 under an oestrogen-inducible promoter were grown without (a, c, e) or with (b,d,f) 0.1 µM β-oestradiol for 4 (a,b), 7 (c,d), and 6 (e,f) days. Bars, 100 µm. g, PFA1 overexpression was able to induce ectopic J0121 GFP expression only in newly grown parts of the root. Two-day-old plants were transferred to the induction medium containing 0.1 μM β-oestradiol and grown vertically for 2 days. Note that ectopic GFP is present only in the distal position relative to the star (*) mark. Bar, 0.1 mm. a-g, Experiments were repeated once with similar results.

Extended Data Fig. 2 Expression patterns of PFA1, PFA2, PFA3 and PFA4.

ad, PFA1pro::PFA-GFP. PFA1pro::PFA1-GFP is preferentially expressed in the pericycle and is not expressed in the lateral root primordia, except for some cells at the shoulder of the primordia (c and d). 6-day-old plants. e-g, PFA2 pro::H2B-tdTomato siganl is present in stage-II lateral root primordia (e), and was decreased in outer two layers of stageV and stage-VII lateral root primordia. 6-day old plants. h-k, PFA3 pro::H2B-tdTomato. Signal was present in the stele cells in the meristem (h,i) and in the mature zone (j). The signal was decreased in outer two layers of a stageV primordium (k). l-o, 6-day-old PFA4 pro::H2B-tdTomato seedlings. l,m, PFA4 was expressed in protoxylem in the root apical meristem. m, Cross section of (l). Weak signals were detected in the central cylinder. a-o, scale bars, 50 µm. a-o, Experiments were repeated once with similar results.

Extended Data Fig. 3 Overexpression of PFA2 confers the potential of auxin-induced cell division outside the pericycle.

a, Numbers of anticlinal cell divisions in the pericycle. b, Number of anticlinal cell divisions in the endodermis, cortex, and epidermis. Plants transformed with pER8::PFA2 were grown in the absence or presence of 0.1 µM β-oestradiol for 4 days, and then treated with 1 μM NAA in the absence (grey bars) or presence (red bars) of 0.1 μM β-oestradiol, respectively, for 10 hours. a,b, Mean ± standard error. Two-tailed unpaired Student’s t-tests were used to calculate the P values. n = X biologically independent plants. The experiment was repeated once with similar results.

Source data

Extended Data Fig. 4 Cumulative effects of the ectopic cell division in PFA1 overexpressing root over prolonged auxin treatment.

pER8::PFA1 plants were germinated and grown in absence (-Oest) and in presence (+Oest) of 0.1 μM β-oestradiol for 4 days. For auxin treatment, -Oest and +Oest plant were treated with an auxin, 1-naphthaleneacetic acid (NAA, 1 μM), in absence (-Oest) or in presence (+Oest) of 0.1 μM β-oestradiol, respectively, for 20 hours. Arrowheads, clusters of nuclei in endodermis, cortex, or epidermis.

Extended Data Fig. 5 Overexpression of PFA1 and PFA2 affected LRP development and lateral root emergence.

a, pER8::PFA1 plants were germinated and grown in absence (-Oest) and in presence (+Oest) of 0.1 μM β-oestradiol for 4 days. For auxin treatment, -Oest and +Oest plant were treated with an auxin, 1-naphthaleneacetic acid (NAA, 1 μM), in absence (-Oest) or in presence (+Oest) of 0.1 μM β-oestradiol, respectively, for 70 hours. Pictures showing the region of approximately 3mm-3.5 mm up from QC. b, pER8::PFA2 plants were germinated and grown in absence (-Oest) and in presence (+Oest) of 0.1 μM β-oestradiol for 4 days. For auxin treatment, -Oest and +Oest plant were treated with an auxin, 1-naphthaleneacetic acid (NAA, 1 μM), in absence (-Oest) or in presence (+Oest) of 0.1 μM β-oestradiol, respectively, for 70 hours. Pictures showing the region of approximately 3mm-3.5 mm up from QC. c, A rounded LRP-like structure formed in PFA1-overexpressors when grown in the absence of auxin treatment. pER8::PFA1 plants were grown in the absence (-Oest) or presence (+Oest) of 0.1 μM β-oestradiol for 8 days. a-c, scale bars, 100 µm.

Extended Data Fig. 6 Mutants of the PFA and PFB genes.

For the gene models, the coding regions of the exons are depicted with filled (blue or magenta) boxes (Translational start ATG is at the left ends). Regions corresponding to the bHLH domains are depicted with the magenta colour. We used T-DNA insertion mutants for PFA1, PFA2, and PFA3, for which the insertion positions are shown with yellow flags. Three T-DNA insertional mutants were crossed to generate the pfa1 2 3 triple mutant. A mutation was introduced into the PFA4 gene in the pfa1 2 3 plant to make the pfa1 2 3 4 mutant. A mutation was introduced into the PFA5 gene of a pfa1 2 3 4 plant to make pfa1 2 3 4 5. Then, a mutation was introduced into the PFA6 gene of a pfa1 2 3 4 5 plant to make pfa1 2 3 4 5 6. The pfa1 2 3 4 5 6 mutant line is maintained as PFA1(-/-) PFA2(-/-) PFA3(-/-) PFA4(-/-) PFA5(-/-) PFA6( + /-) heterozygous plants. We mutated PFB1 and PFB2 in GATA23 pro::NLS-GFP line. All mutants made by CRISPR/Cas9 method (pfa4-1. pfa5-1, pfa6-1, pfb1-1, pfb2-1) carried a frameshift mutation that generated a premature stop codon before the conserved bHLH domains.

Extended Data Fig. 7 pfa1 2 3 and pfa1 2 3 4 mutants show decreased auxin-induced cell divisions but normal lateral root formation under normal growth condition.

a, Densities of anticlinally dividing cells in the pericycle of auxin-treated pfa triple and quadruple mutants. Four-day-old plants were treated with 1 µM NAA for the indicated time and the anticlinally-dividing cells (metaphase and anaphase) were examined. b, When grown in the absence of auxin treatment, densities of the lateral root primordia and emerged lateral roots in the 8-day-old-plants of the pfa triple and quadruple mutants were not different from wildtype. a and b, Data are mean ± standard error. P values were calculated using Dunnet’s two-sided multiple comparisons test against WT was applied for each stage. n = X biologically independent plants. Experiments were repeated once with similar results.

Source data

Extended Data Fig. 8 Expression patterns of PFA5 and PFA6.

a-f, PFA5 pro::H2B-tdTomato in a root tip (a,b) and in a mature region (c-f). b, The cross section at the indicated position in (a). PFA5 signal is present in the xylem precursors. c and d, Mature regions. PFA5 is preferentially expressed in the pericycle and vasculature. e. PFA5 was expressed in the stage I primordium, but is down-regulated in later stages of lateral root primordia (f). g-j, The PFA6 pro::PFA6-GFP seedlings. g,h, Root tip. h, The cross section at the indicated position in (g). The signal is present in XPP and xylem precursor cells. i and j, Images of mature regions. The GFP signal is present in stage-I lateral root primordium (i), but is downregulated in later stages (j). Scale bars in a-c and e-j, 50 µm; in d, 10 µm. a-j, Experiments were repeated once with similar results.

Extended Data Fig. 9 Repression of PFA1 target genes impairs the pericycle function for lateral root initiation.

a, J0121 carrying pER8::PFA1-SRDX germinated and grown in the absence (-Oest) or presence (+Oest) of 0.1 µM β-oestradiol for 4-days. The GFP signal was absent when PFA1-SRDX was overexpressed. b, PFA1-SRDX inhibited formation of lateral roots (LR). Plants were germinated and grown in the absence (-Oest) or presence (+Oest) of 0.1 µM β-oestradiol for 11 days. Scale bar, 1 cm. c, Lateral root densities of PFA1-SRDX plants that were germinated and grown in the absence (control) or presence of indicated concentrations of β-oestradiol for 8 days. Tukey’s two-sided multiple comparisons test was applied for each stage. Different letters indicate statistically significant differences (p < 0.05). d, PFA1-SRDX partially impaired auxin-induced cell division in pericycle. Number of anticlinally dividing cells in PFA1-SRDX plants grown in the absence or presence of 0.1 µM β-oestradiol for 4 days and then treated with 1 µM NAA or 1 µM NAA in the absence(grey column) or presence (red column), respectively, of 0.1 µM β-oestradiol for the indicated durations. Two-tailed unpaired Student’s t-tests was used to calculate the P value. In (c) and (d), Data are presented as mean ± standard error. n = X biologically independent plants. a-d, Experiments were repeated once with similar results.

Source data

Extended Data Fig. 10 PFB2-SRDX caused loss of pericycle function for lateral root initiation.

a and b, J0121 carrying pER8::PFB2-SRDX grown in the absence (a) or presence (b) of 1 µM β-oestradiol for 4-days. The GFP signal was absent when PFB2-SRDX was overexpressed (b). c and d, pER8::PFB2-SRDX (J0121) plants grown in the absence (c) or presence (d) of 1 µM β-oestradiol for 11-days. PFB2-SRDX expressing plants lack lateral roots (d). PFB1 (e–h) or PFB2 (i–l) overexpression affected neither the J0121 GFP expression nor lateral root formation. J0121 plants carrying pER8::PFB1(e-h) or pER8::PFB2(i-l) were grown in the absence (e,g,i,k) or presence (f,h,j,l) of 0.1 µM β-oestradiol for 4-days (e,f,i,j) or 11-days (g,h,k,l). m, Densities of the lateral root primordia and emerged lateral roots in the 8-day-old pfb1 pfb2 double mutants. Mean ± standard error. Two-tailed unpaired Student’s t-tests were used to calculate the P values. n = X biologically independent samples. n-q, Expression patterns of PFB1pro::PFB1-GFP in the mature zone (n, o) and the root meristem (p, q). n, The PFB1pro::PFB1-GFP signal was present in the nuclei of the pericycle (red arrows), endodermis (orange arrow), cortex (blue arrow), and (o) in the epidermis (white arrow). Images in n and o are at different z-positions of the same root region. r-t, Expression patterns of PFB2pro::PFB2-GFP in a mature region (r) and the root meristem (t,s). r, Red arrows, the nuclei of pericycle. Orange arrows, endodermis. Bars, 100 µm (a,b,e,f,i,j,n,o,r), 5 mm (c,d,g,h,k,l) and 50 µm (n,o,s,t). a-t, Experiments were repeated once with similar results.

Source data

Supplementary information

Supplementary Information

Supplementary information for all plasmids made in this study.

Reporting Summary

Supplementary Video 1

A Cytrap plant grown in the presence of 0.1 µM β-oestradiol was overlayed with GM Phytagel containing 0.1 µM β-oestradiol and 1 µM NAA. The CYCB1.1 pro::CYCB1.1–GFP signal was acquired every 20 min. The video starts 190 min after the onset of auxin treatment.

Supplementary Video 2

A Cytrap plant transformed with pER8::PFA1 grown in the presence of 0.1 µM β-oestradiol was overlayed with GM Phytagel containing 0.1 µM β-oestradiol and 1 µM NAA. The CYCB1.1 pro::CYCB1.1–GFP signal was captured every 20 min. The video starts 190 min after the onset of auxin treatment. GFP-positive cells that can be judged to be outside the pericycle are marked with red squares.

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 3

Statistical source data.

Source Data Fig. 4

Images and tables for Fig. 4a.

Source Data Extended Data Fig. 3

Statistical source data.

Source Data Extended Data Fig. 7

Statistical source data.

Source Data Extended Data Fig. 9

Statistical source data.

Source Data Extended Data Fig. 10

Statistical source data.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, Y., Mitsuda, N., Yoshizumi, T. et al. Two types of bHLH transcription factor determine the competence of the pericycle for lateral root initiation. Nat. Plants 7, 633–643 (2021). https://doi.org/10.1038/s41477-021-00919-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41477-021-00919-9

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing