Mutations in tetrapyrrole biosynthesis pathway uncouple nuclear WUSCHEL expression from de novo shoot development in Arabidopsis

Abstract

Plant de novo organogenesis in tissue culture systems has long been exploited to study the plasticity of pluripotency. External application of high cytokinin-to-auxin ratio in cultured medium stimulates greening of tissue and promotes nascent shoot apical meristem (SAM) formation. The stem cell niche in SAM is maintained by a negative feedback loop between CLAVATA and WUSCHEL (WUS) signaling. The fact that cytokinin is able to induce expression of SAM master regulator WUS suggests that the capacity of de novo shoot development is largely dependent on WUS activity. However, the molecular mechanism of WUS expression remains obscure. Here we found that WUS expression during de novo SAM formation is affected by the altered tetrapyrrole metabolism catalyzed in the plastid. Loss-of-function mutations in HEME OXYGENASE/LONG HYPOCOTYL 1 (hy1), Mg-CHELATASE H (chlh), Mg-CHELATASE I1 (chli1), and the regulator of Mg-CHELATASE, GENOME-UNCOUPLED 4 (gun4), result in elevated WUS expression but shoot regeneration efficiency is decreased whereas loss-of-function mutation in PROTOPORPHYRIN IX FERROCHELATASE 2 (fc2) exhibits compromised WUS expression with a reduced number of shoots when mutant root explants are cultured on shoot induction medium. Our genetic results show that nuclear WUS expression is affected by tetrapyrrole intermediate(s) under the varying plastid status stimulated by external cytokinin treatment during de novo organogenesis.

Key Message

We propose novel regulatory mechanism of nuclear WUS expression that is likely modulated by tetrapyrrole intermediate(s) under the varying plastid status stimulated by external cytokinin treatment during de novo organogenesis.

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Abbreviations

ahk3:

Arabidopsis histidine kinase 3

ckh2:

Cytokinin hypersensitive 2

cre1:

Cytokinin response 1

fc2:

Protoporphyrin IX ferrochelatase 2

gun:

Genome uncoupled

hy:

Long hypocotyl

pkl:

Pickle

SIM:

Shoot induction medium

wus:

Wuschel

References

  1. Atta R, Laurens L, Boucheron-Dubuisson E, Guivarc’h A, Carnero E, Giraudat-Pautot V, Rech P, Chriqui D (2009) Pluripotency of Arabidopsis xylem pericycle underlies shoot regeneration from root and hypocotyl explants grown in vitro. Plant J 57:626–644

    CAS  Article  PubMed  Google Scholar 

  2. Chan KX, Phua SY, Crisp P, McQuinn R, Pogson BJ (2016) Learning the languages of the chloroplast: retrograde signaling and beyond. Annu Rev Plant Biol 67:25–53

    CAS  Article  PubMed  Google Scholar 

  3. Cheng J, He CX, Zhang ZW, Xu F, Zhang DW, Wang X, Yuan S, Lin HH (2011) Plastid signals confer Arabidopsis tolerance to water stress. Z Naturforsch C 66:47–54

    CAS  Article  PubMed  Google Scholar 

  4. Chory J, Reinecke D, Sim S, Washburn T, Brenner M (1994) A role for cytokinins in de-etiolation in Arabidopsis (det mutants have an altered response to cytokinins). Plant Physiol 104:339–347

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  5. Cortleven A, Marg I, Yamburenko MV, Schlicke H, Hill K, Grimm B, Schaller GE, Schmulling T (2016) Cytokinin regulates the etioplast-chloroplast transition through the two-component signaling system and activation of chloroplast-related genes. Plant Physiol 172:464–478

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Furuta K, Kubo M, Sano K, Demura T, Fukuda H, Liu YG, Shibata D, Kakimoto T (2011) The CKH2/PKL chromatin remodeling factor negatively regulates cytokinin responses in Arabidopsis calli. Plant Cell Physiol 52:618–628

    CAS  Article  PubMed  Google Scholar 

  7. Huang YS, Li HM (2009) Arabidopsis CHLI2 can substitute for CHLI1. Plant Physiol 150:636–645

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. Inoue T, Higuchi M, Hashimoto Y, Seki M, Kobayashi M, Kato T, Tabata S, Shinozaki K, Kakimoto T (2001) Identification of CRE1 as a cytokinin receptor from Arabidopsis. Nature 409:1060–1063

    CAS  Article  PubMed  Google Scholar 

  9. Kubalová I, Ikeda Y (2017) Chlorophyll measurement as a quantitative method for the assessment of cytokinin-induced green foci formation in tissue culture. J Plant Growth Regul 36:516–521

    Article  CAS  Google Scholar 

  10. Larkin RM (2016) Tetrapyrrole signaling in plants. Front Plant Sci 7:1586. https://doi.org/10.3389/fpls.2016.01586

    Article  PubMed  PubMed Central  Google Scholar 

  11. Larkin RM, Alonso JM, Ecker JR, Chory J (2003) GUN4, a regulator of chlorophyll synthesis and intracellular signaling. Science 299:902–906

    CAS  Article  PubMed  Google Scholar 

  12. Lee DK, Parrott DL, Adhikari E, Fraser N, Sieburth LE (2016) The mobile bypass signal arrests shoot growth by disrupting shoot apical meristem maintenance, cytokinin signaling, and WUS transcription factor expression. Plant Physiol 171:2178–2190

    Article  PubMed  PubMed Central  Google Scholar 

  13. Li W, Liu H, Cheng ZJ, Su YH, Han HN, Zhang Y, Zhang XS (2011) DNA methylation and histone modifications regulate de novo shoot regeneration in Arabidopsis by modulating WUSCHEL expression and auxin signaling. PLoS Genet 7:e1002243. https://doi.org/10.1371/journal.pgen.1002243

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. Meng WJ, Cheng ZJ, Sang YL, Zhang MM, Rong XF, Wang ZW, Tang YY, Zhang XS (2017) Type-B Arabidopsis response regulators specify the shoot stem cell niche by dual regulation of WUSCHEL. Plant Cell 29:1357–1372

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Miller C, Skoog F, Von Saltza M, Strong F (1955) Kinetin, a cell division factor from deoxyribonucleic acid. J Am Chem Soc 77:1392

    CAS  Article  Google Scholar 

  16. Mochizuki N, Brusslan JA, Larkin A, Nagatani A, Chory J (2001) Arabidopsis genomes uncoupled 5 (GUN5) mutant reveals the involvement of Mg-chelatase H subunit in plastid-to-nucleus signal transduction. Proc Natl Acad Sci USA 98:2053–2058

    CAS  Article  PubMed  Google Scholar 

  17. Mochizuki N, Tanaka R, Tanaka A, Masuda T, Nagatani A (2008) The steady-state level of Mg-protoporphyrin IX is not a determinant of plastid-to-nucleus signaling in Arabidopsis. Proc Natl Acad Sci USA 105:15184–15189

    CAS  Article  PubMed  Google Scholar 

  18. Motte H, Vercauteren A, Depuydt S, Landschoot S, Geelen D, Werbrouck S, Goormachtig S, Vuylsteke M, Vereecke D (2014a) Combining linkage and association mapping identifies receptor-like protein kinase1 as an essential Arabidopsis shoot regeneration gene. Proc Natl Acad Sci USA 111:8305–8310

    CAS  Article  PubMed  Google Scholar 

  19. Motte H, Vereecke D, Geelen D, Werbrouck S (2014b) The molecular path to in vitro shoot regeneration. Biotechnol Adv 32:107–121

    CAS  Article  PubMed  Google Scholar 

  20. Ogas J, Cheng JC, Sung ZR, Somerville C (1997) Cellular differentiation regulated by gibberellin in the Arabidopsis thaliana pickle mutant. Science 277:91–94

    CAS  Article  PubMed  Google Scholar 

  21. Perianez-Rodriguez J, Manzano C, Moreno-Risueno MA (2014) Post-embryonic organogenesis and plant regeneration from tissues: two sides of the same coin? Front Plant Sci 5:219. https://doi.org/10.3389/fpls.2014.00219

    Article  PubMed  PubMed Central  Google Scholar 

  22. Peter E, Grimm B (2009) GUN4 is required for posttranslational control of plant tetrapyrrole biosynthesis. Mol Plant 2:1198–1210

    CAS  Article  PubMed  Google Scholar 

  23. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11:118–130

    CAS  PubMed  Google Scholar 

  24. Sugimoto K, Jiao Y, Meyerowitz EM (2010) Arabidopsis regeneration from multiple tissues occurs via a root development pathway. Dev Cell 18:463–471

    CAS  Article  PubMed  Google Scholar 

  25. Susek RE, Ausubel FM, Chory J (1993) Signal transduction mutants of Arabidopsis uncouple nuclear CAB and RBCS gene expression from chloroplast development. Cell 74:787–799

    CAS  Article  PubMed  Google Scholar 

  26. Tanaka R, Kobayashi K, Masuda T (2011) Tetrapyrrole metabolism in Arabidopsis thaliana. Arabidopsis Book 9:e0145. https://doi.org/10.1199/tab.0145

    Article  PubMed  PubMed Central  Google Scholar 

  27. Tsuzuki T, Takahashi K, Inoue S, Okigaki Y, Tomiyama M, Hossain MA, Shimazaki K, Murata Y, Kinoshita T (2011) Mg-chelatase H subunit affects ABA signaling in stomatal guard cells, but is not an ABA receptor in Arabidopsis thaliana. J Plant Res 124:527–538

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  28. Valvekens D, Montagu MV, Van Lijsebettens M (1988) Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85:5536–5540

    CAS  Article  PubMed  Google Scholar 

  29. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:0034

    Article  Google Scholar 

  30. Woodson JD, Perez-Ruiz JM, Chory J (2011) Heme synthesis by plastid ferrochelatase 1 regulates nuclear gene expression in plants. Curr Biol 21:897–903

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  31. Zhang TQ, Lian H, Zhou CM, Xu L, Jiao Y, Wang JW (2017) A two-step model for de novo activation of WUSCHEL during plant shoot regeneration. Plant Cell 29:1073–1087

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  32. Zhao X, Bramsiepe J, Van Durme M, Komaki S, Prusicki MA, Maruyama D, Forner J, Medzihradszky A, Wijnker E, Harashima H, Lu Y, Schmidt A, Guthorl D, Logrono RS, Guan Y, Pochon G, Grossniklaus U, Laux T, Higashiyama T, Lohmann JU, Nowack MK, Schnittger A (2017) Retinoblastoma Related1 mediates germline entry in Arabidopsis. Science. https://doi.org/10.1126/science.aaf6532

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by Grant Agency in the Czech Republic (GAČR17-23702S and GAČR 18-23972Y) and the European Regional Development Fund (ERDF) Project (No. CZ.02.1.01/0.0/0.0/16_019/0000827). We thank European Arabidopsis Stock Centre (NASC) and Tatsuo Kakimoto for providing cre1-2 mutant seeds.

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IK, DZ, AM, and YI carried out experiments and wrote the manuscript.

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Correspondence to Yoshihisa Ikeda.

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Communicated by Jochen Kumlehn.

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Kubalová, I., Zalabák, D., Mičúchová, A. et al. Mutations in tetrapyrrole biosynthesis pathway uncouple nuclear WUSCHEL expression from de novo shoot development in Arabidopsis. Plant Cell Tiss Organ Cult 139, 395–401 (2019). https://doi.org/10.1007/s11240-019-01680-w

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Keywords

  • Arabidopsis thaliana
  • Cytokinin
  • Retrograde signaling
  • Shoot apical meristem
  • Tetrapyrrole biosynthesis
  • WUSCHEL