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.
This is a preview of subscription content, log in via an institution to check access.
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
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
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
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
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
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
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
Huang YS, Li HM (2009) Arabidopsis CHLI2 can substitute for CHLI1. Plant Physiol 150:636–645
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
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
Larkin RM (2016) Tetrapyrrole signaling in plants. Front Plant Sci 7:1586. https://doi.org/10.3389/fpls.2016.01586
Larkin RM, Alonso JM, Ecker JR, Chory J (2003) GUN4, a regulator of chlorophyll synthesis and intracellular signaling. Science 299:902–906
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
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
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
Miller C, Skoog F, Von Saltza M, Strong F (1955) Kinetin, a cell division factor from deoxyribonucleic acid. J Am Chem Soc 77:1392
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
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
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
Motte H, Vereecke D, Geelen D, Werbrouck S (2014b) The molecular path to in vitro shoot regeneration. Biotechnol Adv 32:107–121
Ogas J, Cheng JC, Sung ZR, Somerville C (1997) Cellular differentiation regulated by gibberellin in the Arabidopsis thaliana pickle mutant. Science 277:91–94
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
Peter E, Grimm B (2009) GUN4 is required for posttranslational control of plant tetrapyrrole biosynthesis. Mol Plant 2:1198–1210
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
Sugimoto K, Jiao Y, Meyerowitz EM (2010) Arabidopsis regeneration from multiple tissues occurs via a root development pathway. Dev Cell 18:463–471
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
Tanaka R, Kobayashi K, Masuda T (2011) Tetrapyrrole metabolism in Arabidopsis thaliana. Arabidopsis Book 9:e0145. https://doi.org/10.1199/tab.0145
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
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
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
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
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
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
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.
Author information
Authors and Affiliations
Contributions
IK, DZ, AM, and YI carried out experiments and wrote the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by Jochen Kumlehn.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
11240_2019_1680_MOESM1_ESM.jpg
Supplemental Fig. 1 Semi-quantitative RT-PCR analysis of GUN3 or CHLD transcripts in gun3 or chld T-DNA insertion lines. (a) CHLD mRNA level in wild-type and chld mutant 5-day-old seedlings. (b) GUN3 mRNA level in wild-type and in gun3 mutant 5-day-old seedlings. TUB3 is used as internal control. Supplementary material 1 (JPEG 62 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-019-01680-w