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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 134, Issue 2, pp 277–288 | Cite as

Early expression of WUSCHEL is a marker for in vitro shoot morphogenesis in tobacco and Beta palonga

  • Marufa Sultana
  • Gaurab Gangopadhyay
Original Article
  • 176 Downloads

Abstract

We have studied the role of growth regulators behind in vitro shoot organogenesis and somatic embryogenesis in two plant systems, viz. tobacco (Nicotiana tabacum L. var. Jayasri) and Beta palonga R.K. Basu & K.K. Mukh. We have also correlated the phenomena of de differentiation with the relative expression of WUS (WUSCHEL) gene in a time-dependent manner. The results indicated that early WUS gene expression is a definite marker for in vitro shoot organogenesis in tobacco and Beta both in direct and indirect modes of regeneration. Additionally, we have performed a comparative homology modeling and in silico structural analysis of WUSCHEL proteins of B. palonga, B. vulgaris, and Arabidopsis to find out the commonality of the ligand binding site. The amino acids of the binding sites were identical (Arginine, Tryptophan, Proline, Asparagine, and Tyrosine) in the three materials under study; except two additional amino acids (Isoleucine and Alanine) in B. vulgaris.

Keywords

In vitro morphogenesis Beta Tobacco WUSCHEL Homology modeling Docking qRT-PCR 

Notes

Acknowledgements

We acknowledge the guidance and encouragement of Professor K. K. Mukherjee, Bose Institute. Authors are grateful to the Director of Bose Institute for providing financial and infrastructural support. The financial assistance in form of research fellowship provided by UGC (University Grants Commission), India is acknowledged by the first author (MS). Technical assistance of Mr. Jadab Ghosh and Mrs. Kaberi Ghosh is also duly acknowledged.

Author contributions

MS, GG: conceived and designed the experiments. GG: performed the tissue culture experiments. MS: performed the molecular biology experiments. GG: wrote the paper.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The manuscript complies with the Ethical Rules applicable for Plant Cell Tissue and Organ Culture.

References

  1. Bouchabke´-Coussa O, Obellianne M, Linderme D, Montes E, Maia-Grondard A, Vilaine F, Pannetier C (2013) Wuschel over expression promotes somatic embryogenesis and induces organogenesis in cotton (Gossypium hirsutum L.) tissues cultured in vitro. Plant Cell Rep 32:675–686CrossRefPubMedGoogle Scholar
  2. Bouche N, Bouchez D (2001) Arabidopsis gene knockout: phenotypes wanted. Curr Opinion Plant Biol 4:111–117CrossRefGoogle Scholar
  3. Chatfield SP, Capron R, Severino A, Penttila P-A, Alfred S, Naha H, Provart NJ (2013) Incipient stem cell niche conversion in tissue culture: using a systems approach to probe early events in WUSCHEL dependent conversion of lateral root primordia into shoot meristems. Plant J 73:798–813CrossRefPubMedGoogle Scholar
  4. Detrez C, Sangwan RS, Sangwan-Norreel BS (1989) Phenotypic and karyotypic status of Beta vulgaris plants regenerated from direct organogenesis in petiole culture. Theor Appl Genet 77:462–468CrossRefPubMedGoogle Scholar
  5. Duclercq J, Sangwan-Norreel B, Catterou M, Sangwan RS (2011) De novo shoot organogenesis: from art to science. Trends Plant Sci 16:597–606CrossRefPubMedGoogle Scholar
  6. Gangopadhyay G, Basu S, Mukherjee BB, Gupta S (1997) Effect of salt and osmotic shocks on unadapted and adapted callus lines of tobacco. Plant Cell Tiss Org Cult 49:45–52CrossRefGoogle Scholar
  7. Gangopadhyay G, Bandyopadhyay T, Datta S, Basu D, Mukherjee KK (2003) Agrobacterium-mediated genetic transformation in Indian Spinach (Beta palonga). Plant Cell Biotechnol Mol Biol 4:193–196Google Scholar
  8. Gordon SP, Heisler MG, Reddy GV, Ohno C, Das P et al (2007) Pattern formation during de novo assembly of the Arabidopsis shoot meristem. Development 134:3539–3548CrossRefPubMedGoogle Scholar
  9. Gordon SP, Chickarmane VS, Ohno C, Meyerowitz EM (2009) Multiple feedback loops through cytokinin signaling control stem cell number within the Arabidopsis shoot meristem. Proceed Natl Acad Sci USA 106:16529–16534CrossRefGoogle Scholar
  10. Haecker A, Groß-Hardt R, Geiges B, Sarkar A, Breuninger H, Herrmann M, Laux T (2004) Expression dynamics of WOX genes mark cell fate decisions during early embryonic patterning in Arabidopsis thaliana. Development 131:657–668CrossRefPubMedGoogle Scholar
  11. Holtorf H, Guitton M-C, Reski R (2002) Plant functional genomics. Naturwissenschaften.  https://doi.org/10.1007/s00114-002-0321-3 PubMedCrossRefGoogle Scholar
  12. Ikeuchi M, Ogawa Y, Iwase A, Sugimoto K (2016) Plant regeneration: cellular origins and molecular mechanisms. Development 143:1442–1451CrossRefPubMedGoogle Scholar
  13. Jönsson H, Heisler M, Reddy GV, Agrawal V, Gor V, Shapiro BE, Mjolsness E, Meyerowitz EM (2005) Modeling the organization of the WUSCHEL expression domain in the shoot apical meristem. Bioinformatics 21:232–240. ( https://doi.org/10.1093/bioinformatics/bti1036)CrossRefGoogle Scholar
  14. Lazim MIM, Badruzaman NA, Peng KS, Long K (2015) Quantification of cytokinins in coconut water from different maturation stages of Malaysia’s Coconut (Cocos nucifera L.) varieties. J Food Process Technol 6:515.  https://doi.org/10.4172/2157-7110.1000515 CrossRefGoogle Scholar
  15. Li W, Li Z, Zhai Y, Wang C (2015) A highly efficient castor regeneration system identified through WUSCHEL expression. Chem Eng Trans 46:1393–1398.  https://doi.org/10.3303/CET1546233 CrossRefGoogle Scholar
  16. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Method 25:402–408CrossRefGoogle Scholar
  17. Lowe K, Wu E, Wang N et al (2016) Morphogenic regulators Baby boom and Wuschel improve monocot transformation. Plant Cell 28:1998–2015CrossRefPubMedCentralPubMedGoogle Scholar
  18. Marchler-Bauer A, Shennan L, Anderson JB et al. (2011) CDD: a conserved domain database for the functional annotation of proteins. Nucleic Acids Res 39(Database issue):D225–D229CrossRefPubMedGoogle Scholar
  19. Marchler-Bauer A, Derbyshire MK, Gonzales NR et al. (2015) CDD: NCBI’s conserved domain database. Nucleic Acids Res 43(Database issue):D222–D226CrossRefPubMedGoogle Scholar
  20. Marhavy P, Bielach A, Abas L, Abuzeineh A, Duclercq J, Tanaka H, Parezova M, Petrasek J, JirıFriml J, Kleine-Vehn J, Benkova E (2011) Cytokinin modulates endocytic trafficking of PIN1 auxin efflux carrier to control plant organogenesis. Dev Cell 21:796–804CrossRefPubMedGoogle Scholar
  21. Mayer KF, Schoof H, Haecker A, Lenhard M, Jürgens G, Laux T (1998) Role of WUSCHEL in regulating stem cell fate in the Arabidopsis shoot meristem. Cell 95:805–815CrossRefPubMedGoogle Scholar
  22. Meng WJ, Cheng ZJ, Sang YL, Zhang MM, Rong XF, Wang JW, Tang YY, Zhang XS (2017) Type-B Arabidopsis response regulators specify the shoot stem cell niche by dual regulation of WUSCHEL. Plant Cell 29:1357–1372PubMedPubMedCentralGoogle Scholar
  23. Mitra S, Mukherjee KK (2001) Direct organogenesis in Indian spinach. Plant Cell Tissue Org Cult 67:191–194CrossRefGoogle Scholar
  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  25. Pathi KM, Tula S, Tuteja N (2013) High frequency regeneration via direct somatic embryogenesis and efficient Agrobacterium—mediated genetic transformation of tobacco. Plant Signalling Behav 8:e24354.  https://doi.org/10.4161/psb.24354 CrossRefGoogle Scholar
  26. Phillips GC (2004) In vitro morphogenesis in plants–recent advances. In Vitro Cell Dev Biol - Plant 40:342–345CrossRefGoogle Scholar
  27. Schoof H, Lenhard M, Haecker A, Mayer KF, Jürgens G, Laux T (2000) The stem cell population of Arabidopsis shoot meristem is maintained by a regulatory loop between the CLAVATA and WUSCHEL genes. Cell 100:635–644CrossRefPubMedGoogle Scholar
  28. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissue cultures in vitro. Symp Soc Exp Biol 11:118–131PubMedGoogle Scholar
  29. Somssich M, Je B, Simon R, Jackson D (2016) CLAVATA–WUSCHEL signaling in the shoot meristem. Development 143:3238–3248CrossRefPubMedGoogle Scholar
  30. Su Y-H, Liu Y-B, Zhang X-S (2011) Auxin–cytokinin interaction regulates meristem development. Mol Plant 4:616–625CrossRefPubMedPubMedCentralGoogle Scholar
  31. Thorpe TA (2000) Somatic embryogenesis: morphogenesis, physiology, biochemistry and molecular biology. Korean J Plant Tissue Cult 27:245–258Google Scholar
  32. Verdeil J-L, Hocher V, Huet C, Grosdemange F, Escoute J, Ferriere N, Nicole M (2001) Ultra structural changes in coconut calli associated with the acquisition of embryogenic competence. Ann Bot 88:9–18CrossRefGoogle Scholar
  33. Wahl V, Brand LH, Guo Y-L, Schmid M (2010) The FANTASTIC FOUR proteins influence shoot meristem size in Arabidopsis thaliana. BMC Plant Biol 10:285CrossRefPubMedPubMedCentralGoogle Scholar
  34. Wang J, Tian C, Zhang C, Shi B, Cao X, Zhang T-Q, Zhao Z, Wang J-W, Jiao Y (2017) Cytokinin signalling activates WUSCHEL expression during axillary meristem initiation. Plant Cell 29:1373–1387PubMedPubMedCentralGoogle Scholar
  35. Yang J, Roy A, Zhang Y (2013a) Protein-ligand binding site recognition using complementary binding-specific substructure comparison and sequence profile alignment. Bioinformatics 29:2588–2595CrossRefPubMedPubMedCentralGoogle Scholar
  36. Yang J, Roy A, Zhang Y (2013b) BioLiP: a semi-manually curated database for biologically relevant ligand-protein interactions. Nucleic Acids Res 41(Database issue):D1096–D1103PubMedGoogle Scholar
  37. Yildiz M (2012) The prerequisite of the success in plant tissue culture: high frequency shoot regeneration. INTEC.  https://doi.org/10.5772/51097 CrossRefGoogle Scholar
  38. Zhang Y (2008) I-TASSER server for protein 3D structure prediction. BMC Bioinform.  https://doi.org/10.1186/1471-2105-9-40 CrossRefGoogle Scholar
  39. Zhang N, Huang X, Bao Y, Wang B, Liu L, Dai L, Chen J, An X, Sun Y, Peng D (2015) Genome-wide identification and expression profiling of WUSCHEL-related homeobox (WOX) genes during adventitious shoot regeneration of watermelon (Citrullus lanatus). Acta Physiol Plant 37:224 (12 pages)CrossRefGoogle Scholar
  40. Zhao XY, Su YH, Cheng ZJ, Zhang XS (2008) Cell fate switch during in vitro plant organogenesis. J Integr Plant Biol 50:816–824CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Division of Plant BiologyBose InstituteKolkataIndia

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