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.
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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 32:675–686
Bouche N, Bouchez D (2001) Arabidopsis gene knockout: phenotypes wanted. Curr Opinion Plant Biol 4:111–117
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 73:798–813
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 77:462–468
Duclercq J, Sangwan-Norreel B, Catterou M, Sangwan RS (2011) De novo shoot organogenesis: from art to science. Trends Plant Sci 16:597–606
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 49:45–52
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 4:193–196
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 134:3539–3548
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 106:16529–16534
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 131:657–668
Holtorf H, Guitton M-C, Reski R (2002) Plant functional genomics. Naturwissenschaften. https://doi.org/10.1007/s00114-002-0321-3
Ikeuchi M, Ogawa Y, Iwase A, Sugimoto K (2016) Plant regeneration: cellular origins and molecular mechanisms. Development 143:1442–1451
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)
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
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
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 25:402–408
Lowe K, Wu E, Wang N et al (2016) Morphogenic regulators Baby boom and Wuschel improve monocot transformation. Plant Cell 28:1998–2015
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–D229
Marchler-Bauer A, Derbyshire MK, Gonzales NR et al. (2015) CDD: NCBI’s conserved domain database. Nucleic Acids Res 43(Database issue):D222–D226
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 21:796–804
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 95:805–815
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 29:1357–1372
Mitra S, Mukherjee KK (2001) Direct organogenesis in Indian spinach. Plant Cell Tissue Org Cult 67:191–194
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
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
Phillips GC (2004) In vitro morphogenesis in plants–recent advances. In Vitro Cell Dev Biol - Plant 40:342–345
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 100:635–644
Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissue cultures in vitro. Symp Soc Exp Biol 11:118–131
Somssich M, Je B, Simon R, Jackson D (2016) CLAVATA–WUSCHEL signaling in the shoot meristem. Development 143:3238–3248
Su Y-H, Liu Y-B, Zhang X-S (2011) Auxin–cytokinin interaction regulates meristem development. Mol Plant 4:616–625
Thorpe TA (2000) Somatic embryogenesis: morphogenesis, physiology, biochemistry and molecular biology. Korean J Plant Tissue Cult 27:245–258
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 88:9–18
Wahl V, Brand LH, Guo Y-L, Schmid M (2010) The FANTASTIC FOUR proteins influence shoot meristem size in Arabidopsis thaliana. BMC Plant Biol 10:285
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 29:1373–1387
Yang J, Roy A, Zhang Y (2013a) Protein-ligand binding site recognition using complementary binding-specific substructure comparison and sequence profile alignment. Bioinformatics 29:2588–2595
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–D1103
Yildiz M (2012) The prerequisite of the success in plant tissue culture: high frequency shoot regeneration. INTEC. https://doi.org/10.5772/51097
Zhang Y (2008) I-TASSER server for protein 3D structure prediction. BMC Bioinform. https://doi.org/10.1186/1471-2105-9-40
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)
Zhao XY, Su YH, Cheng ZJ, Zhang XS (2008) Cell fate switch during in vitro plant organogenesis. J Integr Plant Biol 50:816–824
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.
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MS, GG: conceived and designed the experiments. GG: performed the tissue culture experiments. MS: performed the molecular biology experiments. GG: wrote the paper.
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The manuscript complies with the Ethical Rules applicable for Plant Cell Tissue and Organ Culture.
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Communicated by Sergio J. Ochatt.
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Sultana, M., Gangopadhyay, G. Early expression of WUSCHEL is a marker for in vitro shoot morphogenesis in tobacco and Beta palonga. Plant Cell Tiss Organ Cult 134, 277–288 (2018). https://doi.org/10.1007/s11240-018-1421-x
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DOI: https://doi.org/10.1007/s11240-018-1421-x