Abstract
Plastid engineering technique has been established only in Nicotiana tabacum, and the widespread application is severely limited so far. In order to exploit a method to transfer the genetically transformed plastomes already obtained in tobacco into other plant species, somatic cell fusion was conducted between a plastome transformant of tobacco and a cultivar of petunia (Petunia hybrida). A tobacco strain whose plastids had been transformed with aadA (a streptomycin/spectinomycin adenylyltransferase gene) and mdar [a gene for monodehydroascorbate reductase (MDAR)] and a petunia variety, ‘Telstar’, were used as cell fusion partners. An efficient regeneration system from the protoplasts of both the parents, and effectiveness of selection for the aadA gene with spectinomycin were established before the cell fusion. In addition, the influence of UV irradiation on the callus development from the protoplasts and shoot regeneration of tobacco was investigated. Protoplasts were cultured after cell fusion treatment with polyethylene glycol, and asymmetric somatic cybrids were selected using the aadA gene as a marker. Although many shoots of tobacco that had escaped the UV irradiation regenerated, several shoots possessing the morphology of petunia and the resistance to spectinomycin were obtained. Molecular analyses of the petunia type regenerants demonstrated that they had the nuclear and mitochondrial genomes derived from petunia besides the chloroplasts of tobacco transformed with aadA and mdar. Furthermore, it was ascertained that mdar was transcribed in the somatic cybrids. The results indicate the success in intergeneric transfer of transformed plastids of tobacco into petunia.
Similar content being viewed by others
References
Chiang YJ, Stushnoff C, McSay AE (2005) Overexpression of mannitol-1-phosphate dehydrogenase increases mannitol accumulation and adds protection against chilling injury in petunia. J Am Soc Hortic Sci 130:605–610
Dragoeva A, Atanassov I, Atanassov A (1999) CMS due to tapetal failure in cybrids between Nicotiana tabacum and Petunia hybrida. Plant Cell Tissue Organ Cult 55:67–70
Glimelius K, Bonnett HT (1986) Nicotiana cybrids with Petunia chloroplasts. Theor Appl Genet 72:794–798
Guo WW, Prasad D, Cheng YJ, Serrano P, Oeng XX, Grosser JW (2004) Targeted cybridization in citrus: transfer of Satsuma cytoplasm to seedy cultivars for potential seedlessness. Plant Cell Rep 22:752–758
Hayashi S, Tujimura M, Terachi T (2007) Characterization of transplastomic tobacco plants with the genes encoding for the ROS-scavenging enzymes. Proc. International Symposium on Chloroplast Genomics and Genetic Engineering, pp 27
Hinnisdaels S, Bariller L, Mouras A, Sidorov V, Del-Favero J, Veuskens J, Negrutiu I, Jacobs M (1991) Highly asymmetric intergeneric nuclear hybrids between Nicotiana and Petunia: evidence for recombinogenic and translocation events in somatic hybrid plants after ‘‘gamma’’-fusion. Theor Appl Genet 82:609–614
Hou BK, Zhou YH, Wan LH, Zhang ZL, Shen GF, Chen ZH, Hu ZM (2003) Chloroplast transformation in oilseed rape. Transgenic Res 12:111–114
Kao KN, Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126:105–110
Khodakouskaya M, Li Y, Li J, Vaňková R, Malbeck J, McAvoy R (2005) Effects of cor15a-IPT gene expression on leaf senescence in transgenic Petunia x hybrida and Dendranthema x grandiflorum. J Exp Bot 56:1165–1175
Kuchuk N, Sytnyk K, Vasylenko M, Shakhovsky A, Komarnytsky I, Kushnir S, Gleba Y (2006) Genetic transformation of plastids of different Solanaceae species using tobacco cells as organelle hosts. Theor Appl Genet 113:519–527
Mol J, Cornish E, Mason J, Koes R (1999) Novel coloured flowers. Curr Opin Biotechnol 10:198–201
Mulin M, Tran Thahn Van K (1989) In vitro flower formation from thin epidermal cell layers of a partial somatic hybrid between Petunia hybrida (Hort.) and Nicotiana plumbaginifolia (Viv.). Plant Cell Tissue Organ Cult 16:195–206
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Pental D, Hamill JD, Pirrie A, Cocking EC (1986) Somatic hybridization of Nicotiana tabacum and Petunia hybrida. Recovery of plants with P. hybrida nuclear genome and N. tabacum chloroplast genome. Mol Gen Genet 202:342–347
Ruf S, Hermann M, Berger IJ, Carrrer H, Bock R (2001) Stable genetic transformation of tomato plastids and expression of a foreign protein in fruit. Nat Biotechnol 19:870–875
Sidorov VA, Kasten D, Pang SZ, Hajdukiewicz PTJ, Staub JM, Nehra NS (1999) Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker. Plant J 19:209–216
Sikdar SR, Serino G, Chaudhuri S, Maliga P (1998) Plastid transformation in Arabidopsis thaliana. Plant Cell Rep 18:20–24
Svab Z, Hajdukiewicz P, Maliga P (1990) Stable transformation of plastids in higher plants. Proc Natl Acad Sci USA 87:8526–8530
Yamagishi H, Nakagawa S (2004) Somatic hybrids between Arabidopsis thaliana and cabbage (Brassica oleracea). J Japan Soc Hortic Sci 73:319–323
Yamagishi H, Landgren M, Forsberg J, Glimelius K (2002) Production of asymmetric hybrids between Arabidopsis thaliana and Brassica napus utilizing an efficient protoplast culture system. Theor Appl Genet 104:959–964
Zubko MK, Zubko EI, van Zuilen K, Meyer P, Day A (2004) Stable transformation of petunia plastids. Transgenic Res 13:523–530
Acknowledgments
This work was partly supported by the Program for Basic and Applied Researches for Innovations in Bio-oriented Industry (BRAIN). We are grateful to Y. Yamashita for the assistance.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by R. Rose.
Rights and permissions
About this article
Cite this article
Sigeno, A., Hayashi, S., Terachi, T. et al. Introduction of transformed chloroplasts from tobacco into petunia by asymmetric cell fusion. Plant Cell Rep 28, 1633–1640 (2009). https://doi.org/10.1007/s00299-009-0763-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-009-0763-6