Abstract
Flow cytometry and microsatellite analyses were used to evaluate the trueness-to-type of somatic embryogenesis-regenerated plants from six important Spanish grapevine (Vitis vinifera L.) cultivars. Tetraploid plants were regenerated through somatic embryogenesis from all of the cultivars tested with the exception of ‘Merenzao’. In addition, an octoploid plant was obtained in the cv. ‘Albariño’, and two mixoploids in ‘Torrontés’. The most probable origin of these ploidy variations is somaclonal variation. The cv. ‘Brancellao’ presented significantly more polyploids (28.57%) than any other cultivar, but it must be noted that 50% of the adult field-grown ‘Brancellao’ mother plants analysed were mixoploid. Hence, it is probable that these polyploids originated either from somaclonal variation or by separation of genotypically different cell layers through somatic embryogenesis. Microsatellite analysis of somatic embryogenesis-regenerated plants showed true-to-type varietal genotypes for all plants except six ‘Torrontés’ plants, which showed a mutant allele (231) instead of the normal one (237) at the locus VVMD5. There was not a clear relationship between the occurrence of the observed mutant regenerated plants and the callus induction media composition, the developmental stage of the inflorescences, the type of explant used for starting the cultures or the type of germination (precocious in differentiation medium or normal in germination medium) in any of the cultivars tested, except ‘Torrontés’. The mutant plants described herein have been transplanted to soil for future evaluation of putative phenotypic traits of interest. These mutants can be useful both for breeding programs and for functional genomic approaches aimed at increasing knowledge of the biology of grapevine.
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Abbreviations
- BA:
-
6-Benzyladenine
- CIM:
-
Callus induction medium
- CV:
-
Coefficient of variation
- DAPI:
-
4,6-Diamidino-2-phenylindole
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- DM:
-
Differentiation medium
- FCM:
-
Flow cytometry
- GA3:
-
Gibberellic acid
- IAA:
-
Indole acetic acid
- MS:
-
Murashige and Skoog
- nDNA:
-
Nuclear DNA
- NN:
-
Nitsch and Nitsch
- NOA:
-
Naphthoxyacetic acid
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Acknowledgments
This research was funded by Xunta de Galicia (projects PGIDIT06PXIB310171PR and 2007/030). This paper is a contribution of the Interuniversity Research Group in Biotechnology and Reproductive Biology of Woody Plants. E. Rodriguez was supported by Portuguese FCT (SFRH/BD/27467/2006) fellowships. We thank Jaroslav Doležel (Institute Experimental Botany, Olomouc, Czech Republic) for his generous gift of seeds of Solanum lycopersicum cv. ‘Stupicke’, and Joao Loureiro (Universidade de Coimbra, Portugal) for his help with cytometry analyses. The authors also thank Mª José Graña and Julián Benéitez for their valuable help during plant material collection at the Centro de Formación y Experimentación de Viticultura y Enología de Ribadumia (Pontevedra, Spain), a viticultural facility owned by the regional government of Galicia (Spain).
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Prado, M.J., Rodriguez, E., Rey, L. et al. Detection of somaclonal variants in somatic embryogenesis-regenerated plants of Vitis vinifera by flow cytometry and microsatellite markers. Plant Cell Tiss Organ Cult 103, 49–59 (2010). https://doi.org/10.1007/s11240-010-9753-1
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DOI: https://doi.org/10.1007/s11240-010-9753-1