Application of AFLPs to characterize somaclonal variation in anther-derived grapevines

Abstract

The AFLP (amplified fragment length polymorphism) technique was used to characterize genetic variation in grapevine ( Vitis vinifera L.) regenerated from anther culture. 12 plants obtained by direct embryogenesis from cv. Valerien, and twelve plants obtained by indirect embryogenesis from cv. Mission were evaluated by AFLP. For each genotype the results were analyzed in comparison to control, represented by one field-grown plant (anther-donor). In contrast to cv. Mission, where no difference in polymorph pattern was observed in the DNA restricted with Pst I/ Mse I, in cv. Valerien we found bands with different distribution among anther-derived plants and between them and the mother plant. When the DNA samples were digested with Eco RI in combination with methylation sensitive restriction enzymes Msp I or Hpa II, the AFLP products showed a higher polymorphism in cv. Mission with respect to the number of specific bands in correlation with the age of culture at the moment of regeneration from anthers. Direct embryogenesis from anther culture of cv. Valerien was associated with genetic variation induced in a very early stage of in vitro culture, while the indirect somatic embryogenesis, specific for cv. Mission and/or long-term culture was accompanied by changes in the methylation status. There is some evidence that all the analyzed grapevine somaclones regenerated from in vitro -cultured anthers are genetically distinct from the original cultivars.