Meiotic Chromosome Pairing, Isozyme Analyses and Ferritin Expression in a Red Clover Mutant Capable of Somatic Embryogenesis

Red clover genotypes capable of regenerating plantletsin vitrofromnon-meristem-derived callus are rare. A previous study identifieda pair of near isogenic lines which were derived from a singleseed but differed in regenerative ability. The callus-derivedplants of this clone were highly regenerative when reintroducedto culture whereas the epicotyl-derived plants produced non-regenerativecallus cultures. The objective of the present study was to observemeiotic chromosome pairing and to compare isozyme profiles andferritin gene expression in regenerative and non-regenerativeplants and cultures from the clone. Meiotic cells exhibitednormal homologous chromosome pairing. Starch gel zymograms fromglasshouse-grown regenerative (F49R) and non-regenerative (F49M)plants failed to show somaclonal variation for alcohol dehydrogenase,glutamate dehydrogenase, esterase or peroxidase. Isoelectricfocusing of callus cultures from regenerative and non-regenerativeplants revealed that regeneration was accompanied by a reductionin staining intensity and numbers of peroxidase bands comparedto non-regenerative cultures. A unique cathodic peroxidase band(pI 7.6) was associated with non-regenerative cultures. Ferritinexpression was greater in callus than in fresh petiole tissue.Ferritin expression remained high in non-regenerative calluscultures but declined in regenerative cultures as regenerationprogressed.Copyright 1999 Annals of Botany Company Trifolium pratenseL., red clover, ferritin, isozymes, meiosis, peroxidase, somatic embryogenesis.