Enhanced expression of a cold-induced gene coding for a glycine-rich protein in regenerative somaclonal variants of red clover (Trifolium pratense L.)

Cold hardiness and in vitro regeneration are closely related traits in sele cted alfalfa and red clover germplasms. We investigated the cold-induction of three genes, MsnCIA, MsaCIB and MsaCIC, implicated in the cold tolerance of alfalfa, in non-regenerative and regenerative somaclonal variants of re d clover by Northern blot analysis using alfalfa cDNA clones as hybridizati on probes. Only the MsaCIA gene exhibited increased steady-state mRNA level s in cold-acclimated red clover. We observed, however, enhanced-cold induct ion of steady-state levels of MsaCIA gene transcripts in all regenerative s omaclonal variants compared to non-regenerative red clover lines, Moreover, these two traits, regeneration and enhanced cold-induction of MsnCIA, were concomitantly transmitted to progeny suggesting that enhanced expression o f MsaCIA and the regenerative trait are either linked, or the expression of both traits are regulated by a 'master' gene whose expression is modified by somatic embryogenesis.