An unstable anthocyanin mutation recovered from tissue culture of alfalfa (Medicago sativa) : 2. Stable nonrevertants derived from reculture.

An unstable recessive ("mutable") allele, c2-m4, of a locus required for anthocyanin pigmentation in alfalfa (Medicago sativa L.) reverts to a stable functional state at high frequency in vitro. It was previously established that a white-flowered mutant ("WFM") and a white-flowered progeny of WFM ("WHGW3") each carry the unstable allele. More than 20% of plants regenerated from tissue cultures of WFM and WHGW3 are revertant. It is here established that most nonrevertant plants regenerated from cultures of WFM and WHGW3 are stabilized in the recessive condition. Reculture of nonrevertants of WFM and WHGW3 indicated that there are three classes of nonrevertants: (i) Nonrevertants which revert in vitro at a high frequency typical of WFM; (ii) Nonrevertants which revert upon reculture but at significantly lower frequencies than WFM; and (iii) Nonrevertants which do not revert upon reculture. These observations are discussed in terms of transposable element action in vitro.