Establishment and maintenance of friable,embryogenic maize callus and the involvement of L-proline

Friable, embryogenic maize (Zea mays L.), inbred line A188, callus was established and maintained for more than one year without apparent loss of friability or embryogenic potential. Embryoid development was abundant in these cultures and plants were easily regenerated. Frequencies of friable-callus initiation and somatic-embryoid formation increased linearly with addition to N6 medium (C.C. Chu et al. 1975, Sci. Sin. [Peking] 18, 659–668) of up to 25 mM L-proline. Proline additions up to 9 mM to MS medium (inorganic elements of T. Murashige and F. Skoog 1962, Physiol. Plant. 15, 473–497, plus 0.5 mg 1^-1 thiamine hydrochloride and 150 mg 1^-1L-asparagine monohydrate) did not stimulate embryoid formation. A major part of the difference between MS and N6 media could be attributed to their respective inorganic nitrogen components. L-Glutamine was not a satisfactory substitute for L-proline. Of 111 regenerated plants grown to maturity from three independent friable, embryogenic cell lines ranging in age from three to seven months, only four plants were abnormal based on morphology and pollen sterility. Seed was produced by 77% of the regenerated plants.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - MS medium containing inorganic elements of Murashige and Skoog (1962), plus 0.5 ml 1^-1 thiamine hydrochloride and 150 mg 1^-1L-asparagine monohydrate - N6 medium of Chu et al. (1975)Paper No. 13,904, Scientific Journal Series Minnesota Agricultural Experiment Station