Selection of hydroxyproline-resistant proline-accumulating mutants of cauliflower (brassica oleracea var. botrytis)

Summary

A procedure is described by which hydroxyproline-resistant lines could be selected from regenerating curd tissue of cauliflower. Mutagenesis was by N-nitroso-N-ethylurea, supplied as a drop of 0.3 mM solution on each 3 mm diameter curd piece. The mutagen generated numerous morphological and pigment mutations without significantly affecting shoot regeneration from explants. Thirty one resistant shoots were recovered from more than six thousand explants mutagenised on regeneration medium supplemented with 3 mM hydroxyproline, while none was obtained from a similar number of non-mutagenised controls. Out of twenty-three resistant shoots which survived subculture, only one showed consistently elevated levels of endogenous proline. During early shoot culture passages, proline levels were 3.6–4.7 times higher than controls, but this was reduced to 1.6 times after 10–12 culture passages in the absence of hydroxyproline. Possible reasons for this decline are discussed. Leaf strip assays suggest resistant shoots may be chimeras and current efforts are directed towards regenerating solid mutants from resistant sectors. These will then be evaluated for any alteration in frost tolerance.