Androgenetic green plants from winter rye, Secale cereale L., of diverse origin

Twenty varieties and breeders' lines of winter rye, Secale cereale L., including Finnish, Polish, Russian, Swedish and Danish germplasm, were tested for their anther culture ability. To establish a reference point, the initial culture method was based on published results from rye anther culture. Following results from methodological studies, a modified protocol was tested on the same 20 winter rye lines. The modifications to the protocol included growth of mother plants under controlled greenhouse conditions, prolonged cold pretreatment of spikes for 3 weeks at 4 °C, excision of anthers with microspores in the late uninucleate/binucleate developmental stage and induction on liquid medium supplemented with a buoyancy increasing component, Ficoll. With the modified protocol, the total production of green androgenetic plants increased nearly fivefold; it reached an average of 2.75% with the best line yielding 12.5%, while the proportion of green to albino plants increased by 50% to 25.1%. Eighteen rye lines showed green plant regeneration ability. Several genetically different winter rye lines with relatively good anther culture ability, yielding ≥ 1% green plants per 100 anthers, were identified for the use in further methodical studies and for researching doubled haploidy in rye breeding.