Influence of Genotype, Plant Growth Temperature and Anther Incubation Temperature on Microspore Embryo Production in Brassica napus ssp. oleifera

Eleven F₁ hybrid genotypes of winter rape (Brassica napus ssp.oleifera) were used in a study of induction and growth of microspore-derivedembryos. Plants of each genotype were grown in controlled environmentsat either a constant 15°C or a constant 20°C, both witha 16 h photoperiod. Equal numbers of buds, approximately 2.5mm in length, containing uninucleate microspores were harvestedfrom each genotype and either pretreated (14 d at 4°C) ordissected immediately after harvest. Anthers were cultured onliquid medium based upon that of Murashige and Skoog (1962)and containing 8% sucrose, 0.5 mg dm^–3 naphthylaceticacid and 0.05 mg dm^–3 benzylaminopurine. Anthers fromequal samples of buds were incubated at 35°C for 0, 1, 2or 3 d before transfer to 30°C (21 d) and then 25°C.After a total of 42 d incubation, cultures were scored for thepresence of macroscopic embryos (1–2 mm in length) andfor the presence of anthers containing aborted embryoids whichhad not developed further. The results showed first that bud pretreatment completely inhibitedinduction and secondly that anthers of all genotypes had anabsolute requirement for a 35°C treatment (optimal duration2 d) in order to induce embryoid formation. In the great majorityof genotypes plants grown at 15°C provided more productiveanthers than plants grown at 20°C. However, within eachtreatment there were great differences both in the frequencyof anthers showing induced embryoids and of the final yieldof embryos. There was evidence that hybrids with a common parentresponded similarly under certain treatments. This confirmedthe importance of genotypic control for some components of embryoyield. Key words: Brassica napus, Rape, Anther culture, Pollen, Haploid