Doubled haploid production in rocket (Eruca sativa Mill.) through isolated microspore culture

Abstract

Haploid induction in rocket (Eruca sativa Mill.), a novel and increasingly important vegetable, was studied in microspore culture. A procedure based on a high sucrose NLN medium and heat shock treatment resulted in nuclear divisions and embryo induction. The effect of genotype both among seed lots and among single plants was a major factor influencing embryo formation. The addition of activated charcoal was essential for obtaining reproducible results, 0.2 mg l−1 being superior to 1.0 mg l−1. A 24 h heat shock treatment at 32°C doubled the embryogenic response compared to a 48 h treatment. Embryo conversion was only efficient (23%) for embryos that had been cultured on medium with activated charcoal and subcultured on solid B5 medium; pretreatment of embryos with ABA or desiccation for 1–3 weeks inhibited embryo conversion. Analysis of ploidy level revealed that the majority (65.6%) of 489 regenerated plantlets tested were diploid. Breeding programs and genetic studies of rocket are likely to benefit substantially from the established method.