Fast and reliable detection of doubled-haploids in Asparagus officinalis by stringent RAPD-PCR

A stringent RAPD-PCR protocol was developed for reliable distinction at an early stage between somatic and doubled-haploid androgenic regenerates emerging from asparagus anther culture by using decamer primers with a high G/C content and/or primers with a dinucleotide sequence. RAPD-PCR was used to test 87 putative doubled-haploids from seven donor plants of different varieties and breeding lines, with differing genetic backgrounds. All of them differed from the anther donor, which suggests they had the desired doubled-haploid constitution, and demonstrates the effectiveness of the in vitro protocol used. This simple, reliable, fast, and comparatively cheap method eliminates the need for discriminating testcrosses and, thus, can significantly accelerate asparagus breeding programmes.