Efficient Doubled Haploid Production in Perennial Ryegrass (Lolium perenne L.)

Hybrid breeding has contributed significantly to the enormous yield increases that many major crops have undergone during the previous century. Its success relies on the exploitation of heterosis, the superior performance of an F1 hybrid compared to its inbred parents. Attempts to implement hybrid breeding in forage grasses, such as perennial ryegrass (Lolium perenne L.), are hampered by its highly effective self-incompatibility system as well as its sensitivity to inbreeding depression. Homozygous inbred lines are therefore difficult to create using the classical method of repeated selfing. Here, we report an efficient method to obtain homozygous genotypes of perennial ryegrass using doubled haploid (DH) induction. By means of anther culture, completely homozygous lines were obtained within one generation cycle. A highly genotype dependent response was observed for traits such as the number of embryos/calli per 100 cultured anthers and the percentages of green and albino plants regenerated. Transgressive segregation, indicative of heritable and polygenic control of the traits, was also found. Our general aim is to develop a molecular marker system to select for high responsiveness and to facilitate the introgression of this trait into advanced breeding germplasm. Segregating mapping populations will be phenotyped during anther culture and genotyped via a genotyping-by-sequencing (GBS) approach. Family-based association mapping will be used to identify marker-trait associations. In this way, an efficient breeding tool to screen germplasm for DH induction capacity will be developed. Our work will significantly accelerate forage grass breeding and constitute the first step towards efficient production of grass hybrids.