Effect of selection and temperature stress on the production of 2n gametes in Lotus tenuis

Five Lotus tenuis genotypes showing 2n pollen production greater than 5% and producing 4x progenies in 4x(male sterile Lotus corniculatus) × 2x crosses were found in different natural populations. Meiotic analyses of these plants showed some second metaphases with disoriented spindles, i.e. parallel and tripolar spindles which resulted in diads and triads, respectively. Because useful genes for winter growth and resistance to powdery mildew from L. tenuis can be introgressed into L. corniculatus via 2n gametes, a programme of selection was started to increase the occurrence of 2n gamete production in L. tenuis. Controlled crosses were performed between genotypes producing 2n gametes and the progenies were checked for 2n pollen production and meiotic aberration, following the same methodologies as used for the parental plants. Although in some progeny plants the frequency of 2n gamete production increased remarkably, the overall progeny level of 2n gamete production remained unchanged. This suggested that different genes for meiotic aberration exist in natural populations and/or differential seed-setting ability exists between genotypes. Diallel crosses between selected genotypes confinned this hypothesis. Evaluation of 2n pollen production of clones grown under different temperature regimes (warm vs. temperate) showed that, in L. tenuis, the effects of genotype, temperature and genotype × temperature interaction are significant. In particular, high temperature increases the level of 2n gamete production in L. tenuis.