Eco-tilling for haploid inducers in chicory genepool.

This project aims to identify haploid inducer lines in chicory, using eco-tilling to screen for naturally occuring mutations in nine candidate genes putatively involved in haploid induction. We used SMAP design software to create two complementary panels of highly multiplex amplicon sequencing (HiPlex) primers, and screened 35 accessions of chicory (Chicorium intybus var. sativum) and 25 accessions of witloof (Chicorium intybus var. foliosum), for interesting (knock-out) alleles. Sequencing was performed in two subsequent rounds, first by 1-D pool-Seq in three independent pools of up to 10 individuals, and then a second round screen at individual plant level of pools that carried interesting alleles.