Mapping of Necrosis and Ethylene Inducing-Like Peptides (NLPs) Recognition in Brassica napus by Bulk-Segregant Analysis

In this study, bulk segregant analysis (BSA) was used to map the genomic region associated with the recognition of Necrosis & Ethylene-inducing peptide 1-like proteins (NLPs) in Brassica napus. BcNEP2 from Botrytis cinerea was used as a representative NLP in this study. To genetically map regions responsible for NLP-recognition, DNA pools created from a bi-parental cross of NLP-responsive (Ningyou1) and non-responsive (Ningyou7) lines. 960 seeds from Ning1×7 F2 population were sown and grown under controlled environment room (CER) conditions. Each F2 plant was phenotyped for BcNEP2- and flg22-induced ROS production for BSA. BcNEP2-induced ROS production data from the F2 population was normally distributed. Extremes from the tails of the normal distribution representing 5% of the population were selected to create DNA pools. Individual F2 plants were selected to generate four DNA bulks based on their magnitude of BcNEP2- and flg22-induced ROS production and their ability to recognize BcNEP2 molecule. According to their NLP-recognition, Pool1 and Pool2 were created with non-responsive lines and Pool3 and Pool4 were including NLP-responsive lines. Whereas based on the magnitude ROS production Pool1 and Pool3 were selected from low phenotype and Pool2 and Pool4 were representing high phenotype. Leaf tissue from individual F2 plants was used for whole genome sequencing. Leaf samples from each selected line were bulked for DNA isolation to create pools and same quantity of samples were also used for the parental lines. The data represents whole genome sequencing data from Semi-winter oilseed rape Ningyou1, Ningyou7 and the four DNA pools created from (Ning1×7) F2 population based on their magnitude of BcNEP2- and flg22-induced ROS production and their ability to recognize BcNEP2 molecule.