Identification and validation of genomic regions for pod shatter resistance in Brassica rapa using QTL-seq and traditional QTL mapping.

Abstract

Background: Pod shatter resistance is an important trait in Brassica species, significantly impacting the yield and profitability of growers. Identifying genomic regions and understanding genes underlying shatter resistance is a major objective of breeding programs. Brassica rapa, commonly known as rape or field mustard, is an ancestral species of Brassica napus and Brassica juncea - the most widely oilseed crops grown worldwide. In this study, we performed diversity analysis of B. rapa accessions, bulked segregant analysis based quantitative trait locus-sequencing (QTL-seq), and traditional quantitative trait locus (QTL) mapping in an F₂ population to identify genomic regions associated with pod shatter resistance in B. rapa.

Results: A considerable genetic variation for pod shatter resistance, measured as rupture energy (RE), varied from 0.63 to 3.49 mJ^(½) was revealed among 90 accessions of B. rapa. Cluster analysis based on 10,324 DArTseq markers showed that pod shatter-resistant accessions originated from diverse sources. We further investigated the genetic and anatomical bases of variation in pod shatter resistance from two contrasting parental lines, ATC90153 (maternal parent with high RE) and ATC91215 (paternal parent with low RE). Bulked segregant resequencing analysis of parental lines and two pooled samples, prepared from 10 resistant and 10 sensitive lines to pod shatter, identified three genomic regions for shatter resistance on chromosomes A06 and A09. Traditional QTL analysis validated marker-pod shatter resistance associations on chromosomes A06 and A09 in the same F₂ population using a linkage map based on 23,274 DArTseq markers. Physical positions of significantly associated markers and the priori pod dehiscence genes on the B. rapa reference genome sequence suggested BEE1/PEROXIDASE/TCP8 on A06 and ADPG1/SHP1/MYB116 genes on A09 as potential candidates for pod shatter resistance. Sequence comparison of parental lines identified sequence variants (194 SNPs and 74 InDELs on A06, and two SNPs and two InDELs on A09) in the promoter and downstream regions of B. rapa genes within the QTL.

Conclusions: We identified QTLs and priori candidate genes associated with variation in pod shatter resistance on chromosomes A06 and A09 in B. rapa. This study provides potential gene targets to understand molecular mechanisms and improve pod shatter resistance in Brassica crops.