Abstract

Allotetraploid rapeseed (Brassica napus L. AnAnCnCn, 2n=4x=38) is highly susceptible to boron (B) deficiency, a widespread limiting factor that causes severe losses in seed yield. The genetic variation in the sensitivity to B deficiency found in rapeseed genotypes emphasizes the complex response architecture. In this research, a B-inefficient genotype, 'Westar 10' ('W10'), responded to B deficiencies during vegetative and reproductive development with an over-accumulation of reactive oxygen species, severe lipid peroxidation, evident plasmolysis, abnormal floral organogenesis, and widespread sterility compared to a B-efficient genotype, 'Qingyou 10' ('QY10'). Whole-genome re-sequencing (WGS) of 'QY10' and 'W10' revealed a total of 1 605 747 single nucleotide polymorphisms and 218 755 insertions/deletions unevenly distributed across the allotetraploid rapeseed genome (~1130Mb). Digital gene expression (DGE) profiling identified more genes related to B transporters, antioxidant enzymes, and the maintenance of cell walls and membranes with higher transcript levels in the roots of 'QY10' than in 'W10' under B deficiency. Furthermore, based on WGS and bulked segregant analysis of the doubled haploid (DH) line pools derived from 'QY10' and 'W10', two significant quantitative trait loci (QTLs) for B efficiency were characterized on chromosome C2, and DGE-assisted QTL-seq analyses then identified a nodulin 26-like intrinsic protein gene and an ATP-binding cassette (ABC) transporter gene as the corresponding candidates regulating B efficiency. This research facilitates a more comprehensive understanding of the differential physiological and transcriptional responses to B deficiency and abundant genetic diversity in rapeseed genotypes, and the DGE-assisted QTL-seq analyses provide novel insights regarding the rapid dissection of quantitative trait genes in plant species with complex genomes.

Highlights

  • Boron (B) is an essential micronutrient for the normal growth and development of higher plants (Warington, 1923)

  • High-quality clean reads were mapped to the B. napus ‘Darmor-bzh’ transcriptome reference, and the mRNA abundances of the unigenes, which were identified by TopHat and Cufflinks (Trapnell et al, 2012), were normalized by the fragments per kilobase of exon model per million mapped reads (FPKM) (Trapnell et al, 2010)

  • The availability of the complete B. napus (‘Darmor-bzh’) genome reference sequence (Chalhoub et al, 2014) and the advancement of next-generation sequencing (NGS) technologies provide great opportunities to reveal the genetic diversity among various B. napus genotypes and for the genetic enhancement of oilseed rape

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Summary

Introduction

Boron (B) is an essential micronutrient for the normal growth and development of higher plants (Warington, 1923). B deficiency primarily affects developing tissues, with symptoms including the inhibition of root elongation and leaf expansion, and reduced fertility (Shorrocks, 1997; Lordkaew et al, 2011). Failures caused by B deficiencies have occurred in more than 80 countries, involving over 132 crop species (Shorrocks, 1997; Goldbach et al, 2001). Allotetraploid rapeseed (Brassica napus L., AnAnCnCn, ~1130 Mb, 2n=4x=38) originated from spontaneous interspecific hybridization between the diploid progenitors B. rapa (ArAr, ~485 Mb, 2n=2x=20) and B. oleracea (CoCo, ~630 Mb, 2n=2x=18) about 7500 years ago, followed by chromosome doubling, a process known as allopolyploidy (Chalhoub et al, 2014) These polyploidy events resulted in numerous duplicated segments and homoeologous regions within the B. napus genome (Chalhoub et al, 2014), which cause enormous challenges in the localization of quantitative trait loci (QTLs) for agronomic traits; for example, inaccuracies are caused by homologous sequences from different chromosomes and the interactions between homologue genes (Liu et al, 2015)

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