Mapping and Identifying a Candidate Gene (Bnmfs) for Female-Male Sterility through Whole-Genome Resequencing and RNA-Seq in Rapeseed (Brassica napus L.)

Abstract

In oilseed crops, carpel and stamen development play vital roles in pollination and rapeseed yield, but the genetic mechanisms underlying carpel and stamen development remain unclear. Herein, a male- and female-sterile mutant was obtained in offspring of a (Brassica napus cv. Qingyou 14) × (Qingyou 14 × B. rapa landrace Dahuang) cross. Subsequently, F2–F9 populations were generated through selfing of the heterozygote plants among the progeny of each generation. The male- and female-sterility exhibited stable inheritance in successive generations and was controlled by a recessive gene. The mutant kept the same chromosome number (2n = 38) as B. napus parent but showed abnormal meiosis for male and female. One candidate gene for the sterility was identified by simple sequence repeat (SSR) and insertion deletion length polymorphism (InDel) markers in F7–F9 plants, and whole-genome resequencing with F8 pools and RNA sequencing with F9 pools. Whole-genome resequencing found three candidate intervals (35.40–35.68, 35.74–35.75, and 45.34–46.45 Mb) on chromosome C3 in B. napus and candidate region for Bnmfs was narrowed to approximately 1.11-Mb (45.34–46.45 M) by combining SSR and InDel marker analyses with whole-genome resequencing. From transcriptome profiling in 0–2 mm buds, all of the genes in the candidate interval were detected, and only two genes with significant differences (BnaC03g56670D and BnaC03g56870D) were revealed. BnaC03g56870D was a candidate gene that shared homology with the CYP86C4 gene of Arabidopsis thaliana. Quantitative reverse transcription (qRT)-PCR analysis showed that Bnmfs primarily functioned in flower buds. Thus, sequencing and expression analyses provided evidence that BnaC03g56870D was the candidate gene for male and female sterility in the B. napus mutant.

Integrated QTL and eQTL Mapping Provides Insights and Candidate Genes for Fatty Acid Composition, Flowering Time, and Growth Traits in a F2 Population of a Novel Synthetic Allopolyploid Brassica napus

“…In total, 15,098 genes were differentially expressed between Da-Ae and Da-Ol-1. The number of differentially expressed genes between genotypes we identified are higher than other studies ( Teng et al, 2017 ; Li et al, 2018 ) because we included more (five) tissues types (see section “Materials and Methods”). It is also possible that the higher number of differentially expressed genes is because the two parental genotypes are adapted to different eco-environments (winter and spring).…”

Mapping and Identifying a Candidate Gene Plr4, a Recessive Gene Regulating Purple Leaf in Rice, by Using Bulked Segregant and Transcriptome Analysis with Next-Generation Sequencing

“…In the past few decades, molecular markers have played an important role in target gene location and selective breeding. However, the effect of gene mapping in different populations varies, and linkage mapping is difficult to carry out on some groups [30]. The purple leaf characters are regulated by multiple genes [20,26,27], and several related genes are located in the adjacent regions of chromosome 4 including the plr4 gene.…”

Characterization of the BnA10.tfl1 Gene Controls Determinate Inflorescence Trait in Brassica napus L.

“…Paraffin sectioning of the plant SAMs was performed as previously described [24]. The samples were treated with 50% FAA (Formalin-acetic acid-alcohol), then safranine-dyed, rinsed, dehydrated, cleared, infiltrated, embedded, sliced, sealed, and examined with a Nikon microscope (Nikon, Tokyo, Japan) [25]. Scanning electron microscopy (SEM) was performed following the previously described methods [26]: the SAMs were transferred to 2.5% glutaraldehyde and then rinsed, fixed again, and dehydrated.…”