Lza/) applying HISAT2 [61] (http://ccb.jhu.edu/ software/hisat2/index.shtml). The study count worth was determined by HTSeq [62] (https://htseq.readthedocs.io/ en/release_0.11.1/). Fragments per kilobase million (FPKM) values had been calculated to estimate gene expression levels. DEGs involving the two groups have been identified using DESeq [63] based on p 0.05 and |log2 foldchange| 1. Gene ontology (GO) enrichment analysis of your DEGs was performed making use of topGO [64], andqRT-PCR was performed on a BioRad CFX96 real-time system employing a kit from Vazyme Biotechnology Co., Ltd. (Nanjing, China). The reaction conditions were as follows: 95 for 30 s and 40 cycles (95 for 10 s, 56 for 30 s, 72 for 60 s). The 2-Ct approach was utilized to evaluate the relative expression of genes based on the stable expression degree of BnaActin 7 [10]. The primer pairs were designed by Vector NTI Advance 11.five.1 computer software and synthesized by Sangon Biotech (Shanghai, China) (Table 2).Measurement of physiological parameters in rootsThe physiological parameters, which includes soluble protein (PRO), soluble sugar (SUG), malondialdehyde (MDA), proline content material, and phenylalanine ammonia-lyase (PAL), superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities were measured. All measurements have been performed in triplicate and means have been calculated for additional analysis. The proline content was estimated using the method described by predecessors [69]. The contents of PRO, SUG, MDA, PAL, SOD, CAT, and POD had been CXCR1 review measured utilizing kits from Sino Finest Biological Technology Co., Ltd. (Shanghai, China).Wang et al. BMC Genomics(2021) 22:Web page 14 ofAbbreviations SNP: single nucleotide polymorphism; DEGs: differentially expressed genes; FPKM: fragments per kilobase million; GO: Gene ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; PRO: soluble protein; SUG: soluble sugar; MDA: malondialdehyde; PAL: phenylalanine ammonia-lyase; SOD: superoxide dismutase; CAT: catalase; POD: peroxidase; DOX: dioxygenases; LOX3: lipoxygenase three; ADH1: alcohol dehydrogenase 1; RBOH: respiratory burst oxidase homologue; WRKY: WRKY DNA-binding protein; ACO1: ACC oxidase 1; CYP450: cytochrome P450; ABC: ATP binding cassette subfamily; BCAT4: branched-chain aminotransferase four; MPK3: mitogen-activated protein kinase three; CDPK: calcium-dependent protein kinase; ERF2: ethylene-responsive element-binding issue 2; OPCL1: OPC-8:0 CoA ligase3.four.five.six.Supplementary InformationThe online version includes supplementary material offered at https://doi. org/10.1186/s12864-021-07614-1.7.eight. Additional file 1 Table S1. Good quality and annotation of RNA-seq assembly. More file two Table S2. Genes identified by combined GO and KEGG enrichment evaluation. Acknowledgements We are grateful to all of the colleagues in our laboratory, and thank Chongqing Engineering Study Center for supplying the seeds of Brassica napus. Authors’ contributions CC and QYZ conceived the study. LYW, RLW and WL performed the experiments. LYW wrote the original manuscript. JYW, CYL, QYZ and CC Kinesin-7/CENP-E custom synthesis helped to revise the manuscript. HSS, LJM and FY collected samples and measured physiological parameters. All authors have read and agreed to the published version from the manuscript. The author(s) study and approved the final manuscript. Funding This research was supported by grants from the National Important Study and Development Program (2018YFD0100500) and Chongqing Technology Innovation and Application Improvement (cstc2019jscx-msxmX0383). The funding bodies pla.