Lza/) applying HISAT2 [61] (http://ccb.jhu.edu/ software/hisat2/index.shtml). The study count value was determined by HTSeq [62] (https://htseq.readthedocs.io/ en/release_0.11.1/). Fragments per kilobase million (FPKM) values had been IL-10 web calculated to estimate gene expression levels. DEGs between the two groups had been identified employing DESeq [63] determined by p 0.05 and |log2 foldchange| 1. Gene ontology (GO) enrichment evaluation of the DEGs was performed utilizing topGO [64], andqRT-PCR was performed on a BioRad CFX96 real-time method utilizing a kit from Vazyme Biotechnology Co., Ltd. (Nanjing, China). The reaction circumstances have been as follows: 95 for 30 s and 40 cycles (95 for ten s, 56 for 30 s, 72 for 60 s). The 2-Ct method was applied to evaluate the relative expression of genes determined by the steady expression level of BnaActin 7 [10]. The primer pairs were developed by Vector NTI Advance 11.5.1 computer software and synthesized by Sangon Biotech (Shanghai, China) (Table two).Measurement of physiological parameters in rootsThe physiological parameters, like 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 were performed in triplicate and signifies were calculated for further evaluation. The proline content was estimated using the approach described by predecessors [69]. The contents of PRO, SUG, MDA, PAL, SOD, CAT, and POD have been measured applying kits from Sino Greatest Biological Technologies Co., Ltd. (Shanghai, China).Wang et al. BMC Genomics(2021) 22:Web page 14 ofAbbreviations SNP: MEK2 MedChemExpress 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 3; 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 3; CDPK: calcium-dependent protein kinase; ERF2: ethylene-responsive element-binding issue 2; OPCL1: OPC-8:0 CoA ligase3.four.5.6.Supplementary InformationThe on-line version consists of supplementary material accessible at https://doi. org/10.1186/s12864-021-07614-1.7.eight. Further file 1 Table S1. Good quality and annotation of RNA-seq assembly. Added 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 Analysis Center for delivering the seeds of Brassica napus. Authors’ contributions CC and QYZ conceived the study. LYW, RLW and WL carried out the experiments. LYW wrote the original manuscript. JYW, CYL, QYZ and CC 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 with the manuscript. The author(s) study and approved the final manuscript. Funding This analysis was supported by grants in the National Key Investigation and Development Strategy (2018YFD0100500) and Chongqing Technology Innovation and Application Development (cstc2019jscx-msxmX0383). The funding bodies pla.