3.1. Identification of SNAREs in B. napus

To identify all candidate members of the SNARE family in B. napus, we combined three different methods. First, we identified the conserved domains and Pfams in all of the 64 SNAREs from A. thaliana (https://www.arabidopsis.org/, accessed on 23 August 2019) using the online batch CD-search server (https://www.ncbi.nlm.nih.gov/Structure/bwrpsb/bwrpsb.cgi, accessed on 25 August 2019 ). We generated 10 Pfams and 12 conserved domains specific for the SNARE superfamily (Table S2) as previously reported [12]. Then, genome-wide analysis of the Pfams was conducted and proteins containing the above Pfams were considered as SNARE protein candidates. Second, we searched for SNARE conserved domains and Pfams annotation in the B. napus genome annotation resources (http://www.genoscope.cns.fr/brassicanapus/, accessed on 26 October 2021) where proteins annotated with any of the 10 Pfams or the 12 conserved domains were identified as SNARE protein candidates. Third, the sequences of the 64 SNARE proteins in A. thaliana were used for similarity search against B. napus proteome by BLASTp. Finally, all possible BnaSNAREs identified by these three methods were validated by CDD (http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi, accessed on 14 January 2020), Pfam (http://pfam.xfam.org/, accessed on 21 January 2020) and SMART (http://smart.embl-heidelberg.de/, accessed on 16 January 2020) analyses.

Biochemical parameters such as length of sequences, molecular weights, and isoelectric points of BnaSNAREs were calculated by the ProtParam tool (https://web.expasy.org/protparam/, accessed on 19 January 2020) and the subcellular localization was predicted by three different tools: Plant-mPLoc [48], CELLO [49], and pLoc-mEuk [50].