4.1. Genome-Wide Identification of m6A-Related Family in Aegilops tauschii

The whole genome reference and annotated protein sequences of Aegilops tauschii were downloaded from EnsemblPlants database (https://plants.ensembl.org/index.html, accessed on 30 November 2022) and then used as the local protein database. The Hidden Markov Model (HMM) profile of the m6A regulators domains (including PF05063, PF17098 and PF15912) were downloaded from PFAM database (http://pfam.xfam.org/ (accessed on 10 March 2023)) and used as the query to search against local protein database using the HMMER 3.0 with the threshold of E-value < 1 × 10⁻⁵ [29]. Meanwhile, the m6A regulatory proteins of Arabidopsis thaliana and rice were further used as queries to perform BLASTP search against the local protein database (E value < 1 × 10⁻⁵, agreement 50%, coverage > 50%) [30]. The results obtained from above two methods were integrated and removed the redundant manually to identify the putative m6A-related proteins in Aegilops tauschii (Aetm6As). Finally, these putative Aetm6As were submitted to the PFAM (https://pfam.xfam.org (accessed on 10 March 2023)) database and the NCBI-CDD (http://www.ncbi.nlm.nih.gov/cdd/ (accessed on 10 March 2023)) database to confirm the presence and completeness of the conserved domains. Only genes containing conserved and typical domains of m6A writers, erasers and readers were retained as candidates for further analysis. The EXPASy online software (https://www.expasy.org/ (accessed on 10 March 2023)) was used to predict the molecular weight(Mw), length of amino acid(AA) and isoelectric point(pI) of these m6A proteins were investigated. Their subcellular localization was predicted based on the Plant—mPLoc subcellular location (http://www.csbio.sjtu.edu.cn/bioinf/plant-multi/ (accessed on 10 March 2023)) and the Busca online software.