*4.2. Characterization of LiNAC Proteins*

The website ExPasy (https://www.expasy.org/) (accessed on 18 March 2022) predicted and examined the physicochemical characteristics of all LiNAC potential proteins, including molecular weight and theoretical points. TMHMM-2.0, a website that can be accessed online (https://services.healthtech.dtu.dk/service.php?TMHMM-2.0) (accessed on 18 March 2022), examined the transmembrane structure of LiNAC protein. The online program MEME (https://meme-suite.org/meme/tools/meme) (accessed on 20 March 2022) was used to find conserved motifs. The following parameters were utilized: maximum number of motifs = 20; search model = zero or one occurrence per sequence; default values for the remainder. TBtools software v1.098661(Chen, C.J.; Guangzhou, China) was used to display the results [37]. Cell-PLoc2.0 was used to create predictions for subcellular localization (http://www.csbio.sjtu.edu.cn/bioinf/plantmulti/#) (accessed on 20 March 2022).

#### *4.3. Sequence Alignment and Phylogenetic Tree Construction*

Using MUSCLE, the sequences of the found 21 LiNAC proteins and 80 AtNAC protein sequences were aligned. In MEGA 6.0, the outcome was utilized to create a neighborjoining (NJ) phylogenetic tree using 1000 bootstrap repetitions. P-distance and pairwise were the predetermined parameters. Phylogenetic tree were illustrations created with iTOL (https://itol.embl.de/) (accessed on 20 March 2022). Using the Jalview software 2.11.2.4 (Andrew, M.W.; Cambridge, MA, USA) (http://www.jalview.org/)(accessed on 20 March 2022), the amino acid sequences of conserved domains were compared and modified and conserved motif Logos were produced using the WebLogo program (http://weblogo.threeplusone.com) (accessed on 20 March 2022).

#### *4.4. Annotation and Enrichment Analysis in GO and KEGG Databases*

Online software EggNOG-Mapper (http://eggnog-mapper.embl.de/) (accessed on 20 March 2022) [38] and KEGG database (https://www.kegg.jp/) (accessed on 20 March 2022) were used to annotate the GO and KEGG functions of *LiNAC* gene. The results were collated using the eggNOG-mapper Helper function of TBtools v1.098661 and the text files for downstream analysis were output into GO Enrichment and KEGG Enrichment Analysis for enrichment analysis, respectively. Finally, the data were seen and examined using the online charting tool HIPLOT (https://hiplot.com.cn/) (accessed on 20 March 2022).
