*2.1. Identification of NAC Genes in L. indica*

Using AtNAC proteins as queries, 60 *LiNAC* genes were discovered by BLASTP searches in the *L. indica* transcriptome database and 43 putative *LiNAC* genes were discovered using an HMMER3 search in the *L. indica* protein database using the NAC-type NAM model (PF02365). Finally, using SMART and the NCBI Conserved Domain to remove duplicated genes and proteins without NAM-conserved domains, we were able to obtain 21 members of the *LiNAC* family. For further searching, all genes were given the designations *LiNAC1-LiNAC21* in the order of their assembled IDs (Table 1).


, *11*, 2168

*Plants* **2022**

**Table 1.** *LiNAC* gene and protein

characterization.

Proteins encoded by the 21 *LiNAC* genes contained 234 (LiNAC14) to 668 (LiNAC7) amino acids and their molecular weights (MWs) ranged from 26,491.05 (LiNAC14) to 74,336.43 (LiNAC7) Da, with an average molecular weight of 39,677 Da. Their predicted isoelectric point (pI) ranged from 4.42 (LiNAC4) to 9.93 (LiNAC19), with an average value of 6.90. Instability index calculation predicted that 17 (81%) of the NAC proteins were unstable. The LiNAC3, LiNAC4, LiNAC14, and LiNAC15 predicted proteins had instability indices less than 40 and were classified as stable proteins. Aliphatic amino acid indices showed that the thermal stability of the proteins ranged from 55.89 (LiNAC9) to 75.47 (LiNAC14). All LiNACs predicted protein grand average of hydropathicity (GRAVY) values as negative, demonstrating that they were predominantly hydrophilic. The predicted transmembrane structure results showed that LiNAC7 and LiNAC16 proteins had one transmembrane structure at the C-terminal of the 21 LiNAC proteins (Figure S1). Cell-PLoc subcellular localization predictions suggested that almost all LiNAC proteins were located in the Nucleus (Table 1).
