**2. Results**

#### *2.1. Sequence Analysis of* PLP\_deC *Family Members*

HMMER software was used to identify candidate genes in the genomes of *D. o*ffi*cinale* and *P. equestris*. All candidate genes were submitted to Pfam and SMART for verifying the presence of the PLP\_deC domain. The sequences without the conserved domain and the redundant sequences were removed. Finally, a total of 14 *PLP\_deC*-family sequences were obtained in *D. o*ffi*cinale* and *P. equestris*. The basic information of each *PLP\_deC* was listed in Table 1. We found that in *D. o*ffi*cinale*, the molecular weights ranged from 34.48 kDa (DoGAD4) to 79.29 kDa (DoAAD2), with an average molecular weight of 57.64 kDa, and the theoretical isoelectric points ranged from 5.27 (DoHDC1) to 7.52 (DoAAD1), with an average value of 6.12. In *P. equestris*, the molecular weights ranged from 53.44 kDa (PeHDC1) to 57.26 kDa (PeGAD3), with an average molecular weight of 55.53 kDa, while the theoretical isoelectric points ranged from 5.53 (PeGAD1) to 8.55 (PeGAD3), with an average of 6.52. These results showed that most genes have an acidic pI. We analyzed the subcellular localization of 8 and 6 PLP\_deC-family protein sequences in *D. o*ffi*cinale* and *P. equestris* by Target P 1.1. and the results showed the presence of a signal peptide in DoAAD1 and DoAAD3, suggesting their cellular localization in extracellular with a probability of 0.735 and 0.561, The proteins of DoHDC1, PeAAD1, PeAAD2 and PeHDC1 are possibly located in other cellular compartments, e.g. chloroplast transit peptide and mitochondrial, whereas the location of the remaining eight proteins is unknown (Table 2).


**Table 1.** Sequence analysis of pyridoxal phosphate-dependent decarboxylase (*PLP\_deC*) family members.

**Note:** <sup>√</sup> shows conserved domain of Pyridoxal\_deC.


Note: Confidence levels range from 1 to 5, and 1 shows the highest predicted reliability with difference > 0.8; 2: 0.8 > difference > 0.6; 3: 0.6 > difference > 0.4; 4: 0.4 > difference > 0.2; 5: 0.2 > difference. Location: C, M, and S, represent chloroplast, mitochondrial and extracellular, respectively. \_ other location, \* unknown location.
