*3.1. Genome-Wide Identification and Analysis of Hsf and Hsp Gene Family Members in Eggplant*

To search for *Hsf* and *Hsp* genes in eggplant, we used the conserved Hsf and Hsp domain consensus sequences of several proteins as BLASTP queries against the eggplant

genome database (http://eggplant.kazusa.or.jp/, accessed on 6 June 2021). In addition, homology searches using identified protein sequences of *Arabidopsis thaliana* were performed. After automated database searching and a manual review, 24 and 117 genes were identified as members of the *Hsf* and *Hsp* families in eggplant, respectively, whose classification and naming were based on the rules of the *Hsp* gene families from Arabidopsis and tomato, including *sHsp*, *Hsp60*, *Hsp70*, *Hsp90* and *Hsp100*. The *Hsf* and *Hsp* gene families in eggplant were relatively large compared with those in Arabidopsis and those in tomato and rice, respectively. The numbers of identified genes in the *Hsf*, *sHsp*, *Hsp60*, *Hsp70*, *Hsp90* and *Hsp100* families of eggplant were 24, 39, 21, 30, 17 and 10, respectively (Table 1).


**Table 1.** Numbers of *Hsf* and *Hsp* genes in Arabidopsis, eggplant, tomato and rice.

As shown in Supplementary Materials Table S1, the amino acid lengths for Hsfs ranged from 111 (*SmHsfA1c*) to 496 (*SmHsfA1b*), with deduced molecular weights from 12.2 kDa to 54.8 kDa and the predicted isoelectric points of Hsfs were divergent, ranging from 4.60 (*SmHsfA3*) to 9.64 (*SmHsfA1d*). The length of sHsp proteins ranged from 87 (*Sm10.2-sHsp*) to244 amino acids (*Sm27.2-sHsp*) and the predicted molecular weights were between 10.2 kDa (*Sm10.2-sHsp*) and 27.2 kDa (*Sm10.2-sHsp*). In addition, the predicted pI-values of sHsp proteins ranged from 4.56 (*Sm10.2-sHsp*) to 10.49 (*Sm12.7-sHsp*). The amino acids lengths were consistent with the molecular weights of Hsp60s. The amino acid number and molecular weight for Smcpn60-4 was the highest, while that for SmCpn60- 7.3 was the lowest and the predicted pI-values ranged from 5.26 (*SmCpn60-a1*) to 10.29 (*SmCpn60-7.3*). The deduced length of the Hsp70 proteins ranged from 85 (*SmmtHsc70-3*) to 914 (*SmHsp70-18*) amino acids and the highest- and lowest-molecular-weight SmHsp70s were *SmHsp70-18* (103.1 kDa) and *SmHsp70-5* (11.5 kDa), respectively, while the pI values ranged from 4.52 (*SmmtHsc70-3*) to 9.35 (*SmHsp70-19*). The length of Hsp90 proteins ranged from 137 (*SmHsp90-4.4*) to 782 (*SmHsp90-6*) amino acids, the predicted molecular weights of Hsp90s were between 16.2 kDa (*SmHsp90-4.4*) and 89.6 kDa (*SmHsp90-7.1*) and the predicted isoelectric points ranged from 4.78 (*SmHsp90-5*) to 9.55 (*SmHsp90-2.1*). The longest amino acids lengths and highest molecular weights in Hsp100s were *SmHsp100- ClpB1*, with 979 amino acids and 110.2 kDa, respectively. In contrast, the smallest was *SmHsp100-ClpC3*; the predicted isoelectric points ranged from 5.38 (*SmHsp100-ClpB3*) to 9.07 (*SmHsp100-ClpC1*) and these proteins were distributed from the alkaline to acidic.
