4.4. The Bioinformatics Analysis of VvACO1 and PpACO1

The protein sequences of VvACO1 and PpACO1 were obtained from the NCBI (https://www.ncbi.nlm.nih.gov/guide/) (accessed on 10 November 2021) and Phytozome 12 (https://phytozome.jgi.doe.gov/pz/portal.html#) (accessed on 1 September 2021). Furthermore, the homologous protein sequences of VvACO1 and PpACO1 from the other 6 non-climacteric and climacteric species, including FvACO1 (Fragaria vesca, {"type":"entrez-protein","attrs":{"text":"XP_004293334.1","term_id":"470114261","term_text":"XP_004293334.1"}}XP_004293334.1), PaACO1 (Prunus avium, {"type":"entrez-protein","attrs":{"text":"XP_021823230.1","term_id":"1220073462","term_text":"XP_021823230.1"}}XP_021823230.1), CcACO (Citrus clementina, {"type":"entrez-protein","attrs":{"text":"XP_006449820.1","term_id":"567915013","term_text":"XP_006449820.1"}}XP_006449820.1), SlACO1 (Solanum lycopersicum, {"type":"entrez-protein","attrs":{"text":"NP_001234024.2","term_id":"902763266","term_text":"NP_001234024.2"}}NP_001234024.2), MdACO (Malus domestica, {"type":"entrez-protein","attrs":{"text":"XP_008348143.1","term_id":"658025488","term_text":"XP_008348143.1"}}XP_008348143.1) and PbACO (Pyrus × bretschneideri, {"type":"entrez-protein","attrs":{"text":"XP_009365121.1","term_id":"694377671","term_text":"XP_009365121.1"}}XP_009365121.1), were obtained using the Blast function of the NCBI online website (https://blast.ncbi.nlm.nih.gov/Blast.cgi) (accessed on 15 September 2021). Sequence analyses of VvACO1 and PpACO1 were conducted using DNANMAN 8.0 software (accessed on 17 September 2021), including ORF sequence identification and multiple sequence alignment. A phylogenetic tree was constructed using the Maximum Likelihood method with MEGA 5.0 software (accessed on 18 September 2021), with a validation parameter of Bootstrap = 1000. The protein conservative domains were analyzed using the SMART online program (http://smart.embl-heidelberg.de/) (accessed on 1 October 2021). The conserved motifs of the proteins were identified using MEME 5.5.5 online software (http://meme-suite.org/tools/meme) (accessed on 10 October 2021). The protein secondary structures were predicted using PRABI (https://npsa-pbil.ibcp.fr/cgi-bin/secpred_sopma.pl) (accessed on 13 October 2021) and PSIPRED 4.0 (http://bioinf.cs.ucl.ac.uk/psipred/) (accessed on 14 October 2021), and the protein tertiary structures were predicted using SWISS-MODEL online software (https://swissmodel.expasy.org/interactive) (accessed on 17 October 2021). The cis-elements in the promoter regions were predicted using the PlantCARE online program (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) (accessed on 25 October 2021).