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| Identification and Analysis of PPO Gene Family Members in Lotus (Nelumbo nucifera) |
| LI Ting-Ting1, LI Yu-Rui1, YAN Min-Li1, ZHANG Hai-Xing1, WANG Rui-Hong1, KE Wei-Dong2, GUO Hong-Bo1,* |
1 College of Chemistry and Pharmacy, Northwest A&F University, Yangling 712100, China;
2 Institute of Vegetable, Wuhan Agricultural Academy, Wuhan 430065, China |
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Abstract Polyphenol oxidase (PPO) widely exits in plants, animals and insects with multigene family members. It is the key enzyme causing browning for fruits and vegetables. It will be an effective means to reduce PPO gene expression and enzyme activity through grasp of its family members and their function. In order to further understand the function of PPO gene family members in lotus (Nelumbo nucifera), bioinformatics methods were used to identify members of the PPO gene family: PPO1, PPO4, aureusidin synthase 1 (AS-1) and AS-2. Its physicochemical properties, transmembrane region, signal peptide, subcellular localization, gene structure, conserved motif, secondary and tertiary structures were predicted, and PPO activity and gene response in different tissues, growth time and induction factors were detected by kit and qPCR, respectively. The results showed that the length of the CDS region of the 4 genes was between 1 740 and 1 833 bp, the number of encoded amino acids was 579 to 610, the molecular weight was 66.21 to 68.65 kD, and the isoelectric point was 5.87 to 8.59, all of which belonged to hydrophilic unstable proteins. PPO1 and PPO4 had no transmembrane region and no signal peptide, which were non-secretory proteins. AS-1 had no signal peptide but one transmembrane region. AS-2 was a secreted protein with one transmembrane region. PPO1 and PPO4 were located in the chloroplast, AS-1 was located in the endoplasmic reticulum, and AS-2 was located in the cytoplasm. PPO1 and PPO4 had no introns but both had 12 conserved motifs. AS-1 and AS-2 had one intron and 10 conserved motifs, respectively. The 4 proteins were very similar in structure, and the random coil was the main structural element of the proteins. The α-helix, β-turn and extension chain were dispersed throughout the polypeptide chain. The activity of PPO and expression of PPO1 and PPO4 were the highest in relatively young tissue buds, those of AS-1 was the highest in roots, and those of AS-2 was the highest in leaves. The activity of PPO and expression of PPO1, PPO4, and AS-1 were increased under different induction treatments, but AS-2 had no significant change; with the change of growth time, the PPO activity tended to increase first and then decrease. The expression of PPO1, PPO4 and AS-1 increased rapidly in around 30 d, while AS-2 did not change significantly. This study laid the foundation for further exploration of the characteristics and functions of the lotus PPO gene family.
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Received: 17 July 2021
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Corresponding Authors:
*hbguo@nwsuaf.edu.cn
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