Cloning and Bioinformatics Analysis of Chlorophyll Degrading Gene PPH from Curcuma alismatifolia

Abstract

To obtain key enzyme gene PPH in chlorophyll degradation process of Curcuma alismatifolia , on the basis of obtaining a large amount of transcriptome information by sequencing the full-length transcriptome, we had screened and analyzed these transcriptome information and obtained 2 PPH genes which named PPH1 and PPH2 . The PPH1 gene (GenBank: MT077178) has a full-length cDNA sequence of 1 795 bp in length, an open reading frame of 1437 bp (from 138 to 1 574 bp), and encode a sequence with 478AA amino acid. The PPH2 gene (GenBank: MT077179) has a full-length cDNA sequence of 1393bp, an open reading frame of 1227bp (from 70 to 1296bp), and encode a sequence with 408AA amino acid. Using Blast, Translate tool (ExPASy), Clustal Omega, Find Conserved Domains (NCBI), ProtParam, TMHMM Server, SOPMA, SWISS-MODEL, ClustalX (1.81), MEGA4.1 and so on. Their amino acid composition, physical and chemical properties, conserved domains, secondary structures, tertiary crystal structures, and molecular phylogeny were predicted and analyzed. The nucleotide and protein amino acid sequences of PPH1 and PPH2 have high homology with these PPH genes of other species, and both of them contain a conserved region PLN02578 with hydrolase characteristic. Molecular phylogenetic analysis showed that a small cluster of PPH1 and PPH2 were closest to Musa acuminate PPH (XP_018677219.1), but far away from dicotyledons. This study provided a molecular basis for improving color of Curcuma alismatifolia sterile bracts by genetic transformation in the future.