ABSTRACT
ObjectiveThe biosynthetic pathways of benzylisoquinoline alkaloids（BIAs） in Nelumbo nucifera are of great theoretical and economic value. In this paper， N. nucifera O-methyltransferase（NnOMT） and N. nucifera N-methyltransferase（NnNMT） gene families were identified and analyzed by bioinformatics in order to facilitate the biosynthetic pathway of BIAs in N. nucifera. MethodBased on the whole genome of N. nucifera， UniPort and National Center for Biotechnology Information（NCBI） databases were used to identify the NnOMT and NnNMT gene families of N. nucifera， and analyze their physicochemical properties and subcellular localization， then TBtools， MEME， MEGA 11.0， FigTree 1.4.4 and other tools were used to analyze the phylogeny， sequence characteristics， gene structure， functional annotation and cis-acting elements of NnOMT and NnNMT genes identified in the previous stage. ResultA total of 61 NnOMT and NnNMT genes were identified in this paper， the number of amino acids encoded by these genes ranged from 168 aa to 580 aa， the isoelectric point ranged from 4.76 to 9.16， and the relative molecular weight ranged from 18 699.52 Da to 64 934.53 Da， most of which showed acidic and mostly hydrophilic proteins. There were 10 conserved motifs， Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis enriched a total of 12 pathways， including metabolism， biosynthesis of phenylpropane and isoquinoline alkaloids， etc. And Visualization of Gene Ontology（GO） enrichment results showed that 61 NnOMT and NnNMT genes were annotated to 32 items， which included 16 molecular functions［such as reduced nicotinamide adenine dinucleotide（NADH） activity and exopeptidase activity］ and 16 biological processes（such as metabolic process of carbon tetrachloride， anaerobic carbon tetrachloride metabolic process and responses to exogenous biological stimuli）. There were a variety of cis-acting elements in the promoter regions of NnOMT and NnNMT genes， mainly promoter and enhancer regions element， light responsive element and methyl jasmonate responsive element. ConclusionIn this study， a comprehensive bioinformatics analysis of 61 NnOMT and NnNMT genes is carried out based on the genome data of N. nucifera， which lays a foundation for research on the gene structure and function of NnOMT and NnNMT gene families， and provides a reference for biosynthetic pathway elucidation of BIAs in N. nucifera.
ABSTRACT
ObjectiveCYP71 gene family is one of the CYP71 clans belonging to cytochrome P450， which plays an important role in secondary metabolites， especially terpenoid biosynthesis. To understand the characteristics of CYP71 family of diploid Perilla frutescens and predict its function， this study identified and systematically analyzed the family by bioinformatics. MethodOn the basis of the whole genome of diploid P. frutescens PC99， the conserved domains of CYP71 family of diploid P. frutescens were screened， and the sequence characteristics， gene structure， chromosome location， phylogeny and cis-acting elements were analyzed by National Center for Biotechnology Information （NCBI）， TBtools， MEME， Molecular Evolutionary Genetics Analysis （MEGA）， Cytoscape and other tools. ResultA total of 68 CYP71 genes were identified from diploid P. frutescens， which were unevenly distributed on 34 chromosomes and belonged to two subfamilies. They encoded 481-530 amino acids and contained 10 conserved motifs， with the isoelectric point of 5.70-9.03 and the molecular weight of 54 217.07-60 031.79 Da. The enrichment analysis and functional annotation analysis revealed 11 enriched pathways and 114 categories， and the genes were mainly annotated in biological processes. There were many cis-acting elements in the promoter region of CYP71， mainly light-responsive and methyl jasmonate-responsive elements. Protein-protein interaction analysis indicated that CYP71 protein had multiple functions such as terpene cyclase activity. ConclusionThis study lays a foundation for the functional study of CYP71 family， and provides a reference for the biosynthesis of monoterpenes in P. frutescens and the directional cultivation of excellent varieties.