Cloning and functional study of lysophosphatidic acid acyltransferase gene in Perilla frutescens.

Lysophosphatidic acid acyltransferase (LPAAT) is a key enzyme in plant triacylglycerol biosynthesis. Investigating the function of Perilla frutescens LPAAT gene (PfLPAAT) in oil biosynthesis can help to reveal the molecular mechanism of plant oil accumulation. In this study, RT-PCR was used to obtain PfLPAAT, and the basic physical and chemical properties, transmembrane domain, and subcellular localization of the putative protein of PfLPAAT was analyzed by bioinformatics methods, and phylogenetic analysis of homologous proteins was conducted. The qRT-PCR was used to analyze the relative expression level of PfLPAAT in different tissues and developmental stages of seeds in Perilla. The expression vector pCAMBIA1303-PfLPAAT was constructed and transformed into Arabidopsis thaliana by floral dip method, and the oil content and fatty acid composition of transgenic Arabidopsis seeds were analyzed. The results showed that the PfLPAAT sequence was 1149 bp in length and encoded 382 amino acids. The theoretical isoelectric point of the putative protein was 9.60, and the molecular mass was 43.02 kD. Bioinformatics indicated that PfLPAAT functioned in endoplasmic reticulum and belonged to the PLN02380 superfamily. The qRT-PCR revealed that PfLPAAT expressed in all tissues of Perilla, and the highest level was observed in leaves and seeds of 15 days after flowering. Compared with wild-type Arabidopsis, the overexpression of PfLPAAT in Arabidopsis improved seed oil content significantly, with a range of 2.04% to 13.40%. Fatty acid composition analysis demonstrated that the content of oleic acid, linoleic acid, linolenic acid, and arachidic acid increased significantly, while the content of palmitic acid, stearic acid, arachidic acid, arachidonic Lysophosphatidic acid acyltransferase (LPAAT) is a key enzyme in plant triacylglycerol biosynthesis. Investigating the function of Perilla frutescens LPAAT gene (PfLPAAT) in oil biosynthesis can help to reveal the molecular mechanism of plant oil accumulation. In this study, RT-PCR was used to obtain PfLPAAT, and the basic physical and chemical properties, transmembrane domain, and subcellular localization of the putative protein of PfLPAAT was analyzed by bioinformatics methods, and phylogenetic analysis of homologous proteins was conducted. The qRT-PCR was used to analyze the relative expression level of PfLPAAT in different tissues and developmental stages of seeds in Perilla. The expression vector pCAMBIA1303-PfLPAAT was constructed and transformed into Arabidopsis thaliana by floral dip method, and the oil content and fatty acid composition of transgenic Arabidopsis seeds were analyzed. The results showed that the PfLPAAT sequence was 1149 bp in length and encoded 382 amino acids. The theoretical isoelectric point of the putative protein was 9.60, and the molecular mass was 43.02 kD. Bioinformatics indicated that PfLPAAT functioned in endoplasmic reticulum and belonged to the PLN02380 superfamily. The qRT-PCR revealed that PfLPAAT expressed in all tissues of Perilla, and the highest level was observed in leaves and seeds of 15 days after flowering. Compared with wild-type Arabidopsis, the overexpression of PfLPAAT in Arabidopsis improved seed oil content significantly, with a range of 2.04% to 13.40%. Fatty acid composition analysis demonstrated that the content of oleic acid, linoleic acid, linolenic acid, and arachidic acid increased significantly, while the content of palmitic acid, stearic acid, arachidic acid, arachidonic [ABSTRACT FROM AUTHOR]