Genome-Wide Analysis of KCS Gene Family in Ginkgo biloba L. and Functional Identification of KCS7 in Oleic Acid Synthesis

银杏叶中KCS基因家族的全基因组分析及KCS7在油酸合成中的功能鉴定

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Abstract

Background: β-ketolipoyl coenzyme A synthase (KCS) is an essential limiting catalyst involved in carbon chain elongation during fatty acid biosynthesis, characterized by strict substrate specificity. C18:1 (oleic acid) plays a vital role in cell membranes and is essential for nutrient storage and stress defense. There are indications of significant accumulation and rapid synthesis of C18:1 during the early growth stages of Ginkgo biloba L. episperm. The KCS gene family in G. biloba has yet to be analyzed, and the role of KCS in oleic acid synthesis remains unexplored. Methods: In this study, this issue was investigated using transcriptomic and metabolomic data, bioinformatics analysis to screen a key gene from the KCS gene family, and dual validation using yeast and Arabidopsis thaliana expression systems to probe its function. Results: A total of 11 members of the GbKCS gene family were identified, and the dynamics of these genes were analyzed during exocarp development in the G. biloba genome. Among them, the gene designated GbKCS7 showed a highly direct association with the content of C18:1. Heterologous expression of GbKCS7 in yeast increased C18:1N12 and C18:1 content by 3.18-fold and 2.07-fold, respectively. Overexpression of GbKCS7 in Arabidopsis showed that C18:1 was increased by 27.70% and 31.43% in GbKCS7-OE-1 and GbKCS7-OE-2 strains, correspondingly, in juxtaposition to the non-transgenic plants. In addition, the content of VLCFAs increased to varying degrees. Conclusions: These outcomes offer important insights for investigating the role of KCS genes in fatty acid synthesis to further improve G.biloba resistance.

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