Genome-wide Identification of the FAT Gene Family and Functional Analysis of CeFATA in Cyperus esculentus

doi:10.13560/j.cnki.biotech.bull.1985.2025-1395

Abstract

Abstract:

Objective Fatty acyl-ACP thioesterase (FAT) plays a key role in plant oil synthesis. This study aimed to characterize FAT family genes in yellow nutsedge (Cyperus esculentus L.) and to provide a theoretical basis and technical support for breeding high-oil cultivars and improving lipid metabolism. Method Based on the whole-genome data of yellow nutsedge, the CeFAT gene family was systematically identified and subjected to sequence and phylogenetic analyses. The subcellular localization of CeFATA was determined, and the function of CeFATA was evaluated using yeast and tobacco genetic transformation systems. Result Three CeFAT genes, CeFATA, CeFATB1, and CeFATB2, were identified. They showed similar gene structures and physicochemical properties, and were located on three different chromosomes. Phylogenetic analysis indicated that CeFATA was most closely related to OsFATA from rice, whereas CeFATB1 and CeFATB2 were closely related to GmFATB from soybean. RT-qPCR analysis showed that CeFAT genes were highly expressed at five developmental stages of the tuber, with CeFATA showing the highest expression level. Subcellular localization confirmed that CeFATA was located in chloroplasts. Overexpression of CeFATA increased total lipid content in Saccharomyces cerevisiae by 3.1% and exhibited substrate preference for C18:1. In transgenic tobacco, CeFATA overexpression increased total fatty acid contents in leaves and seeds by 4.74% and 8.4%, accompanied by a significant decrease in starch content, a decrease soluble sugar content in leaves, an increase soluble sugar content in seeds, and no significant change in protein content. Conclusion CeFATA increases total lipid and oleic acid contents in yeast and tobacco, promotes the export of oleic acid from plastids, and redirects carbon flux from carbohydrate biosynthesis toward oil biosynthesis.

Key words: Cyperus esculentus L., fatty acyl-ACP thioesterase (FAT), subcellular localization, fatty acid biosynthesis, functional analysis

LI Shu-ting, WANG Ying, DING Jia-hao, HU Rong-fang, LI Chen, SONG Ya-nan, XUE Jin-ai. Genome-wide Identification of the FAT Gene Family and Functional Analysis of CeFATA in Cyperus esculentus[J]. Biotechnology Bulletin, 2026, 42(8): 162-172.

Figures/Tables 9

Fig. 1 Chromosomal localization of CeFATs genes in C. esculentus (A) and synteny analysis with three plant species (Arabidopsis, soybean, and rice) (B)

Fig. 2 Phylogenetic tree analysis of CeFATs and FAT proteins from different speciesCe: Cyperus esculentusis; Pd: Phoenix dactylifera;Cn: Cocos nucifera; Ma: Musa acuminata; Mb: Musa balbisiana; At: Arabidopsis thaliana; Os: Oryza sativa; Gm: Glycine max; Ha: Helianthus annuus; Nt: Nicotiana tabacum; Si: Sesamum indicum;Ac: Ananas comosus

Fig. 3 Analysis of FAT family protein structure and amino acid composition in C. esculentus

Fig. 4 Cis-acting element analysis of CeFATs genes in C. esculentus

Fig. 5 Transcriptome expression profiling (A) and RT-qPCR analysis (B) of CeFATs at different tuber developmental stages in C. esculentusDifferent lowercase letters indicate significant differences (P<0.05). The same below

Fig. 6 Subcellular localization of CeFATA protein

Fig. 7 Total fatty acid content and growth curve of transgenic yeast under exogenous fatty acid additionA: Total fatty acid content; B: Growth curves of empty-vector yeast with four exogenous fatty acids; C: Growth curves of transgenic yeast with four exogenous fatty acids

Fig. 8 Fatty acid contents of transgenic yeast

Fig. 9 Physiological and biochemical indices of transgenic tobacco linesA: Total lipid content in leaves. B: Protein content in leaves. C: Starch content in leaves. D: Soluble sugar content in leaves. E: Total lipid content in seeds. F: Protein content in seeds. G: Starch content in seeds. H: Soluble sugar content in seeds. WT: Wild tobacco; EV: Empty vector tobacco; CeFATA: Transgenic tobacco lines. DW: Dry weight

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