Abstract: Cyperus esculentus L. is an emerging oil crop and one of the few known crops capable of synthesizing large amount of oil in underground tubers. Elucidating the genetic mechanisms underlying tuber oil biosynthesis is of great significance for molecular breeding of C.esculentus.Acetyl‑CoA carboxylase（ACCase），a key rate‑limiting enzyme catalyzing the first committed step of fatty acid biosynthesis，plays an important role in oil synthesis. In this study，using the cultivar Yuyousha No.2 as the experimental material，members of C.esculentus ACCase gene family（CeACCases） were identified based on genomic data. Their physicochemical properties，subcellular localization，phylogenetic relationships，conserved motifs，and promoter cis‑acting elements were systematically analyzed using bioinformatics methods.The expression patterns of each member in different tissues were examined by qRT‑PCR.The key gene CeACCase2 was cloned，and its subcellular localization and heterologous overexpression functional verification in Arabidopsis thaliana were performed.The results showed that a total of 13 CeACCases members were identified from C.esculentus.Their encoded proteins were mainly localized in chloroplasts，with significant differences in physicochemical properties. Phylogenetic and conserved motif analyses divided the family members into five subfamilies，among which the homomeric members possessed the most complete conserved motifs.The promoter regions were enriched with oil synthesis‑related cis‑acting elements. qRT‑PCR analysis revealed that CeACCases members exhibited relatively high expression levels in tubers.Subcellular localization results indicated that the CeACCase2 protein was mainly localized in chloroplasts. Overexpression of CeACCase2 in Arabidopsis significantly increased seed oil content by 13.1 percentage ponits compared with the wild type. In conclusion，this study confirms that CeACCase2 plays a key role in oil biosynthesis，providing theoretical support for elucidating the genetic basis of the tuber oil biosynthesis pathway in C.esculentus.

Key words: Cyperus esculentus, Oil synthesis, Acetyl?CoA carboxylase（ACCase）, Subcellular localization, Functional verification

摘要： 油莎豆（Cyperus esculentus L.）是一种新兴油料作物，是目前已知少数能在地下茎豆中大量合成油脂的作物之一，解析其茎豆油脂合成的遗传机制对油莎豆分子育种具有重要意义。乙酰辅酶A羧化酶（Acetyl‑CoA carboxylase，ACCase）作为脂肪酸合成途径中第一步反应的关键限速酶，在油脂合成中发挥着重要作用。以豫油莎2 号为试材，基于基因组数据鉴定油莎豆乙酰辅酶A 羧化酶基因家族（CeACCases）成员，利用生物信息学方法系统分析其理化性质、亚细胞定位、系统进化关系、保守基序及启动子顺式作用元件；通过qRT-PCR检测各成员在不同组织的表达模式；并对关键基因CeACCase2进行克隆、亚细胞定位及拟南芥异源过表达功能验证。结果表明，从油莎豆中共鉴定到13个CeACCases成员，其编码蛋白质主要定位于叶绿体，理化性质差异明显。系统发育和保守基序分析将该家族成员划分为5个亚家族，其中同质型成员具有最为完整的保守基序；启动子区富集油脂合成相关顺式作用元件。qRT-PCR分析显示，CeACCases成员在茎豆中均具有较高的表达量。亚细胞定位结果表明，CeACCase2蛋白主要定位于叶绿体；在拟南芥中过表达CeACCase2可使种子油脂含量较野生型显著提高13.1百分点。综上，证实CeACCase2在油脂合成中发挥关键作用，为解析油莎豆茎豆油脂合成途径的遗传基础提供了理论支持。

关键词: 油莎豆, 油脂合成, 乙酰辅酶A羧化酶, 亚细胞定位, 功能验证