生物技术通报2026，Vol.42Issue(10)：88-97,10.DOI:10.13560/j.cnki.biotech.bull.1985.2026-0470
燕麦溶血磷脂酸酰基转移酶AsLPAAT5基因克隆与功能分析
Cloning and Functional Characterization of the Lysophosphatidic Acid Acyltransferase Gene AsLPAAT5 in Avena sativa
摘要
Abstract
[Objective]Lysophosphatidic acid acyltransferase(LPAAT)is a key rate-limiting enzyme in triacylglycerol(TAG)assembly pathway.This study aimed to investigate the function of the oat(Avena sativa L.)AsLPAAT5 gene in lipid biosynthesis and accumulation,providing new insights for improving oil yield and quality in oat and other crops.[Method]The AsLPAAT5 gene(Gene ID:AVESA.00001b.r1.1Ag0003121.1)was cloned from oat,and its structural characteristics were analyzed using bioinformatics tools.The enzymatic activity and biological function of AsLPAAT5 were evaluated using the Escherichia coli LPAAT-deficient mutant strain SM2-1,Saccharomyces cerevisiae wild-type strain INVSc1,and Arabidopsis thaliana transformation systems.[Result]The protein encoded by AsLPAAT5 possessed the typical conserved domains of LPAAT.AsLPAAT5 was expressed in roots,stems,leaves,panicles,and seeds at 9,17,and 25 days after flowering,with the highest expression level detected in seeds at 25 days after flowering.Functional complementation assays in E.coli confirmed that AsLPAAT5 can restore membrane lipid biosynthesis in the mutant strain SM2-1,demonstrating LPAAT enzymatic activity.Overexpression of AsLPAAT5 in S.cerevisiae resulted in a significant 2.11％increase in total fatty acid content compared with wild-type yeast,along with a 14.25％increase in C16:1 content.Overexpression of AsLPAAT5 in A.thaliana increased thousand-seed weight by 2.2％–2.8％,enhanced total oil content by 1.9％–2.0％,and significantly elevated C18:2 content by 25.49％–26.0％.[Conclusion]AsLPAAT5 is an endoplasmic reticulum-localized enzyme with typical LPAAT catalytic activity.Heterologous expression of AsLPAAT5 significantly promotes total lipid accumulation in hosts and exerts a specific regulatory effect on the enrichment of unsaturated fatty acids,particularly C18:2.关键词
燕麦/溶血磷脂酸酰基转移酶/油脂合成/亚油酸/功能分析Key words
Avena sativa L./lysophosphatidic acid acyltransferase/oil biosynthesis/linoleic acid/functional analysis引用本文复制引用
胡荣芳,丁家豪,马菊花,谭辉进,宋亚楠,薛金爱..燕麦溶血磷脂酸酰基转移酶AsLPAAT5基因克隆与功能分析[J].生物技术通报,2026,42(10):88-97,10.基金项目
国家自然科学基金项目(31401430),山西省基础研究计划(202103021224170),山西省高等学校科学研究优秀成果培育项目(J242042025),山西农业大学生物育种工程项目(YZGC101),山西农业大学其他纵向科研项目(2024QT043) （31401430）
