CrLEC1基因调控莱茵衣藻油脂合成的功能解析  

Functional analysis of CrLECI gene regulating lipid synthesis in Chlamydomonas reinhardti

作　　者：张浩东 李亚军[1,2] 谭德冠 张家明[1,2] 马帅 ZHANG Haodong;LI Yjun;TAN Deguan;ZHANG Jiaming;MA Shuai(Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences/Hainan Institute of Tropical Agricultural Resources,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China;Hainan Key Laboratory of Tropical Microbe Resources,Haikou 571101,China)

机构地区：[1]中国热带农业科学院热带生物技术研究所中国热带农业科学院海南热带农业资源研究,海南海口571101 [2]海南省热带微生物资源重点实验室,海南海口571101

出　　处：《生物技术》2024年第3期295-303,共9页Biotechnology

基　　金：海南省自然科学基金项目(321MS08);中国热带农业科学院基本科研业务费创新团队项目(1630052020020)。

摘　　要：[目的]通过解析转录因子LEC1(Leafy cotyledon 1)在微藻油脂生物合成中的调控功能,为建立利用基因工程培育高含油量和高生物量微藻的育种策略提供基础。[方法]采用无缝克隆法构建含CrLEC1基因的过表达载体;采用玻璃珠转化法进行莱茵衣藻(Chlamydomonas reinhardtii)CC849转化;采用尼罗红荧光法测定转基因藻株的油脂含量。[结果]成功构建了pCam1304::LEC1::GFP载体,获得137个阳性转化的藻株,经检测有16个为转基因藻株。发现转基因藻株6-15的生长不受氮胁迫的影响,且在缺氮或含氮培养条件下其生长状况均较野生型藻株好;在缺氮培养条件下,转基因藻株6-15、6-9、5-22、5-25、5-34的油脂含量均显著高于野生型藻株。[结论]CrLEC1基因正调控莱茵衣藻的油脂合成。[Objective]To provide a basis for establishing breeding strategies for cultivating high oil content and high biomass microalgae through genetic engineering,the regulatory function of the transcription factor LEC1(Leaf cotyledon 1)in microalgae oil biosynthesis will be analyzed.[Method]An overexpression vector containing CrLECl gene were constructed by seamless cloning method,Chlamydomonas reinhardti CC849 cells were transformed by glass bead transformation method,and the oil content of transgenic algae strains were determined by Nile red fluorescence assay.[Result]JThe plasmid pCaml304::LECl::GFP were successfully constructed.137 positive transformed algae strains were obtained,of which 16 transgenic algae strains were detected.The growth of transgenic algae strains 6-15 was not affected by nitrogen stress,and their growth status was better than that of wild-type algae strains under nitrogen deficient or nitrogen containing culture conditions.Under nitrogen deficient conditions,the oil content of transgenic algae strains 6-15,6-9,5-22,5-25 and 5-34 was significantly higher than that of wild-type algae strains.[Conclusion]CrLEC gene was positively regulating lipid synthesis in Chlamydomonas reinhardti.

关 键 词：莱茵衣藻 转化 过量表达 转录因子 LEC1 油脂 合成 氮胁迫 

分 类 号：Q949.211[生物学—植物学]