山西农业大学学报（自然科学版）2018，Vol.38Issue(3)：23-29,7.DOI:10.13842/j.cnki.issn1671-8151.201712034
杜氏盐藻Ds FA D2基因的鉴定及缺氮胁迫下表达分析
Identification of DsFAD2 gene in Dunaliella Salina and its expression analysis under nitrogen-defi-ciency stress
摘要
Abstract
[Objective]w-6 fatty acid desaturase 2(FA D2)is responsible for the conversion of oleic acid(18:1)to linole-ic acid(18:2),which is the first key enzyme for polyunsaturated fatty acid synthesis.This study investigated the func-tion of DsFAD2 in the accumulation of oil synthesis in Dunaliella Salina.[Methods]Dunaliella salina strain(YC-011) was isolated from Yuncheng Salt Lake,and was selected for investigation.High-fidelity PCR was used to clone Ds-FA D2 gene from Dunaliella salina.Several bioinformatics tools were employed to characterize the physical-chemical features,transmembrane regions,3D structure,and conserved domains of DsFAD2 protein.Real-time PCR was used to detect the expression pattern of DsFA D2 gene under nitrogen(N)stress.[Results]the DsFAD2 protein possessed five transmembrane regions and was located in the endoplasmic reticulum.The typical structures of the membrane-bound proteins were detected in the DsFAD2 protein sequence.In addition,DsFAD2 contained three conserved histi-dine domains and an aromatic amino acid sequence,which were unique in the FAD2 protein family.Phylogenetic analy-sis revealed that DsFAD2 was the closest to Chlamydomonas reinhardtii CrFAD2 and Volvox carteri VcFAD2.qRT-PCR analysis showed that N-deficiency stress significantly induced the expression of DsFA D2 in algal cells.The ex-pression of DsFA D2 was 2.8-fold higher in the treated cells than that of the control.[Conclusion]DsFAD2 was in-volved in the N-stress response of D.salina,and led to the increase of polyunsaturated fatty acid synthesis and stress resistance ability in the algal cells.This study provided a scientific reference for the further analysis of molecular mech-anism on underlying lipid accumulation and N-stress response in microalgal cells.关键词
杜氏盐藻(Dunaliellasalina)/ω-6脂肪酸去饱和酶2(FAD2)/不饱和脂肪酸/缺氮胁迫Key words
Dunaliella salina/Omega-6 fatty acid desaturase 2(FA D2)/Unsaturated fatty acids/Nitrogen-deficiency stress分类
农业科技引用本文复制引用
宋亚楠,安茜,岳敏,赵熙宁,刘宝玲,薛金爱,李润植..杜氏盐藻Ds FA D2基因的鉴定及缺氮胁迫下表达分析[J].山西农业大学学报（自然科学版）,2018,38(3):23-29,7.基金项目
国家自然科学基金(31401430) （31401430）
国家农业部"948"项目(2014-Z39) （2014-Z39）
山西省煤基重点科技攻关项目(FT-2014-01) （FT-2014-01）
山西省重点研发计划重点项目(201603D312005) （201603D312005）
