qRT-PCR分析莱茵衣藻氮、硫同化相关基因的表达  

Analysis of the Genes Expression Related to Nitrogen and Sulfur Assimilation by qRT-PCR in Chlamydmonas reinhardtii

作　　者：李平[1,2] 费小雯[3] 李兴涵 邓晓东[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所,海南海口571101 [3]海南医学院理学院,海南海口571101

出　　处：《热带作物学报》2014年第12期2403-2409,共7页Chinese Journal of Tropical Crops

基　　金：海南省重大科技专项(No.ZDZX2013023-1-27);海南省自然科学基金(No.313077);海南省工程技术研究中心专项(No.GCZX2011006;GCZX2012004;GCZX2013004);中央级公益性科研院所基本科研业务费(No.ITBB110507;130305)

摘　　要：实验发现,缺氮、缺硫的环境有利于莱茵衣藻油脂的积累,说明与氮、硫同化相关的基因也与油脂合成有着密切的关系.分析了氮、硫同化通路相关基因,选择氮同化相关的6个基因（NAR3、NAR1.1、NII1、NIA1、NIT2、AOX1）以及硫同化相关的6个基因（SULP1、SIR1、SLT1、ARS1、SULTR1、ATS1）进行qRT-PCR分析.对莱茵衣藻CC124以及CC425藻株分别进行缺氮、缺硫处理,Trizol法连续提取4d的RNA作为模板,进行实时荧光定量PCR检测基因的表达水平.结果发现,这12种基因均较对照上调表达,其中,莱茵衣藻氮、硫运载体基因上调表达非常显著.缺氮条件下NAR3、NAR1.1较正常最高上调表达200倍;缺硫条件下SLT1较对照有1 000～2000倍的mRNA表达量,SULTR2也最高能增高450倍的表达量.The results showed that the accumulation of oil in the Chlamydmonas reinhardtii strains increased without nitrogen and sulfur.It proved that genes associated with nitrogen and sulfur synthesis have the close relationship with the assimilation of oil.In this work we chose six genes （NAR3、NAR1.1、NII1、NIA1、NIT2、AOX1） related to nitrogen assimilation and six genes （SULP1、SIR1、SLT1、ARS1、SULTR1、ATS1） related to sulfur assimilation to analysis mRNA expression by qRT-PCR.Experimented on Chlamydmonas reinhardtii CC124 and CC425 strains with treatment of nitrogen and sulfur free,extracted 4 consecutive days mRNA as a template using Trizol method,and detected gene expression levels by qRT-PCR.The results showed that the mRNA expression levels of the 12 genes higher than the normal ones,especially the transporter genes.Up-regulation of the expression of NAR3 NAR1.1 is 200-fold higher than the normal under the condition of nitrogen deficiency; Under the condition of sulfur starvation,the up-regulation of the expression of SULTR2 is 450-fold the highest level and SLT1 is the highest one which is 1 000～2 000-fold.

关 键 词：莱茵衣藻 缺氮 缺硫 QRT-PCR 

分 类 号：Q78[生物学—分子生物学]