金线莲应答高温胁迫的转录组学特征分析

Anoectochilus roxburghii(Wall.)Lindl.in Response to Heat Stress:Transcriptomic Analysis

为了研究高温胁迫下金线莲转录组表达特征,分析其热激响应机制。以正常培养和高温胁迫(45o C)的金线莲(NYJ2)为材料,利用Illumina HiSeqTM2000平台进行测序,通过Trinity软件进行De novo组装。结果共获得75688个unigene,有28323个unigene在Nr、KOG、KEGG和Swisspro数据库中得到功能注释,注释率为37.42%。其中,17532个unigene在KOG数据库中可归类到25个功能家族;10108个unigene被KEGG数据库注释到128个代谢途径中;17485个unigene被GO数据库的生物过程、分子功能和细胞组成三大功能注释到47个条目中。777个unigene在高温胁迫前后差异表达显著,其中胁迫后上调表达为362个,下调表达为415个,获得响应热胁迫的基因24个,包括热激蛋白的基因1个,PSⅠ和PSⅡ相关的基因15个、叶绿体rbcL基因3个、PLD基因2个,CAT编码基因、GAPDH基因、CYP编码基因各1个。本研究从转录组水平分析了金线莲对热胁迫的响应,这些数据将为功能基因的鉴定奠定基础,为培育耐高温金线莲品系提供了参考依据。

The aims are to study the transcriptome expression characteristics of Anoectochilus roxburghii(Wall.)Lindl.under heat stress and to analyze its heat shock response mechanism.A.roxburghii(NYJ2)under normal culture and high temperature stress(45o C)were used as materials and sequenced by using Illumina HiSeqTM2000 platform,and De novo assembly was performed by Trinity software.As a result,a total of 75,688 unigenes were obtained,and 28,323 unigenes were annotated in Nr,KOG,KEGG and Swisspro database,with an annotation rate of 37.42%.Among them,17,532 unigenes were annotated into 25 functional categories of KOG database;10,108 unigenes were annotated into 128 metabolic pathways by the KEGG databases;and 17,485 unigenes were annotated in the GO database,which were divided into 47 functional categories through biological process,molecular function and cell composition.There were 777 differentially expressed unigenes before and after heat stress,of which 362 unigenes were up-regulated and 415 unigenes were down-regulated after stress.24 genes were found in response to heat stress,including 1 heat shock protein gene,15 genes related to PSⅠand PSⅡ,3 chloroplast rbcL genes,2 PLD genes,1 CAT-coding gene,1 GAPDH gene and 1 CYP-coding gene.This research analyzed the response of A.roxburghii to heat stress at transcriptome level.These data will lay a foundation for identification of functional genes,and provide a reference basis for the cultivation of A.roxburghii heat-resistant strains.