摘要
【目的】探索菜心响应高温胁迫的转录组分析及基因挖掘,为筛选响应高温胁迫的菜心耐热基因及耐热菜心分子育种提供参考依据。【方法】以菜心耐热品种四九-19号和热敏品种特青迟心4号2个耐热性差异明显的菜心品种为试验材料,经高温(42℃)胁迫处理后,对植株地上部和地下部共24个样本基因进行转录组测序,筛选出差异表达基因(DEGs),利用COG、GO、KEGG、KOG、Pfam、Swiss-Prot、eggNOG和Nr生物数据库中对DEGs进行功能注释,筛选响应高温胁迫的热激转录因子和热激蛋白,同时利用实时荧光定量PCR验证重要基因表达情况,并分析其启动子区顺式作用元件。【结果】通过RNA-Seq在24个样本中共获得207.18 Gb的Clean bases和47462个DEGs;8个功能注释数据库均得到注释的仅2663个DEGs,Nr数据库中得到注释的DEGs最多,为47324个,占比99.71%;4个样本比较组中剔除重复DEGs,获得15761个DEGs。GO功能注释结果显示,DEGs富集到生物过程、细胞组分和分子功能三大类;KEGG信号通路富集结果表明,植物激素信号转导、碳代谢和氨基酸的生物合成信号通路最为富集。在|log2Fold Change|≥8且P<0.001的高DEGs筛选标准下,4个比较组有141个DEGs共表达,38个基因在四九-19号菜心特有表达,54个基因在特青迟心4号菜心特有表达;采用Nr、Pfam和Swiss-Prot等数据库对2个菜心品种共有表达和特有表达的共233个DEGs进行功能注释分析,获得响应高温胁迫9个热激转录因子基因和25个热激蛋白基因;最终筛选出10个与响应高温胁迫高度相关的DEGs;10个DEGs的实时荧光定量PCR验证结果显示,基因表达趋势与RNA-Seq数据结果一致;通过启动子区顺式作用元件分析结果显示,10个DEGs存在多种响应环境和激素信号的顺式作用元件。【结论】筛选获得10个与响应高温诱导相关且显著上调的重要候选基因,其启动子区域含有多种响应环境和激素信号的顺�
【Objective】The purpose of the study was to explore the transcriptome analysis and gene mining of flowering Chinese cabbage in response to high temperature stress,and to provide a reference for screening heat-resistant genes of flowering Chinese cabbage in response to high temperature stress and molecular breeding of heat-resistant flowering Chinese cabbage.【Method】Heat-resistant variety Sijiu-19 flowering Chinese cabbage and heat-sensitive variety Teqingchixin No.4 flowering Chinese cabbage,two greatly different heat-resistant varieties of flowering Chinese cabbage,were used as experimental materials.After high temperature(42℃)stress treatment,transcriptome sequencing(RNASeq)technology was 766 conducted on genes of 24 samples in the shoots and roots of the plants.The differential expression genes(DEGs)were screened,and functional annotation to DEGs was performed using COG,GO,KEGG,KOG,Pfam,Swiss-Prot,eggNOG and Nr biological databases,in order to screen heat shock transcription factors and heat shock proteins in respond to high temperature stress.At the same time,real-time fluorescence quantitative PCR was used to verify important genes of expression and to analyze the cis acting elements of the promoter region.【Result】A total of 207.18 Gb of clean bases and 47462 DEGs were obtained from 24 samples using RNA-Seq.Only 2663 DEGs were annotated in all 8 functional annotation databases,while the Nr database had the highest number of annotated genes,which were 47324 DEGs,accounting for 99.71%of all genes.Employing eliminating duplicate DEGs from four comparison groups,15761 DEGs were obtained.The GO functional annotation results showed that DEGs were enriched into three major categories:biological processes,cellular components,and molecular functions.The KEGG signaling pathway enrichment results indicated that the plant hormone signal transduction,carbon metabolism and biosynthesis of amino acids signaling pathways were the most enriched.Under high screening criteria for DEGs with|log2 Fold Change|≥8 an
作者
江定
李光光
袁凡崇
雷世康
张华
戴修纯
郑岩松
JIANG Ding;LI Guang-guang;YUAN Fan-chong;LEI Shi-kang;ZHANG Hua;DAI Xiu-chun;ZHENG Yan-song(Guangzhou Academy of Agricultural Sciences,Guangzhou,Guangdong 510335,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2024年第3期766-783,共18页
Journal of Southern Agriculture
基金
广东省科技计划项目(2022B0202080001)
广州市科技计划项目(202201010854)。
关键词
菜心
高温胁迫
转录组测序
差异表达基因(DEGs)
基因挖掘
flowering Chinese cabbage
high temperature stress
transcriptome sequencing
differential expression genes(DEGs)
gene mining
