海马齿根系响应盐胁迫的转录组分析

Analysis of the root transcriptomes in Sesuvium portulacastrum respond to salt stress

  • 摘要: 【目的】对盐胁迫下海马齿根系进行转录组测序分析,挖掘海马齿根系耐盐相关基因,为揭示海马齿耐盐的分子机制提供参考。【方法】利用Illumina测序技术对0 mmol/L NaCl (对照组)和400 mmol/L NaCl胁迫处理(盐胁迫处理组)下的海马齿根系进行转录组测序分析,从中筛选出差异表达基因,选取13个基因进行实时荧光定量PCR (qRTPCR)检测,以验证转录组数据的可靠性。【结果】在海马齿根系转录组中共鉴定出305145个转录本,平均长度为622 bp,其中,对照组有146177个长度>300 bp的转录本,盐胁迫处理组有72173个长度>300 bp的转录本;共有65535条Unigenes在Nr、GO、Swiss-Prot、COG和KEGG五大数据库注释成功,占Unigenes总数的52.36%。对照组和盐胁迫处理组共有65535个差异Unigenes,其中,有182个热休克蛋白基因。对照组和盐胁迫处理组间共有24042个差异表达基因,从中选取13个基因进行qRT-PCR检测,结果显示,9个基因表达上调,其余4个基因表达下调,与转录组测序结果一致。24042个差异表达基因中,共有10106个显著差异基因富集到129条代谢通路,其中富集程度排名前10的代谢途径为核糖体、次级代谢生物合成、RNA转运、内吞作用、剪接体、甘油磷脂代谢、内质网加工、吞噬、醚脂类代谢和植物-病原体相互作用,参与盐胁迫相关的硫代谢、脯氨酸积累、活性氧(ROS)代谢、与盐胁迫相关的钙信号通路和过氧化氢代谢等途径的差异基因上调。【结论】在盐胁迫下海马齿差异表达基因如小分子量热激蛋白基因、抗氧化酶相关基因及与离子交换相关基因发挥了重要调控作用。

     

    Abstract: 【Objective】To conduct transcriptome sequencing analysis on Sesuvium portulacastrum under salt tolerance,and to find out genes related to salt tolerance in the root of S. portulacastrum,so as to provide reference for studying molecular mechanism of S. portulacastrum.【Method】 Illumina sequencing technology was applied to compare and analyze the transcriptomesand the differentially expressed genes(DEGs)related to salt tolerance inroots of S. portulacastrum under 0 mmol/L NaCl(control group)and 400 mmol/L NaCl salt stress(salt stress treatment group), respectively. Thirteen DEGs were selected to verify the reliability of transcriptome data by using quantitative real-time PCR(qRT-PCR) analysis.【Result】A total of 305145 transcripts with an average length of 622 bp were identified in the roots of S. portulacastrum transcriptome. In the control group,146177 transcripts were greater than 300 bp;in the salt treatment group, 72173 transcripts were greater than 300 bp. A total of 65535 unigenes were successfully annotated in the five databases of Nr,GO,Swiss-Prot,COG and KEGG,accounting for 52.36% of the total unigenes and including 182 heat shock protein genes. A number of 24042 DEGs between the control group and the salt stress treatment group were identified. Thirteen candidate genes were selected for qRT-PCR analysis,and the result showed that 9 genes were up-regulated and 4 genes were down-regulated,which was consistent with transcriptome sequencing analysis. Among the 24042 DEGs, 10106 significant DEGs were enriched in 129 metabolic pathways. The top 10 enriched metabolic pathways were ribosome,biosynthesis of secondary metabolites,RNA transport,endocytosis,spliceosome,glycerophospholipid metabolism,protein processing in endoplasmic reticulum,phagocytosis,ether lipid metabolism and plant-pathogen interaction path. Differential genes in sulfur metabolism,proline accumulation,eactive oxygen species (ROS)metabolism,calcium signaling pathway and hydrogen peroxide metabolism related to salt stress were up-regulated.【Conclusion】 Under salt stress, DEGs in S. portulacastrum,such as small molecular heat shock protein genes,genes related to anti-oxidation and genes related to ion exchange,play an important role in regulation.

     

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