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本研究分别选取了不同生长时期、不同温度培养以及低硅条件培养的玛氏骨条藻(Skeletonema marinoi),以Illumina Hiseq 2000平台进行转录组高通量测序分析,共获得39,098个转录本。通过NR注释、GO注释和KEGG通路注释等一系列生物信息学手段对该转录组进行了基因注释和代谢途径分析。在此基础上重点分析了玛氏骨条藻的氮代谢途径,发现共存在20种酶和蛋白及6条相关代谢途径。对38个编码这些酶的基因序列比对结果表明,其与假微型海链藻同源基因具有较高的一致性。同时对这些样品进行数字基因表达谱分析,获得不同生长时期氮代谢途径中酶和蛋白的编码基因的差异表达情况:与对数期相比,在稳定期参与硝酸盐同化过程的两种酶(硝酸盐还原酶、亚硝酸还原酶)的基因和参与氨氮代谢的两种酶(氨甲酰磷酸合成酶和氨基甲酸激酶)的基因均有明显的上调表达,这是玛氏骨条藻对所处的不同生长状态下氮元素存在形态的变化所做出的分子响应。由转录组测序构建的代谢途径及相关基因的差异表达信息有助于对玛氏骨条藻氮代谢关键酶基因调控过程的解析,为进一步研究赤潮藻氮素营养限制过程中相关基因的表达调控模式奠定了基础,可为深入了解赤潮藻对营养元素的利用提供依据。
In this study, Skeletonema marinoi cultured at different growth stages, different temperature and low silicon conditions were selected and subjected to high-throughput sequencing using the Illumina Hiseq 2000 platform. A total of 39,098 transcripts were obtained. A series of bioinformatics tools, such as NR annotation, GO annotation and KEGG pathway annotation, were used to annotate and metabolize this transcriptome. On this basis, we focused on the analysis of the nitrogen metabolism of Skeletonema pressii and found that there are 20 kinds of enzymes and proteins and 6 related metabolic pathways. The alignment of the 38 genes encoding these enzymes showed that they were highly homologous to the homologous genes of Thalassiosira pseudonana. At the same time, these samples were subjected to digital gene expression profiling to obtain differentially expressed genes of enzymes and proteins in nitrogen metabolism pathway at different growth stages: compared with the logarithmic phase, two enzymes participating in the nitrate assimilation process in the stationary phase Nitrate reductase, nitrite reductase) genes and two genes involved in ammonia metabolism (carbamyl phosphate synthase and carbamate kinase) were significantly up-regulated Of the different growth conditions under the presence of nitrogen elements in the molecular changes made by the response. The differential expression information of metabolic pathway and related genes constructed by transcriptome sequencing is helpful for the analysis of the regulation of nitrogen metabolism key enzyme gene of Skeletonema laniata so as to further study the regulation mode of the expression of related genes Laid the foundation for further understanding of red tide algae to provide the basis for the use of nutrients.