NaCl胁迫下黑果枸杞转录组测序分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0492

摘要/Abstract

摘要： 研究黑果枸杞在不同浓度盐胁迫下基因表达谱变化情况,为进一步研究黑果枸杞抗盐分子机制奠定研究基础。对0（CK）、50、250 mmol/L NaCl溶液胁迫的黑果枸杞组培苗的根和叶在胁迫时间为0、1、12 h时分别取样,采用转录组测序（RNA-Seq）技术进行测序分析。结果表明,转录组测序共产生 222.49 Gb 原始数据,拼接出 Unigenes 86 037条,注释到 7大功能数据库（GO、KEGG、KOG、NR、Pfam、Swiss-Prot和egg NOG）上的 Unigenes 总数为46 594个,占总Unigenes 的54.76%,还有38 929个Unigenes在这些数据库中没有得到注释。通过 GO 分类和 KEGG Pathway 富集性分析,分别归于 51 个 GO 类别和 211条代谢途径。差异表达基因分析显示,黑果枸杞叶片和根的上调基因和下调基因数随着NaCl浓度的增大和处理时间的延长均呈增加趋势,叶片中的上调基因数（7 514）小于下调基因数（9 032）,根中的上调基因数（12 347）大于下调基因数（11 559）。在黑果枸杞盐胁迫下转录组中发现28 325个SSR位点,最多的为单核苷酸 SSR,占 70.47%。综合分析表明,黑果枸杞对盐胁迫的反应是一个多基因参与、多个生物过程协同调控的过程,基因表达量的变化可能是基因调控的主要方式。

关键词: 黑果枸杞, 盐胁迫, 转录组测序, 功能注释

Abstract: This work aims to study the changes of gene expression profiles of Lycium ruthenicum Murr. under salt stress with different concentrations,and to provide a basis for further studying the molecular mechanism of salt resistance of L. ruthenicum. First,the roots and leaves of tissue culture seedlings of L ruthenicum stressed by NaCl at the concentrations of 0（CK）,50 and 250 mmol/L were sampled at stress duration of 0,1 and 12 h,respectively. Then,the sequence analysis was conducted using transcriptome sequencing technique（RNA-Seq）. The results showed that a total of 222.49 Gb raw data were generated by transcriptome sequencing,from which 86 037 Unigenes were obtained. The 46 594 Unigenes,accounting for 54.76% of the total Unigenes,were annotated in the 7 functional databases（GO,KEGG,KOG,NR,Pfam,Swiss-Prot and egg NOG）,while the rest 38 929 Unigenes were not annotated in the known genes databases. These Unigenes were classified into 51 GO categories and 211 metabolic pathways using GO classification and KEGG Pathway cluster analysis. Analysis of differentially expressed genes revealed that the up-regulated genes and down-regulated genes in the leaves and roots of L. ruthenicum increased with the NaCl concentration as well as treatment time increasing. The up-regulated genes（7 514）in leaves were less than down-regulated（9 032）genes,while the up-regulated genes（12 347）in roots were more than down-regulated genes（11 559）. Total 28 325 SSR（Simple Sequence Repeats）loci were discovered in L. ruthenicum under salt stress transcriptome,most of which were single nucleotide SSR and accounted for 70.47% of the total. Comprehensive analysis showed that the responses of L. ruthenicum to salt stress involved many genes and were regulated by several collaborative biological processes. The changes of gene expression level might be the major approach in gene regulation.

Key words: Lycium ruthenicum Murr., salt stress, transcriptome sequencing, function annotation

马彦军, 段慧荣, 魏佳, Richard John Tiika, 单立山, 马瑞. NaCl胁迫下黑果枸杞转录组测序分析[J]. 生物技术通报, 2020, 36(2): 100-109.

MA Yan-jun, DUAN Hui-rong, WEI Jia, RICHARD John Tiika, SHAN Li-shan, MA Rui. Transcriptome Sequencing Analysis of Lycium ruthenicum Murr. Under NaCl Stress[J]. Biotechnology Bulletin, 2020, 36(2): 100-109.

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