Liu Xuexia, Fan Wenqiang, Jiao Huihui, Gao Han, Tang Jianning, Zhu Jinzhong, Yue Sijun, Zheng Rui
Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia, Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, College of Life Science, Ningxia University, Yinchuan 750021, Ningxia, China.
Wolfberry Industry Development Center, Yinchuan 750021, Ningxia, China.
Sheng Wu Gong Cheng Xue Bao. 2023 Jul 25;39(7):3015-3036. doi: 10.13345/j.cjb.220821.
To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in L. and analyzing their expression regulation mechanism.
为了探索与不同品种枸杞果实中活性成分生物合成相关的差异表达基因（DEGs），并揭示活性成分差异的分子机制，我们利用Illumina NovaSeq 6000高通量测序技术，对‘宁杞1号’和‘宁杞7号’果实的绿果期、转色期和成熟期进行转录组测序。随后，我们比较了两个品种果实不同发育阶段相关基因的表达谱。结果显示，共获得811818178条clean reads，产生了121.76 Gb的有效数据。在‘宁杞1号’和‘宁杞7号’的绿果期、转色期和成熟期分别获得2827、2552和2311个DEGs，其中2153、2050和1825个基因在包括基因本体论（GO）、京都基因与基因组百科全书（KEGG）和蛋白质直系同源簇（KOG）在内的六个数据库中得到注释。在GO数据库中，绿果期、转色期和成熟期的1307、865和624个DEGs分别在生物过程、细胞成分和分子功能中富集。在KEGG数据库中，三个发育阶段的DEGs主要集中在代谢途径、次生代谢物生物合成和植物-病原体相互作用。在KOG数据库中，三个发育阶段分别注释了1775、1751和1541个DEGs。在PubMed数据库中搜索注释基因发现，在绿果期、转色期和成熟期分别挖掘到18、26和24个与活性成分合成相关的DEGs。这些基因参与类胡萝卜素、黄酮类、萜类、生物碱、维生素代谢途径等。通过RT-qPCR验证了7个DEGs，结果与转录组测序一致。本研究从转录水平为不同枸杞品种活性成分含量差异提供了初步证据。这些证据可能有助于进一步探索枸杞中活性成分生物合成的关键基因，并分析其表达调控机制。
