转录组测序技术揭示参与松萝酸生物合成的聚酮合酶基因
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摘要
松萝酸是地衣产生的独特化合物,具有抗菌,抗病毒以及抗肿瘤活性。然而,目前对松萝酸生物合成还不够清楚。本研究采用转录组测序技术首先获得产生松萝酸时期皮革肾岛衣地衣型真菌转录组数据,通过生物信息学分析获得转录组中全部的聚酮合酶基因(包括还原型聚酮合酶;非还原型聚酮合酶以及真菌III聚酮合酶)。再利用荧光定量PCR和液质质谱分析聚酮合酶基因表达情况与松萝酸产量情况,通过关联分析得出参与松落酸生物合成的聚酮合酶基因。结果显示:转录组分析获得22条聚酮合酶基因;其中18天能够在DNA检测到;通过结构域分析确定8条聚酮合酶基因用于荧光定量PCR检测;用荧光定量PCR和液质质谱关联分析得出其中一条聚酮合酶(NPPKS1)(KS-AT-DH-ACP-ACP-MT-TE)基因表达与松萝酸产量变化一致。证明NPPKS1参与松萝酸生物合成。
引文
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Luo H,Yamamoto Y,Jeon H S,et al.2011,Production of anti-Helicobacter pylori metabolite by the lichen-forming fungus Nephromopsis pallescens[J].The Journal of Microbiology,49(1):66-70.
Singh N,Nambiar D,Kale R K,et al.2013,Usnic acid inhibits growth and induces cell cycle arrest and apoptosis in human lung carcinoma A549 cells[J].Nutrition and cancer,65(sup1):36-43.
Song Y,Dai F,Zhai D,et al.2012,Usnic acid inhibits breast tumor angiogenesis and growth by suppressing VEGFR2-mediated AKT and ERK1/2 signaling pathways[J].Angiogenesis,15(3):421-432.
Stocker-W?rg?tter E.2015 Biochemical Diversity and Ecology of Lichen-Forming Fungi:Lichen Substances,Chemosyndromic Variation and Origin of Polyketide-Type Metabolites(Biosynthetic Pathways)[M]//Recent Advances in Lichenology.Springer India,:161-179.
Wang Y,Wang J,Cheong Y H,et al.2014,Three new non-reducing polyketide synthase genes from the lichen-forming fungus Usnea longissima[J].Mycobiology,42(1):34-40.
Yaegashi J,Oakley B R,Wang C C C.2014,Recent advances in genome mining of secondary metabolite biosynthetic gene clusters and the development of heterologous expression systems in Aspergillus nidulans[J].Journal of industrial microbiology&biotechnology,41(2):433-442.