Elshobary Mostafa E, Osman Mohamed E, Abo-Shady Atef M, Komatsu Emy, Perreault Hélène, Sorensen John, Piercey-Normore Michele D
Department of Biological Sciences, University of Manitoba, Winnipeg, Manitoba R3T 2N2; and Department of Botany, University of Tanta, Egypt
Department of Botany, Faculty of Science, University of Tanta, Egypt.
Mycologia. 2016 Jul-Aug;108(4):646-56. doi: 10.3852/15-263. Epub 2016 Apr 18.
Lichen secondary metabolites (polyketides) are produced by the fungal partner, but the role of algal carbohydrates in polyketide biosynthesis is not clear. This study examined whether the type and concentration of algal carbohydrate explained differences in polyketide production and gene transcription by a lichen fungus (Cladonia rangiferina). The carbohydrates identified from a free-living cyanobacterium (Spirulina platensis; glucose), a lichen-forming alga (Diplosphaera chodatii; sorbitol) and the lichen alga that associates with C. rangiferina (Asterochloris sp.; ribitol) were used in each of 1%, 5% and 10% concentrations to enrich malt yeast extract media for culturing the mycobiont. Polyketides were determined by high performance liquid chromatography (HPLC), and polyketide synthase (PKS) gene transcription was measured by quantitative PCR of the ketosynthase domain of four PKS genes. The lower concentrations of carbohydrates induced the PKS gene expression where ribitol up-regulated CrPKS1 and CrPKS16 gene transcription and sorbitol up-regulated CrPKS3 and CrPKS7 gene transcription. The HPLC results revealed that lower concentrations of carbon sources increased polyketide production for three carbohydrates. One polyketide from the natural lichen thallus (fumarprotocetraric acid) also was produced by the fungal culture in ribitol supplemented media only. This study provides a better understanding of the role of the type and concentration of the carbon source in fungal polyketide biosynthesis in the lichen Cladonia rangiferina.
地衣次生代谢产物(聚酮化合物)由真菌共生体产生,但藻类碳水化合物在聚酮化合物生物合成中的作用尚不清楚。本研究调查了藻类碳水化合物的类型和浓度是否能解释地衣真菌(石蕊)聚酮化合物产量和基因转录的差异。从自由生活的蓝细菌(钝顶螺旋藻;葡萄糖)、地衣形成藻类(乔氏双球藻;山梨醇)以及与石蕊共生的地衣藻类(星状绿藻属;核糖醇)中鉴定出的碳水化合物,分别以1%、5%和10%的浓度用于富集麦芽酵母提取物培养基,以培养真菌共生体。通过高效液相色谱法(HPLC)测定聚酮化合物,并通过对四个聚酮合酶(PKS)基因的酮合成酶结构域进行定量PCR来测量聚酮合酶基因转录。较低浓度的碳水化合物诱导了PKS基因表达,其中核糖醇上调了CrPKS1和CrPKS16基因转录,山梨醇上调了CrPKS3和CrPKS7基因转录。HPLC结果显示,较低浓度的碳源增加了三种碳水化合物的聚酮化合物产量。仅在添加核糖醇的培养基中,真菌培养物也产生了一种来自天然地衣叶状体的聚酮化合物(富马原岛衣酸)。本研究更好地理解了碳源的类型和浓度在石蕊地衣真菌聚酮化合物生物合成中的作用。