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丝状真菌次级代谢产物的功能与合成调控研究进展
Research advance in functional and synthetic regulation of secondary metabolites of filamentous fungi
({{custom_author.role_en}}), {{javascript:window.custom_author_en_index++;}}真菌是具有高度多样性的微生物资源,其产生的次级代谢产物具有很多重要的作用,如紫外线防护,自身发育以及防御外部侵害,而这些次级代谢产物的生物合成需要多个基因参与调控。本文主要综述了真菌次级代谢产物的药用价值,在真菌发育过程中的生态功能以及合成调控机制。
Fungi are highly diverse organisms producing secondary metabolites with many important functions such as UV protection, self-development and defense against external aggression. The biosynthesis of these secondary metabolites requires participation of multiple genes. In this paper the medicinal value of secondary metabolites of filamentous fungi, the ecological functions during fungal development and the mechanisms of synthetic regulation are reviewed.
真菌 / 次级代谢产物 / 生态功能 / 表观遗传调控 {{custom_keyword}} /
fungi / secondary metabolites / ecological function / epigenetic regulation {{custom_keyword}} /
图1 FgMetR蛋白结构域组成、氨基酸序列比对及系统发育树分析 A：FgMetR的蛋白结构域组成；B：禾谷镰孢菌与代表性真菌中MetR同源蛋白的BRLZ结构域的多序列比对；C：禾谷镰孢菌与代表性真菌中MetR同源蛋白系统发育树Fig. 1 FgMetR protein domain prediction, alignment of amino acid sequences and phylogenetic tree analysis. A: FgMetR protein domain architecture; B: Multiple sequence alignment of BRLZ domains in FgMetR orthologs from representative fungal species; C: Phylogenetic tree analysis of FgMetR orthologs from representative fungal species. |
图2 FgMetR基因敲除突变体与互补体的构建 A：FgMetR基因敲除与检测策略示意图；probe表示Southern blot探针位置，M、K1、K2分别表示B图中PCR扩增片段的位置，X表示Southern blot中酶切基因组使用的限制性内切酶XhoⅠ；B：PH-1菌株和敲除转化子PCR检测；C：PH-1菌株和敲除转化子Southern blot检测；D：敲除体和互补转化子的PCR检测Fig. 2 Construction of FgMetR gene knockout mutants and complemented strains. A: Schematic diagram of FgMetR gene knockout and detection strategy; the probe label indicates the location of the probe used in the Southern blot anlysis; the labels M, K1 and K2 indicate the location of PCR fragments in Fig. 1B; the label X markes the location of restriction enzyme XhoⅠused for genomic digestion in Southern blot anylasis; B: PCR analysis of FgMetR deletion transformants; C: Southern blot analysis of FgMetR deletion transformants and PH-1 strain; D: PCR detection analysis of a FgMetR complemented strain and a FgMetR gene knockout strain. |
图3 不同菌株的菌落生长和色素形成情况 A：不同菌株在PDA培养基上生长4 d的菌落形态；B：不同菌株在PDB培养基里生长2 d后的色素形成情况；C：不同菌株在PDA培养基上培养4 d后的菌落直径，误差线表示3次独立实验的标准误，不同字母表示经LSD检验存在显著性差异(P<0.05)；下同；D：不同菌株在试管中的PDA培养基上生长4 d后的气生菌丝生长情况Fig. 3 Colony growth morphology and pigmentation of different strains. A: Colony morphology of different strains cultured on PDA plates for 4 days; B: Two-day-old PDB cultures of different strains; C: Colony diameters of different strains cultured on PDA plates for 4 days; Error bars indicate the standard error of three independent assays; Different letters above bars indicate significant difference using LSD test (P<0.05); Similarly hereinafter; D: Aerial hyphae of each strain in tubes cultured on PDA plates for 4 days. |
图5 分生孢子产量测定和有性生殖过程观测 A：各菌株在CMC培养基中培养5 d后的分生孢子产量；B：各菌株在胡萝卜培养基上诱导有性生殖20 d后产子囊壳、子囊及子囊孢子情况Fig. 5 Conidiation determination and observation of sexual reproduction. A: Conidial production of each strain after incubation in CMC medium for 5 d; B: Observation of the morphology of perithecia, asci and ascospore is produced on carrot medium after 20 days of sexual induction. |
图6 致病力检测实验 接种各菌株14 d后小麦胚芽鞘典型症状(A)、病斑长度(B)以及接种14 d后小麦穗的典型症状(C)和侵染小穗数(D)Fig. 6 Pathogenicity test. The representative symptoms (A) and lesion length (B) on wheat cleoptiles inoculated by different strains in 14 d post-inoculation (dpi), and the representative symptoms (C) and the number of infected spikelets (D) on wheat head invaded by different strains in 14 days. |
图7 各测试菌株对戊唑醇的敏感性测定 A：各菌株在含不同浓度的戊唑醇的PDA培养基中培养4 d后的菌落形态；B：戊唑醇对各菌株的EC50值Fig. 7 Determination of the sensitivity of each test strain to tebuconazole. A: Colony morphology of each strain cultured for 4 days on PDA plates with different concentrations of tebuconazole; B: EC50 values of the tebuconazole to each strain. |
图8 各测试菌株对咪鲜胺的敏感性测定 A：各菌株在含不同浓度的咪鲜胺的PDA培养基中培养4 d后的菌落形态；B：咪鲜胺对各菌株的EC50值Fig. 8 Determination of the sensitivity of each test strain to prochloraz. A: Colony morphology of each strain cultured for 4 days on PDA plates with different concentrations of prochloraz; B: EC50 values of the prochloraz to each strain. |
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PDF(836 KB)
图1 FgMetR蛋白结构域组成、氨基酸序列比对及系统发育树分析 A：FgMetR的蛋白结构域组成；B：禾谷镰孢菌与代表性真菌中MetR同源蛋白的BRLZ结构域的多序列比对；C：禾谷镰孢菌与代表性真菌中MetR同源蛋白系统发育树图2 FgMetR基因敲除突变体与互补体的构建 A：FgMetR基因敲除与检测策略示意图；probe表示Southern blot探针位置，M、K1、K2分别表示B图中PCR扩增片段的位置，X表示Southern blot中酶切基因组使用的限制性内切酶XhoⅠ；B：PH-1菌株和敲除转化子PCR检测；C：PH-1菌株和敲除转化子Southern blot检测；D：敲除体和互补转化子的PCR检测图3 不同菌株的菌落生长和色素形成情况 A：不同菌株在PDA培养基上生长4 d的菌落形态；B：不同菌株在PDB培养基里生长2 d后的色素形成情况；C：不同菌株在PDA培养基上培养4 d后的菌落直径，误差线表示3次独立实验的标准误，不同字母表示经LSD检验存在显著性差异(P<0.05)；下同；D：不同菌株在试管中的PDA培养基上生长4 d后的气生菌丝生长情况图4 FgMetR参与硫代谢调控 在MM-S培养基及含不同硫源的MM-S培养基上培养4 d的菌落形态图5 分生孢子产量测定和有性生殖过程观测 A：各菌株在CMC培养基中培养5 d后的分生孢子产量；B：各菌株在胡萝卜培养基上诱导有性生殖20 d后产子囊壳、子囊及子囊孢子情况图6 致病力检测实验 接种各菌株14 d后小麦胚芽鞘典型症状(A)、病斑长度(B)以及接种14 d后小麦穗的典型症状(C)和侵染小穗数(D)图7 各测试菌株对戊唑醇的敏感性测定 A：各菌株在含不同浓度的戊唑醇的PDA培养基中培养4 d后的菌落形态；B：戊唑醇对各菌株的EC50值图8 各测试菌株对咪鲜胺的敏感性测定 A：各菌株在含不同浓度的咪鲜胺的PDA培养基中培养4 d后的菌落形态；B：咪鲜胺对各菌株的EC50值
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