Chooi Yit-Heng, Krill Christian, Barrow Russell A, Chen Shasha, Trengove Robert, Oliver Richard P, Solomon Peter S
Plant Sciences Division, Research School of Biology, The Australian National University, Canberra, Australian Capital Territory, Australia
Separation Science and Metabolomics Laboratory, Murdoch University, Murdoch, Western Australia, Australia.
Appl Environ Microbiol. 2015 Jan;81(1):177-86. doi: 10.1128/AEM.02745-14. Epub 2014 Oct 17.
Parastagonospora nodorum is a pathogen of wheat that affects yields globally. Previous transcriptional analysis identified a partially reducing polyketide synthase (PR-PKS) gene, SNOG_00477 (SN477), in P. nodorum that is highly upregulated during infection of wheat leaves. Disruption of the corresponding SN477 gene resulted in the loss of production of two compounds, which we identified as (R)-mellein and (R)-O-methylmellein. Using a Saccharomyces cerevisiae yeast heterologous expression system, we successfully demonstrated that SN477 is the only enzyme required for the production of (R)-mellein. This is the first identification of a fungal PKS that is responsible for the synthesis of (R)-mellein. The P. nodorum ΔSN477 mutant did not show any significant difference from the wild-type strain in its virulence against wheat. However, (R)-mellein at 200 μg/ml inhibited the germination of wheat (Triticum aestivum) and barrel medic (Medicago truncatula) seeds. Comparative sequence analysis identified the presence of mellein synthase (MLNS) homologues in several Dothideomycetes and two sodariomycete genera. Phylogenetic analysis suggests that the MLNSs in fungi and bacteria evolved convergently from fungal and bacterial 6-methylsalicylic acid synthases.
小麦颖枯病菌是一种影响全球小麦产量的病原菌。先前的转录分析在小麦颖枯病菌中鉴定出一个部分还原型聚酮合酶(PR-PKS)基因SNOG_00477(SN477),该基因在侵染小麦叶片期间高度上调。相应的SN477基因被破坏导致两种化合物的产生丧失,我们将这两种化合物鉴定为(R)-苹果菌素和(R)-O-甲基苹果菌素。利用酿酒酵母酵母异源表达系统,我们成功证明SN477是产生(R)-苹果菌素所需的唯一酶。这是首次鉴定出负责(R)-苹果菌素合成的真菌聚酮合酶。小麦颖枯病菌ΔSN477突变体在对小麦的毒力方面与野生型菌株没有任何显著差异。然而,200μg/ml的(R)-苹果菌素抑制了小麦(普通小麦)和桶状苜蓿(蒺藜苜蓿)种子的萌发。比较序列分析确定在几个座囊菌纲和两个粪壳菌纲属中存在苹果菌素合酶(MLNS)同源物。系统发育分析表明,真菌和细菌中的MLNS从真菌和细菌的6-甲基水杨酸合酶趋同进化而来。