Chung Kuang-Ren, Liao Hui-Ling
Citrus Research and Education Center, and Department of Plant Pathology, Institute of Food and Agricultural Sciences (IFAS), University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.
Microbiology (Reading). 2008 Nov;154(Pt 11):3556-3566. doi: 10.1099/mic.0.2008/019414-0.
Elsinochromes are nonhost-selective, light-activated, polyketide-derived toxins produced by many phytopathogenic Elsinoë species. We recently showed that the polyketide synthase-encoding gene EfPKS1 is essential for elsinochrome biosynthesis in the citrus scab fungus Elsinoë fawcettii. Sequence analysis beyond the EfPKS1 gene identified nine putative ORFs: four genes, designated RDT1, TSF1, PRF1 and ECT1, all encode polypeptides likely to have biosynthetic or efflux functions; five additional genes, OXR1 and EfHP1 to EfHP4, encode hypothetical proteins of unknown function. Northern-blot analysis revealed that expression of these genes in E. fawcettii was not completely correlated with accumulation of elsinochromes under nitrogen limitation, alkaline pH or high concentrations of glucose. Targeted disruption of the TSF1 gene, encoding a putative transcriptional activator, yielded fungal mutants unable to produce elsinochromes, and defective in both conidiation and expression of RDT1, EfPKS1, PRF1 and EfHP1, whereas expression of RDT1, TSF1, PRF1 and ECT1 was nearly abolished in EfPKS1-disrupted mutants. By contrast, expression of OXR1, EfHP2 and EfHP3 was not affected by disrupting either EfPKS1 or TSF1. Taken together, the results indicate that in addition to polyketide synthase, the products of TSF1 and other adjacent genes may also play a crucial role in elsinochrome production.
痂囊腔菌素是由许多植物病原痂囊腔菌属物种产生的非寄主选择性、光激活、聚酮化合物衍生的毒素。我们最近发现,编码聚酮合酶的基因EfPKS1对柑橘痂囊腔菌Elsinoë fawcettii中痂囊腔菌素的生物合成至关重要。对EfPKS1基因以外的序列分析鉴定出9个推定的开放阅读框:4个基因,命名为RDT1、TSF1、PRF1和ECT1,均编码可能具有生物合成或外排功能的多肽;另外5个基因,OXR1和EfHP1至EfHP4,编码功能未知的假定蛋白质。Northern杂交分析表明,这些基因在E. fawcettii中的表达与在氮限制、碱性pH或高浓度葡萄糖条件下痂囊腔菌素的积累并不完全相关。对编码假定转录激活因子的TSF1基因进行靶向破坏,产生了无法产生痂囊腔菌素的真菌突变体,并且在分生孢子形成以及RDT1、EfPKS1、PRF1和EfHP1的表达方面存在缺陷,而在EfPKS1破坏的突变体中,RDT1、TSF1、PRF1和ECT1的表达几乎被消除。相比之下,OXR1、EfHP2和EfHP3的表达不受EfPKS1或TSF1破坏的影响。综上所述,结果表明,除了聚酮合酶外,TSF1和其他相邻基因的产物可能在痂囊腔菌素的产生中也起着关键作用。