Institute for Biotechnology in Plant Production, University of Natural Resources and Life Sciences, Konrad Lorenz Straße 20, 3430 Tulln, Austria.
Mol Plant Pathol. 2013 Oct;14(8):772-85. doi: 10.1111/mpp.12048. Epub 2013 Jun 6.
Fusarium head blight, caused by Fusarium graminearum, is a devastating disease of wheat. We developed near-isogenic lines (NILs) differing in the two strongest known F. graminearum resistance quantitative trait loci (QTLs), Qfhs.ndsu-3BS (also known as resistance gene Fhb1) and Qfhs.ifa-5A, which are located on the short arm of chromosome 3B and on chromosome 5A, respectively. These NILs showing different levels of resistance were used to identify transcripts that are changed significantly in a QTL-specific manner in response to the pathogen and between mock-inoculated samples. After inoculation with F. graminearum spores, 16 transcripts showed a significantly different response for Fhb1 and 352 for Qfhs.ifa-5A. Notably, we identified a lipid transfer protein which is constitutively at least 50-fold more abundant in plants carrying the resistant allele of Qfhs.ifa-5A. In addition to this candidate gene associated with Qfhs.ifa-5A, we identified a uridine diphosphate (UDP)-glycosyltransferase gene, designated TaUGT12887, exhibiting a positive difference in response to the pathogen in lines harbouring both QTLs relative to lines carrying only the Qfhs.ifa-5A resistance allele, suggesting Fhb1 dependence of this transcript. Yet, this dependence was observed only in the NIL with already higher basal resistance. The complete cDNA of TaUGT12887 was reconstituted from available wheat genomic sequences, and a synthetic recoded gene was expressed in a toxin-sensitive strain of Saccharomyces cerevisiae. This gene conferred deoxynivalenol resistance, albeit much weaker than that observed with the previously characterized barley HvUGT13248.
镰刀菌穗腐病由禾谷镰刀菌引起,是小麦的一种毁灭性疾病。我们开发了在两个最强的已知禾谷镰刀菌抗性数量性状位点(QTL),Qfhs.ndsu-3BS(也称为抗性基因 Fhb1)和 Qfhs.ifa-5A 上存在差异的近等基因系(NILs),它们分别位于 3B 染色体的短臂和 5A 染色体上。这些表现出不同抗性水平的 NIL 被用于鉴定在 QTL 特异性方式下因病原体和模拟接种样本之间而显著变化的转录物。在接种禾谷镰刀菌孢子后,16 个转录物对 Fhb1 的反应有显著差异,352 个转录物对 Qfhs.ifa-5A 的反应有显著差异。值得注意的是,我们鉴定了一种脂质转移蛋白,它在携带 Qfhs.ifa-5A 抗性等位基因的植物中至少丰富 50 倍。除了与 Qfhs.ifa-5A 相关的候选基因外,我们还鉴定了一个尿嘧啶二磷酸(UDP)-糖基转移酶基因,命名为 TaUGT12887,与携带 Qfhs.ifa-5A 抗性等位基因的系相比,该基因在携带两个 QTL 的系中对病原体的反应呈正差异,表明该转录物依赖于 Fhb1。然而,这种依赖性仅在已经具有更高基础抗性的 NIL 中观察到。从可用的小麦基因组序列中重新构建了 TaUGT12887 的完整 cDNA,并在对毒素敏感的酿酒酵母菌株中表达了合成的重编码基因。该基因赋予脱氧雪腐镰刀菌烯醇抗性,尽管比先前表征的大麦 HvUGT13248 观察到的抗性弱得多。