Department of Biotechnology, Corteva Agriscience, Johnston, Iowa, USA.
Oxford, Pennsylvania, USA.
Mol Plant Pathol. 2023 Jul;24(7):758-767. doi: 10.1111/mpp.13267. Epub 2022 Sep 30.
Northern corn leaf blight, caused by the fungal pathogen Exserohilum turcicum, is a major disease of maize. The first major locus conferring resistance to E. turcicum race 0, Ht1, was identified over 50 years ago, but the underlying gene has remained unknown. We employed a map-based cloning strategy to identify the Ht1 causal gene, which was found to be a coiled-coil nucleotide-binding, leucine-rich repeat (NLR) gene, which we named PH4GP-Ht1. Transgenic testing confirmed that introducing the native PH4GP-Ht1 sequence to a susceptible maize variety resulted in resistance to E. turcicum race 0. A survey of the maize nested association mapping genomes revealed that susceptible Ht1 alleles had very low to no expression of the gene. Overexpression of the susceptible B73 allele, however, did not result in resistant plants, indicating that sequence variations may underlie the difference between resistant and susceptible phenotypes. Modelling of the PH4GP-Ht1 protein indicated that it has structural homology to the Arabidopsis NLR resistance gene ZAR1, and probably forms a similar homopentamer structure following activation. RNA sequencing data from an infection time course revealed that 1 week after inoculation there was a threefold reduction in fungal biomass in the PH4GP-Ht1 transgenic plants compared to wild-type plants. Furthermore, PH4GP-Ht1 transgenics had significantly more inoculation-responsive differentially expressed genes than wild-type plants, with enrichment seen in genes associated with both defence and photosynthesis. These results demonstrate that the NLR PH4GP-Ht1 is the causal gene underlying Ht1, which represents a different mode of action compared to the previously reported wall-associated kinase northern corn leaf blight resistance gene Htn1/Ht2/Ht3.
北方玉米叶枯病由真菌病原菌 Exserohilum turcicum 引起,是玉米的主要病害。50 多年前,人们首次鉴定出第一个主要的抗 E. turcicum race 0 位点 Ht1,但其潜在基因仍未知。我们采用基于图谱的克隆策略来鉴定 Ht1 致病基因,发现它是一个卷曲螺旋核苷酸结合、富含亮氨酸重复(NLR)基因,我们将其命名为 PH4GP-Ht1。转基因测试证实,将天然的 PH4GP-Ht1 序列引入易感玉米品种可导致对 E. turcicum race 0 的抗性。对玉米嵌套关联作图基因组的调查显示,易感 Ht1 等位基因的基因表达非常低或没有。然而,过表达易感 B73 等位基因并没有导致抗性植株,表明序列变异可能是抗性和易感表型之间的差异基础。PH4GP-Ht1 蛋白的建模表明,它与拟南芥 NLR 抗性基因 ZAR1 具有结构同源性,并且可能在激活后形成类似的五聚体结构。感染时间过程的 RNA 测序数据显示,与野生型植物相比,PH4GP-Ht1 转基因植物在接种后 1 周时真菌生物量减少了三倍。此外,PH4GP-Ht1 转基因植物比野生型植物具有更多的接种反应差异表达基因,并且与防御和光合作用相关的基因富集。这些结果表明,NLR PH4GP-Ht1 是 Ht1 的致病基因,与先前报道的壁相关激酶北方玉米叶枯病抗性基因 Htn1/Ht2/Ht3 相比,它代表了一种不同的作用模式。