Swedish University of Agricultural Sciences, Department of Plant Biology, Uppsala BioCenter, Linnean Center for Plant Biology, P.O. Box 7080, S-75007 Uppsala, Sweden.
J Exp Bot. 2024 Jul 10;75(13):4148-4164. doi: 10.1093/jxb/erae185.
Plant defense responses to the soil-borne fungus Verticillium longisporum causing stem stripe disease on oilseed rape (Brassica napus) are poorly understood. In this study, a population of recombinant inbred lines (RILs) using the Arabidopsis accessions Sei-0 and Can-0 was established. Composite interval mapping, transcriptome data, and T-DNA mutant screening identified the NITRATE/PEPTIDE TRANSPORTER FAMILY 5.12 (AtNPF5.12) gene as being associated with disease susceptibility in Can-0. Co-immunoprecipitation revealed interaction between AtNPF5.12 and the MAJOR LATEX PROTEIN family member AtMLP6, and fluorescence microscopy confirmed this interaction in the plasma membrane and endoplasmic reticulum. CRISPR/Cas9 technology was applied to mutate the NPF5.12 and MLP6 genes in B. napus. Elevated fungal growth in the npf5.12 mlp6 double mutant of both oilseed rape and Arabidopsis demonstrated the importance of these genes in defense against V. longisporum. Colonization of this fungus depends also on available nitrates in the host root. Accordingly, the negative effect of nitrate depletion on fungal growth was less pronounced in Atnpf5.12 plants with impaired nitrate transport. In addition, suberin staining revealed involvement of the NPF5.12 and MLP6 genes in suberin barrier formation. Together, these results demonstrate a dependency on multiple plant factors that leads to successful V. longisporum root infection.
植物对土壤真菌黄萎病菌（Verticillium longisporum）引起油菜（Brassica napus）茎条纹病的防御反应知之甚少。在这项研究中，利用拟南芥品系 Sei-0 和 Can-0 建立了一个重组自交系（RILs）群体。复合区间作图、转录组数据和 T-DNA 突变体筛选将硝酸盐/肽转运蛋白家族 5.12（AtNPF5.12）基因鉴定为与 Can-0 易感性相关的基因。共免疫沉淀显示 AtNPF5.12 与 MAJOR LATEX PROTEIN 家族成员 AtMLP6 之间存在相互作用，荧光显微镜证实了这种相互作用发生在质膜和内质网中。CRISPR/Cas9 技术被应用于油菜和拟南芥中 NPF5.12 和 MLP6 基因的突变。油菜 npf5.12 mlp6 双突变体和拟南芥中真菌生长的增加表明这些基因在防御黄萎病菌方面的重要性。这种真菌的定殖也依赖于宿主根中可用的硝酸盐。因此，在硝酸盐转运受损的 Atnpf5.12 植物中，硝酸盐耗尽对真菌生长的负面影响较小。此外，亚表皮素染色显示 NPF5.12 和 MLP6 基因参与了亚表皮素屏障的形成。综上所述，这些结果表明，黄萎病菌成功感染根需要依赖多种植物因素。
