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PubMed2025

Gene Editing and Protein Tagging in the Oomycete Phytophthora infestans Using CRISPR-Cas12a.

Molecular genetic tools such as CRISPR-Cas gene editing systems are invaluable for understanding gene and protein function and revealing the details of a pathogen's life and disease cycles. Here we present protocols for genome editing in Phytophthora infestans, an oomycete with global importance as a pathogen of potato and tomato. Using a vector system that expresses variants of Cas12a from Lachnospiraceae bacterium and its guide RNA from a unified transcript, we first present a method for editing genes through the non-homologous end-joining (NHEJ) pathway. We then describe an application of homology-directed repair (HDR), in which Cas12a is used to fuse a protein-coding gene with a fluorescent or epitope tag. Both methods should be adaptable to many oomycetes other than P. infestans.

Gene Editing↗
PubMed2026

Diverse haplotypes at a complex Solanum americanum locus confer resistance to Phytophthora infestans and P. capsici.

Plants encounter diverse pathogens and have evolved a two-layered innate immune system to detect pathogen molecules and activate defense mechanisms that restrict infection. Most cloned plant Resistance (R) genes encode NLR immune receptors. NLR genes are often found in clusters of paralogs with sequence and copy number variation; whether these NLR clusters evolve in response to single or multiple pathogens has been unclear. We report here the isolation of a Phytophthora capsici resistance gene, Rpc2, along with a novel P. infestans resistance gene, Rpi-amr5, from two Solanum americanum accessions. These orthologous genes reside in the Rpi-amr1 cluster, which has previously been associated with resistance to P. infestans. By screening RXLR effector libraries of P. infestans and P. capsici, we identified multiple effectors recognised by both NLRs. Our findings highlight the complexity of NLR clusters and evolution driven by interactions with multiple pathogens. This work will underpin efforts to elevate resistance against Phytophthora pathogens and enhances our understanding of NLR evolution.

Journal Article↗
PubMed2026

Shining Light on Late Blight.

In this retrospective on my journey in science, I shine light on the ins and outs of the late blight pathogen Phytophthora infestans. While studying plant pathology, I became fascinated by the molecular mechanisms that govern the development of cells and organisms and ended up unraveling intimate plant-microbe interactions. I have been fortunate to work in an inspiring environment and with committed coworkers. I am proud of what we achieved as a team. By digging into the biology of Phytophthora and exploring genomes and pathogenicity mechanisms, we uncovered a treasure trove of novelties and peculiarities that offer ample opportunities for designing pathogen-informed control strategies.

Plant Diseases↗
PubMed2026

Centuries of Potato Late Blight: Tracking Global Epidemics and Managing Future Outbreaks.

Phytophthora infestans killed the potato crop in Ireland in 1845, leading to widespread famine and the death of more than one million people. Historic herbarium specimens from the famine era were used to understand the pathogen's biology and track its global spread, providing a valuable resource for research. Historic outbreaks in the United States and Europe were caused by the FAM-1 lineage, whereas the US-1 lineage spread later. The famine lineage was basal in the phylogeny and ancestral to modern US-1, Mexican, and globally aggressive lineages. An admixture between the famine lineage and the Andean species Phytophthora andina was revealed, indicating a South American origin of the disease. Temporal changes in the presence and abundance of virulence genes were observed in historic compared to modern genomes. Expansion in effector abundance occurred as new genotypes emerged in the mid-twentieth century. Disease surveillance and genotyping on a global scale have helped to inform disease management.

Solanum tuberosum↗
PubMed2021

Spray-induced gene silencing for disease control is dependent on the efficiency of pathogen RNA uptake.

Recent discoveries show that fungi can take up environmental RNA, which can then silence fungal genes through environmental RNA interference. This discovery prompted the development of Spray-Induced Gene Silencing (SIGS) for plant disease management. In this study, we aimed to determine the efficacy of SIGS across a variety of eukaryotic microbes. We first examined the efficiency of RNA uptake in multiple pathogenic and non-pathogenic fungi, and an oomycete pathogen. We observed efficient double-stranded RNA (dsRNA) uptake in the fungal plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani, Aspergillus niger and Verticillium dahliae, but no uptake in Colletotrichum gloeosporioides, and weak uptake in a beneficial fungus, Trichoderma virens. For the oomycete plant pathogen, Phytophthora infestans, RNA uptake was limited and varied across different cell types and developmental stages. Topical application of dsRNA targeting virulence-related genes in pathogens with high RNA uptake efficiency significantly inhibited plant disease symptoms, whereas the application of dsRNA in pathogens with low RNA uptake efficiency did not suppress infection. Our results have revealed that dsRNA uptake efficiencies vary across eukaryotic microbe species and cell types. The success of SIGS for plant disease management can largely be determined by the pathogen's RNA uptake efficiency.

Ascomycota↗
PubMed2026

Balancing growth and immunity of potato by humidity-dependent expression of a late blight resistance gene.

Inducible expression of resistance genes is an effective approach to balance plant growth and immunity, thus facilitating the development of disease-resistant crop cultivars. While pathogen-responsive and immunity-related promoters have been adopted for this purpose, alternative design strategies remain to be explored. High relative humidity (RH) has been recognized as a crucial permissive environmental condition for the occurrence of devastating plant diseases including tomato and potato late blight. Here, we identified humidity-activated cis-regulatory elements (HAEs) in Solanum lycopersicum through an integrative analysis of transcriptomics and chromatin accessibility data. Sequence homology-inferred HAEs in S. tuberosum can predict humidity-elicited changes in downstream gene expression. Transgenic S. tuberosum lines expressing a late blight resistance gene driven by an artificial humidity-inducible promoter containing a natural S. tuberosum HAE were generated. These transgenic lines exhibited comparable late blight resistance levels to the lines overexpressing the same resistance gene in controlled zoospore inoculation bioassays, while avoiding growth suppression and tuber yield penalties in common garden experiments. Our findings highlight the importance of plant cis-regulatory elements in the transcriptional responses to high RH and provide a proof-of-concept for a humidity-inducible environment-responsive resistance gene deployment strategy to engineer disease-resistant crop cultivars without compromising growth and yield.

Phytophthora infestans↗