HighlightsWe have constructed Z. tritici ku70 and ku80 null mutants.Gene targeting frequency in the ku null strains is greater than 85%.Deletion of KU70 and KU80 does not affect in vitro growth or pathogenicity.Sulfonylurea resistance was established as a new positive selection marker in Z. tritici.Ternary vectors were constructed to enable yeast recombinational cloning in Z. tritici.

HighlightsWe generate a pilot library of 32 Z. tritici over-expression strains.We develop a high throughput technique for in vitro phenotypic screening.This protocol can probe both hyphal and budding Z. tritici morphologies.A putative transcription factor impedes hyphal biosynthesis when over-expressed.Genome wide functional analysis will enable discovery of novel virulence attributes.

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In this article, we offer a contribution to the emerging debate on the role of citizen participation in food system policy making. A key driver is a recognition that solutions to complex challenges in the food system need the active participation of citizens to drive positive change. To achieve this, it is crucial to give citizens the agency in processes of designing policy interventions. This requires authentic and reflective engagement with citizens who are affected by collective decisions. One such participatory approach is citizen assemblies, which have been used to deliberate a number of key issues, including climate change by the UK Parliament’s House of Commons (House of Commons., 2019). Here, we have undertaken analysis of a citizen food assembly organized in the City of York (United Kingdom). This assembly was a way of hearing about a range of local food initiatives in Yorkshire, whose aim is to both relocalise food supply and production, and tackle food waste. These innovative community-based business models, known as ‘food hubs’, are increasing the diversity of food supply, particularly in disadvantaged communities. Among other things, the assembly found that the process of design and sortation of the assembly is aided by the involvement of local stakeholders in the planning of the assembly. It also identified the potential for public procurement at the city level, to drive a more sustainable sourcing of food provision in the region. Furthermore, this citizen assembly has resulted in a galvanizing of individual agency with participants proactively seeking opportunities to create prosocial and environmental change in the food system.

Libraries of protein-encoding sequences can be generated by identification of open reading frames (ORFs) from a genome of choice that are then assembled into collections of plasmids termed ORFeome libraries. These represent powerful resources to facilitate functional genomic characterization of genes and their encoded products. Here, we report the generation of an ORFeome for Zymoseptoria tritici, which causes the most serious disease of wheat in temperate regions of the world. We screened the genome of strain IP0323 for high confidence gene models, identifying 4,075 candidates from 10,933 predicted genes. These were amplified from genomic DNA, were cloned into the Gateway entry vector pDONR207, and were sequenced, providing a total of 3,022 quality-controlled plasmids. The ORFeome includes genes predicted to encode effectors (n = 410) and secondary metabolite biosynthetic proteins (n = 171) in addition to genes residing at dispensable chromosomes (n = 122) or those that are preferentially expressed during plant infection (n = 527). The ORFeome plasmid library is compatible with our previously developed suite of Gateway destination vectors, which have various combinations of promoters, selection markers, and epitope tags. The Z. tritici ORFeome constitutes a powerful resource for functional genomics and offers unparalleled opportunities to understand the biology of Z. tritici. [Formula: see text] Copyright © 2019 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Libraries of protein-encoding sequences can be generated by identification of open reading frames (ORFs) from a genome of choice that are then assembled into collections of plasmids termed ORFeome libraries. These represent powerful resources to facilitate functional genomic characterization of genes and their encoded products. Here, we report the generation of an ORFeome for Zymoseptoria tritici, which causes the most serious disease of wheat in temperate regions of the world. We screened the genome of strain IP0323 for high confidence gene models, identifying 4075 candidates from 10,933 predicted genes. These were amplified from genomic DNA, cloned into the Gateway® Entry Vector pDONR207, and sequenced, providing a total of 3022 quality-controlled plasmids. The ORFeome includes genes predicted to encode effectors (n = 410) and secondary metabolite biosynthetic proteins (n = 171), in addition to genes residing at dispensable chromosomes (n= 122), or those that are preferentially expressed during plant infection (n = 527). The ORFeome plasmid library is compatible with our previously developed suite of Gateway® Destination vectors, which have various combinations of promoters, selection markers, and epitope tags. The Z. tritici ORFeome constitutes a powerful resource for functional genomics, and offers unparalleled opportunities to understand the biology of Z. tritici.