The John Innes Centre Publications Repository contains details of all publications resulting from our researchers.
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The creation of this publications repository was funded by BBSRC.
Characterisation of the CfEcp2 gene, encoding for recognition of the conserved fungal effector Ecp2 in Solanum pimpinellifolium and Nicotiana paniculata
ID: 55707read more
Members of the Capnodiales class of fungi have evolved contrasting lifestyles on plants, ranging from biotrophy, through to endotrophy and necrotrophy. Many of these fungi contain homologs of the effector protein called Ecp2. The ‘currant’ tomato (Solanum pimpinellifolium) resistance (R) gene Cf-Ecp2 recognises Ecp2 produced by the pathogenic fungus, Cladosporium fulvum. In this study, Cf-Ecp2 was fine-mapped to the Orion locus, which contains eight Homologs of Cladosporium resistance gene Cf-9 (Hcr9s), four of which share 100% sequence identity within their open reading frame. The Cf-Ecp2 locus exemplifies the complex nature of many R gene loci brought about by successive rounds of tandem duplication. This extensive duplication renders R gene loci complex and difficult to resolve. As a consequence, the sequencing of a BAC minimal tiling path across the Cf-Ecp2 locus required use of both short read and long read sequencing technologies, with MinION providing vital scaffolding reads. A transposon mutagenesis experiment generated two deletion mutants. The mutants had lost the ability to recognise Ecp2 along with the OR2A (2A) gene from the Cf-Ecp2 locus. Wild-type tomato (Solanum lycopersicum) Cf0 stable transformants, overexpressing 2A via the cauliflower mosaic virus 35S promoter, partially recapitulated the S. pimpinellifolium CfEcp2 phenotype. Lack of penetrance of the phenotype in the transgenic plants was attributed to the functional interference of 2A overexpression. The non-host, Nicotiana paniculata TW99, also recognises Ecp2 from C. fulvum. N. paniculata CfEcp2 was characterised for its ability to recognise many homologs of Ecp2, including those from Mycosphaerella fijiensis (causal agent of Black Sigatoka on banana) and the wilt- causing fungi, Fusarium oxysporum and Verticillium dahliae. The ability of Cf-Ecp2 to code for the recognition of an effector from many plant pathogens provides an exciting opportunity to engineer resistance to such pathogens in important crops.
Current Opinion in Systems Biology (3) 7-14
Publisher's version: 10.1016/j.coisb.2017.02.003
ID: 55842read more
The application of mathematical modelling to chromatin-mediated
gene regulation is gaining momentum, but is still
surprisingly rare. Here we review examples in which the
combination of quantitative experimentation and mathematical
modelling has given mechanistic insights into the processes
involved. Examples include recruitment of epigenetic regulators,
the establishment and maintenance of epigenetic
memory, the dynamic cell cycle – dependent changes in
chromatin binding of epigenetic regulators, and the contribution
of 3D genome architecture to cell identity. The successful
combination of theory and experiment requires tractable
experimental systems in which quantitative measurements and
precise perturbations are possible. The advent of single cell
technologies and genome editing presents an unprecedented
opportunity for combining quantitative experiments, precise
perturbation and modelling, that in future will enable new
epigenetic data to be embedded in a coherent theoretical
Isolation and Characterization of a Double Stranded DNA Megavirus Infecting the Toxin-Producing Haptophyte Prymnesium parvum.
Viruses (9) 40
Publisher's version: 10.3390/v9030040
ID: 55836read more
Prymnesium parvum is a toxin-producing haptophyte that causes harmful algal blooms globally, leading to large-scale fish kills that have severe ecological and economic implications. For the model haptophyte, Emiliania huxleyi, it has been shown that large dsDNA viruses play an important role in regulating blooms and therefore biogeochemical cycling, but much less work has been done looking at viruses that infect P. parvum, or the role that these viruses may play in regulating harmful algal blooms. In this study, we report the isolation and characterization of a lytic nucleo-cytoplasmic large DNA virus (NCLDV) collected from the site of a harmful P. parvum bloom. In subsequent experiments, this virus was shown to infect cultures of Prymnesium sp. and showed phylogenetic similarity to the extended Megaviridae family of algal viruses.