Gene × environment tests discriminate the new EU2 evolutionary lineage of Phytophthora ramorum and indicate that it is adaptively different

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Abstract:

A new evolutionary lineage of the destructive introduced tree pathogen Phytophthora ramorum, EU2 lineage, was recently discovered attacking larch and other hosts in Northern Ireland and south west Scotland, UK. Sixteen ‘medium × agar concentration × incubation temperature’ stress environments were tested to find a rapid and repeatable method to discriminate the known EU2 lineage from the EU1, NA1 and NA2 lineages in culture, in particular from the EU1 already prevalent across the UK; and to investigate whether EU2 might be adaptively different. At 28°C on both Carrot agar and V8 juice agar, the mean radial growth rates of all four lineages were significantly different, with NA2 > EU2 > EU1 > NA1. At this temperature, EU2 colonies were not only phenotypically distinct from EU1 and all other lineages but on average grew three times as fast as EU1. This indicates that EU2 is adaptively different from EU1. Twelve days growth in the environment ‘V8A/2% agar/28°C gave excellent discrimination of all four lineages in three repeat experiments, including clear discrimination of EU2 from EU1. Each lineage exhibited a distinctive colony pattern. The utility of this test environment was examined further by screening fresh UK isolates of unknown lineage from new larch outbreak sites alongside standard isolates. The lineage assignments predicted were corroborated by gene sequencing and RFLP profiling. These results also revealed that the EU2 lineage was present at several new larch sites in south west Scotland, whereas isolates from geographically adjacent areas such as the Isle of Mull, north west Scotland, the Isle of Man and north west England were all of EU1 lineage.

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