Pepperwood Preserve Case Study

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Figure 1: Pepperwood Preserve study system for U. californica for susceptibility of P. ramorum depicted by climate and topography aerial map. No image rights are owned and the map was created by Celeste Dodge. Two red boxes in bottom right of the image indicates the approximate location of the two study sites.
In October of 2016, bay leaves were collected at Pepperwood Preserve, Sonoma County at a study system composed of mixed evergreen forest that interfaces with nearby serpentine grasslands and oak woodlands. The site has a south-west aspect that receives evening sunlight, and has varying degrees of slope. Branches were collected at varying degrees of reachable height below below 3-meters and represent a limited aspect of U. californica
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californica trees were selected for branch testing at two nearby areas sharing similar ecosystem characteristics, for a total of 36 trees sampled. Branches selected had a minimum of two years of leaf growth and asymptomatic leaves, which were cut with pruners and transferred into clear plastic bags. The bags were stored at 4°C for 7 days in the dark.
Preparation of P. ramorum zoospores at [2.0x 10^5 zoospores ml-1] began by placing P. ramorum into an aqueous solution with hemp seeds as a nutrient source. Material was transferred over to a V8 agar plate which was left to grow for 15 days at room temperature with light. Afterward, 10 mL of sterile soil water was added to the agar plate, which was stored for two more days at room temperature without light. The agar plate was then stored for a single day at 4°C in the dark. Zoospores with soil water were removed from the agar plate.
Inoculation
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Since, leaf moisture content is an internal measure, it thereby, is unlikely to directly influence how zoospores were transported external to the leaf surface. Farther research on the influence of water in this pathosystem should focus on relative humidity (RH) and leaf surface condensation, which are more additional moisture metrics that could influence P. ramorum spread in field environments. Leaf moisture content was >50% for all that were leaves tested, which suggest water is an unlikely limiting reagent, if involved whatsoever, in the mechanisms influence the magnitude of necrosis in bay

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