FIGURE LEGENDS:
Figure 1: Herbarium specimens are often mounted with roots intact and
nodules (inset) can be found on even the oldest specimens. The nodule
pictured here was take from specimen 324040 collected in 1962 from Walla
Walla, Washington. Photo credit: Water Fertig (plant specimen) and Layla
Taylor-Spriggs (inset)
Figure 2: We had no growth on any of our negative controls (A) and
historic leaf controls (B). We had leaf endophytes grow from our fresh
controls, however only for a subset of the media types (C). We had low
probability of growth for plates streaked with crushed nodules from 1950
(D) and no probability of growth on plates streaked with nodules from
1974 (E) and 1986 (F). Probability of growth was high for more recent
specimen nodules including 2004 (G) and 2015 (H). Probability of growth
was nearly 100% with contemporary nodules (I).
Figure 3: Sanger sequencing (done on 42% of our isolates) revealed
isolates from four genera isolated from our 1950 sample. We identified
isolates from 12 genera in our two nodules taken from the 2004 specimen.
We found isolates from three genera from our nodule taken from a 2015
specimen.