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JS initially hypothesized that, as beetles increased in
elevation, biotic factors such as predation and parasitism would give way
to abiotic factors such as temperature in controlling the beetles' populations
(Smiley 1986). One aspect of the beetles' natural history was particularly
interesting in this regard. The larvae of the beetles secreted large amounts
of a pungeantly aromatic white liquid, which would probably be active in
repelling predators. A literature review revealed that, in European species
of Chrysomela, this liquid was derived from salicylate precursor
chemicals in the willow food (Pasteels et al 1983).
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JS assembled a team of students from UC Irvine and, in 1982,
began an in-depth study of the biology of these organisms. The first season's
results revealed marked differences in the timing of spring emergence of
the beetles, with late spring emergence from under late melting snow packs.
They also revealed that beetle growth rates were not affected by removal
of secretion, as predicted by Pasteels' et. al. theories of the sequestration
process. In 1983 and 1984 the study was expanded to specifically address
aspects of the elevation gradient. A survey of salicylate chemistry revealed
strong changes with increasing elevation, as well as improved survival on
plants with greater amounts of salicylates (Smiley et al 1985). Studies
also revealed two Chrysomela-specific hunting predators, Symmorphus
cristatus, a eumenid hole-nesting wasp (Hymenoptera: Vespidae) and
Parasyrphus melanderi, a hover fly whose larvae feed on Chrysomela
eggs and larvae (Diptera:Syrphidae). In addition,
in 1984 and 85, lower-elevation willow beetle populations were compared
with upper populations, including transplant experiments. The results indicated
differences in survival and growth rate between elevations, as well as providing
evidence for local adaptation (Smiley and Rank 1986).
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In 1985 NR began in depth studies of egg-laying and food
plant choice behavior in C. aeneicollis, and discovered a marked
preference for plants rich in salicylates. He generally did not record higher
survivorship on those plants, however. He speculated that the specialist
hunting predators were keying in on the salicylate rich plants, nullifying
any advantage for the beetles of feeding on those plants. (Rank 1992, 1994)
He verified this prediction for syrphid fly females, who preferentially
oviposit on salicylate-rich plants (Rank and Smiley 1994). In 1988, NR began
what has since become a long-term study of the population genetic structure
of the willow beetles. Analyzing beetles from numerous populations in the
eastern Sierra Nevada, he found ample evidence of differentiation at all
scales: between-drainage, between-elevation (site), and down to between
plant within site. The pattern of differentation suggested gene flow between
populations, as well as evidence for selection for particular genotypes
at particular elevations (Rank 1992).
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In 1987 the numbers of willow beetles began to decline,
corresponding with a widespread drought in California, which lasted until
1993. In the summer of 1994 the populations began to rebound, in some areas
spreading out from the highest elevation sites which may have served as
refugia. By 1995 populations were healthy, and remained so until 2003 and
2004, when some populations began to decline. The patterns of beetle abundance
roughly correspond with annual precipitation, mainly in the form of winter
snow.
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In 1996, NR resampled the newly expanded beetle populations
to look for genetic change. The subsequest analysis, done in collaboration
with ED (Rank and Dahlhoff, 2003), showed strong evidence for directional
selection at the PGI locus, possibly based on cooler temperatures and interactions
with heat shock proteins (HSP's). This began a productive collaboration,
involving many students from both institutions, specifically into the evolution
of thermal tolerance in willow beetles and more broadly, into the behavior,
ecology and evolution of the beetles (Dahlhoff and Rank, 2000, Neargarter
et al 2003, McMillan et al 2005).
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In 1997 JS began intensive observations on Symmorphus
hole nesting wasps in Big Pine Creek. He verified that Symmorphus only take
3rd instar C. aeneicollis larvae, and observed many interactions
between wasps, nest holes, and beetle prey (Sears et al 2001). The possibility
that Symmorphus could nearly or completely extirpate local populations
of C. aeneicollis was confirmed. Observations along the Big Pine
Creek gradient have also confirmed that the wasps are colonizing upper elevation
sites for the first time since the study started. (Nakatani 2004). This
corresponds to a more complex upward shift in beetle populations.