10.5061/DRYAD.NK98SF7SS
Kahilainen, Aapo
0000-0001-9180-6998
University of Helsinki
Oostra, Vicencio
0000-0002-1273-1906
University of Liverpool
Somervuo, Panu
University of Helsinki
Minard, Guillaume
University of Lyon System
Saastamoinen, Marjo
University of Helsinki
Alternative developmental and transcriptomic responses to host plant water
limitation in a butterfly metapopulation
Dryad
dataset
2021
FOS: Biological sciences
European Research Council
https://ror.org/0472cxd90
637412
Kone Foundation
https://ror.org/05jwty529
201802795
2021-10-10T00:00:00Z
2021-10-10T00:00:00Z
en
https://doi.org/10.1101/2021.02.24.432453
https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE159376
215236 bytes
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CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
The dataset is from a study examining the effects of host plant water
stress on the developmental and transcriptomic responses of its specialist
Lepidopteran herbivore. The study combines host plant metabolic profiling
with development assays and full-transcriptome sequencing of herbivore
larvae. First, we profiled metabolic differences between well-watered and
water-limited ribwort plantain (Plantago lanceolata) using proton nuclear
magnetic resonance spectroscopy (1H-NMR). Second, we tested how
performance of developing Glanville fritillary (Melitaea cinxia) larvae
was affected by host plant water limitation experienced at different
larval developmental stages. Third, we examined larval gene regulatory
responses to water limited host plants by sequencing full transcriptomes
of 77 female larvae (RNA seq). Finally, to examine intrapopulation
variation in the responses of the larvae, we compared the phenotypic and
transcriptomic responses across full-sib families originating from
different parts of the metapopulation. In this dataset, we provide data
for the P. lanceolata metabolite responses to water limitation and
developmental responses of the M. cinxia larvae to feeding on water
limited P. lanceolata. The transcriptomic data are available from
NCBI's Gene Expression Omnibus, with the accession number GSE159376.
We collected seeds from a natural population in the Åland Islands (60.196°
Lat., 20.704° Lon.) and after germination planted 360 plants in 0.75 litre
pots (two saplings each). We reared the plants for three months in
controlled greenhouse conditions (ca. 40 ml water / pot daily, 15L:9D
photoperiod with 26:18 °C temperature cycle) before initiating the water
limitation treatment. We exposed 240 plants to a water limitation
treatment in which daily watering was reduced by 50% compared to controls
(20ml per pot). Each morning prior to watering (9-10 am), we randomly
harvested P. lanceolata leaves from control and water limited plants and
cut them into 2.25cm2 pieces, discarding the basal and tip parts of the
leaves. We used these pieces to feed larvae during the experiment (see
below), and selected a random subset of six pieces from both treatments
for metabolomics assays. For the assays, we recorded the fresh biomass of
each piece of leaf, snap froze the pools in liquid nitrogen and stored
them in -80 °C. We then freeze dried the samples for 48 hours after which
we measured their dry weight, estimated relative water content in the
sample [i.e. (fresh mass – dry mass) / fresh mass], ground the dried pool
samples in dry ice using a sterile pestle and prepared the finely ground
tissue powder for proton nuclear magnetic resonance spectrometry (1H-NMR)
following the protocol described by Kim et al. (2010). The 1H-NMR spectra
of the P. lanceolata pool samples was recorded at the Finnish Biological
NMR Center (Institute of Biotechnology, University of Helsinki,
http://www.biocenter.helsinki.fi/bi/) using a Bruker Avance III HD NMR
spectrometer (Bruker BioSpin, Germany) operated at 1H frequency of 850.4
MHz equipped with a cryogenic probe head. The 1H-NMR spectra were aquired
at a 25 °C temperature with a “zgpr” pulse sequence. After obtaining the
spectra, we corrected the baseline, used automatic peak detection, binned
the observed peaks into 0.04 ppm bins for all pool samples and normalized
them according to the TSP signal variation and dry mass of the sample
using MNOVA v.10.0.2 software (Mestrelab research S.L., Spain). We further
annotated the peaks within the binned intervals based on signals of pure
compounds for Aucubin, Catalpol and Verbascoside or from published
characteristic signals (Agudelo-Romero et al., 2014; Gogna et al., 2015;
Kim et al., 2010). M. cinxia larval families, treatments and developmental
assays We created experimental full-sib larval family groups by mating
individuals originating from local populations across the natural Åland
islands metapopulation. From nine mated females, we picked two larval
groups of a minimum of eighty larvae each to enter the experiment. On the
day after hatching, we divided the larval groups into four smaller groups
of twenty larvae each and placed them on separate petri dishes (9 cm
diameter, 1.5 cm deep) lined with filter paper. We then randomly assigned
each of the dishes to one of four different treatments mimicking different
temporal exposures to drought stressed host plants. In addition to a
control treatment, in which we fed the larvae with control reared host
plants only, the larvae experienced water limited host plant at different
stages during pre-diapause development: (1) at late pre-diapause
development during 3rd and 4th larval instars, (2) at early pre-diapause
development during 1st and 2nd larval instars, and (3) throughout their
pre-diapause development from the 1st to the 5th larval instar. In all
treatments, we fed the larvae daily with pieces of host plant leaf tissue
corresponding to the treatment. We provided ad libitum food such that – in
order to avoid feeding on old leaf tissue with potentially altered
phytochemistry – the larvae consumed most but not all of the leaf tissue
during the next 24 hours after provisioning. During the experiment, we
recorded development time, body mass at diapause and mortality during
development. Once the last larva on a petri dish had entered diapause, we
allowed them to spend another four days in normal rearing temperature and
photoperiod, after which we measured their body mass and placed them in
climate chambers (+5 °C, 95% air humidity) for diapause. We allowed the
larvae to diapause for six months, after which we woke them up and
recorded overwintering mortality.
The data are accompanied by a README.txt file describing all data fields
in all data files. Note that although larval development time and body
mass related data is provided at the individual level, the records do not
have individual level IDs, Therefore the development time and diapause
body mass data cannot be combined with each other to examine their
association within each individual. This is because the larvae spend their
pre-diapause development and overwintering in full-sib larval groups
making it very challenging to track individual larvae across their
development. Note also that the data also contains some outlier cases we
suggest removing before any analyses. Larvae in the control treatment of
the first replicate larval group of family F-5 (clutch ID 17) had a
mortality of 55%. As the larvae in this group were developing poorly in
general, we concluded it to be an outlier case, potentially suffering from
a disease or some other unknown agent. For transparency we provide these
cases with the data, but recommend excluding this larval group from any
further analyses. In all data files regarding M. cinxia larvae, records
for this group have been marked as outlier cases with an "x" in
the data column "outlier".