Proteome data on the microbial microbiome of grasshopper feces

We present proteome data from the microbiota (feces) after a diet shift from a natural diverse to a monocultural meadow with Dactylis glomerata. The abundant grasshopper species, Chorthippus dorsatus, was taken from the wild and kept in captivity and were fed with Dactylis glomerata for five days. For phytophagous insects, the efficiency of utilization of hemicellulose and cellulose depends on the gut microbiota. Shifts in environmental and management conditions alter the presence and abundance of plant species which may induce adaptations in the diversity of gut microbiota. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD005126.


a b s t r a c t
We present proteome data from the microbiota (feces) after a diet shift from a natural diverse to a monocultural meadow with Dactylis glomerata. The abundant grasshopper species, Chorthippus dorsatus, was taken from the wild and kept in captivity and were fed with Dactylis glomerata for five days. For phytophagous insects, the efficiency of utilization of hemicellulose and cellulose depends on the gut microbiota. Shifts in environmental and management conditions alter the presence and abundance of plant species which may induce adaptations in the diversity of gut microbiota. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD005126. &

Value of the data
Protein assessment of the microbiota of the grasshopper species Chorthippus dorsatus. Metaproteome from the grasshoppers provides the basis for more functional analyses of the grasshopper microbiota.
Relevant information for the grasshoppers ecology on the basis its microbiota.

Data
We present the first dataset of this relevant type of phytophagous insects since there is so far no metaproteome dataset on the gut microbiota of grasshoppers available. To detect the diet dependent metabolic adaptation, a shift in diet from a diverse food source to the single-species food Dactylis glomerata was performed.

Grasshopper culture and feces sampling
We selected Chorthippus dorsatus that was at that time the most dominant grasshopper species on a ruderal meadow that has been under this type of land use for 20 years. The meadow was dominated by grasses, namely Poa pratensis, Dactylis glomerata, Festuca pratensis, and Bromus sterilis, which are all potential food plants for the generalist herbivores in grasslands [1][2][3][4]. Three days before the start of the experiment, we caught female and male grasshoppers with sweep nets and kept them separately in cages with a mixed grass diet from the meadow until the start of the experiment. In August 2014, 2 male and 2 female grasshoppers were transferred to a cage without food for 1 day to synchronize for gut content. On the following day D. glomerata, which was grown in a climate chamber under standardized and controlled soil and moisture conditions for about 2.5 months, was added to the cage. Excrements were sampled before (d0) and after adding D. glomerata at day 1, 3, and 5 (day 1, day 3 and day 5) for one week and frozen at À 80°C.

Protein extraction and sample preparation
After the sampling of feces (day 0, 1, 3, 5 and 6), three feces were pooled and were considered as one replicate. For protein extraction, to the feces 4 glass beads (3 mm, Carl Roth GmbH), two spatula tips of zirconium beads (0.1 mm diameter, Biospec.) and 800 mL of lysis buffer (500 mM NaCl, 50 mM Tris-HCl, pH8, 50 mM EDTA, 4% (w/v) SDS) were added. Feces were disrupted by FastPrep (3 Â 1 min, 5.5 ms À 1 , MP Biomedicals) and incubated for 15 min at 95°C. After centrifugation (14,000 rpm, 5 min, 4°C), the supernatant was taken. To the pellets 300 mL of lysis buffer were added and the procedure with FastPrep and heating was repeated exclusively for the pellet. The supernatant was precipitated over night at À 20°C with acetone (2.5 fold of ice-cold acetone). Protein pellets were harvested by centrifugation (14,000 rpm). Dried pellets were dissolved in 20 mL of SDS sample buffer (2% w/v SDS, 2 mM beta-mercaptoethanol, 4% v/v glycerol, 40 mM Tris-HCl pH 6.8, 0.01% w/v bromophenol blue), heated to 90°C for 4 min and separated by SDS polyacrylamide gel electrophoresis. Proteins were stained in gel with Coomassie G-250 (Merck). Gel was cut into small pieces (band per samples), destained, dehydrated and proteolytically cleaved overnight at 37°C trypsin (Promega) [5]. Extracted peptides were desalted using C18 ZipTip column (Merck Millipore). Peptide lysates were resuspended in 0.1% formic acid and injected to liquid chromatography mass spectrometry (LC-MS/MS).

Mass spectrometric measurement and data analysis
Mass spectrometry was performed using Orbitrap Fusion (Thermo Fisher Scientific) coupled to a TriVersa NanoMate (Advion) as described [6]. To assess the protein functions of the different bacterial  Chromatin structure and dynamic is present at time point (d0) with 34%, but disappeared at the later time points. Carbohydrate transport and metabolism increases from d0 (21.7%) to d1 (53%) and then slowly decrease to d6 (44%). Amino acid transport and metabolism has also increase of the abundance from d0 (10%) to d6 (44%).