This study was untargeted, i.e., no metabolites were identified.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse-phase"],"chromatography_protocol":["Samples were subjected to ultra-high performance liquid chromatography (UHPLC: Dionex Ulti-Mate 3000, Thermo Fisher Scientific, San José, CA, USA) coupled to quadrupole time-of-flight mass spectrometry (QTOF-MS/MS: compact, Bruker Daltonics, Bremen, Germany).
Chromatographic separation of samples (4 μL) was performed at 45 °C on a Kinetex XB-C18 column (150 × 2.1 mm, 1.7 μm, with guard column, Phenomenex), using the following gradient involving eluent A [H2O with 0.1% formic acid (FA; Sigma-Aldrich)] and eluent B [acetonitrile: LC-MS grade (Thermo Fisher Scientific); with 0.1% FA] at a flow rate of 0.5 mL/min: 2 to 30% B within 20 min, to 75% B within 9 min, followed by cleaning and equilibration of the column.
"],"publication":["Biochemical changes in Robinia pseudoacacia leaflets in dependence of leaf mining, plant age and location. 10.1186/s12870-026-08364-6. PMID:41691214"],"submitter_name":["Rabea Schweiger"],"submitter_affiliation":["Department of Chemical Ecology, Bielefeld University"],"organism_part":["leaflet"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["For chemical analyses, the samples were milled, processed and analyzed in random order. Metabolic fingerprinting of (semi-)polar metabolites was performed as described in (Schweiger et al. 2021) with some modifications. Leaf powder (10 mg dw) was extracted in an ice-cold ultrasonic bath for 15 min, using 450 μL ice-cold 90% methanol (LC-MS grade, Thermo Fisher Scientific, Loughborough, UK) with 10 mg/L hydrocortisone (Sigma-Aldrich, Steinheim, Germany) as internal standard. Twelve blanks without plant material were prepared in addition. Supernatants were filtered (0.2 μm, Phenomenex, Torrance, CA, USA).
REFERENCES:
Schweiger R, Castells E, Da Sois L, Martínez-Vilalta J, Müller C. Highly species-specific foliar metabolomes of diverse woody species and relationships with the leaf economics spectrum. Cells. 2021;10:644.
"],"organism":["Robinia pseudoacacia"],"data_transformation_protocol":["The T-ReX 3D algorithm (MetaboScape 2021b, Bruker Daltonics) was used for m/z axis recalibration and for picking of metabolic features, each being described by a retention time (RT) and m/z value, in MS mode. Features with a minimum peak height of 1,000 and at least 15 data points were included. All features likely belonging to the same metabolite were collected in so-called buckets, using a correlation coefficient threshold of 0.8 and allowing [M–H]–, [M + Cl]–, [M+HCOOH–H]–, [2 M–H]– and [M–H–H2O]– ions along with their corresponding isotopes and charge states. For quantification, from each bucket the feature with the highest intensity was used, omitting features within or close to the injection peak (i.e., those with RT < 1.25 min). Peak heights were divided by those of hydrocortisone ([M+HCOOH–H]–), the averaged blanks were subtracted and peaks were divided by the sample mass.
"],"study_factor":["Herbivory","Tree age","Tree ID","Location"],"submitter_email":["rabea.schweiger@uni-bielefeld.de"],"metabolights_link":["https://www.ebi.ac.uk/metabolights/MTBLS14099"],"sample_collection_protocol":["Study locations
The samples were collected from R. pseudoacacia growing in Dnipro city and its vicinity in the Ukrainian North Steppe subzone, which has a temperate continental climate. Four study locations were selected (BotG, Botanical Garden; May, Mayorka; MonIs, Monastyrsky Island; LomFP, Lomyvsky Forest-Park), which are characterized by different environmental conditions and in which R. pseudoacacia trees serve distinct functions, either as ornamental plantations in green recreational zones or to protect against soil erosion.
The formal taxonomic identification of R. pseudoacacia was carried out by Dr. Iryna Ivanko. Voucher specimens of this species have been deposited in the Herbarium of Oles Honchar Dnipro National University (DSU), Ukraine, voucher numbers 123,017–123,119.
Sampling
We sampled biomass from urban trees for which the exact planting origin cannot be determined; no seeds or reproductive material were used. As these trees are components of municipal urban green infrastructure, no special permits were required for sample collection. Leaflets of R. pseudoacacia plants were collected during the vegetation season in September 2024. In the study region, the studied leaf miner species are usually in their second generation (personal observation). For the sampling, trees younger than 10 years with similar morphological and biometric characteristics were selected per location. In the Botanical Garden, samples were taken in addition from trees of 10–25 years and trees older than 25 years. Trees were selected based on similar characteristics within age class. Tree cores were taken at a height of 1.3 m of the stems to estimate the age of each tree. At each site, leaf samples of 10 trees were taken, except for Monastyrsky Island, where only five trees of the respective category were available and these were sampled twice. In total, there were 16 groups.
All trees showed symptoms of herbivore damage by either one or both miner species, P. robiniella (P) and M. robiniella (M). While both herbivore species consume mesophyll and produce blotch mines, the spatial feeding patterns slightly differ. Larvae of P. robiniella feed digitate mines closer to the adaxial leaf side, while larvae of M. robiniella form elongate-oval mines at the abaxial leaf side (Davis & De Prins 2011, Needham et al. 1928). Trees infested with P and trees infested with P and M were selected for sampling. Within these trees, terminal leaflets of one of three categories were collected: without damage symptoms (“uninfested”, U), infested with P or infested with M. For the category U, the remaining parts of the compound leaves were uninfested, while for the categories P and M other leaflets of the compound leaves were uninfested or infested by the same herbivore species. Leaf samples were taken under cloudless conditions from the lower third of the crown of southern exposure at approximately the same sampling height (1.7–1.9 m) across trees. First, 20 terminal leaflets per tree from entire compound leaves of the same infestation category were harvested using scissors, washed with tap water and dried on paper sheets at ambient temperature within about 5–7 min. Then, only the upper section of the terminal leaflet of each compound leaf was cut with a scalpel, avoiding the lower section which carried the mines in case of infested leaflets. Leaflet sections from 20 leaves were pooled in one paper bag, representing one sample. All stages of the sampling were carried out with gloves and the instruments used (scissors, scalpel) were cleaned with ethanol between samples. The leaflets were air-dried at ambient conditions in the dark for 6–7 weeks and further dried in a drying oven at 40 °C for 4 d. A similar approach had been previously used by other researchers to analyze various phytochemicals from R. pseudoacacia leaves that had been air-dried at room temperature (Uzelac et al. 2023). Sampling in liquid nitrogen or direct quenching in solvents was logistically not possible in the present study.
REFERENCES
Davis DR, De Prins J. Systematics and biology of the new genus Macrosaccus with descriptions of two new species (Lepidoptera, Gracillariidae). ZooKeys. 2011; 98:29–82.
Needham J, Frost SW, Tothill BH. Leaf-mining insects. Baltimore. The Williams & Wilkins company. 1928.
Uzelac M, Sladonja B, Sola I, Dudas S, Bilic J, Famuyide IM, McGaw LJ, Eloff JN, Mikulic-Petkovsek M, Poljuha D. Invasive alien species as a potential source of phytopharmaceuticals: phenolic composition and antimicrobial and cytotoxic activity of Robinia pseudoacacia L. leaf and flower extracts. Plants. 2023;12(14):2715.
"],"omics_type":["Metabolomics"],"study_design":["Metabolomics","Allocation","Mass Spectrometry","leaf miner","Herbivory","ultra high performance liquid chromatography","Gracillariidae","Nitrogen","Carbon","untargeted metabolite profiling"],"curator_keywords":["Metabolomics","Allocation","Mass Spectrometry","leaf miner","Herbivory","ultra high performance liquid chromatography","Gracillariidae","Nitrogen","Carbon","untargeted metabolite profiling"],"mass_spectrometry_protocol":["Via a T-piece, part of the sample stream was subjected to negative electrospray ionization and QTOF-MS/MS, with the following settings in MS mode: 5 Hz spectra rate, centroid mode, mass-to-charge (m/z) range 50–1,300, end plate offset 500 V, capillary voltage 3000 V, N2 as nebulizer (3 bar) and dry gas (12 L/min, 275 °C), low mass m/z 90, quadrupole ion energy 4 eV, collision energy 7 eV. The AutoMSMS mode was used for fragmentation (MS/MS), with N2 as collision gas and ramping isolation widths and collision energies along with the precursor m/z. Prior to each sample, sodium formate solution was pumped into the QTOF for m/z recalibration. Some samples had to be excluded due to technical issues, reducing the samples sizes of the LC-MS dataset to n = 7–10.
"],"pubmed_title":["Biochemical changes in Robinia pseudoacacia leaflets in dependence of leaf mining, plant age and location."],"pubmed_authors":["Sytnyk Svitlana S, Schweiger Rabea R, Holoborodko Kyrylo K, Müller Caroline C"],"additional_accession":[]},"is_claimable":false,"name":"Biochemical changes in Robinia pseudoacacia leaflets in dependence of leaf mining, plant age and location","description":"Plant responses to leaf-mining insects are still poorly studied, especially in woody species. Robinia pseudoacacia is widely cultivated in Ukraine, where it is not native. There, the trees are facing increasing damage from the likewise introduced leaf miner species Parectopa robiniella and Macrosaccus robiniella. We examined the effects of infestation by these species on foliar carbon (C) and nitrogen (N) content and on metabolic fingerprints of trees of different age and at different locations, expecting context dependency in induction responses. Uninfested or miner-infested leaflets were collected from infested trees of three age classes at one location and from trees of one age class at four distinct locations. C and N content were mostly determined by location, with minor effects of tree age and herbivory, for N in interaction. Among the 3,121 metabolic features detected with UHPLC-QTOF-MS/MS, 1,087 were quantitatively modulated by herbivory in at least one age–location combination. Metabolic responses to the two leaf miner species partially overlapped. The magnitude and direction of metabolic shifts varied with both tree age and location. The differences in leaf biochemistry and in responses to herbivory between tree age classes and locations may reflect physiological constraints, past environmental conditions, age-dependent defense strategies, resource availabilities and allocation as well as local edaphic and microclimatic conditions, but potentially also different damage levels. That the leaflets of R. pseudoacacia exhibited context-dependent biochemical responses to the leaf miners highlights the importance of accounting for plant ontogenetic stage and local environmental heterogeneity when evaluating plant–leaf miner interactions.
","dates":{"publication":"2026-03-20","submission":"2026-03-20"},"accession":"MTBLS14099","cross_references":{"pubmed":["41691214"]}}