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Optimization of a solid phase microextraction (SPME) method for analyzing litter volatiles

ABSTRACT: Leaf metabolomes are constantly changing in response to biotic and abiotic conditions, alongside internal physiological processes. Additionally, the diversity of plant species in a surrounding community influences the leaf metabolome. However, it is rarely considered how these changes might affect the chemical composition of litter emissions, which can mediate interactions with the decomposer community. To study these effects, reproducible methods for trapping litter volatiles are essential, particularly for analyzing larger sample numbers and after longer storage periods. This study optimized and validated a solid-phase microextraction (SPME) method for analyzing small amounts of litter from Quercus petraea within a biodiversity ecosystem functioning experiment. The SPME optimization focused on the fiber material, extraction temperature, and duration of sampling. The final optimized method, using a PDMS/DVB fiber and 60-minute extraction at 70 degrees celsius, demonstrated a high reproducibility, with %RSD values ranging from 0.073 to 0.119% (intraday) and 0.115 to 0.169% (interday) for retention time, and 4.41 to 7.72% (intraday) and 7.67 to 11.0% (interday) for peak area. This makes it a reliable and reproducible approach for the collection and analysis of litter volatiles. After validation, this method was applied to test the hypothesis that oak leaf litter volatile profiles are influenced by the surrounding biodiversity. The results showed that the main compound classes in oak leaf litter were fatty acids and terpenoids. Furthermore, plot diversity level was negatively correlated with terpenoid emission rates. By detecting these changes in volatile composition, we gained further insights into how tree diversity affects litter chemistry, which may also impact the decomposer network.

INSTRUMENT(S): SCION TQ

ORGANISM(S): Quercus Petraea (ncbitaxon:38865)

SUBMITTER: Gabriela A. S. Escaliante

PROVIDER: MSV000097746 | MassIVE | Mon Apr 28 04:47:00 BST 2025

REPOSITORIES: MassIVE

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Project description:In the production of seedless table grapes, it is conventional to use plant growth regulators including gibberellins. Little is known about the differences in aroma volatiles between seedless-treated grapes and nontreated (seeded) grapes. Therefore, in this study, the aroma volatile profiles of seedless-treated and nontreated ‘Shine Muscat’ grape berries during ripening were compared using gas chromatography-mass spectrometry. Measurements of volatiles during ripening showed 202 peaks in the seedless-treated and nontreated whole grape berries. According to two-way analysis of variance, the number of volatiles with differences between seedless-treated and nontreated berries and/or between ripening stages was 123, whereas those with no differences between treatments and between ripening stages was 79. Two-way hierarchical clustering analysis for the 123 volatiles showed that seedless-treated berries at the early ripening stages were separated from the other berries, and the seedless-treated and nontreated berries at the post-ripening stage were classified into the same cluster. At the early ripening stage, more lipoxygenase-pathway volatiles were produced in the seedless-treated berries than in the non-treated ones. Linalool compounds increased in both seedless-treated and nontreated berries with ripening. Gene expression profile comparisons using principal component analysis of RNA-sequencing data showed that the seedless-treated berries ripened earlier than the nontreated berries at the early ripening stage. The number of differentially expressed genes in the seedless-treated berries decreased during ripening. Using weighted gene co-expression network analyses, 12 modules and 24 modules were detected in berry skin and flesh, respectively. The correlation analysis revealed that 33 volatiles correlated with four modules in the skin and 50 volatiles correlated with nine modules in the flesh. Most of the volatiles correlated with these modules were those that showed significant differences between treatments and/or ripening stages by two-way analysis of variance. The differences in the aroma volatile profiles between seedless-treated and nontreated berries decreased as harvest was delayed, suggesting that delaying the harvest may make it possible to bring the aroma of seedless-treated ‘Shine Muscat’ berries closer to the original aroma of the seeded berries.

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Optimization of a solid phase microextraction (SPME) method for analyzing litter volatiles

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