Laser spectroscopy steered 13 C-labelling of plant material in a walk-in growth chamber.

RATIONALE: Carbon-13 (13 C)-labelled plant material forms the basis for experiments elucidating soil organic carbon dynamics and greenhouse gas emissions. Quantitative field-scale tracing is only possible if plants are labelled homogeneously in large quantities. By using a laser spectrometer to automatically steer the isotopic ratio in the chamber, it is possible to obtain large amounts of homogeneously labelled plant material. METHODS: Ninety-six maize plants were labelled for 25 days until tassel formation in a 15 m3 walk-in growth chamber with a continuous air δ13 C-CO2 value of 400‰. A Los Gatos Research laser absorption spectrometer controlled the ambient δ13 C-CO2 value in the chamber through steering of the mass flow controllers with 13 C-enriched and natural abundance CO2 gas. RESULTS: Laser absorption spectroscopy steering kept the δ13 C value of chamber air between 368 and 426‰. The resulting 1 kg dry matter of 13 C-labelled shoots showed an average δ13 C value of 384‰ and accuracy of 8‰ (half width of the 95% confidence interval). Only the oldest leaves showed larger heterogeneity. The growth chamber eliminated variability between plants. The δ13 C value of the stabile material did not differ significantly from that of bulk material. CONCLUSIONS: Laser spectroscopy controlled 13 C labelling of plants in a walk-in growth chamber successfully kept the isotopic ratio of the CO2 in the chamber air constant. Therefore, large quantities of material were labelled homogeneously at the inter- and intra-plant level, thus establishing a method to provide high-quality input for quantitative isotopic tracer studies.

Uptake of (14)C-atropine and/or its transformation products from soil by wheat (Triticum aestivum var Kronjet) and their translocation to shoots.

Plant uptake of toxins and their translocation to edible plant parts are important processes in the transfer of contaminants into the food chain. Atropine, a highly toxic muscarine receptor antagonist produced by Solanacea species, is found in all plant tissues and can enter the soil and hence be available for uptake by crops. The absorption of atropine and/or its transformation products from soil by wheat (Triticum aestivum var Kronjet) and its distribution to shoots was investigated by growing wheat in soil spiked with unlabeled or (14)C-labeled atropine. Radioactivity attributable to (14)C-atropine and its transformation products was measurable in plants sampled at 15 d after sowing (DAS) and thereafter until the end of experiment. The highest accumulation of (14)C-atropine and/or its transformation products by plants was detected in leaves (between 73 and 90% of the total accumulated) with lower amounts in stems, roots, and seeds (approximately 14%, 9%, and 3%, respectively). (14)C-Atropine and/or its transformation products were detected in soil leachate at 30, 60, and 90 DAS and were strongly adsorbed to soil, with 60% of the applied dose adsorbed at 30 DAS, plateauing at 70% from 60 DAS. Unlabeled atropine was detected in shoots 30 DAS at a concentration of 3.9 ± 0.1 µg kg(-1) (mean ± SD). The observed bioconcentration factor was 2.3 ± 0.04. The results suggest a potential risk of atropine toxicity to consumers.

Total body nitrogen and total body carbon as indicators of body protein and body lipids in the melon fly Bactrocera cucurbitae: effects of methoprene, a juvenile hormone analogue, and of diet supplementation with hydrolyzed yeast.

The application of methoprene, and providing access to diet including hydrolyzed yeast, are treatments known to enhance mating success in the male melon fly Bactrocera cucurbitae Coquillett (Diptera: Tephritidae), supporting their use in mass rearing protocols for sterile males in the context of sterile insect technique (SIT) programmes. The objective of the present laboratory study was to investigate the effect of methoprene application and diet supplementation with hydrolyzed yeast (protein) on the turnover of body lipids and protein to confirm the feasibility of their application in melon fly SIT mass-rearing programmes. While females had access to a diet that included hydrolyzed yeast (protein), males were exposed to one of the following treatments: (1) topical application of methoprene and access to diet including protein (M+P+); (2) only diet including protein (M-P+); (3) only methoprene (M+P-) and (4) untreated, only sugar-fed, control males (M-P-). Total body carbon (TBC) and total body nitrogen (TBN) of flies were measured at regular intervals from emergence to 35 days of age for each of the different treatments. Nitrogen assimilation and turnover in the flies were measured using stable isotope ((15)N) dilution techniques. Hydrolyzed yeast incorporation into the diet significantly increased male body weight, TBC and TBN as compared to sugar-fed males. Females had significantly higher body weight, TBC and TBN as compared to all males. TBC and TBN showed age-dependent changes, increasing until the age of sexual maturity and decreasing afterwards in both sexes. Methoprene treatment did not significantly affect TBC or TBN. The progressive increase with age of TBC suggests that lipogenesis occurs in adult male B. cucurbitae, as is the case in other tephritids. Stable isotope dilution was shown to be an effective method for determining N uptake in B. cucurbitae. This technique was used to show that sugar-fed males rely solely on larval N reserves and that the N uptake rate in males with access to diet including hydrolyzed yeast was higher shortly after emergence and then stabilized. The implications of the results for SIT applications are discussed.