❤❤❤ Garden Leaf Sweeper Research Paper

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Garden Leaf Sweeper Research Paper



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The eggs are deposited directly on the leaves or stem in early summer and hatch after a few weeks. Both larvae and adults feed on the plant, which can sometimes lead to quite severe damage. Both G. For the pheromone collection experiments, adults of G. Before trials, the beetles were stored in ventilated plastic boxes in the laboratory and fed with fresh leaves from their respective host plant when needed. All plants used were potted and kept in the common garden or green house prior to the tests. Bartelt et al. Hence, all five species of Galerucella included in this study were used with one of their respective host plants: G. The pheromone collection was performed from late May to early July A few days before the tests, all plants were checked thoroughly for any herbivorous insects and kept in the green house until the time for the experiments.

Plants in bloom or plants afflicted by herbivores or showing fresh cuts or injuries were not used in the experiments. To distinguish volatiles emitted by the beetles from those released by the plants, three experimental setups were used for each beetle—host plant pair: 1 10 male beetles per plant [ G. The leaves of the mechanically damaged plants were perforated using a preparation needle 1.

As far as possible, the same amount of damage was inflicted to each plant, in total perforation holes per plant. This roughly mimicked the amount of damage inflicted by 10 feeding beetles during the time for volatile collection 24 h. Approximately the same leaf biomass was used for each setup and a new plant was prepared for each experiment. Important considerations have to be taken into account regarding the use of SPME technique for collection of constantly produced volatiles and interpretation of data. For this purpose, volatiles were collected and analysed in exactly the same way, using the same fibre type, the same headspace volume, controlled, stable temperatures during sampling and addition of internal standard to monitor unsaturation of fibres.

Under these circumstances, SPME is an extremely sensitive, user-friendly, and excellent technique. However, comparison of quantities of different compounds in the same sample without the use of labelled standards is not possible because of different affinities of compounds on the fibre and because of different vapour pressures of the target compounds based on SPME guidelines, J Chem Ecol. After purification of the fibre, the wall of the polyester bag was pierced by the needle of the syringe, and the purified fibre was exposed to a headspace.

The male beetles or larvae were put on the plant just before enclosing it in the polyester bag and allowed to feed during the time for volatile collection 24 h. After collection, the fibre was retracted into the needle, removed from the bag and placed into a GC injector for desorption of volatiles. Helium was used as the carrier gas with an inlet pressure of 10 psi. Identity of dimethylfuran-lactone in the samples was confirmed by comparison of mass spectral data and GC-retention times of natural product with the corresponding data of synthetic standard and those published by Bartelt et al. Relative amount of the target compound was expressed as an area under a peak using arbitrary units.

The synthetic scheme of the Galerucella spp. The synthetic pathway to the dimethylfuran-containing macrolide pheromone 1 consists of preparing 3,4-dimethylfuran 4 , attaching suitable hydroxyl and acyl side chains at position 2 and 5, and finally closing the lactone cycle. This synthesis follows the main part the paths described by Bartelt et al. Synthetic scheme of Galerucella pheromone dimethylfuran-lactone: 12,dimethyl-5,dioxabicyclo[9. Behavioural responses of Galerucella spp. This setup was arranged so that four olfactometer trials could be run simultaneously.

Before trials, one beetle was allowed to acclimatize inside the olfactometer for 5 min. After the trials, the sex of each beetle was determined through dissection of the genital parts. Beetles that were inactive for more than 5 min during the trials were excluded from the study. The olfactometer studies were started by investigating the behavioural responses of G. Each arm zone of the olfactometer was connected to a ml plastic syringe, with the plunger withdrawn to allow air to be drawn through the arena. Next, the behavioural responses of the beetles to a blend of the pheromone and odours from their respective host plants were investigated. These studies were performed on G. The experimental procedure followed the protocol described above, but with different odour treatments; 1 pheromone vs.

For the host plant treatments to cm branches were cut from potted plants of L. Plants in bloom or plants afflicted by herbivores were not used in the tests. The branches were cut so that each branch had on average 5—7 leaves to keep the leaf biomass similar amongst the trials. As Galerucella beetles usually respond stronger to damaged plants compared to non-damaged plants, five leaves on each branch were mechanically damaged one hole per leaf using a preparation needle 1. Relative amounts of dimethylfuran-lactone trapped from male beetles of the five Galerucella species, as well as those of the internal standard, were log transformed and analysed for variance using ANOVA. Zero values were not included in the analyses.

The analyses were carried out using the computer program package Statistica, version In the statistical analyses of the behavioural data, the replicate was the individual beetle and the analyses were performed on the number of recordings each beetle individual spent in each arm zone during the min session. In the first part of the behavioural test responses to the synthetic pheromone all five Galerucella species were included and in the second part involving host plant odours G. Wilcoxon signed rank tests were used to analyse whether the beetles visited one arm zone significantly more than the other. Recordings made when the beetles were located in the central zone of the arena were excluded, as these recordings provided less information about attraction towards any stimulus.

Analyses of behavioural data were performed using R 2. The aggregation pheromone dimethylfuran-lactone of G. The same pheromone was produced in significantly lower amounts also by males of G. In the experiment using larvae instead of adult beetles, no compounds were found that could be distinguished from the volatiles emitted from the plants. Selected ion chromatogram of records of volatiles collected from plants with feeding Galerucella beetles as well as from synthetic dimethylfuran-lactone. The values on the Y axis are quantities peak areas corresponding to the amount of compound analysed, related to the abundance of ions formed.

Differences were found amongst the five Galerucella species in their behavioural responses to the synthetic pheromone dimethylfuran-lactone. Similar to previous studies, both males and females of G. There was no evidence for G. Behavioural responses in Galerucella spp. Data were analysed using Wilcoxon signed rank tests. Error bars indicate standard error of the mean. In the behavioural studies involving host plant odours, males of G. When given a choice between the blend and L.

Somewhat different patterns were observed for G. Moreover, both males and females of G. Behavioural responses of a G. Males of the closely related chrysomelid species G. In the study presented here, the same pheromone is shown to be emitted in lower amounts also by a third species, G. The response for G. Accordingly, individuals of G. When comparing the blend to the pheromone alone, females of G. Thus, even though G. One reason for the low pheromone emission found in G. Although Fragaria sp. There is a possibility that the level of pheromone produced differs when the beetles are feeding on different plant species. The same is true for G. However, it is possible that males of G. Although dimethylfuran-lactone is the only pheromone detected and tested in this study, additional pheromone components may be present.

Putative aggregation pheromones in G. In curculionid beetles, previous studies suggest that the chemical structural relationships of aggregation pheromones are in accordance to the subfamily taxonomy Ambrogi et al. However, this is an area that is still poorly investigated. In this study, no compounds have yet been detected that could function as putative pheromone components in G. Whether this is due to true absence of volatiles or too low emission of compounds released by the beetles in relation to volatiles released by the plants cannot be assessed in this study.

To conclude, pheromone production and response in Galerucella seem to be connected to the phylogenetic relatedness between the beetle species, where the three most closely related species included in this study both produce and respond to the same pheromone dimethylfuran-lactone. Interestingly, these three species are also attacked by the same parasitoid Asecodes parviclava , whereas the two congeners, G. To better understand the pheromone evolution in Galerucella , additional ecological factors apart from reproductive isolation ought to be taken into account.

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VCH, New York. Synth Commun — Adaptations by herbivores to natural enemies exploiting prey semiochemistry. Trends Plant Sci — Chem Cent J 5 1 Step 2: Internal review is undertaken by the Research Office. Step 3: Feedback is provided to applicants and, if necessary, applications reopened for amendment. Step 4: Applicants resubmit applications before the NRF final deadline. The administrator will obtain institutional approval from the Deputy Dean for Research and forward this to the funder.

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