Do Enemies of Herbivores Influence Plant Growth and Chemistry? Evidence from a Seminatural Experiment

It is predicted that enemies of insect herbivores may influence the effects of herbivores on their host plants by affecting the choice of plant genotypes. To examine the effect of predators, we conducted two experiments, each with a different caterpillar species (Junonia coenia and Pyrrharctia isabella). Under seminatural conditions, we provided a choice between two genotypes of plantain (Plantago lanceolata) with different levels of iridoid glycosides and used Podisus maculiventris stinkbugs as predators. There were four treatments: no herbivores and no predators, low density of herbivores and no predators, high density of herbivores and no predators, and high density of herbivores plus predators. The caterpillars had little effect on plant growth but did influence the iridoid glycoside concentration. For the Junonia experiment, the concentration of iridoid glycosides was less for plots with a low density of caterpillars (with no predators) compared to the other treatments of caterpillar density. In the Pyrrharctia experiment, catalpol was induced by a high density of caterpillars (with no predators). There were no increases in total iridoid glycosides associated with either herbivore species. The presence of predators had no effect on plant growth or total iridoid glycoside pattern. The lack of effect by predators seems to reflect the relatively large variation in iridoid glycoside concentration among leaf ages, and the herbivores ability to respond to that variation, such that the difference in iridoid glycoside concentrations in the plant genotypes was less important.     

Epoxidation of n-hexene and cyclohexene over titanium-containing catalysts

TS-1, Ti-beta and Ti-MCM-41 molecular sieves have been prepared by direct hydrothermal synthesis method and applied to the epoxidation of n-hexene and cyclohexene with H₂O₂ under mild conditions. Ti-beta with extremely low Al content was synthesized by using a seed method to suppress the formation of diol produced by Brønsted acid sites present in Ti-beta. It was also found that a large amount of by-products (1-ol and 1-one) formed over hydrophilic Ti-MCM-41. We further modified Ti-MCM-41 by silylation with bis(trimethylsilyl) trifluoroacetamide (BSTFA). Among these catalysts, the Ti-beta with low content of Al enhanced the yield of epoxide and suppressed the formation of diol markedly. The silylated Ti-MCM-41 reduced the formation of by-products and promoted the yields of epoxide significantly. Based on experimental results, a reaction mechanism with two parallel and competitive reactions was proposed.     

Visualizing a Plant Defense and Insect Counterploy: Alkaloid Distribution in <Emphasis Type="Italic">Lobelia</Emphasis> Leaves Trenched by a Plusiine Caterpillar

Insects that feed on plants protected by latex canals often sever leaf veins or cut trenches across leaves before feeding distal to the cuts. The insects thereby depressurize the canals and reduce latex exudation at their prospective feeding site. How the cuts affect the distribution and concentration of latex chemicals was not known. We modified a microwave-assisted extraction technique to analyze the spatial distribution of alkaloids in leaves of Lobelia cardinalis (Campanulaceae) that have been trenched by a plusiine caterpillar, Enigmogramma basigera (Lepidoptera: Noctuidae). We produced sharp two dimensional maps of alkaloid distribution by microwaving leaves to transfer alkaloids to TLC plates that were then sprayed with Dragendorff’s reagent to visualize the alkaloids. The leaf prints were photographed and analyzed with image processing software for quantifying alkaloid levels. A comparison of control and trenched leaves documented that trenching reduces alkaloid levels by approximately 50% both distal and proximal to the trench. The trench becomes greatly enriched in alkaloids due to latex draining from surrounding areas. Measurements of exudation from trenched leaves demonstrate that latex pressures are rapidly restored proximal, but not distal to the trench. Thus, the trench serves not only to drain latex with alkaloids from the caterpillar’s prospective feeding site, but also to isolate this section, thereby preventing an influx of latex from an extensive area that likely extends beyond the leaf. Microwave-assisted extraction of leaves has potential for diverse applications that include visualizing the impact of pathogens, leaf miners, sap-sucking insects, and other herbivores on the distribution and abundance of alkaloids and other important defensive compounds.     

Plant Cyanogenesis of <Emphasis Type="Italic">Phaseolus lunatus</Emphasis> and its Relevance for Herbivore–Plant Interaction: The Importance of Quantitative Data

Quantitative experimental results on the antiherbivorous effect of cyanogenesis are rare. In our analyses, we distinguished between the total amount of cyanide-containing compounds stored in a given tissue [cyanogenic potential (HCNp)] and the capacity for release of HCN per unit time (HCNc) from these cyanogenic precursors as a reaction to herbivory. We analyzed the impact of these cyanogenic features on herbivorous insects using different accessions of lima beans (Phaseolus lunatus L.) with different cyanogenic characteristics in their leaves and fourth instars of the generalist herbivore Schistocerca gregaria Forskål (Orthoptera, Acrididae). Young leaves exhibit a higher HCNp and HCNc than mature leaves. This ontogenetic variability of cyanogenesis was valid for all accessions studied. In no-choice bioassays, feeding of S. gregaria was reduced on high cyanogenic lima beans compared with low cyanogenic beans. A HCNp of about 15 μmol cyanide/g leaf (fresh weight) with a corresponding HCNc of about 1 μmol HCN released from leaf material within the first 10 min after complete tissue disintegration appears to be a threshold at which the first repellent effects on S. gregaria were observed. The repellent effect of cyanogenesis increased above these thresholds of HCNp and HCNc. No repellent action of cyanogenesis was observed on plants with lower HCNp and HCNc. These low cyanogenic accessions of P. lunatus were consumed extensively—with dramatic consequences for the herbivore. After consumption, locusts showed severe symptoms of intoxication. Choice assays confirmed the feeding preference of locusts for low over high cyanogenic leaf material of P. lunatus. The bioassays revealed total losses of HCN between 90 and 99% related to the estimated amount of ingested cyanide-containing compounds by the locusts. This general finding was independent of the cyanogenic status (high or low) of the leaf material.     

Effect of hydrophobic polystyrene microphases on temperature-responsive behavior of poly(N-isopropylacrylamide) hydrogels

Reversible addition-fragmentation chain transfer polymerization was employed to prepare the crosslinked poly(N-isopropylacrylamide)-graft-polystyrene networks (PNIPAAm-g-PS). Due to the immiscibility of PNIPAAm with PS, the crosslinked PNIPAAm-g-PS copolymers displayed the microphase-separated morphology. While the PNIPAAm-g-PS copolymer networks were subjected to the swelling experiments, it is found that the PS block-containing PNIPAAm hydrogels significantly exhibited faster response to the external temperature changes according to swelling, deswelling, and reswelling experiments than the conventional PNIPAAm hydrogels. The improved thermo-responsive properties of hydrogels have been interpreted on the basis of the formation of the specific microphase-separated morphology in the hydrogels, i.e., the PS blocks pendent from the crosslinked PNIPAAm networks were self-assembled into the highly hydrophobic nanodomains, which behave as the microporogens and thus promote the contact of PNIPAAm chains and water. The self-organized morphology in the hydrogels was further confirmed by photon correlation spectroscopy (PCS). The PCS shows that the linear model block copolymers of PNIPAAm-g-PS networks were self-organized into micelle structures, i.e., the PS domains constitute the hydrophobic nanodomains in PNIPAAm-g-PS networks.     

Granulation of hydrophobic powders

Granulation of hydrophobic powders is frequently required in the pharmaceutical industry. The structural complexity of new drug molecules mean that is increasingly common for entire classes of drug compounds to be highly hydrophobic. This creates considerable difficulty in understanding, controlling and trouble-shooting these industrial granulation processes.There have been many recent advances in granulation theory. Essential to this theory is that wetting and spreading of the fluid through the powder particles is a prerequisite for good granulation. The possibility of a fine, hydrophobic powder spreading over the surface of the liquid during nucleation has been identified theoretically based on surface chemistry and as a potential nucleation mechanism. Recently, investigation confirmed that nucleation can occur by spreading of the sub-micron particles around the template drop to form a “liquid marble”. The hollow granule structure formed by the “solid-spreading” nucleation mechanism suggests the possibility of using the controlled, open granule structure to manufacture designer structured agglomerates with advantageous properties, including controlled granule structure and size and excellent ideal flow and handling properties. This paper describes single drop solid-spreading nucleation experiments where single fluid droplets are placed onto loosely packed powder beds of hydrophobic powders and the formation of the powder shell observed. Experimental results and observations for some model systems are presented, together with a preliminary framework for liquid marble formation.     

Host Recognition by the Specialist Hoverfly <Emphasis Type="Italic">Microdon mutabilis</Emphasis>, a Social Parasite of the Ant <Emphasis Type="Italic">Formica lemani</Emphasis>

The larva of the hoverfly Microdon mutabilis is a specialist social parasite of the ant Formica lemani that is adapted to local groups of F. lemani colonies but mal-adapted to colonies of the same species situated only a few hundred meters away. At a study site in Ireland, F. lemani shares its habitat with four other ant species. All nest under stones, making the oviposition choice by M. mutabilis females crucial to offspring survival. In this study, we tested the hypothesis that, as an extreme specialist, M. mutabilis should respond to cues derived from its host rather than from its microenvironment, a phenomenon that has hitherto only been addressed in the context of herbivorous insects and their parasitoids. In behavioral assays, M. mutabilis females reacted to volatiles from F. lemani colonies by extending their ovipositors, presumably probing for an oviposition substrate. This behavior was not observed toward negative controls or volatiles from colonies of Myrmica scabrinodis, the host ant of the closely related Microdon myrmicae. Coupled gas chromatography-electroantennography (GC-EAG) that used antennal preparations of M. mutabilis located a single physiologically active compound within an extract of heads of F. lemani workers. Coupled GC-mass spectrometry (GC-MS) tentatively identified the compound as a methylated methylsalicylate. GC co-injection of the extract with authentic samples showed that of the four possible isomers (methyl 3-, 4-, 5-, and 6-methylsalicylate), only methyl 6-methylsalicylate co-eluted with the EAG-active peak. Furthermore, the response to methyl 6-methylsalicylate was four times higher than to those of the other isomers. Coupled GC-EAG and GC-MS also revealed physiological responses to two constituents, 3-octanone and 3-octanol, of the M. scabrinodis alarm pheromone. However, the behavioral trials did not reveal any behavior that could be attributed to these compounds. Results are discussed in the context of four phases of host location behavior, and of the characteristics, which volatile cues should provide to be useful for an extreme specialist such as M. mutabilis.     

THE MODIFIED PGR EQUATION OF STATE: ASYMMETRIC MIXTURE VLE REPRESENTATIONS AND PREDICTIONS

A set of mixing rules was proposed for the modified Park-Gasem-Robinson (PGR) equation of state (EOS) to extend its predictions to mixtures. The phase behavior predictive capability of this segment-segment interaction model was evaluated for selected binary asymmetric mixtures involving ethane, carbon dioxide, and hydrogen in normal paraffins. The predicted bubble point pressures for the ethane + n-paraffin and carbon dioxide + n-paraffin binaries were compared to those of the Peng-Robinson (PR), simplified perturbed hard-chain theory (SPHCT), and original PGR equations. The a priori predictive capability of the modified PGR EOS is significantly better than that of the PR, SPHCT, and original PGR equations of state for ethane binaries with absolute-average percent deviation (%AAD) of 5%. However, this EOS produces comparable representations for ethane binaries (%AAD of 1.9%) and carbon dioxide binaries (%AAD of 2.0). For hydrogen binaries, the modified PGR EOS showed much better representations (%AAD of 1.7) than the original PGR equation and was comparable to the PR equation.     

Evaluation of solvent accessibility epitopes for different dehydrogenase inhibitors

Knowledge about the orientation of ligands or inhibitors bound to a protein is vital for the development of new drugs. It was recently shown that solvent accessibility epitopes for protein ligands can be mapped by transferring magnetization from water molecules to the ligand to derive the ligand orientation. This is based on the fact that NMR signals of ligands arising from magnetization transferred from solvent molecules via the protein have a different sign from those arising from direct magnetization transfer from bulk water. Herein we critically evaluate the applicability of solvent accessibility mapping to derive binding orientations for ligands of two dehydrogenases (AKR1C3 and HSD17beta1) with very different binding pockets, including complexes in which the ligand is buried more deeply inside the protein. We also evaluate the possibility of using co-solvents, such as DMSO, for magnetization transfer.     

Oligodeoxynucleotides containing 3-bromo-3-deazaadenine and 7-bromo-7-deazaadenine 2'-deoxynucleosides as chemical probes to investigate DNA-protein interactions

We describe the design and proof of concept of a pair of chemical probes for investigating DNA-protein interactions-specifically, the incorporation of 7-bromo-7-deazaadenine and 3-bromo-3-deazaadenine 2'-deoxynucleosides (Br(7)C(7)dA and Br(3)C(3)dA) into oligodeoxynucleotides (ODNs)-and their utility. Whereas the bromo substituent of the Br(7)C(7)dA unit in an ODN duplex acts sterically to inhibit binding with NF-kappaB, which interacts with the duplex in its major groove, the bromo substituent of the Br(3)C(3)dA unit acts sterically to inhibit binding with RNase H, which interacts with the duplex in its minor groove. In addition, the utilization of ODNs containing 7-deazaadenine and 3-deazaadenine 2'-deoxynucleosides (C(7)dA and C(3)dA), together with the pair of chemical probes, afforded valuable information on the requirement for nitrogen atoms located in either the major or minor grooves. Accordingly, we were able to show the utility of ODNs containing Br(7)C(7)dA, Br(3)C(3)dA, C(7)dA, and C(3)dA for the investigation of DNA-protein interactions.