Chemical phenotype matching between a plant and its insect herbivore

Two potential outcomes of a coevolutionary interaction are an escalating arms race and stable cycling. The general expectation has been that arms races predominate in cases of polygenic inheritance of resistance traits and permanent cycling predominates in cases in which resistance is controlled by major genes. In the interaction between Depressaria pastinacella, the parsnip webworm, and Pastinaca sativa, the wild parsnip, traits for plant resistance to insect herbivory (production of defensive furanocoumarins) as well as traits for herbivore “virulence” (ability to metabolize furanocoumarins) are characterized by continuous heritable variation. Furanocoumarin production in plants and rates of metabolism in insects were compared among four midwestern populations; these traits then were classified into four clusters describing multitrait phenotypes occurring in all or most of the populations. When the frequency of plant phenotypes belonging to each of the clusters is compared with the frequency of the insect phenotypes in each of the clusters across populations, a remarkable degree of frequency matching is revealed in three of the populations. That frequencies of phenotypes vary among populations is consistent with the fact that spatial variation occurs in the temporal cycling of phenotypes; such processes contribute in generating a geographic mosaic in this coevolutionary interaction on the landscape scale. Comparisons of contemporary plant phenotype distributions with phenotypes of herbarium specimens collected 9–125 years ago from across a similar latitudinal gradient, however, suggest that for at least one resistance trait—sphondin concentration—interactions with webworms have led to escalatory change.

Correlated effects of sperm competition and postmating female mortality

Adaptations in one sex may impair fitness in the opposite sex. Experiments with Drosophila melanogaster have shown that seminal fluid from the male accessory gland triggers a series of postmating responses in the female, including increased egg laying rate and lower remating propensity, but that accessory gland proteins also increase female death rate. Here, we tested the relationships among the longevity of females mated to males from 51 chromosome-extracted D. melanogaster lines, male-mating ability, and sperm-competitive ability. We found significant differences in longevity of females mated to males of different genotypes, and all mated females showed a higher death rate than control virgin females shortly after mating. Both the age-independent mortality parameter (the intercept of the female's survival function) and the slope of the mortality rate curve were significantly correlated with the proportion of progeny sired by the first male to mate relative to tester males (sperm-defense ability, P1). No significant correlation was found between the proportion of progeny sired by the second-mating male relative to tester males (sperm-offense ability, P2) and any mortality parameter. Our results support the hypothesis of a tradeoff between defensive sperm-competitive ability of males and life-history parameters of mated females.

Manipulation of the phenolic chemistry of willows by gall-inducing sawflies

The ability to induce galls on plants has evolved independently in many insect orders, but the adaptive significance and evolutionary consequences of gall induction are still largely unknown. We studied these questions by analyzing the concentrations of various plant defense compounds in willow leaves and sawfly galls. We found that the galls are probably nutritionally beneficial for the sawfly larvae, because the concentrations of most defensive phenolics are substantially lower in gall interiors than in leaves. More importantly, changes in chemistry occur in a similar coordinated pattern in all studied willow species, which suggests that the insects control the phenolic biosynthesis in their hosts. The resulting convergence of the chemical properties of the galls both within and between host species indicates that the role of plant chemistry in the evolution of host shifts may be fundamentally less significant in gallers than in other phytophagous insects.

Feeding specialization and host-derived chemical defense in Chrysomeline leaf beetles did not lead to an evolutionary dead end

Combination of molecular phylogenetic analyses of Chrysomelina beetles and chemical data of their defensive secretions indicate that two lineages independently developed, from an ancestral autogenous metabolism, an energetically efficient strategy that made the insect tightly dependent on the chemistry of the host plant. However, a lineage (the interrupta group) escaped this subordination through the development of a yet more derived mixed metabolism potentially compatible with a large number of new host-plant associations. Hence, these analyses on leaf beetles document a mechanism that can explain why high levels of specialization do not necessarily lead to “evolutionary dead ends.”

Three-dimensional reconstruction of the recombinant type 3 ryanodine receptor and localization of its amino terminus

Recombinant type 3 ryanodine receptor (RyR3) has been purified in quantities sufficient for structural characterization by cryoelectron microscopy and three-dimensional (3D) reconstruction. Two cDNAs were prepared and expressed in HEK293 cells, one encoding the wild-type RyR3 and the other encoding RyR3 containing glutathione S-transferase (GST) fused to its amino terminus (GST-RyR3). RyR3 was purified from detergent-solubilized transfected cells by affinity chromatography using 12.6-kDa FK506-binding protein in the form of a GST fusion as the affinity ligand. Purification of GST-RyR3 was achieved by affinity chromatography by using glutathione-Sepharose. Purified recombinant RyR3 and GST-RyR3 proteins exhibited high-affinity [3H]ryanodine binding that was sensitive to activation by Ca2+ and caffeine and to inhibition by Mg2+. 3D reconstructions of both recombinant RyR3 and GST-RyR3 appeared very similar to that of the native RyR3 purified from bovine diaphragm. Comparison of the 3D reconstructions of RyR3 and GST-RyR3 revealed that the GST domains and, hence, the amino termini of the RyR3 subunits are located in the “clamp” structures that form the corners of the square-shaped cytoplasmic region of homotetrameric RyR3. This study describes the 3D reconstruction of a recombinant ryanodine receptor and it demonstrates the potential of this technology for characterizing functional and structural perturbations introduced by site-directed mutagenesis.

Myosin Vb Is Associated with Plasma Membrane Recycling Systems

Myosin Va is associated with discrete vesicle populations in a number of cell types, but little is known of the function of myosin Vb. Yeast two-hybrid screening of a rabbit parietal cell cDNA library with dominant active Rab11a (Rab11aS20V) identified myosin Vb as an interacting protein for Rab11a, a marker for plasma membrane recycling systems. The isolated clone, corresponding to the carboxyl terminal 60 kDa of the myosin Vb tail, interacted with all members of the Rab11 family (Rab11a, Rab11b, and Rab25). GFP-myosin Vb and endogenous myosin Vb immunoreactivity codistributed with Rab11a in HeLa and Madin-Darby canine kidney (MDCK) cells. As with Rab11a in MDCK cells, the myosin Vb immunoreactivity was dispersed with nocodazole treatment and relocated to the apical corners of cells with taxol treatment. A green fluorescent protein (GFP)-myosin Vb tail chimera overexpressed in HeLa cells retarded transferrin recycling and caused accumulation of transferrin and the transferrin receptor in pericentrosomal vesicles. Expression of the myosin Vb tail chimera in polarized MDCK cells stably expressing the polymeric IgA receptor caused accumulation of basolaterally endocytosed polymeric IgA and the polymeric IgA receptor in the pericentrosomal region. The myosin Vb tail had no effects on transferrin trafficking in polarized MDCK cells. The GFP-myosin Va tail did not colocalize with Rab11a and had no effects on recycling system vesicle distribution in either HeLa or MDCK cells. The results indicate myosin Vb is associated with the plasma membrane recycling system in nonpolarized cells and the apical recycling system in polarized cells. The dominant negative effects of the myosin Vb tail chimera indicate that this unconventional myosin is required for transit out of plasma membrane recycling systems.

Effect of interleukin 12 on tumor induction by 3-methylcholanthrene.

Interleukin (IL)-12 has strong antitumor activity in transplantable tumor systems in the mouse. The present study was designed to determine whether tumor induction by 3-methylcholanthrene (3-MC), a carcinogenic hydrocarbon, can be inhibited by IL-12. BALB/cBy mice were injected subcutaneously with 25 micrograms or 100 micrograms of 3-MC and treated with 100 ng, 10 ng, or 1 ng of IL-12 for 5 days a week for 18 weeks, with a schedule of 3 weeks on and 1 week off. In mice injected with 25 micrograms of 3-MC, treatment with 100 ng of IL-12 delayed tumor appearance and reduced tumor incidence. Tumor appearance was also delayed in mice injected with 100 micrograms of 3-MC and treated with 100 ng of IL-12, but the final tumor incidence was the same as in non-IL-12-treated mice. In contrast to the characteristically round, hard, well-circumscribed, and protruding tumor induced by 3-MC, a percentage of tumors induced in IL-12-treated mice had atypical characteristics: flat, soft, and invasive. Atypical tumors had a longer latent period and were more frequently seen in mice injected with 100 micrograms of 3-MC and treated with 100 ng of IL-12. Interferon gamma, IL-10, and tumor necrosis factor could be induced throughout the treatment period by IL-12, indicating that repeated injections of IL-12 do not induce a state of tachyphylaxis. High production of interferon gamma by CD8 T cells and a TH2-->TH1 or TH0 shift in the cytokine secretion profile of CD4 T cells were also seen in the IL-12-treated mice. IL-12 provides a powerful new way to explore the defensive role of the immune system in tumorigenesis.

Primary structure and expression of a pathogen-induced protease (PR-P69) in tomato plants: Similarity of functional domains to subtilisin-like endoproteases.

A 69-kDa proteinase (P69), a member of the pathogenesis-related proteins, is induced and accumulates in tomato (Lycopersicon esculentum) plants as a consequence of pathogen attack. We have used the polymerase chain reaction to identify and clone a cDNA from tomato plants that represent the pathogenesis-related P69 proteinase. The nucleotide sequence analysis revealed that P69 is synthesized in a preproenzyme form, a 745-amino acid polypeptide with a 22-amino acid signal peptide, a 92-amino acid propolypeptide, and a 631-amino acid mature polypeptide. Within the mature region the most salient feature was the presence of domains homologous to the subtilisin serine protease family. The amino acid sequences surrounding Asp-146, His-203, and Ser-532 of P69 are closely related to the catalytic sites (catalytic triad) of the subtilisin-like proteases. Northern blot analysis revealed that the 2.4-kb P69 mRNA accumulates abundantly in leaves and stem tissues from viroid-infected plants, whereas the mRNA levels in tissues from healthy plants were undetectable. Our results indicate that P69, a secreted calcium-activated endopeptidase, is a plant pathogenesis-related subtilisin-like proteinase that may collaborate with other defensive proteins in a general mechanism of active defense against attacking pathogens.

Chemical egg defense in a green lacewing (Ceraeochrysa smithi)

The green lacewing Ceraeochrysa smithi (Neuroptera, Chrysopidae), like other members of its family, lays its eggs on stalks, but it is unusual in that it coats these stalks with droplets of an oily fluid. The liquid consists of a mixture of fatty acids, an ester, and a series of straight-chain aldehydes. Relative to the eggs of a congeneric chrysopid that lacks stalk fluid, the eggs of C. smithi proved well protected against ants. Components of the fluid, in an assay with a cockroach, proved potently irritant. Following emergence from the egg, C. smithi larvae imbibe the stalk fluid, thereby possibly deriving nutritive benefit, defensive advantage, or both.

Evolutionary consequences of food chain length in kelp forest communities.

Kelp forests are strongly influenced by macroinvertebrate grazing on fleshy macroalgae. In the North Pacific Ocean, sea otter predation on macroinvertebrates substantially reduces the intensity of herbivory on macroalgae. Temperate Australasia, in contrast, has no known predator of comparable influence. These ecological and biogeographic patterns led us to predict that (i) the intensity of herbivory should be greater in temperate Australasia than in the North Pacific Ocean; thus (ii) Australasian seaweeds have been under stronger selection to evolve chemical defenses and (iii) Australasian herbivores have been more strongly selected to tolerate these compounds. We tested these predictions first by measuring rates of algal tissue loss to herbivory at several locations in Australasian and North Pacific kelp forests. There were significant differences in grazing rates among sea otter-dominated locations in the North Pacific (0-2% day-1), Australasia (5-7% day-1), and a North Pacific location lacking sea otters (80% day-1). The expectations that chronically high rates of herbivory in Australasia have selected for high concentrations of defensive secondary metabolites (phlorotannins) in brown algae and increased tolerance of these defenses in the herbivores also were supported. Phlorotannin concentrations in kelps and fucoids from Australasia were, on average, 5-6 times higher than those in a comparable suite of North Pacific algae, confirming earlier findings. Furthermore, feeding rates of Australasian herbivores were largely unaffected by phlorotannins, regardless of the compounds' regional source. North Pacific herbivores, in contrast, were consistently deterred by phlorotannins from both Australasia and the North Pacific. These findings suggest that top-level consumers, acting through food chains of various lengths, can strongly influence the ecology and evolution of plantherbivore interactions.

Illusory contours activate specific regions in human visual cortex: evidence from functional magnetic resonance imaging.

The neural basis for perceptual grouping operations in the human visual system, including the processes which generate illusory contours, is fundamental to understanding human vision. We have employed functional magnetic resonance imaging to investigate these processes noninvasively. Images were acquired on a GE Signa 1.5T scanner equipped for echo planar imaging with an in-plane resolution of 1.5 x 1.5 mm and slice thicknesses of 3.0 or 5.0 mm. Visual stimuli included nonaligned inducers (pacmen) that created no perceptual contours, similar inducers at the corners of a Kanizsa square that created illusory contours, and a real square formed by continuous contours. Multiple contiguous axial slices were acquired during baseline, visual stimulation, and poststimulation periods. Activated regions were identified by a multistage statistical analysis of the activation for each volume element sampled and were compared across conditions. Specific brain regions were activated in extrastriate cortex when the illusory contours were perceived but not during conditions when the illusory contours were absent. These unique regions were found primarily in the right hemisphere for all four subjects and demonstrate that specific brain regions are activated during the kind of perceptual grouping operations involved in illusory contour perception.

Induced oleoresin biosynthesis in grand fir as a defense against bark beetles.

Grand fir (Abies grandis) saplings and derived cell cultures are useful systems for studying the regulation of defensive oleoresinosis in conifers, a process involving both the constitutive accumulation of resin (pitch) in specialized secretory structures and the induced production of monoterpene olefins (turpentine) and diterpene resin acids (rosin) by nonspecialized cells at the site of injury. The pathways and enzymes involved in monoterpene and diterpene resin acid biosynthesis are described, as are the coinduction kinetics following stem injury as determined by resin analysis, enzyme activity measurements, and immunoblotting. The effects of seasonal development, light deprivation, and water stress on constitutive and wound-induced oleoresinosis are reported. Future efforts, including a PCR-based cloning strategy, to define signal transduction in the wound response and the resulting gene activation processes are delineated.

Alterações dos estados de ânimo presentes de jovens atletas de futebol em função do decurso temporal durante um período competitivo

São escassos os estudos que analisam o contínuo temporal dos estados de ânimo ao longo de um período competitivo esportivo. Embora os estados de ânimo pareçam estáveis ao longo do tempo, diferentes estímulos e contextos presentes modificam a intensidade e a valência desses estados. Além disso, há fenômenos psicológicos como decaimento, em que traços de informação perdem sua ativação devido, principalmente, à passagem do tempo e a expectativa, que é a espera pela ocorrência de um evento em um determinado tempo. O objetivo desse estudo foi examinar as alterações dos estados de ânimo em jovens atletas de futebol, separados por posição e função, que ocorreram num período competitivo, em função do decurso temporal. Assim, processos como decaimento dos estados de ânimo e a influência da expectativa pela ocorrência jogo foram analisados, bem como a influência do contexto nas variações dos estados de ânimo dos atletas. Participaram deste estudo 18 jovens atletas (média de 15,4 anos ± 0,266) de um clube de futebol que estava disputando um campeonato estadual. Para o acesso aos estados de ânimo, foi utilizada a versão reduzida da Lista de Estados de Ânimo Presentes (LEAP), juntamente com um formulário de instruções de preenchimento, aplicada minutos antes de alguns treinamentos e jogos. Foram calculados os valores de presença de cada Fator da LEAP em cada evento para cada participante. Os dados foram coletados em três tipos de Eventos: antes do último treino antecedente ao jogo (Treino-Pré), antes do jogo (Pré-jogo) e antes do primeiro treino subsequente ao jogo (Treino-Pós). Os 18 jogadores foram divididos em dois grupos: Ações Defensivas (AD) e Ações Ofensivas (AO). Foram encontrados padrões de alteração dos estados de ânimo, representados pelos Fatores II (Fadiga), VII (Interesse) e XII (Serenidade) da LEAP, em função do decurso temporal, permitindo a análise dos processos de decaimento desses estados de ânimo e a influência da expectativa nessas alterações. Também foi encontrado que alguns estados de ânimo diferiram seus padrões de alteração de acordo com um intervalo temporal (Fatores IV Limerência/Empatia e; VII Interesse), bem como tiveram valores de presença diferentes na comparação entre esses intervalos. Além disso, os Fatores III (Esperança), V (Fisiológico) e XI (Receptividade) apresentaram padrões de alteração em função do decurso temporal em diferentes intervalos temporais. Variáveis contextuais, como o resultado das partidas e a competição esportiva em si, também foram influentes nessas alterações. Fadiga, esperança, empatia, estados ligados à propriocepção, interesse, receptividade e serenidade foram os estados de ânimo presentes durante todo o estudo. Ressalta-se a importância de incluir a temporalidade como variável influente nos modelos de variação de processos neurobiológicos, sobretudo nas investigações acerca de aspectos subjetivos como os estados de ânimo.

Estudo semântico-lexical do códice oitocentista Memoria sobre o Plano de Guerra Offensiva e Deffensiva da Capitania de Matto Grosso

Esta tese, com o intuito de contribuir para uma reflexão em torno da história da formação da língua portuguesa no Brasil, propõe como objetivo geral realizar um estudo do léxico no município de Cáceres-MT, tendo como base a discussão sobre manutenção, tendência à manutenção, desuso, tendência ao desuso e neologismo semântico de unidades lexicais extraídas de um manuscrito oitocentista. Os objetivos específicos são os seguintes: (i) compreender a história social da Capitania de Mato Grosso e do município de Cáceres, a partir das informações constantes no manuscrito Memoria, e aspectos que envolvam as condições de produção do documento e a biografia do autor; (ii) levantar o léxico do manuscrito, com recorte nos substantivos e adjetivos para servir de base na seleção das unidades lexicais a serem testadas in loco, e investigar a acepção registrada no documento das unidades lexicais, caracterizando, assim, o léxico do período oitocentista; (iii), fazer um cotejo lexicográfico abrangendo dicionários gerais dos séculos XVIII ao XXI; (iv) testar e identificar, a partir do corpus oral constituído por meio de pesquisa de campo na região urbana cacerense, o grau de manutenção, tendência à manutenção, desuso, tendência ao desuso e neologismo semântico em relação às unidades lexicais e suas respectivas acepções registradas no manuscrito. Dessa forma, toma-se como corpus de língua escrita de análise o manuscrito oitocentista Memoria sobre o plano de guerra offensiva e deffensiva da Capitania de Matto Grosso e, a partir das unidades lexicais selecionadas e extraídas dele, realizou-se a pesquisa de campo para o recolhimento do corpus de língua oral. Antes dessa recolha, tendo como base teórico-metodológica as disciplinas de Dialetologia e de Geolinguística, selecionou-se a localidade (município de Cáceres - MT) e os informantes (total de dezesseis); elaborou-se o questionário semântico-lexical, considerando fundamentalmente a proposta apresentada pelo Comitê Nacional do Projeto ALiB (2001); e realizou-se a pesquisa de campo e as transcrições das entrevistas. Para análise de natureza semântico-lexical dos corpora, recorreu aos estudos lexicográficos e lexicológicos. Tomando por base os resultados do estudo realizado, constatou-se que na realidade linguística do informante cacerense encontram-se unidades que já integravam o léxico oitocentista da língua portuguesa escrita no Brasil, ou seja, há uma memória semântico-lexical que se mantém no sistema lexical, provavelmente, devido às condições sócioculturais do município de Cáceres, Mato Grosso, cuja população, em grande parte, por quase duzentos anos, viveu na área rural. Todavia, vislumbrou-se um certo equilíbrio entre a manutenção do léxico oitocentista sem deixar de lado a inovação e o mecanismo polissêmico constitutivo do léxico.

Role Expectations and State Socialization: Germany's Rediscovery of the Use of Force 1990-1995

This dissertation engages the question of why German political elites accepted the use of force during the 1990s and started to commit the country's armed forces to multilateral peacekeeping missions. Previous governments of the Federal Republic had opposed foreign deployment of the military and Germany was characterized by a unique strategic culture in which the efficacy of military force was widely regarded as negative. The rediscovery of the use of force constituted a significant reorientation of German security policy with potentially profound implications for international relations. I use social role theory to explain Germany's security policy reorientation. I argue that political elites shared a national role conception of their country as a dependable and reliable ally. Role expectations of the international security environment changed as a result of a general shift to multilateral intervention as means to address emerging security problems after the Cold War. Germany's resistance to the use of force was viewed as inappropriate conduct for a power possessing the economic and military wherewithal of the Federal Republic. Elites from allied countries exerted social pressure to have Germany contribute commensurate with capabilities. German political elites adapted role behavior in response to external expectations in an effort to preserve the national role conception of a dependable and reliable ally. Security policy reorientation to maintain Germany's national role conception was pursued by conservative elites who acted as 'role entrepreneurs'. CDU/CSU politicians initiated a process of role adaptation to include the use of force for non-defensive missions. They persuaded Social Democrats and Alliance 90/Green party politicians that the maintenance of the country's role conception necessitated a reorientation in security policy to accommodate the changes in the security environment.