Rhizobium leguminosarum genes required for expression and transfer of host specific nodulation

The contributions of various nod genes from Rhizobium leguminosarum biovar viceae to host-specific nodulation have been assessed by transferring specific genes and groups of genes to R. leguminosarum bv. trifolii and testing the levels of nodulation on Pisum sativum (peas) and Vicia hirsuta. Many of the nod genes are important in determination of host-specificity; the nodE gene plays a key (but not essential) role and the efficiency of transfer of host specific nodulation increased with additional genes such that nodFE < nodFEL < nodFELMN. In addition the nodD gene was shown to play an important role in host-specific nodulation of peas and Vicia whilst other genes in the nodABCIJ gene region also appeared to be important. In a reciprocal series of experiments involving nod genes cloned from R. leguminosarum bv. trifolii it was found that the nodD gene enabled bv. viciae to nodulate Trifolium pratense (red clover) but the nodFEL gene region did not. The bv. trifolii nodD or nodFEL genes did significantly increase nodulation of Trifolium subterraneum (sub-clover) by R. leguminosarum bv. viciae. It is concluded that host specificity determinants are encoded by several different nod genes.     

Induction of hepatic drug metabolizing enzymes by DL-methionine in rats

A single intraperitoneal injection of DL-methionine (500 mg/kg body wt.) to adult male Wistar rats was shown to significantly induce all the components of the hepatic microsomal mixed function oxidase system such as NADPH cytochrome C reductase activity, cytochromes P-450 and b₅, as well as activities of drug metabolizing enzymes such as aminopyrine demethylase and uridine 5′ -diphosphate-glucuronosyltransferase. Combined administration of nicotinamide (250 mg/kg body wt.) and DL-methionine (500 mg/kg body wt.) was shown to bring about an additional increase (25-30%) in the activities of these enzymes as compared to their induction on independent administration of the two endobiotics. In rats bearing Yoshida sarcoma (ascites) tumour as well as in normal rats injected with serum from tumour bearing animals, the decreased activities of hepatic mixed function oxidases could be restored to their normal levels by administration of DL-methionine (500 mg/kg body wt.) to these rats. Whereas actinomycin D (1 mg/kg body wt.) had no effect on the increased incorporation of [¹⁴C] labelled leucine into microsomal proteins following administration of nicotinamide, the enhanced incorporation of the label following DL-methionine administration was completely inhibited by the same dose of actinomycin D. Administration of cycloheximide (0·5 mg/kg body wt.) to rats could completely inhibit the increased incorporation of [¹⁴C] leucine into hepatic microsomal proteins following independent administration of nicotinamide and DL-methionine. Similar inhibitory pattern with actinomycin D and cycloheximide was also demonstrated in case of induction of NADPH cytochromeC reductase activity by both these endobiotics.     

Host plant recognition in monophagous weevils: specificity in feeding responses of Ceutorhynchus constrictus and the variable effect of sinigrin

Host plant relations of the monophagous weevil Ceutorhynchus constrictus Marsh. (Coleoptera: Curculionidae: Ceutorhynchinae) feeding on garlic mustard, Alliaria petiolata (Bieb.) Cavara & Grande (Cruciferae) were studied in the laboratory. Most other crucifers were rejected in choice tests using garlic mustard as a reference plant, but Brassica nigra, Sinapis alba and Thlaspi arvense were as acceptable as the host plant. Flowering plants of Descurainia sophia were acceptable while young plants of this species were not. The most important feeding stimulants in extracts of garlic mustard were uncharged, water soluble compounds. The most abundant glucosinolate in garlic mustard, sinigrin, was a feeding stimulant, too. However, the feeding stimulatory activity of sinigrin was only expressed in the presence of still unidentified uncharged compounds from garlic mustard leaves. Host plant relations in monophagous crucifer-feeding insects is discussed in relation to the distinctness of glucosinolate patterns found in their host plants.

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Zusammenfassung Ceutorhynchus constrictus Marsh. (Coleoptera: Chrysomelidae: Ceutorhynchinae) ist ein monophager Rüsselkäfer, der an Knoblauchhederich frisst. Das Wirtswahl-Verhalten dieses Käfers ist im Labor untersucht worden. Die meisten Crucifiren waren im Wahlversuche nicht akzeptiert, wenn Knoblauchhederich als Vergleichspflanze vorhanden war. Von Brassica nigra, Sinapis alba, und Thlaspi arvense wurden im Vergleich gleiche Mengen verzehrt wie von der Wirtspflanze. Blühende Descurainia sophia Pflanzen wurden, im Gegensatz zu Jungpflanzen der gleichen Art, angenommen. Die wichtichsten Phagostimulanten in Extrakten von Knoblauchhederich-Blättern waren ungeladene, wasserlösliche Substanzen. Das häufigste Glukosinolat im Knoblauchhederich, Sinigrin, war auch ein Phagostimulant. Doch war die phagostimulierende Wirkung von Sinigrin nur in Kombinationen mit noch nicht identifizierten, ungeladenen Substanzen aus Knoblauchhederich-Blätter nachweisbar. Wirtspfanzen-Beziehungen von monophagen Insekten werden diskutiert im Zusammenhang mit der Eigenart des Glukosinolat-Inhaltes ihrer Wirtspflanzen.

     

Performance of nymphal and adult Nezara viridula on selected alternate host plants

Laboratory studies with the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), feeding on seeds or fruits of some alternate host plants, demonstrated differences in nymphal survival and development time, and body weight at adult emergence. Lowest mortalities (25%) were observed for nymphs on fruits of dakota mustard [Brassica kaber (DC.)] and siberian motherwort (Leonurus sibiricus L.). On fruits of castor bean (Ricinus communis L.) nymph mortality was ca. 60%, and on mature seeds of L. sibiricus ca. 75%. Total development time from second instar to adult ranged from ca. 26 days on B. kaber to ca. 42 days on R. communis. Fresh body weight at adult emergence was greatest for nymphs on L. sibiricus and lowest on R. communis. Adult survival after 30 days and mean longevity was greater on L. sibiricus than on the remaining foods, being drastically reduced on R. communis. In general, reproductive performance on all foods was similar, except R. communis, on which no female reproduced. Female weight gain of N. viridula was generally greater on L. sibiricus On R. communis, adults lost weight during their first week and showed the lowest weight gain at the end of day 29. Field observations coupled with these laboratory studies suggest that in the Warta area (northern Paraná state, Brazil, 23° 11 S), L. sibiricus and B. kaber are important alternate food plants in the seasonal phenology of N. viridula. R. communis, however, is of poor nutritional quality, serving primarily as a refuge.

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Résumé L'alimentation au laboratoire de N. viridula avec les graines et les fruits de diverses plantes a entraîné différentes mortalités larvaires, différentes vitesses de développement et des différences dans les poids des adultes à l'émergence. Les mortalités les plus faibles (25%) on été observées avec les fruits de Brassica kaber DC. et de Leonurus sibiricus L.Avec les fruits de Ricinus communis L., la mortalité larvaire était de 60%; sur graines mûres de L. sibiricus, elle était de 75%. La durée du développement du second stade à l'adulte a varié de 26 jours sur B. kaber à 42 jours sur R. communis. Le poids frais des adultes à l'émergence était le plus élevé avec L. sibiricus et le plus faible avec de R. communis. La longévité des adultes au-delà de 30 jours était la plus élevée avec L. sibiricus, et était fortement réduite avec R. communis. Dans l'ensemble, la reproduction était de même importance avec les différents aliments, à l'exception de R. communis avec lequel les femelles ne se reproduisaient pas. Le gain de poids des femelles était généralement supérieur avec L. sibiricus; avec R. communis les adultes ont perdu du poids pendant la première semaine et ont présenté le plus faible gain à la fin du 29ème jour.Des observations dans la nature, associées à ces expériences de laboratoire, laissent supposer que dans la zone de Warta (au Nord de l'état de Paraná, 23° 11 S, au Bresil), L. sibiricus et B. kaber sont des plantes hôtes alternatives importantes dans la phénologie saisonnière de N. viridula. Cependant, R. communis, de faible valeur alimentaire, sert avant tout de refuge.

     

Controlling fungi that colonize eggs of the western corn rootworm in the laboratory

Females of the polyphagous, ectoparasitoid, Exeristes roborator (F.) (Hymenoptera: Ichneumonidae), learned an olfactory stimulus associated with an artificial host microhabitat in the laboratory. In a two-choice olfactometer, females previously given hosts only in association with one stimulus showed a greater tendency to visit exclusively, and spend time in, chambers containing a source of that stimulus, than control females. Learning of olfactory cues could act in conjunction with learning of visual cues to allow the parasitoid to identify accurately microhabitats that experience has taught it contain hosts. It could also allow the parasitoid to identify these resources when visual cues are not available.

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Résumé Nous avons supposé que les femelles polyphages de l'ichneumonide ectoparasite, E. roborator (F.), pourraient apprendre les stimuli olfactifs liés au microhabitat artificiel dans lequel elles avaient attaqué leur hôte. 4 lots de femelles ont été placés dans des conditions différentes: le microhabitat des hôtes du premier lot était du Parafilm imprégné d'odeur de pomme; celui des hôtes du second était un Parafilm non traité. Les hôtes des lots témoins 3 et 4 étaient présentés respectivement avec les deux microhabitats et sans aucun microhabitat. Placées dans un olfactomètre statique contenant du Parafilm traité à la pomme et du Parafilm vierge, des femelles du lot 1 ont eu tendance à ne visiter que l'enceinte contenant du Parafilm traité à la pomme, elles ont passé environ 75% de leur temps dans cette enceinte. A l'opposé, les femelles des 3 autres lots ont eu tendance à ne visiter que l'enceinte sans odeur de pomme, et elles n'ont passé que 34 à 46% de leur temps dans l'enceinte traitée à la pomme. Les différences entre les réactions du lot 1 et des autres lots sont significatives, mais entre les lots 2, 3 et 4, elles ne le sont pas. Une expérience témoin confirme que les préférences des femelles du lot 1 ne sont pas innées; E. roborator a donc appris à reconnaitre les stimuli olfactifs du microhabitat sentant la pomme. L'apprentissage des signaux olfactifs liés au microhabitat de l'hôte pourrait s'effectuer seul ou en relation avec l'apprentissage de signaux visuels permettant au parasitoïde d'identifier avec précision les microhabitats d'un hôte convenable.

     

Temporal changes in the egglaying behaviour of the Hessian fly

Responses of mated female Hessian flies were investigated by analysing the behaviour of individual flies in wheat and oats. The behavioural repertoire of such females included: flying, alighting on leaves, arching of the body so that the tip of the abdomen touched the leaf surface, antennation, movements of the tip of the abdomen across the leaf at right angles to leaf veins, sitting with the ovipositor straight but still extended, and sitting with the ovipositor telescoped into the body. After alighting, females on wheat showed a higher frequency of transitions from arching to antennation and a lower frequency of transitions from arching to abdomen straight than females on oats. During the first 5 min of observations, individuals released into arenas with wheat arched and antennated 2–3 times more frequently than females released into oats. Time allotted to behaviours also differed; during the first 5 min, females in wheat spent 50 percent more time arching, whereas females in oats spent 50 percent more time sitting. Females in wheat also stayed longer and laid 4 times more eggs than females in oats. Temporal changes in egglaying were monitored by quantifying hourly rates of egglaying in no-choice assays for several hours following mating at 9:00 am. During the first and second hours post-mating, egglaying occurred infrequently. However, during the third hour post-mating (11:00 am to noon) females on wheat laid 5 times more eggs than females on oats. Rates of egglaying decreased on wheat but increased on oats during the fourth hour, and then during the fifth hour, decreased on both wheat and oats. Changes in egglaying responses were also evident when behaviours of individual females were measured 1–3 h vs. 3–7 h post-mating. Females deprived of host plants and released into wheat or oats later in the day showed higher frequencies of arching and antennation and laid more eggs before leaving the arena.

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Résumé Les réactions de femelles préalablement accouplées de Mouches de Hesse ont été examinées en analysant le comportement de femelles isolées sur blé et sur avoine. Le répertoire comportemental de ces femelles comprenait: le vol, l'atterrissage sur feuille, la flexion du corps de sorte que l'extrémité de l'abdomen touchât la surface de la feuille, l'antennation, les mouvements de l'extrémité de l'abdomen sur la feuille à angle droit des nervures, le repos avec la tarière droite et encore dévaginée, le repos avec la tarière télescopée à l'intérieur du corps. Sur blé plus que sur avoine, les femelles après atterrissage ont présenté une fréquence plus élevée de passage de la flexion à l'antennation que de la flexion à l'abdomen droit. Durant les 5 premières minutes d'observation, les individus libérés dans des enceintes avec blé fléchirent et antennèrent 2 à 3 fois plus que ceux libérés sur avoine. Les durées des différentes séquences différaient aussi: sur blé, pendant les 5 premières minutes, les femelles passèrent plus de 50% du temps à fléchir, tandis que sur avoine elles passèrent plus de 50% du temps en repos. Les femelles restèrent aussi plus longtemps sur les feuilles de blé et y pondirent 4 fois plus d'oeufs que sur avoine.Les femelles de M. destructor ont montré une plasticité du seuil d'acceptation. Pendant les premières heures de ponte, elles ont été très sélectives et refusèrent, ou ne pondirent que quelques oeufs sur avoine, mais acceptèrent volontiers le blé. La discrimination s'est poursuivie tant que les femelles ont eu accès au blé en même temps qu'à l'avoine. Cependant, quand les femelles ont été privées de blé pendant plusieures heures, l'acceptation de l'avoine a augmenté. Cet accroissement de l'acceptation a eu lieu à peu près au moment où les femelles sur blé pondaient leurs derniers oeufs.

     

Trauma, degenerative disease, and other pathologies among the Gombe chimpanzees

The well-known and extremely well-documented chimpanzees from Gombe National Park were analyzed for presence of skeletal pathologies. Of the 15 animals available for study, 11 were old and complete enough to permit systematic analysis. Of these, 10 showed some evidence of skeletal pathological involvement. The most common type of lesion seen resulted from trauma. Those chimps with the most fractures (Old Female, 3; Flo, 4; Hugo, 8) are consistently the oldest individuals in the sample. In addition to accidental falls, the most common cause of trauma was from interpersonal violence, resulting in bite wounds (see in two individuals) and fractures (see in three individuals). Conversely to trauma, degenerative disease was exceedingly rare in this population, found in no large intervertebral joints (N = 344) and only two major synovial joints (N = 186). In fact, the complete lack of osteophytosis, even in older individuals, stands in stark contrast to the situation seen in modern humans, perhaps in our species reflecting a biomechanical cost of bipedality.     

Microgeographic distribution of immature Ixodes dammini ticks correlated with that of deer

In order to determine whether the small-scale distribution of immature Ixodes dammini Spielman et al. corresponds closely to the activity patterns of white-tailed deer, Odocoileus virginianus (Zimmerman), these relationships were examined in a site on Long Island, New York, U.S.A. We first determined the extent and temporal pattern of adult ticks feeding on deer by examining twenty-three resident deer tranquilized during September-December 1985. I. dammini adults infested deer throughout this fall period, most abundantly during October and November. With radio-telemetry collars attached to deer we determined the relative frequency that they occupied 0.25 ha quadrats of the study site. During the following summer, we examined white-footed mice, Peromyscus leucopus (Rafinesque), that inhabited these quadrats and removed immature ticks from each. 8975 larval and 163 nymphal I. dammini were removed from 208 mice trapped in forty-three such quadrats. The frequency of deer using these quadrats was positively correlated with both the number of larval and of nymphal ticks per mouse. These results suggest that risk of I. damminiborne zoonotic disease may be decreased by locally reducing deer density in sites that experience intense human activity.     

Deficient glycosylation of arylsulfatase A in pseudo arylsulfatase-A deficiency

Summary Deficient arylsulfatase-A activity is diagnostic of a neurodegenerative human lysosomal storage disease, metachromatic leukodystrophy. Paradoxically, similar enzyme deficiency also occurs in normal individuals, who are known as being pseudo arylsulfatase-A deficient. We showed previously that this phenotype is associated with a structural gene mutation that produces an exceptionally labile enzyme. We now report on the nature and consequence of this mutation. When the mutant arylsulfatase-A is deglycosylated by endoglycosidase H, only one smaller molecular species was generated, instead of the two from the normal enzyme. This is consistent with the loss of one of the two N-linked oligosaccharide side chains known to be present on the wild-type enzyme. Quantitative analysis of mannose and leucine incorporation showed that the mutant enzyme incorporated two- to tenfold less mannose than the normal enzyme on a molar basis. This deficient glycosylation was specific to arylsulfatase-A. Another lysosomal enzyme not affected in this mutation, beta-hexosaminidase, was glycosylated normally in the mutant cells. The remaining single oligosaccharide side chain released from the mutant arylsulfatase-A by pronase digestion was normally processed to complex and high-mannose forms. However, the high-mannose side chains contained 30% fewer phosphorylated residues than those of the normal enzyme. Nevertheless, this reduced level of phosphorylation did not prevent targeting of the mutant enzyme to the lysosomes, a process normally mediated through phosphorylated mannose residues. In conclusion, pseudo arylsulfatase-A deficiency is a unique human mutation associated with reduced glycosylation and phosphorylation of a lysosomal enzyme with the loss of one of the two carbohydrate side chains. The mutation results in greatly reduced enzyme stability, thus indicating a role for oligosaccharides in maintaining enzyme stability within the degradative environment of the lysosomes. However, the residual catalytic activity or subcellular targeting of the mutant enzyme was not affected. These properties probably account for the benign clinical presentation of pseudo arylsulfatase-A deficiency.Abbreviations PD Pseudo arylsulfatase-A Deficiency - ARA Arylsulfatase-A     

The morphology of allograft rejection in Styela plicata (Urochordata: Ascidiacea)

Summary This study identifies three discrete processes responsible for the rejection of tunic tissue transplanted between individuals of the solitary ascidian Styela plicata. The first stage of rejection is characterized by the destruction of blood vascular components within incompatible allografts. In the second phase, dense boundaries of extracellular material are deposited between grafts and the surrounding host tunic, effectively amputating the transplanted tissues. Finally, detached transplants undergo a gradual necrosis which results in the total degeneration of extracellular graft matrices. Of these three phases, the initial cellular depletion of allografts is responsible for the immunological specificity that is characteristic of histocompatibility in S. plicata. The subsequent amputation and necrosis of extracellular graft matrices are taken to be non-specific consequences of the initial cellular reactivity.