Attraction of the bark beetle Tomicus piniperda to Scots pine trees in relation to tree vigor and attack density

The attraction of Tomicus piniperda (L.) (Coleoptera: Scolytidae) to Scots pine trees of low and higher vigor with and without previous attacks was monitored with sticky traps. Somewhat higher numbers of beetles were caught on low vigor trees than on trees of higher vigor, indicating differences in olfactory stimuly between the two classes of trees.Many more beetles were caught on trees attacked by T. piniperda and on trees with simulated bark beetle galleries (manually drilled holes) than on control trees with no or only a few attacks. The strong attraction of beetles to attacked trees is attributed to the beetles responding to host volatiles released from the galleries.

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Zusammenfassung Der Anflug von Tomicus piniperda (L.) (Coleoptera: Scolytidae) an Kiefern in schlechtem oder besserem Gesundheitszustand, mit oder ohne vorherigen Befall, wurde mit Hilfe von Leimfallen untersucht. An Bäumen in schlechtem Zustand wurden etwas mehr Käfer gefangen als an Bäumen in besserem Gesundheitszustand. Das deutet auf Unterschiede in Geruchsreizen zwischen den beiden Klassen des Baumzustands hin.An Kiefern, die von T. piniperda befallen waren oder simulierte Einbohrungen (von Hand gebohrte Löcher) hatten, wurden viel mehr Käfer gefangen als an Kontrollbäumen ohne oder mit sehr geringem Befall. Der starke Anflug von Käfern an befallene Bäume wird gedeutet als Reaktion auf Duftstoffe des Wirts, die aus den Borkenkäfergängen abgegeben werden.

     

Monoterpenes governing host selection in the bark beetles Hylurgops palliatus and Tomicus piniperda

Under field conditions, both bark beetles, Hylurgops palliatus Gyll. and Tomicus piniperda L. (Col., Scolytidae) preferred flight barrier traps baited with ethanol and oleoresin of their favoured host, Picea abies L., and Pinus sylvestris L., respectively, compared to traps baited with ethanol and various monoterpenes. However, replacing the host-specific oleoresin with -pinene and the pine-characteristic terpinolene enhanced trap catches of both species. Response of H. palliatus increased with myrtenol, but decreased with (+)-trans-verbenol, while T. piniperda was attracted by both terpene alcohols.

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Zusammenfassung Die Lockwirkung wirtsspezifischer Terpenmuster auf die sekundären Rindenbrüter Hylurgops palliatus und Tomicus piniperda wird im Freiland untersucht. Beide Arten fliegen bevorzugt Fallen an, die mit Ethanol und dem Rohharz ihrer Wirtsbäume, Picea abies bzw. Pinus sylvestris, beködert sind. Der Ersatz der Rohharze durch eine Mischung aus -Pinen und dem für Kiefernharz charakteristischen Terpinolen erhöht die Fangzahlen beider Arten deutlich. Die Präferenzen für Monoterpene deuten unterschiedlich enge Wirtsbindungen an: Bei breitem Wirtsspektrum bevorzugt H. palliatus wenig spezifische Terpene und hohe Ethanol-Konzentrationen, der zur Monophagie an Frischholz neigende Waldgärtner T. piniperda jedoch geringe Ethanol-Konzentrationen und spezifische Terpenkombinationen. H. palliatus wird von dem Terpenalkohol (-)-Myrtenol deutlich angelockt, während das von aggressiven Scolytiden bekannte Pheromon (+)-trans-Verbenol seinen Anflug im Gegensatz zu T. piniperda zu hemmen scheint.

     

Kairomonal responses of Tomicus destruens (Col., Scolytidae) to host volatiles α-pinene and ethanol

The Mediterranean pine shoot beetle Tomicus destruens is one of the most damaging bark beetles attacking Mediterranean pine forests in southern Europe and north Africa. We studied the attractiveness of the host volatiles α-pinene and ethanol at a range of release rates, alone or in combination, to T. destruens , in order to develop an attractive lure for the management of this beetle. T. destruens was attracted slightly to the host volatile α-pinene, but a strong synergistic effect was found in the attraction towards monoterpene when ethanol was added to the bait. The highest catches of T. destruens were obtained by the optimal blend releasing 300 mg/day of α-pinene and 900 mg/day of ethanol. In contrast to data reported for the related species T. piniperda , Mediterranean pine shoot beetles were clearly attracted to baits releasing ethanol alone (1350 mg/day). trans -Verbenol, which was also added to host volatiles in some tests, did not affect the response. The use of the attractive blend proposed would have a low impact on the natural enemy population of Thanasimus formicarius because of asynchronies in flight periods. Other non-target insects, such as the facultative predator or competitor Oxipleurus nodieri , were also significantly attracted. These results allow the development of an operative lure for the effective monitoring of T. destruens , although improvements by the addition of other host volatiles should be studied.     

Bruchid resistance of transgenic azuki bean expressing seed α-amylase inhibitor of common bean

Various species of bruchid beetles including Callosobruchus chinensis, C. maculatus and C. analis cause postharvest damage of azuki bean seeds, an important East Asian grain legume. The -amylase in the midguts of these insects is inhibited by the -amylase inhibitor (AI) present in common bean seeds. Transformation of azuki bean with the AI gene driven by the promoter of phytohemagglutinin results in high levels of AI in the seeds and the complete block of bruchid development on the seeds. Zabrotes subfasciatus, a South and Central American bruchid that is a storage pest of common bean, develops normally on the transgenic azuki bean.     

Behavioral responses of a leaf beetle to injury-related changes in its salicaceous host

Summary Adults of the leaf beetle Plagiodera versicolora Laich, avoided previously injured shoots of Salix alba Tristis in favor of nearby uninjured ones. The response was rapid and persisted for five days of study. During brief periods of observation, the vast majority of beetles engaged in behaviors other than locomotion such as feeding and resting. Furthermore, the proportions of beetles walking were similar on injured and uninjured shoots. It appears that under these experimental conditions previous injury did not greatly increase searching behavior of the beetles. Leaves on previously injured shoots received less subsequent herbivory than uninjured ones. Findings of these experiments are consistent with patterns of herbivory observed in the field and known reproductive responses of beetles to diets of injured leaves. These results provide new information toward understanding relatively low levels of herbivory on individual willow leaves associated with the feeding of P. versicolora.     

Short communication: Thermostability of α-amylase produced by Bacillus sp. E2—a thermophilic mutant

An -amylase from a hyper-producing strain of Bacillus (sp. E2) was stable at 70°C for 30 min but was quickly inactivated at higher temperatures. In the presence of 10mm Ca²⁺ and starch (20% w/v), however, the enzyme was stable at 90°C for 10 min and after 30 min at 100°C still retained 26% of its initial activity.     

Properties of α-galactosidase II2 from Vicia faba seeds

-Galactosidase II² (MW 43 390) from resting Vicia faba L. seeds had been shown to possess d-glucose/d-mannose-specific lectin activity. Inhibition studies with monosaccharides and an examination of the effects of heat and pH on the catalytic and lectin activities of the enzyme indicate that the enzyme substrate and the lectin haptens bind at different sites on the protein. d-Mannosebinding has been investigated by equilibrium dialysis and spectrophotometrically. Both methods yield K_a values of approx. 3·10³ M^-1 for the interaction and there would appear to be two mannosebinding sites per molecule of enzyme protein. The lectin properties of V. faba -galactosidase II² have been discussed in relation to both V. faba lectin (favin) and other legume -galactosidases.Abbreviations con A concanavalin A - CM-cellulose carboxymethyl cellulose - MW molecular weight - PNPG p-nitrophenyl -d-galactoside - SDS sodium dodecyl sulphate - PAGE polyacrylamide-gel electrophoresis     

Protein export elements from Lactococcus lactis

Summary Broad-host-range plasmids carrying -amylase or -lactamase reporter genes lacking a signal sequence were used to select export elements from Lactococcus lactis chromosomal DNA that could function as signal sequences. Fragments containing such elements were identified by their ability to direct the export of the reporter proteins in Escherichia coli. Several of the selected export elements were also active in Bacillus subtilis and L. lactis, although the efficiencies depended strongly on the host organism and reporter gene used. The export elements AL9 and BL1 were highly efficient in L. lactis in the expression and secretion of at least two heterologous proteins (Bacillus licheniformis -amylase and E. coli TEM--lactamase). AL9 even permitted growth of this organism on starch as the sole carbon source. Nucleotide sequence analysis of five selected fragments indicated that these encode oligopeptides with the major characteristics of typical signal peptides. The putative expression signals had a limited similarity to previously described expression signals for E. coli, B. subtilis and L. lactis. Differences in both expression and export efficiency are likely to underlie the host-specific effects.     

Identification of a locality in snake venomα-neurotoxins with a singificant compositional similarity to marine snailα-conotoxins: Implications for evolution and structure/activity

Summary -neurotoxins from elapid snake venoms and-conotoxins from marine snails bind specifically and with high affinity to nicotinic cholinoceptors. Although both types of toxin are polypeptides, there is more than a fourfold difference in size between the two and no clear sequence homology is evident. A systematic computer search of the three-dimensional structure of erabutoxin b (an-neurotoxin from the false sea snakeLaticauda semifasciata) was performed to identify the locality that most closely matched the amino acid compositions of the smaller-conotoxins (from the marine snailsConus magus andConus geographus). The area of greatest similarity centered on residue position 25 of erabutoxin b, a locale that is conserved throughout the snake-neurotoxins and their homologues. Six Proteins unrelated to erabutoxin b were compared to the-conotoxins to show that the extent of the erabutoxin b/-conotoxin match was too high to be coincidental. Homologues of erabutoxin b, namely-cobratoxin fromNaja naja siamensis and cytotoxin V^II4 fromNaja mossambica mossambica, were also analyzed. The extent of the matching with the-conotoxins decreased in the series erabutoxin b>-cobratoxin>cytotoxin V^II4, and this also relates the order of similarity to the pharmacological properties of the-conotoxins.The-conotoxin-like area of the snake-neurotoxins is peripheral to the site previously considered important for binding to the cholinoceptor, even though it seems to represent the focus of evolutionary convergence between the two types of neurotoxin. The area of resemblance does, however, have strong associations with the conformational behavior of the snake toxins. Hence, the outcome of this study has important consequences for the current ideas on snake-neurotoxin structure/activity relationships and the evolutionary origins of neurotoxicity.     

Influence of Medium and Long Range Interactions in (α/β)8 Barrel Proteins

The residue-residue contacts and the role of medium and long rangeinteractions in 36 (/)₈ barrel proteins have beenanalysed. The influence of long range contacts in the formation ofphysico-chemically similar clusters, and the preference of amino acidresidues towards long range contacts have also been studied. Theresults reveal a nearly uniform level of medium and long rangecontacts in most of the proteins. The residues Gln and Ala havehighest medium range contacts and the residue Pro has the lowestmedium range contacts. The residue Cys has the highest long rangecontact followed by other hydrophobic residues namely Val, Ile andLeu. In the physico-chemically similar clusters identified in theseproteins, 25–40 percent residues are influenced by long rangecontacts, and the residues Cys, Ile, Val and Met are the mostpreferred ones.     

Effect of inhibitors of gibberellin biosynthesis on the induction of α-amylase in embryoless caryopses of Hordeum vulgare cv. Himalaya

The induction of -amylase by exogenously supplied gibberellin A₁ (GA₁) and GA₄ in embryoless caryopses of Hordeum vulgare (cv. Himalaya) was determined indirectly by measuring reducing sugars released from the endosperm. The presence of the inhibitors of GA biosynthesis, 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride (Amo 1618), Ancymidol, 2-chloroethyl trimethyl ammonium chloride (CCC) or (R,S)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,3-triazolyl)pentan-3-ol (PP333) did not inhibit -amylase production by either GA₁ or GA₄.Abbreviations Amo-1618 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride - CCC 2-chloroethyl trimethyl ammonium chloride - cv. cultivar - GA gibberellin - GC gas chromatography - GC-MS combined gas chromatography-mass spectrometry - PP333 (R,S)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,3-triazolyl) pentan-3-01     

Production of α-amylase by Bacillus amyloliquefaciens in batch and continuous culture using a defined synthetic medium

Summary Using a totally defined synthetic medium the effect of lactose and nitrogen on cell physiology and -amylase production by Bacillus amyloliquefaciens B155 were investigated. Results showed cell growth and -amylase production patterns to be similar regardless of the limiting nutrient and suggested stationary phase gene control of -amylase production as opposed to a direct response to nutrient limitation.     

Novel replication mutant of microvirid phage α3 deleted in the complementary strand origin

Summary The bacteriophage 3 origin of complementary strand DNA synthesis (—ori) contains two potential secondary loop structures (I and II), which have been implicated as direct recognition sites for host Escherichia coli DnaG protein. To elucidate to what extent such structures are essential, we introduced a nucleotide deletion within the —ori region, by nuclease digestion of 3 replicative form DNA. A mutant, delB, thus constructed had a 121 nucleotide deletion within the —ori region and was completely lacking in the two putative hairpin loops, I and II. The delB mutant formed smaller plaques on the host E. coli C and had a longer latent period, but the mean burst size at 37° C was almost the same (400 phages) as that of the wild type. In contrast to the parental phage, growth of the mutant depends on host dnaB and dnaC functions. These results indicate that the prototype secondary structures in the 3 origin of complementary strand synthesis are dispensable for delB and that the 3 mutant has an additional replication origin whose function is dependent on DnaB and DnaC proteins, rather than on DnaG protein alone.     

Action spectra and functional antenna sizes of Photosystems I and II in relation to the thylakoid membrane organization and pigment composition

The functional organization of competent photosynthetic units in developing thylakoids from intermittent-light grown pea as well as in the unstacked, stacked and phosphorylated stacked thylakoids from its mature chloroplasts was characterized by polarographic measurements of action spectra, reaction centre contents and optical cross-sections for PS I-mediated O2 uptake and PS II-mediated O2 evolution. The minimum antenna sizes of 60 and 37 chlorophyll a molecules for PS I and PS II, respectively, were determined in developing thylakoids with a ratio of Chl a/Chl b>50. In mature chloroplasts, the embedded light-harvesting chlorophyll a/b-binding (LHC) protein complexes increased the PS I and PS II effective antenna sizes by 3–6 times depending on the thylakoid membrane organization. In unstacked thylakoids, a randomization of PS I, PS II and LHC II led to the most uniform spectral distribution of light harvesting between the two photosystems but caused the maximal difference of their antenna sizes to be 370 and 100 Chls for the competent PS I and PS II units, respectively. Following the Mg2+-induced stacking of thylakoids, opposite complementary changes of the action spectra, antenna sizes and Chl a/Chl b ratios indicated a redistribution of a LHC II pool of 100 Chl ( a + b) molecules from PS I to PS II. Unlike to the stroma-exposed PS II in unstacked thylakoids, the granal PS II units of 200 Chls demonstrated an additional 2-fold increase of the effective antenna size due to energy transfer within PS II dimers under strong background illumination, which closed >90% of reaction centres. Protein phosphorylation of the stacked thylakoids induced a significant inactivation of the O2-evolving PS II centres but did not cause complementary changes of the action spectra and antenna sizes of the competent PS I and PS II. In this case, light harvesting parameters of the O2-evolving PS II units were nearly unaffected, whereas the obvious relative increase of the PS I activity at 650 nm and its decrease at >700 nm both in the action spectrum and optical cross-section measurements might suggest a substitution of PS I units in the O2-reducing fraction by another distinct fraction of -type which in turn is not the same to PS I units in unstacked thylakoids.     

Long-range physical mapping of the alpha-amylase-1 (alpha-Amy-1) loci on homoeologous group 6 chromosomes of wheat

Summary Long-range physical maps of the small multigene family of the malt -amylase genes (-Amy-1) located on the long arms of wheat chromosomes 6A (the -Amy-A1 locus) and 6B (-Amy-B1) were generated by pulsed-field gel electrophoresis analysis. By using three methylation-sensitive rare-cutter restriction endonucleases, NotI, NruI and MluI, and an -Amy-1 cDNA probe and four gene-specific genomic probes from the -Amy-B1 locus, the size of the -Amy-B1 locus was estimated to be about 700 kb and of the -Amy-B1 locus to be about approximately 4300 kb. These two maps indicate clustering of GC-rich and C-methylation-sensitive restriction enzyme recognition sites. At least five regions reminiscent of CpG islands are apparent in -Amy-B1, and three in -Amy-A1. Correlation between recombination frequency and physical distance within the -Amy-B1 locus suggests that 1 cM approximates to 1 Mb in physical distance.     

Using Pair-Coupled Amino Acid Composition to Predict Protein Secondary Structure Content

The pair-coupled amino acid composition is introduced to predict the secondary structure contents of a protein. Compared with the existing methods all based on singlewise amino acid composition as defined in a 20D (dimensional) space, this represents a step forward to the consideration of the sequence coupling effect. The test results indicate that the introduction of the pair-coupled amino acid composition can significantly improve the prediction quality. It is anticipated that the concept of the pair-coupled amino acid composition can be used to simplify the formulation of sequence coupling (or sequence order) effects and to study many other features of proteins as well.     

On the use of phase partition to isolate vacuoles from cultured cells

Vacuoles isolated from cultured rose-cell protoplasts by controlled lysis were subjected to partition in polyethylene glycol-dextran two-phase systems. The vacuoles showed a strong affinity for the polyethylene glycol phase, like plasma membrane and unlike other membraneous organelles. After phase partition at 4°C, dilution of the polyethylene glycol destroyed a large proportion of the vacuoles. After phase partition at room temperature (ca. 20°C), the vacuoles in the polyethylene glycol phase were relatively stable to dilution. Two-phase partition may be useful for purification of intact vacuoles and possibly of tonoplast membranes. Minor proportions of acid phosphatase and -mannosidase were associated with these vacuoles.