Specific domains of plant defensins differentially disrupt colony initiation,cell fusion and calcium homeostasis in Neurospora crassa

MsDef1 and MtDef4 from Medicago spp. are small cysteine‐rich defensins with potent antifungal activity against a broad range of filamentous fungi. Each defensin has a hallmark γ‐core motif (GXCX_3–9C), which contains major determinants of its antifungal activity. In this study, the antifungal activities of MsDef1, MtDef4, and peptides derived from their γ‐core motifs, were characterized during colony initiation in the fungal model, Neurospora crassa. These defensins and their cognate peptides inhibited conidial germination and accompanying cell fusion with different potencies. The inhibitory effects of MsDef1 were strongly mediated by the plasma membrane localized sphingolipid glucosylceramide. Cell fusion was selectively inhibited by the hexapeptide RGFRRR derived from the γ‐core motif of MtDef4. Fluorescent labelling of this hexapeptide showed that it strongly bound to the germ tube plasma membrane/cell wall. Using N. crassa expressing the Ca²⁺ reporter aequorin, MsDef1, MtDef4 and their cognate peptides were each shown to perturb Ca²⁺ homeostasis in specific and distinct ways, and the disruptive effects of MsDef1 on Ca²⁺ were mediated by glucosylceramide. Together, our results demonstrate that MsDef1 and MtDef4 differ markedly in their antifungal properties and specific domains within their γ‐core motifs play important roles in their different modes of antifungal action.     

B1 Cell IgE Impedes Mast Cell-Mediated Enhancement of Parasite Expulsion through B2 IgE Blockade

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Oral toxicity and impact on fecundity of three insecticidal proteins on the gregarious ectoparasitoid Eulophus pennicornis (Hymenoptera: Eulophidae)

Abstract 1 The effects of three insecticidal transgene proteins on selected life parameters of the ectoparasitoid Eulophus pennicornis were investigated. 2 When incorporated into the diet of the adult wasp, the lectins GNA (snowdrop lectin) and Con A (jackbean lectin) significantly reduced longevity at doses of 0.1% w/v and above. At a dose rate of 0.1% w/v, GNA and Con A reduced mean longevity to approximately 13 and 10 days, respectively, compared with average control lifespans of approximately 20 days. The trypsin inhibitor from cowpeas, CpTI, had no marked effect on longevity. 3 Both lectins reduced reproductive fitness of parasitoids when dosed before exposure to hosts. The 1.0% dose reduced fecundity by over 35% for GNA and 70% for Con A. Although reduced fecundity may have been a function of shorter lifespans, smaller egg loads in female wasps provided evidence to suggest that higher lectin doses may have interfered with egg maturation processes. 4 Both lectins were readily detected in whole body extracts of the parasitoid and were seen to persist within their bodies for at least 24 h after cessation of feeding on lectin‐containing diets. 5 The results would indicate that the ingestion of the lectins, either through host feeding or through the consumption of nectar, may pose a hazard to parasitic wasps if present at sufficiently high concentrations.     

SPECIES OF OCEANIC DINOFLAGELLATES IN THE GENERA DISSODINIUM AND PYROCYSTIS: INTERCLONAL AND INTERSPECIFIC COMPARISONS OF THE COLOR AND PHOTON YIELD OF BIOLUMINESCENCE1

We have examined aspects of the bioluminescence of 5 clones of Dissodinium, 1 clone of Pyrocystis acuta, 4 clones of Pyrocystis fusiformis, and 5 clones of Pyrocystis noctiluca. All clones produced the same color bioluminescence with an intensity peak near 474 nm. The in vivo emission spectra of these clones agreed with those previously determined, for 4 other species of marine dinoflagellates. The amount of light emitted by the dinoflagellates in scotophase when mechanically stimulated to exhaustion was determined for most of the clones. The largest species, P. noctiluca and P. fusiformis, emitted 37–89 × 10⁹ photons cell^?1 and 23–62 × 10⁹ photons cell^?1, respectively, about a thousand, times as much light as Gonyaulax species. Pyrocystis acuta emitted 3–6 × 10⁹ photons cell^?1. Three of the 5 clones of Dissodinium were bioluminescent. The range for 3 clones was 5–13 × 10⁹ photons cell^?1. All 5 clones of Dissodinium are morphologically distinct. Both the clones of Dissodinium and Pyrocystis produced much higher numbers of photons per cell nitrogen (ca. 7–50 times) than Gonyaulax polyedra or Pyrodinium bahamense. The data suggested that enzyme turnover occurred in the reactions producing light during mechanical stimulation of Dissodinium and Pyrocystis species.     

THE MARINE DIATOM ETHMODISCUS REX: ITS MORPHOLOGY AND OCCURRENCE IN THE PLANKTON OF THE SARGASSO SEA1,2

Living cells of the diatom Ethmodiscus rex (Rat-Iray) Wiseman & Hendey 1953 were found in the plankton of the southern Sargasso Sea. Apparently, this is the first report, of E rex from the plankton of the Atlantic Ocean. Scanning electron microscopy of peroxide-cleaned frustules revealed, some new morphological features for this species. When viewed, from inside the frustule, the puncta appear as rimmed pits. From outside the frustule, they appear to he shallow depressions with a small opening at the bottom. The so-called mucous tubules in the center of the valve were seen from the out side to be elongate slits and from the inside as obliquely directed flattened cylinders which cap the tubes.     

PHOTOINHIBITION OF MECHANICALLY STIMULABLE BIOLUMINESCENCE IN THE HETEROTROPHIC DINOFLAGELLATE PROTOPERIDINIUM DEPRESSUM (PYRROPHYTA)1

Photoinhibition of mechanically stimulable bioluminescence (MSL) in the heterotrophic dinoflagellate Protoperidinium depressum Bailey was investigated using samples collected from the Massachusetts and southern Texas coasts. The times for both photoinhibition of MSL (ca. 10 min) and dark recovery from photoinhibition of MSL (ca. 45 min) in this species were similar to those reported for autotrophic dinoflagellates. The degree of photoinhibition of MSL was a linear function of the logarithm of photon flux density (PFD). The threshold PFDs for the photoinhibition of MSL were 0.02, 0.6, and 21 μmol photons · m^?2· s^?1 for broad-band blue, green, and red light, respectively. These PFDs are lower than those required for photoinhibition of MSL by the autotrophic dinoflagellates Pyrocystis lunula and Ceratium fusus. We speculate that photosynthetic pigments in autotrophic dinoflagellates shield the photoreceptor that causes photoinhibition of MSL, thus lowering the sensitivity of these dinoflagellates to light. When field-collected P. depressum were kept in the laboratory without growth for a week, photoinhibition of MSL's sensitivity to light increased progressively along with 1) a decrease in its bioluminescence capacity (BCAP), 2) a decrease in the ratio of MSL to BCAP (MSL/BCAP), and 3) a decrease in the orange pigmentation (probably carotenoid) of the dinoflagellate. The action spectrum for photoinhibition of MSL in P. depressum was characterized primarily with a broad peak in the blue extending into the green. We suggest that carotenoid was not a photoreceptor for the photoinhibition of MSL in P. depressum because the peak of the action spectrum was too broad and extended too far into the green part of the spectrum, and because the orange pigment present decreased as photoinhibition of MSL became more sensitive to light.     

A modified DNA barcode approach to define trophic interactions between native and exotic pentatomids and their parasitoids

The establishment of invasive Halyomorpha halys (Stål) outside of its native range may impact native species assemblages, including other pentatomids and their scelionid parasitoids. This has generated interest in defining species diversity and host‐parasitoid associations in this system to better understand the impact of invasive alien species on trophic interactions in invaded regions. Information on scelionid–pentatomid associations in natural habitats is lacking, and species‐level identification of these associations can be tenuous using rearing and dissection techniques. Naturally occurring pentatomid eggs were collected in areas where H. halys has established in Canada and were analysed using a modified DNA barcoding approach to define species‐level trophic interactions. Identification was possible for >90% of egg masses. Eleven pentatomid and five scelionid species were identified, and trophic links were established. Approximately 70% of egg masses were parasitized; parasitism and parasitoid species composition were described for each species. Telenomus podisi Ashmead was the dominant parasitoid and was detected in all host species. Trissolcus euschisti Ashmead was detected in several host species, but was significantly more prevalent in Chinavia hilaris (Say) and Brochymena quadripustulata (Fabricius). Trissolcus brochymenae Ashmead and Tr. thyantae Ashmead were recorded sporadically. Parasitism of H. halys was 55%, and this species was significantly less likely to be parasitized than native pentatomids. The scelionid species composition of H. halys consisted of Te. podisi, Tr. euschisti and Tr. thyantae. Although these species cannot develop in fresh H. halys eggs, we demonstrate that parasitoids attempt to exploit this host under field conditions.     

The tropical white rot fungus, <Emphasis Type="Italic">Lentinus squarrosulus</Emphasis> Mont.: lignocellulolytic enzymes activities and sugar release from cornstalks under solid state fermentation

Lentinus squarrosulus Mont., a high temperature tolerant white rot fungus that is found across sub-Saharan Africa and many parts of Asia, is attracting attention due to its rapid mycelia growth and potential for use in food and biodegradation. A solid state fermentation (SSF) experiment with L. squarrosulus (strain MBFBL 201) on cornstalks was conducted. The study evaluated lignocellulolytic enzymes activity, loss of organic matter (LOM), exopolysaccharide content, and the release of water soluble sugars from degraded substrate. The results showed that L. squarrosulus was able to degrade cornstalks significantly, with 58.8% LOM after 30 days of SSF. Maximum lignocellulolytic enzyme activities were obtained on day 6 of cultivation: laccase = 154.5 U/L, MnP = 13 U/L, peroxidase = 27.4 U/L, CMCase = 6.0 U/mL and xylanase = 14.5 U/mL. L. squarrosulus is a good producer of exopolysaccharides (3.0–5.13 mg/mL). Glucose and galactose were the most abundant sugars detected in the substrate during SSF, while fructose, xylose and trehalose, although detected on day zero of the experiment, were absent in treated substrates. The preference for hemicellulose over cellulose, combined with the high temperature tolerance and the very fast growth rate characteristics of L. squarrosulus could make it an ideal candidate for application in industrial pretreatment and biodelignification of lignocellulosic biomass.     

Long term testicular ischemia-reperfusion injury-induced apoptosis: Involvement of survivin down-regulation

Testicular torsion is associated with damage to the testicular tissue as a result of ischemia-reperfusion injury (IRI) and induction of apoptosis leading to progressive damage to spermatogenesis. Survivin is suggested to be an important regulator in the control of the mitochondrial apoptotic pathway, although its role in torsion-induced IRI is unknown. Therefore, we sought to evaluate testicular survivin expression after long term IRI induced by testicular torsion. Survivin expression was measured by real-time PCR in 6-12 month old New Zealand white rabbits divided into three groups (4 animals/group): group (A) sham control, group (B) ischemia alone for 60 min and group (C) ischemia for 60 min followed by reperfusion for 6 months. Germ cell apoptosis was evaluated by TUNEL assay, Bax/Bcl-2 ratio and DNA fragmentation. The Johnsen score was used to assess testicular morphological damage, while lipid peroxidation was used as an indicator for oxidative stress. Survivin expression was detected in all testicular tissue samples. The rate of survivin expression after IRI was significantly higher (p < 0.05) compared with ischemic only and sham control testes. Its expression in IRI samples was inversely correlated with the significant increase (p < 0.05) in apoptosis, oxidative levels and spermatogenic damage. In conclusion, down-regulation of testicular survivin expression after long term IRI to the testis and its association with apoptosis induction suggests its involvement in the regulation of this apoptotic pathway. These findings also identify survivin as a potential new target for the prevention of germ cell death during testicular torsion.