Genome-wide RNAi analysis of growth and viability in Drosophila cells

A crucial aim upon completion of whole genome sequences is the functional analysis of all predicted genes. We have applied a high-throughput RNA-interference (RNAi) screen of 19,470 double-stranded (ds) RNAs in cultured cells to characterize the function of nearly all (91%) predicted Drosophila genes in cell growth and viability. We found 438 dsRNAs that identified essential genes, among which 80% lacked mutant alleles. A quantitative assay of cell number was applied to identify genes of known and uncharacterized functions. In particular, we demonstrate a role for the homolog of a mammalian acute myeloid leukemia gene (AML1) in cell survival. Such a systematic screen for cell phenotypes, such as cell viability, can thus be effective in characterizing functionally related genes on a genome-wide scale.     

Untersuchungen zur Übereinstimmung des Auftretens zytoplasmatischer Einschlüsse mit dem immunelektronen-mikroskopisch nachgewiesenen Kartoffel-Y-Virus in künstlich infizierten Meristempflanzen-Subkulturen

Zusammenfassung Meristempflanzen-Subkulturen der Kartoffelsorten ‘Vorw?rts’ und ‘Schwalbe’ wurden mit KYV künstlich infiziert und in verschiedenen Infektionsstadien elektronenmikroskopisch untersucht. Neben der Untersuchung des Zytoplasmas von Mesophyllzellen an Ultradünnschnitten wurde Pflanzensaft für die immunelektronenmikroskopische Dekorationsmethode und die Tauchmethode pr?pariert. Es konnte eine zeitliche übereinstimmung des Auftretens von ‘Pinwheels’ im Zytoplasma und immunelektronenmikroskopischer Nachweisbarkeit des KYV beobachtet werden. W?hrend in der Sorte ‘Vorw?rts’ (geringe relative Resistenz gegenüber KYV) bereits 21 Tage nach der Infektion KYV nachgewiesen wurde, war der Nachweis in der Sorte ‘Schwalbe’ (hohe relative Resistenz gegenüber KYV) erst nach einer zus?tzlichen Pfropfinfektion m?glich. Sowohl die Dekorationsmethode als auch der zytoplasmatische Nachweis eignen sich für eine Diagnose des KYV in Pflanzen mit geringer Viruskonzentration.

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Summary PVY-free subcultures of meristem plants of the potato varieties Vorw?rts (relatively low resistance to PVY) and Schwalbe (relatively high resistance to PVY) were infected in vitro withMyzus persicae (Sulz.). Ultra-thin sections were cut from both infected and non-infected plants 11, 21, 29 and 49 days after inoculation and at the same time sap was prepared according to modifications of the immune electron microscopic Decoration method of Milne & Luisoni (1977) and of the Touch method of Brandes (1957). The investigation established the coincidence of ‘Pinwheels’ in the cytoplasm of infected plants and the presence of PVY particles identified by immune electron microscopy, thus confirming the relationship between the appearance of such ‘Pinwheels’ and an infection with PVY. Depending on their level of resistance to the virus, PVY was detectable in different varieties at varying stages of infection (Table 1). Whereas in cv. Vorw?rts, PVY could be detected 21 days after infection, in cv. Schwalbe it was only possible three weeks after a supplementary graft inoculation. Both the Decoration method and cytoplasmic detection are suitable for the diagnosis of PVY in plants with a low virus concentration. The Touch method fails because of its lack of differentiation potential thus preventing the acquisition of additional information.

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Résumé Des plantes indemnes de virus Y de la pomme de terre (PVY) obtenues à partir de repiquages de cultures de méristème de la variété Vorw?rts (faiblement résistante au PVY) et de la variété Schwalbe (fortement résistante au PVY) ont été contaminées au stade in vitro à l'aide deMyzus persicae (Sulz.). Des échantillons de feuilles provenant de plantes saines et de plantes contaminées ont été observés avec le microscope électronique, 11, 21, 29 et 49 jours après la contamination. Des coupes ultrafines ont été obtenues ainsi que du jus de plantes préparés, d'une part à l'aide de la méthode immuno-microscopie électronique de décoration selon Milne & Luisoni (1977) et d'autre part, selon une méthode d'immersion modifiée d'apres Brandes (1957). Les examens ont démontré une concordance temporelle entre l'apparition de ‘Pinwheels’ dans le cytoplasme des plantes contaminées et les particules du PVY décelées par immuno-microscopie électronique. La relation entre l'apparition de ‘Pinwheels’ et l'infection par le PVY a ainsi été démontrée. Selon les résistances variétales, le PVY a été observé à différents stades d'infection (tableau 1). Le PVY a été observé 21 jours après la contamination pour la variété Vorw?rts et seulement 3 semaines après une infection supplémentaire par greffage pour la variété Schwalbe. Pour réaliser un diagnostic du PVY dans les plantes à faible concentration de virus, tant la méthode de décoration que le test du cytoplasme peuvent être appliqués. La méthode par immersion n'a pas apporté d'information supplémentaire en raison de l'absence de moyens de différenciation.

     

Application of Soluble Chitosan in vitro and in the Greenhouse to Increase Yield and Seed Quality of Potato Minitubers

The aim of this work was to investigate whether the application of soluble chitosan in potato micropropagation can improve microplant quality in vitro, help acclimatisation ex vitro, and increase yield and seed quality of minitubers. Potato cv. Désirée microplants were treated in vitro with soluble chitosan added to the semisolid tissue culture medium in different concentrations. Microplants were subsequently transferred to the greenhouse and sprayed with chitosan solutions or remained unsprayed. Untreated microplants were also established ex vitro and sprayed with chitosan, or left unsprayed as a control. Morphological and physiological parameters of plant growth were assessed in vitro and ex vitro. Plantlet quality was evaluated using a ranking system. Minitubers derived from greenhouse plantlets were planted in the field in subtropical conditions, and growth and yield parameters evaluated. The chitosan concentration most beneficial to the in vitro growth of microplants varied between years. In treatments with the best in vitro growth, minituber number and yield in the greenhouse was also increased. Foliar chitosan application at the acclimatisation phase stabilised the effect of in vitro treatment on yield parameters. The seed quality of minitubers derived from chitosan treatments in vitro alone and in combination with foliar treatment at acclimatisation was improved, giving rise to field plants with increased tuber numbers and yields. The combination of chitosan in vitro and foliar application during the acclimatisation phase ex vitro had a greater effect on minituber seed quality than the concentration applied in vitro.     

Contamination of a boatyard for maintenance of pleasure boats

Purpose

The object of this study was to study a boat maintenance facility by investigating the degree of contamination and assessing how leachate water from soil affects organisms from three trophic levels.

Materials and methods

Surface and subsurface (20-cm depth) soil samples were collected in a typical boatyard (200 boats, 12,000 m²) at a 70- (station A), 90- (station B), 120- (station C) and 160-m (station D) distance from the shoreline. Three replicate samples, ～10 m apart, were taken at stations A, B and C, respectively, and one replicate was taken at station D (i.e. altogether 20 samples with 10 at surface and subsurface, respectively). The total copper (Cu), lead (Pb), tin (Sn) and zinc (Zn) concentrations were determined for all replicates. Pooled samples from the respective stations were used for analysis of organotin compounds, irgarol and polyaromatic hydrocarbons. Leachate waters were produced from the pooled samples and used for toxicity testing with the bacterium Vibrio fischeri, the macroalga Ceramium tenuicorne and the crustacean Nitocra spinipes.

Results and discussion

Very high concentrations of Cu, Pb, Zn were detected, with maximum values of 16,300, 6,430 and 18,600 mg/kg dw, respectively. Organic hazardous compounds were found in high concentrations with maximum values of 37, 27 and 16 mg/kg dw for tributytin (TBT), dibutyltin (DBT) and triphenyltin (TPhT), respectively. All pollutants exceeded existing guidance values for both sensitive land use and less sensitive land use by several factors, in both surface and subsurface soil. The least and worst cases of total amount of TBT (12 000 m² and 0.2 m depth) were estimated to be 10 and 122 kg of TBT. Leachates were shown to be toxic in all three test organisms.

Conclusions

Several known hazardous pollutants were found in boatyard maintenance areas and they exceeded recommended guidance values by several factors. Leachates were shown to be toxic to test organisms of several trophic orders. This underlines that boat maintenance facilities in general should be better regulated to minimize further exposure to humans and spread of contaminants in the environment. The amounts of contaminants accumulated in these areas call for investigations of how remediation should be performed.