Systemic infection of black currant flowers by Botrytis cinerea and its possible involvement in premature abscission of fruits

Emasculated flowers of several black currant cultivars were pollinated and then inoculated with dry conidia of Botrytis cinerea in the field and glasshouse. The infection of pistils was examined by U.V. fluorescence microscopy and the incidence of premature flower abscission recorded. Conidia germinated in the stigmatic fluid in all cultivars and hyphae spread symptomlessly throughout the style to infect the pericarp and ovules. Of six cultivars inoculated in the field, cv. Ojebyn was the most, and cv. Ben More the least resistant to flower shedding. Natural infection of stigmas by B. cinerea was common in the field and a high proportion of apparently healthy non-inoculated flowers which abscissed were found to contain infected ovules. Fewer flowers abscissed if inoculations were made 6 days after pollination. Symptomless or latent infection of black currant flowers by B. cinerea may be a contributory cause of premature abscission of developing fruits, or ‘running-off’, recorded in these experiments.     

Overwintering of Sphaerotheca mors-uvae on black currant and gooseberry

The ability of Sphaerotheca mors-uvae to perennate as cleistocarps, and as mycelium in buds was examined during the winters of 1965-6, 1966-7 and 1967-8 in relation to its two principal hosts, gooseberry and black currant. Cleistocarps on black currant leaves were examined from August 1965 to April 1966 and from July 1966 to March 1967. In 1965 cleistocarps were first observed on the leaves on 5 August; in 1966 on 11 July. These continued to develop through August and September and by October approximately 70% contained well-defined ascospores. The ascospore content remained generally at this level until February 1966 and November 1966; then the numbers of cleistocarps with ascospores fell and by April 1966 and March 1967 few such cleistocarps remained. From 21 March 1966 and 15 February 1967, but not otherwise, discharge of ascospores from the overwintered cleistocarps was readily obtained in laboratory tests. The viability and infectivity of the ascospores was demonstrated by allowing them to discharge on to leaf discs of black currant in the laboratory and also on to leaf discs and plants in the field. Sporulating colonies of S. mors-uvae developed within 8 days. Cleistocarps from shoots of black currant were examined from 4 August 1966 to 9 March 1967, and from 27 July 1967 to 1 January 1968. They developed in a similar manner to those on black currant leaves and by September in both 1966 and 1967 over 60% contained ascospores. This level was not maintained; the number of cleistocarps with ascospores fell gradually and by 8 December 1966 and 1 January 1968 few remained. Only in one laboratory test (21 November 1967) were ascospores discharged from a sample of these cleistocarps. Cleistocarps from shoots of gooseberry were examined from July 1966 to March 1967, and from August 1967 to January 1968. The pattern of ascospore development and subsequent decline in number of cleistocarps with ascospores was similar to that observed for black currant shoots. No discharge of ascospores could be demonstrated in laboratory tests. Evidence that S. mors-uvae perennates in buds of gooseberry was obtained by dissecting buds and by inducing buds on surface-sterilized shoots to burst under conditions which precluded chance infection. Field observations also suggested that bud infection occurred on gooseberry. Similar experiments failed to demonstrate the fungus in buds of black currant, and there was no indication of bud infection of this host in the field.     

Transference of resistance to black-currant gall mite, Cecidophyopsis ribis, from gooseberry to black currant

Field and insectary tests confirmed that the black-currant gall mite (Cecidophyopsis ribis) is unable to survive on gooseberry and red currant. A dominant gene Ce, controlling resistance to the gall mite, has been transferred from gooseberry to black currant. Resistant, large-fruited, self-fertile black currants of commercial potential have been obtained in the third backcross. One accession of Ribes bracteosum and three of R. americanum proved field susceptible to the gall mite, but twenty-four accessions of other Ribes species remained free from galled buds for at least 3 years in an infection plot.     

Improvement of the antitumor activity of black currant polysaccharide by an enzymatic treatment

A polysaccharide-rich substance isolated from black currant, named cassis polysaccharide (CAPS), was partially digested with beta-galactosidase from Aspergillus oryzae and its immunostimulatory activity was investigated. The in vitro cytokine-inducing effect of CAPS on RAW264 cells was gradually decreased along with lowering of the average MW of CAPS. In vivo, partially digested CAPS with a mean MW of approximately 20,000 showed the most potent antitumor activity against Ehrlich carcinoma in mice.     

The ecology of cold hardiness in different populations of the black currant gall mite, Cecidophyopsis ribis

Population samples of C. ribis from eastern Norway, western Norway, and England, and cuttings of 5 black currant varieties were exposed for different periods to -18.5°±0.5° C and mites also to 6°. Mite mortality, mite emigration, egg production at 6°, and bud burst of the cuttings, were observed.Marked differences were found between the mite populations both in their ability to produce eggs at 6°, and in cold hardiness at -18.5°. Mortality during cold treatment was inversely related to mite density in different buds as well as in different zones of each bud. Mite emigration was greatly facilitated when the side of the bud touched the twig.When comparing cold hardiness of the mites with that of black currant cuttings, cold treatment of the cuttings as a control measure appeared to be of little promise against the Norwegian populations of C. ribis. With the English population, however, a reasonable safety margin seemed to exist between exposures giving complete kill of the mites, and those which damage the cuttings.

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Zusammenfassung Populationsproben von C. ribis von Rygge, Ost-Norwegen; Hjelmeland, West-Norwegen; und Reading, England, wurden für verschiedene Zeiten Temperaturen von -18.5±0,5° und 6° ausgesetzt. Unbefallene Stecklinge der folgenden fünf Schwarze Johannisbeer-Sorten wurden in ähnlicher Weise mit Kälte von -18.5° behandelt: Bang up, Booskop Giant, Brødtorp, Silvergieter's Zwarte und Wellington XXX. Mortalität, Abwanderung und Eiproduktion der Milben bei 6° und das Knospenaufbrechen der Johannisbeer-Reiser wurden beobachtet.Die Milben von Rygge waren am meisten, die von Reading am wenigsten kälteresistent, während Milben von Hjelmeland eine mittlere Kälteresistenz aufwiesen (Fig. 1). Milben von Rygge, und in geringerem Maße die von Hjelmeland, produzierten bei 6° eine beträchtliche Anzahl Eier, während die Eiproduktion der englischen Population bei dieser Temperatur fast vollständig zum Stillstand gelangte (Tabelle III).Die Mortalität während der Kälte war umgekehrt proportional zur Milbendichte in verschiedenen Knospen (Fig. 2) sowohl wie in verschiedenen Zonen jeder Knospe (Tabelle I). Niedrige Wintertemperaturen wirken demnach nicht als Regulationsfaktor auf die Populationsdynamik dieser Art.Die Wirkung der Kältebehandlung auf die Milbenabwanderung ist in Tabelle II dargestellt. Die Abwanderung der Milben wurde sehr erleichtert, wenn eine Seite der Knospe den Zweig berührte (Fig. 4).Mit einer möglichen Ausnahme von Booskop Giant und Brødtorp konnten die Sorten der Schwarzen Johannisbeeren 16 Tage lang -18,5° ohne sichtbaren Schaden für den Knospenaustrieb unterworfen werden. Milben aus Ost-Norwegen überlebten 16 Tage Kältebehandlung in allen und Milben aus West-Norwegen in einer der Prüfungen. Kältebehandlung der Stecklinge scheint deshalb als Bekämpfungsmaßnahme gegen die norwegischen Populationen von C. ribis wenig erfolgversprechend. In den englischen Milbenpopulationen wurde dagegen nach 9 oder 10 Tagen 100% Mortalität erzielt. Dies scheint einen erträglichen Sicherheitszeitraum zwischen Behandlungen, die vollständige Abtötung der Milben ergeben, und solchen, die den Stecklingen schaden, offenzulassen.

     

Chromosomally-determined induced tolerance to copper in Escherichia coli

Pre-exposure of Echerichia coli to sub-lethal concentrations of cupric sulphate induced copper tolerance: pre-exposed (habituated) organisms were essentially unaffected by concentrations of Cu²⁺ which completely prevented colony formation by non-habituated ones. The observed copper tolerance was not dependent on the selection of copper-resistant mutants but resulted from a phenotypic change in the organisms during the pre-exposure period.     

Specific neonatally induced tolerance to Mls locus determinants

Neonatal injection of CBA/HT6T6 (H-2k, Mlsb) mice with adult, Mls-incompatible (CBA/J [H-2k, Mlsd] X CBA/HT6T6)F1 spleen cells results in the abrogation of cell proliferation and interleukin 2 (IL 2) production in bulk mixed lymphocyte cultures, when spleen cells from the inoculated mice are tested at 6 to 8 wk of age with stimulator cells expressing the Mlsd of the tolerizing inoculum. In limiting dilution assays, this tolerant state was manifested in a 25- to 550-fold (280-fold average) decrease in the frequency of precursors of Mlsd-responsive IL 2-producing T cells. Tolerance was specific in that the frequencies of precursors of IL 2-producing cells responding to Con A, allogeneic H-2d, and self-Ia were not affected. The observed low frequency of Mls-responsive cells was due neither to extensive chimerism resulting in the dilution of Mlsd-responsive cells by the nonresponsive F1 cells of the inoculum, nor to the action of suppressor cells. These findings indicate that neonatal injection of Mls-incompatible spleen cells produces a state of specific tolerance by a clonal deletion or inactivation mechanism. This specific tolerance supports the view that 1) the Mls locus encodes or regulates the expression of defined alloantigenic determinants and 2) Mls-incompatible responder mice have specific receptors for Mls determinants on clonally distributed IL 2-producing responder T cells.     

Immunostimulatory effects of a polysaccharide-rich substance with antitumor activity isolated from black currant (Ribes nigrum L.)

The fruit juice of black currant was found to contain a polysaccharide-rich substance, which was designated cassis polysaccharide (CAPS), with macrophage-stimulating activity. Especially, its interleukin (IL)-1beta-inducing activity was remarkably high, compared with other fruit juice preparations. CAPS was found to consist of rhamnose, mannose, arabinose, galactose, xylose, and glucose in a molar ratio of 11.3:0.9:54.1:29.8:2.0:1.9. CAPS turned out to be partitioned into a soluble component (CAPS-l.m.) and a precipitable component (CAPS-h.m.) with mean MWs of 80,000 and 600,000 respectively in 45% (v/v) ethanol solution. At least in vitro, CAPS-l.m. rather than CAPS-h.m. appeared to play an important role in macrophage activation. Oral administration of black currant juice and CAPS to Ehrlich carcinoma-bearing mice retarded the growth of the solid tumor by 45% and 51% respectively. CAPS administration had a stimulatory effect on the release of IL-2, IL-10, interferon-gamma, and IL-4 from splenocytes in comparison with PBS treatment in tumor-bearing mice. The IL-4 level was, however, still lower than that exhibited by a group of normal mice. CAPS showed a certain cytotoxicity directly against tumor cells.     

The cellular mechanism of maintenance of neonatally induced tolerance to H-2 class I antigens

We used limiting dilution analysis protocols to investigate the mechanism by which in vitro cytotoxic T lymphocyte (CTL) hyporeactivity is maintained in adult mice that had been neonatally tolerized to major histocompatibility complex-encoded antigens. Class I molecules, presented on donor cells having an H-2 K or D region haplotype difference from recipients, readily induce tolerogen-specific CTL hyporeactivity. All attempts to identify any in vitro effects of active suppressive cells operative in the maintenance of this hyporeactivity have been unsuccessful. We conclude that this cytotoxic deficiency is the consequence of in vivo mediated clonal inactivation of the precursors of tolerogen-specific CTL. A presentation and evaluation of the assumptions inherent in this conclusion are made. In contrast to class I molecules, class II molecules, presented on donor cells having an H-2 I region haplotype difference from recipients, are unable to induce tolerogen-specific CTL hyporeactivity, even when injected neonatally at high doses. This inability of class II molecules to induce CTL tolerance parallels the considerable difficulty of inducing helper T lymphocyte tolerance to class II molecules.     

Extraction of cadmium and tolerance of three annual cut flowers on Cd-contaminated soils

To evaluate the production potential and Cd removal by three flower crops, viz.: marigold (Tagetes erecta), chrysanthemum (Chrysanthemum indicum) and gladiolus (Gladiolus grandiflorus), an experiment was conducted on differentially contaminated soils (DTPA-Cd 0.6-68.4 mg kg(-1)). Biotoxicity of Cd lead to reductions in growth and flower yield of marigold at DTPA-Cd >or= 7.9 mg kg(-1) soil, while the productivity of chrysanthemum and gladiolus was sustained up to 21.2 mg kg(-1). DTPA-Cd for 50% yield reduction (C(50)) was 85, 106 and 215 mg kg(-1) soil for marigold, chrysanthemum and gladiolus, respectively, that indicates a better Cd-tolerance in gladiolus. The uptake of Cd increased with contents in soils and the maximum accumulation occurred in leaves. Among the economic parts, gladiolus spikes accumulated the highest Cd (7.2) followed by flowers of marigold (6.5) and chrysanthemum (4.0 mg kg(-1)). But, because of higher biomass, the total Cd removal was the maximum with chrysanthemum (8.3) followed by gladiolus (6.0) and the minimum (2.6 mg m(-2)) with marigold. Gladiolus with highest tolerance and Cd-content in saleable part holds potential to clean up the moderately contaminated soils.     

Mechanism of tolerance to VI-antigen of Salmonella typhi induced by cyclophosphane

High dose Vi-antigen treatment and injection of cyclophosphamide 46 to 48 hours later induced in mice a state of immunological unresponsiveness remaining stable in adoptive transfer. Only low amounts of the antigen were revealed in the blood and spleen of tolerant animals 2 to 3 weeks after the tolerogenic treatment. No T-suppressors were found in the spleen of tolerant mice--the cells of tolerant mice failed to suppress the immune response of normal lymphocytes when transferred together to the irradiated recipients, or to induce tolerance in normal mice. Normal spleen cells restored partially the immune responsiveness in tolerant animals. The results obtained suggest that cyclophosphamide tolerance was due to deletion or the long-term inactivation of the immunocompetent cells.     

Neutrophil tolerance to oxidative stress induced by hypoxia/reoxygenation

Repetitive episodes of hypoxia/reoxygenation induce cellular adaptations resulting in a tolerance process against oxidative stress. We studied the effects of chronic episodes of hypoxia/reoxygenation on neutrophil antioxidant defenses, neutrophil oxidative capability, and oxidative damage induced in neutrophils and plasma. Seven professional apnea divers participated in the study. Blood samples were taken under basal conditions, after a diving apnea session, and under basal conditions after five consecutive days of diving apnea sessions (basal post-diving). Chronic episodes of hypoxia/reoxygenation increased malondialdehyde (MDA), carbonyl derivates and creatine kinase (CPK) in plasma. Neutrophil catalase (CAT) levels were higher in basal post-diving. Neutrophil oxidative burst was maintained after diving, although the maximum response was delayed in basal post-diving. Neutrophil thioredoxin reductase (TR) activity increased in basal post-diving, and glutathione reductase (GR) activity was maintained. Chronic, repetitive episodes of diving apnea induce neutrophil adaptations in order to delay the oxidative burst response and to facilitate protein reduction. Diving apnea could be a good model to study tolerance to the oxidative stress generated by hypoxia/reoxygenation.     

Immunologic tolerance to dinitrophenylated human gamma globulin induced via colostrum

Immunologic unresponsiveness or tolerance was induced in neonatal mice via colostrum by injection of dinitrophenylated human gamma globulin (DNP-HGG) into the mother on the day of birth. Unresponsiveness persisted in the neonates for at least 21 weeks. This longlasting tolerance appeared to be the result of an unresponsiveness to the carrier determinants. Hapten-specific B-cell tolerance was assessed in mice receiving high- or low-epitope-density tolerogen and it was observed that the low-epitope-density tolerogen (DNP₁HGG) resulted in carrier-specific tolerance only. Although mice tolerized with the high-epitope-density conjugates were found to be slightly hyporesponsive in their in vivo B-cell responses, their in vitro hapten-specific responses were normal. This tolerant state induced via colostrum was compared to tolerance induced in utero. This earlier contact with tolerogen resulted in more profound alterations in hapten-specific B-cell responses. An additional interesting finding was that the colostrally induced tolerant state was transmitted to the next generation.     

Absence of suppression in natural and induced tolerance to F antigen

The role of suppression in natural and induced tolerance to F antigen was investigated in two sets of experiments. In the first, CBA mice were submitted to pretreatments which decrease suppression and the antibody response to self- or allo-F type was investigated. The second set of experiments involved the transfer of spleen cells from tolerized or from naturally tolerant mice into normal mice which were then primed with allo-F, as well as the co-transfer of tolerant and primed lymphocytes into normal mice, to test whether tolerant lymphocytes present suppressor cells. The results indicate that the immune response against allo-F antigen is normally kept in a low level by a suppressive mechanism, and that F-specific suppressor T cells are absent from tolerant mice.Abbreviations used in this paper ATx adult thymectomy - BSS buffered salt solution - CFA Freund's complete adjuvant - CY cyclophosphamide - F.1 type-1 F antigen - F.2 type-2 F antigen - PBS phosphate-buffered saline - RIA radioimmunoassay - Th T helper cell - Ts T suppressor cell     

Field experiments on the spread of black currant reversion virus and its gall mite vector (Phytoptus ribis Nal.)

In two experiments the spread of reversion virus from a row of systemi-cally infected black currant bushes heavily infested by the gall mite vector (Phytoptus ribis Nal.) was predominantly in the direction of the winds prevailing during the dispersal period. On each side of the sources there was a curvilinear decrease of galled buds and of virus infection as distance increased. In another experiment a central source of mites and virus was surrounded by concentric hexagons comprising alternate rows of healthy and virus-infected bushes. At leaf-fall, galls were forty times more numerous on virus-infected than on healthy bushes; plants in the sector downwind developed the most galls and those upwind the least. On both healthy and virus-infected bushes in each sector, the incidence of galls decreased with increasing distance from the source. The gradients of infestation were steeper on healthy than on virus-infected bushes, especially in sectors upwind from the source. In some sectors the infestation gradients were distorted because many of the virus-infected bushes were so heavily infested that most of the buds became galled. The spread of virus to initially healthy plants decreased from 100 to 75% near the source, to zero at the periphery. More bushes became infected downwind from the source than upwind. In each experiment more bushes developed galls than later produced symptoms of virus infection, the incidence of which was positively correlated with the number of galls recorded the previous winter.