Biocontrol options for Cirsium arvense with indigenous fungal pathogens

As a first step in the development of a mycoherbicide for biological control of Cirsium arvense indigenous fungal pathogens that had been isolated from diseased hosts were tested both singly and in combination on potted plants under environmental conditions. Disease symptoms (necroses, chloroses and macerations) and parameters of growth and development were evaluated to determine the potential for weed control. During 1998, Phoma destructiva, Phoma hedericola, a Mycelia sterilia and Puccinia punctiformis were inoculated singly. With the exception of the obligate biotroph P. punctiformis (local infections), the single inoculations reduced the measured parameters. A combined inoculum of P. punctiformis and Ph. hedericola was less effective than Ph. hedericola alone. In 1999, Ph. hedericola, Ph. destructiva, Ph. nebulosa and the Mycelia sterila were applied both singly and as a combined inoculum. Of all the tested inocula, the mixture of four pathogens had the greatest potential for biocontrol, significantly reducing reproductive capacity of the plants. These results demonstrate the importance of taking advantage of synergisms in developing a mycoherbicide.     

Fireblight monitoring in Lithuania1

Lithuania has requested that its whole territory should be recognized by the EU as a protected zone for Erwinia amylovora . Fireblight monitoring was performed in 1998/2002 with the aim of detecting and identifying the bacterium, and of determining its distribution in the country. The study consisted of periodic surveys (at least twice a year) of nurseries, orchards, collective farms and host plants, growing individually or in small groups, as well as the surrounding zone within a radius of 250 m. Tests, under conditions of quality control, were applied to host plants with and without symptoms, using detection methods such as ELISA and immunofluorescence (with polyclonal antibodies), semi-selective plating and pathogenicity.     

Détection moléculaire spécifique de la région vir du plasmide pTi d'Agrobacterium tumefaciens dans les sols et plants au Maroc

Le meilleur moyen de contrôle du crown gall ( Agrobacterium tumefaciens ) est la prévention et donc la détection de la bactérie pathogène dans les sols prévus pour les pépinières. Pour cela nous avons essayé une approche moléculaire utilisant la PCR pour caractériser le plasmide pTi par le couple des amorces spécifiques F14/F44 dans les sols de diverses régions marocaines. Cette technique de détection représente un indicateur potentiel d'avertissement sur l'état d'infection des sols et plants par A. tumefaciens qui permet ainsi de déclencher la lutte biologique par le NoGall, produit commercial australien, hébergeant l'antagoniste K₁₀₂₆.     

UV-absorbing compounds and susceptibility of weedy species to UV-B radiation

Stratospheric ozone (O₃) depletion has led to increased terrestrial ultraviolet‐B (UV‐B) radiation (290–320 nm). Leaves exposed to this radiation produce UV‐absorbing compounds in the epidermal cells, which protect plants from UV‐B damage. To determine the role of UV‐absorbing compounds in the UV‐B sensitivity of weeds (common chickweed (Stellaria media), downy brome (Bromus tectorum), green smartweed (Polygonum scabrum), redroot pigweed (Amaranthus retroflexus), spotted cat’s‐ear (Hypochoeris radicata), and stork’s‐bill (Erodium cicutarium)) seedlings were exposed to 0, 4 (field ambient), 7 (18% O₃ depletion) and 11 (37% O₃ depletion) kJ m^?2 d^?1 of biologically effective UV‐B radiation in a greenhouse. Ultraviolet‐absorbing compounds were extracted from the second true‐leaf (0.5 cm² samples) with methanol : distilled water : HCl (79 : 20 : 1) in an 85°C water bath for 15 min, and the absorbance of the extracts measured at 300 nm. The shoot dry biomass was recorded to determine the susceptibility to UV‐B radiation. Common chickweed was the most sensitive and green smartweed the least sensitive weed to UV‐B radiation. The latter accumulated more UV‐absorbing compounds and this accumulation occurred earlier compared with common chickweed. As UV‐B_BE radiation levels increased from 0 to 11 kJ m^?2 d^?1, the green smartweed shoot biomass did not decline. However, the biomass of all five susceptible species declined despite an increase in the UV‐absorbing compounds in response to increased UV‐B radiation. Therefore, formation of a ‘UV‐screen’ in these species is not sufficient to fully prevent UV‐B damage. When the concentration of UV‐absorbing compounds in the six species was plotted against their susceptibility to UV‐B radiation, no relationship was observed. Thus, while the accumulation of UV‐absorbing compounds may be a major factor in the protection of certain species against UV‐B radiation and may offer some degree of defence in other species, it does not explain UV‐B susceptibility differences in weedy species in general.     

Non-target toxicology of a new mosquito larvicide, trypsin modulating oostatic factor

Trypsin modulating oostatic factor (TMOF), a peptide hormone originally isolated from the ovaries of adult Aedes aegypti, is currently under commercial development as a new pesticide chemistry with a novel mode of action for the control of larval mosquitoes. The objective of the current research is to evaluate potential risks of the use of TMOF as an insecticide on non-target organisms. TMOF (YDPAP₆) was degraded in vitro (as determined by HPLC and LC/MS) to DPAP₆, PAP₆, and then AP₆ by leucine aminopeptidase, a pancreatic enzyme found in the digestive system of vertebrates. The rate of degradation of TMOF and PAP₆ was significantly greater than that of DPAP₆, while no metabolism of AP₆ was found. TMOF technical insecticide was produced on a commercial scale by recombinant yeast (heat-killed before application). The technical TMOF when administered in a single dose by gavage to male and female mice at 2000 mg dry weight/kg body weight produced no negative effects as compared to controls up to 12 days after treatment. When male and female mallard ducks were treated by gavage with 1250 mg dry weight of technical TMOF/kg body weight each day for 5 days, again no toxic effects were noted through 35 days after the last treatment. TMOF technical insecticide was also applied to the shaved skin of male and female rabbits at the rate of 2000 mg/kg for 1-2 days, with no effect. The end point observations in these in vivo experiments were mortality; changes in growth rate, behavior, body structure, and color; and possible lesions observed during necropsy. Finally, Daphnia incubated with technical TMOF in rearing water at the level of 1.0 × 10⁶ yeast cells/ml (10 mg/ml) also demonstrated no negative effects on mortality, growth, molting, time to first brood, and production of viable neonates. It appears from these studies that TMOF can be degraded by vertebrate digestive proteases and technical TMOF is not toxic to the non-target organisms examined.     

PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes.     

Fireblight in Békés County (Hungary) in 1996/20021

Fireblight (Erwinia amylovora) appeared in Hungary in 1996. Most damage occurred on apple, pear, quince and medlar, and also on the ornamentals Pyracantha, Sorbus, Cotoneaster and Crataegus. In 1996–2006, an official programme for elimination of infected parts of plants started in Békés county. This mainly concerned trees in towns and villages, since there are few pome‐fruit orchards in the county. Work teams under official direction pruned back or cut down trees. In total, some 13 000 trees were pruned back and nearly 11 000 were cut down. In addition, 21 villages were subjected to special phytosanitary measures. Infection decreased considerably between 1996 and 2002, but over 90% of the inhabited areas in the county remained subject to special measures, because of the very dispersed occurrence of fireblight.     

Effectiveness of plant extracts of Paeonia suffruticosa and Hedera helix against diseases caused by Phytophthora infestans in tomato and Pseudoperonospora cubensis in cucumber

Journal of Plant Diseases and Protection - The effects of ethanolic extracts (20 % per fresh weight (Fw) or 6 % per dry weight) of 20 different plant species were tested against late blight...     

Footsteps from Insect Larvae Damage Leaf Surfaces and Initiate Rapid Responses

Plant resistance to insect herbivory involves gene expression in response to wounding and the detection of insect elicitors in oral secretions (Kessler and Baldwin, 2002, Ann. Rev. Plant/ Biol. 53: 299–328). However, crawling insect larvae stimulate the synthesis of 4-aminobutyrate within minutes and imprints of larval footsteps can be visualized within seconds through superoxide production or transient increases in chlorophyll fluorescence (Bown et al., 2002, Plant Physiol. 129: 1430–1434). Here cryo-scanning electron microscopy was used to demonstrate that larval feet, which are equipped with a perimeter row of hook-like crochets, damage leaf tissue and result in larval footprints. Staining for cell death shows that areas of wounding correspond to footsteps detected through increased chlorophyll fluorescence. Superoxide production in response to footsteps was inhibited by diphenyleneiodonium, an inhibitor of the plasma membrane NADPH oxidase enzyme. Inhibition of superoxide production, however, did not eliminate the detection of cell death. The results demonstrate that larval footsteps damage leaf tissue, and initiate rapid local responses which are not dependent on herbivory or oral secretions. It is proposed that superoxide production at the wound site prevents opportunistic pathogen infection.