Cacao leaf procyanidins increase locally and systemically in response to infection by Moniliophthora perniciosa basidiospores

Flavan-3-ol monomers and oligomers from dimers to decamers were quantitatively determined by HPLC-FLD in leaves of four Theobroma cacao varieties. Cacao flush leaves had higher procyanidin content than mature leaves. Moniliophthora perniciosa basidiospore infection results in an increase in cacao leaf procyanidins in the inoculated leaves, as well as in the uninoculated, adjacent, older leaves on an actively growing branch, resulting in both a local and systemic increase in procyanidin content in response to infection. Procyanidin oligomers increased significantly in leaves of the less susceptible genotype Scavina 6 upon infection, and total procyanidin levels were higher than in Cacao Comum a more disease susceptible genotype. Selection of genotypes with high constitutive and/or inducible content of procyanidins in young flush leaves may be a strategy to develop witches’ broom disease resistant cultivars.     

Competition Effects Among Isolates of Fusarium culmorum Differing in Aggressiveness and Mycotoxin Production on Heads of Winter Rye

Fusarium culmorum is a phytopathogenic, toxigenic fungus causing seedling diseases, foot rot and head blight of cereals. For estimating competition effects in mixtures of two single-spore isolates, two winter rye single crosses were tested with either four isolates individually or four 1 : 1 mixtures of the same isolates in six field environments. Two isolates (FC46, FC64) were highly aggressive deoxynivalenol (DON) and 3-acetyl DON-producers, the other two (FC30, FC71) were medium aggressive nivalenol-producers. Rye heads were inoculated during flowering with conidia of pairs of isolates expressing similar (FC46 + FC64, FC30 + FC71) or contrary (FC46 + FC71, FC30 + FC64) levels of aggressiveness and similar or different concentrations and chemotypes of mycotoxins, respectively. Head blight rating and yield components relative to the non-inoculated plots were recorded as aggressiveness traits. Additionally, mycotoxin concentrations were measured in the rye grain. Random pathogen samples were re-isolated from heads at the onset of symptom development and analysed by molecular markers (RAPD–PCR) in one environment. Aggressiveness of the isolate mixtures was significantly lower than that of the isolates applied individually on both rye genotypes. Similarly, mycotoxin concentrations were significantly lower in the mixtures in seven out of eleven comparisons. Among the re-isolates, the component genotypes of a mixture significantly deviated from the inoculated 1 : 1 ratio when a particular isolate (FC46) was present in the mixture. This isolate displayed a superior competitive ability irrespective of the aggressiveness or mycotoxin profile of the mixing partner illustrating that pathogenic fitness is caused by additional factors that have not, as yet, been identified.     

An assessment of the relative resistance of three hawthorn species to three strains of <Emphasis Type="Italic">Erwinia amylovora</Emphasis> using three different inoculation methods

Fire blight is an important disease of hawthorn plants. In this study, the level of susceptibility of three hawthorn species (Crataegus monogyna, Crataegus laevigata, Crataegus persimilis) to the bacterium Erwinia amylovora was investigated. The results showed that all species were highly susceptible to this pathogen. In addition, the relative virulence of three different E. amylovora strains on the above species was examined. Variability among the strains was found, with strain 3 being the most virulent and strain 1 the least.     

Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens

An isolate of Trichoderma harzianum Rifai from an infected cacao pod produces and secretes nonanoic (pelargonic) acid into a liquid culture medium. Nonanoic acid (NA) was very inhibitory to spore germination and mycelial growth of two cacao pathogens, Crinipellis perniciosa Stahel and Moniliophthora roreri Cif. H.C. Evans. It was highly active causing 75% inhibition of spore germination in an in vitro assay at a rate as low as 0.09 μM for M. roreri and 0.92 μM for C. perniciosa. Mycelial growth was comparatively less sensitive to inhibition, but still there was a 75% reduction in growth with 0.62 μM in M. roreri and 151 μM NA in C. perniciosa. In contrast, NA did not affect Trichoderma mycelial growth or spore germination at concentrations that were inhibitory to the pathogens. 6-pentyl-α-pyrone was also produced and secreted into the medium by T. harzianum, however; it was not antagonistic to the cacao pathogens. Although a number of metabolites produced by Trichoderma spp. have been identified in the past, this is the first report of NA production and secretion by any Trichoderma. The results suggest that NA may play a role in the successful use of some Trichoderma spp. isolates in the biocontrol of fungal diseases of plants.     

Biochemical analysis of a chlorfenapyr-selected resistant strain of Tetranychus urticae Koch

Tetranychus urticae Koch has recently developed resistance to chlorfenapyr in Australia and Japan, but no attempt has yet been made to describe the biochemical mechanisms involved in chlorfenapyr resistance. In this study a laboratory-selected chlorfenapyr-resistant strain was investigated. Resistance to chlorfenapyr was associated with a strong increase in esterase activity and P450 mono-oxygenase (MO) activity but a decrease in 3,3',5,5'-tetramethylbenzidine (TMBZ) peroxidation activity. Differences in esterase activities between susceptible and resistant strains increased with increasing carbon number of the aliphatic side-chain of the nitrophenol substrate. A 4.4-fold increase in the O-deethylation of 7-ethoxy-4-trifluoromethyl coumarin (7-EFC) mediated by P450 MOs was detected. Remarkably, the resistant strain showed only half of the total TMBZ peroxidation activity found in the susceptible strain. The activity of these enzymes was further determined on different crosses and back-crosses of both strains. Results indicated that activities correlated with chlorfenapyr susceptibility and could be considered as biochemical markers. Esterase isozymes of both strains and their crosses were separated with isoelectric focusing (IEF) and visualised after activity staining. It was clear that two distinct zones of enhanced esterase activity were present in the chlorfenapyr-resistant strain (EST 11, pI = 4.88 and EST 16, pI = 4.71). EST 11 was identified with inhibitors as a carboxylesterase. The relative presence and intensity of these esterase zones changed in the different crosses and could be seen as a marker for chlorfenapyr resistance. Glutathione-S-transferase and glucose-6-phosphate dehydrogenase activities were not significantly different between strains. A twofold decrease in TMBZ peroxidase activity in the resistant strain could reflect decreased activation of chlorfenapyr. On the basis of these results the involvement of P450 MOs and esterases in the activation and detoxification of chlorfenapyr in T. urticae is challenged and discussed.     

Natural products as pesticides: Two examples of stereoselective synthesis

The strategy and synthetic efforts leading to efficient stereoselective syntheses of thiangazole, a tris-thiazolinyl-oxazole metabolite isolated from Polyangium spp., and of hydantocidin, a spironucleoside metabolite isolated from Streptomyces hygroscopicus, are discussed.     

The rapid and accurate determination of germ tube emergence site by Blumeria graminis conidia

The development of appressoria by germinating Blumeria graminis conidia depends on its germ tubes making contact with the host surface. Low angle, low temperature scanning electron microscopy showed that 80% or more of first-formed germ tubes of f. spp. hordei, tritici and avenae conidia emerged from close to the host leaf surface, and so made contact with it allowing them to become functional primary germ tubes. Light microscopy of f. sp.hordei confirmed this result and, in addition, showed that germ tubes frequently emerged close to, and contacted, various hydrophobic and hydrophilic artificial substrata. Geometric models of conidium-substratum interfaces were developed and a “surface point method” was derived to predict the frequency with which contact would result if germ tube emergence was a random phenomenon. However, observed contact frequencies were far higher (c. three to eight times) than predicted. Thus, the germ tube emergence site was determined as a response to substratum contact. In part, this appeared to be a non-specific response. Nevertheless, germ tube contact frequencies were greater on the curved surface of leaf epidermal cells than on planar surface, suggesting that specific recognition of leaf surface characteristics may promote directional emergence. The area of contact required to stimulate directional germ tube emergence was very small: contact with a microneedle tip or with a spiders’ suspension thread was sufficient for many conidia. Similarly, on leaves, the only contact is between the tips of a limited number of conidial wall projections and the edges of epicuticular leaf wax plates. Micromanipulation to roll conidia so that their original site of contact with a leaf was rotated away from it, led to the majority of first-formed germ tubes growing away from the leaf, i.e. emerging close to the site of original contact. The experiments indicated that the site of germ tube emergence is determined within 1 min of deposition. This implicated the release of conidial extracellular materials in recognition of the conidium-leaf surface contact site.     

Aerial applications of pesticides in the United Kingdom: 1978 to 1998

The aerial application of pesticides increased rapidly following the end of the Second World War, as, along with the development of a wider range of synthetic pesticides, more planes and pilots became available. It also became clear that new and previously inaccessible areas could be sprayed from the air. By the 1980s there were around 100 planes in the United Kingdom (UK) capable of spraying pesticides. However, increasing environmental pressure brought about the introduction of tougher legislation to control pesticide usage. The Food and Environment Protection Act 1985 and the Control of Pesticides Regulations 1986 (as amended) introduced tight controls over the application of pesticides from the air. Aerial application of pesticides has been monitored each year by the UK Ministry of Agriculture, Fisheries and Food (MAFF) since 1978, during which time the area treated has decreased significantly. This has been partly as a result of changes in legislation but also due to technological improvements in conventional crop‐protection methods. The area treated from the air in the UK has never risen above 2.5% of the area treated from the ground and in 1998 had fallen to less than 0.04% of the total area treated with pesticides. It is likely that this figure will continue to fall in the future. By 1998, the only pesticides to be sprayed from the air comprised fungicides, herbicides and insecticides. As a group, herbicides comprised the greatest area of treatment and the only target to show an increase in area treated in recent years has been bracken (Pteridium aquilinum (L) Kuhn). © Crown copyright 2000. Reproduced with the permission of Her Majesty's Stationery Office.     

Comparative inhibition of the juvenile hormone esterases from Trichoplusia ni,Tenebrio molitor, and Musca domestica

Twenty-seven compounds were screened as potential inhibitors of juvenile hormone esterases. Of these compounds O-ethyl-S-phenyl phosphoramidothiolate provided the best inhibition for the cabbage looper, Trichoplusia ni (Hubner), and the yellow mealworm, Tenebrio molitor L., while the juvenile hormone esterases of the house fly, Musca domestica L., were best inhibited by a juvenoid carbamate (1-(m-phenoxy-N-ethyl carbamate)-3,7-dimethyl-7-methoxy-2E-octene). The inhibition patterns of T. ni and T. molitor are similar, while those of M. domestica are relatively different. Further studies on the juvenile hormone and α-napthyl acetate esterases of T. ni showed that they could be differentially inhibited. Diisopropyl phosphorofluoridate and an alkyl trifluoromethyl ketone selectively inhibit the hydrolysis of α-naphthyl acetate and juvenile hormone, respectively, while O-ethyl-S-phenyl phosporamidothiolate inhibits both enzymes. The juvenile hormone esterases of T. ni also appear to be unique enzymes that are selective for juvenile-hormone-like molecules. The in vivo inhibition of T. ni juvenile hormone esterases by O-ethyl-S-phenyl phosphoramidothiolate slows the in vivo hydrolysis of juvenile hormone and results in delayed pupation and malformed larvae that resemble larval-pupal intermediates. Thus, the esterases involved in juvenile hormone metabolism appear to be important in juvenile hormone regulation.