Comparative growth and pathogenicity of geographical isolates of Sclerotinia sclerotiorum on lentil genotypes

Isolates of Sclerotinia sclerotiorum were collected from infected lentil plants from 2 agro-ecological zones of Syria and used to study their comparative growth on culture media and pathogenicity on different lentil genotypes. The growth studies were carried out on Potato Dextrose Agar (PDA) growth media under laboratory conditions. Mycelial radial growth and sclerotial production were the parameters used to compare the isolates. Pathogenicity studies were carried out with selected isolates on 10 lentil genotypes, infected as detached shoots and as whole potted-plants in the plastic house. The isolates showed considerable variation in cultural characteristics through mycelial growth, mycelial pigmentation and sclerotial production in the media plates. There were significant differences in the growth and sclerotial production of most of the isolates, but no apparent correlation between mycelial growth and sclerotial production among the isolates. Genotype by isolate interactions was significant for the isolates tested for pathogenicity. These interactions, however, appeared to be caused by differences in virulence of the isolates and did not suggest the occurrence of distinct pathogenic races of the pathogen isolates.

Host-delivered RNAi: An effective strategy to silence genes in plant parasitic nematodes

Root-knot nematodes (Meloidogyne spp.) are obligate, sedentary endoparasites that infect many plant species causing large economic losses worldwide. Available nematicides are being banned due to their toxicity or ozone-depleting properties and alternative control strategies are urgently required. We have produced transgenic tobacco (Nicotiana tabacum) plants expressing different dsRNA hairpin structures targeting a root-knot nematode (Meloidogyne javanica) putative transcription factor, MjTis11. We provide evidence that MjTis11 was consistently silenced in nematodes feeding on the roots of transgenic plants. The observed silencing was specific for MjTis11, with other sequence-unrelated genes being unaffected in the nematodes. Those transgenic plants able to induce silencing of MjTis11, also showed the presence of small interfering RNAs. Even though down-regulation of MjTis11 did not result in a lethal phenotype, this study demonstrates the feasibility of silencing root-knot nematode genes by expressing dsRNA in the host plant. Host-delivered RNA interference-triggered (HD-RNAi) silencing of parasite genes provides a novel disease resistance strategy with wide biotechnological applications. The potential of HD-RNAi is not restricted to parasitic nematodes but could be adapted to control other plant-feeding pests.

Impact of mirid (Creontiades spp.) (Hemiptera: Miridae) pest management on Helicoverpa spp. (Lepidoptera: Noctuidae) outbreaks: The case for conserving natural enemies

Creontiades spp. (Hemiptera: Miridae) are sucking pests that attack buds, flowers and young pods in mungbeans, Vigna radiata (L.), causing these structures subsequently to abort. If left uncontrolled, mirids can cause 25-50% yield loss. Traditional industry practice has involved prophylactic applications of dimethoate to control mirids at budding and again a week later. The present trial was initiated to highlight the dangers of such a practice, in particular the risk of a subsequent Helicoverpa spp. lepidopteran pest outbreak. A single application of dimethoate halved the population of important natural enemies of Helicoverpa spp., and caused an above-threshold outbreak of Helicoverpa spp. within 11 days. This shows that even a moderate (e.g. 50%) reduction in natural enemies may be sufficient to increase Helicoverpa spp. populations in mungbeans. As a result, prophylactic sprays should not be used for the control of mirids in mungbeans, and dimethoate should be applied only when mirids are above the economic threshold. Indoxacarb was also tested to establish its effect on Helicoverpa spp., mirids and natural enemies. Indoxacarb showed potential for Helicoverpa spp. control and suppression of mirids and had little impact on natural enemies.

Fruit maturity and soundness relevant to feeding choice by fruit-piercing moths (Lepidoptera: Noctuidae) in citrus crops in northeast Australia

Fruit-piercing moths are significant pests of a range of fruit crops throughout much of the world's tropics and subtropics. Feeding damage by the adult moths is most widely reported in varieties of citrus. In the years 2003 and 2004, fruit-piercing moth activity was observed regularly at night in citrus crops in northeast Australia, to determine the level of maturity (based on rind colour) and soundness of fruit attacked. 'Navelina' navel and 'Washington' navel orange, grapefruit and mixed citrus crops were assessed, and fruit was rated and placed into five categories: green, colouring, ripe, overripe and damaged. There were no statistical differences in the percentage of fruit attacked in each category across crops. However, within the individual crops significant proportions of green 'Navelina' fruit (58.7%) and green mixed citrus (57.1%) were attacked in 2004. Among all the crops assessed, 25.1% of moth feeding occurred on overripe or damaged fruit. Crops started to be attacked at least 8 weeks before picking, but in two crops there were large influxes of moths (reaching 27 and 35 moths/100 trees, respectively) immediately before harvest. Moth activity was most intense between late February and late March. Eudocima fullonia (Clerck) represented 79.1% of all moths recorded on fruit, with Eudocima materna (L.), Eudocima salaminia (Cramer) and Serrodes campana (Guen.) the only other species observed capable of inflicting primary damage. Our results suggest that growers should monitor moth activity from 8 weeks before harvest and consider remedial action if moth numbers increase substantially as the crop matures or there is a history of moth problems. The number of fruit pickings could be increased to progressively remove ripe fruit or early harvest of the entire crop contemplated if late influxes of moths are known.

Review of Nezara viridula (L.) management strategies and potential for IPM in field crops with emphasis on Australia

Nezara viridula (L.) is a cosmopolitan, polyphagous heteropteran that causes economic damage to many crop species. At present, control of N. viridula in Australia and other countries relies heavily upon insecticides, most of which are disruptive to beneficial insects, constituting a constraint on integrated pest management (IPM). Much research has been conducted into non-chemical control methods for N. viridula. This paper reviews the potential for and limitations of sterile insect technique, classical, inundative and conservation biological control, and trap cropping. None of these techniques appear to be adequate for control of N. viridula when used alone but there is scope for these non-chemical approaches to be adopted for use in integrated management of this pest. A proposal is given for one such integrated approach for future development. It includes biopesticides, trap crops and carefully targeted habitat manipulation to enhance arthropod natural enemies as well as area-wide management and grower education.

Analysis of the serological variability of Lettuce mosaic virus using monoclonal antibodies and surface plasmon resonance technology

A panel of 19 monoclonal antibodies (mAbs) was used to study the immunological variability of Lettuce mosaic virus (LMV), a member of the genus Potyvirus, and to perform a first epitope characterization of this virus. Based on their specificity of recognition against a panel of 15 LMV isolates, the mAbs could be clustered in seven reactivity groups. Surface plasmon resonance analysis indicated the presence, on the LMV particles, of at least five independent recognition/binding regions, correlating with the seven mAbs reactivity groups. The results demonstrate that LMV shows significant serological variability and shed light on the LMV epitope structure. The various mAbs should prove a new and efficient tool for LMV diagnostic and field epidemiology studies.

Developing African mahogany (Khaya senegalensis) germplasm and its management for a sustainable forest plantation industry in northern Australia: Progress and needs.

The demonstrated wide adaptability, substantial yield potential and proven timber quality of African mahogany (Khaya senegalensis) from plantings of the late 1960s and early 1970s in northern Australia have led to a resurgence of interest in this high-value species. New plantations or trials have been established in several regions since the early 1990s -in four regions in north Queensland, two in the Northern Territory and one in Western Australia. Overall, more than 1500 ha had been planted by early 2007, and the national annual planting from 2007-2008 as currently planned will exceed 2400 ha. Proceedings of two workshops have summarised information available on the species in northern Australia, and suggested research and development (R&D) needs and directions. After an unsustained first phase of domestication of K. senegalensis in the late 1960s to the early 1970s, a second phase began in northern Australia in 2001 focused on conservation and tree improvement that is expected to provide improved planting stock by 2010. Work on other aspects of domestication is also described in this paper: the current estate and plans for extension; site suitability, soils and nutrition; silviculture and management; productivity; pests and diseases; and log and wood properties of a sample of superior trees from two mature plantations of unselected material near Darwin. Some constraints on sustainable plantation development in all these fields are identified and R&D needs proposed. A sustained R&D effort will require a strategic coordinated approach, cooperative implementation and extra funding. Large gains in plantation profitability can be expected to flow from such inputs.

Completion of the genome sequence of Lettuce necrotic yellows virus, type species of the genus Cytorhabdovirus

We completed the genome sequence of Lettuce necrotic yellows virus (LNYV) by determining the nucleotide sequences of the 4a (putative phosphoprotein), 4b, M (matrix protein), G (glycoprotein) and L (polymerase) genes. The genome consists of 12,807 nucleotides and encodes six genes in the order 3′ leader-N-4a(P)-4b-M-G-L-5′ trailer. Sequences were derived from clones of a cDNA library from LNYV genomic RNA and from fragments amplified using reverse transcription-polymerase chain reaction. The 4a protein has a low isoelectric point characteristic for rhabdovirus phosphoproteins. The 4b protein has significant sequence similarities with the movement proteins of capillo- and trichoviruses and may be involved in cell-to-cell movement. The putative G protein sequence contains a predicted 25 amino acids signal peptide and endopeptidase cleavage site, three predicted glycosylation sites and a putative transmembrane domain. The deduced L protein sequence shows similarities with the L proteins of other plant rhabdoviruses and contains polymerase module motifs characteristic for RNA-dependent RNA polymerases of negative-strand RNA viruses. Phylogenetic analysis of this motif among rhabdoviruses placed LNYV in a group with other sequenced cytorhabdoviruses, most closely related to Strawberry crinkle virus.

Borate protection of softwood from Coptotermes acinaciformis (Isoptera: Rhinotermitidae) damage: Variation in protection thresholds explained

Laboratory and field data reported in the literature are confusing with regard to “adequate” protection thresholds for borate timber preservatives. The confusion is compounded by differences in termite species, timber species and test methodology. Laboratory data indicate a borate retention of 0.5% mass/mass (m/m) boric acid equivalent (BAE) would cause >90% termite mortality and restrict mass loss in test specimens to ≤5%. Field data generally suggest that borate retentions appreciably >0.5% m/m BAE are required. We report two field experiments with varying amounts of untreated feeder material in which Coptotermes acinaciformis (Froggatt) (Isoptera: Rhinotermitidae) responses to borate-treated radiata (Monterey) pine, Pinus radiata D. Don, were measured. The apparently conflicting results between laboratory and field data are explained by the presence or absence of untreated feeder material in the test environment. In the absence of untreated feeder material, wood containing 0.5% BAE provided adequate protection from Coptotermes sp., whereas in the presence of untreated feeder material, increased retentions were required. Furthermore, the retentions required increased with increased amounts of susceptible material present. Some termites, Nasutitermes sp. and Mastotermes darwiniensis Froggatt, for example, are borate-tolerant and borate timber preservatives are not a viable management option with these species. The lack of uniform standards for termite test methodology and assessment criteria for efficacy across the world is recognized as a difficulty with research into the performance of timber preservatives with termites. The many variables in laboratory and field assays make “prescriptive” standards difficult to recommend. The use of “performance” standards to define efficacy criteria (“adequate” protection) is discussed.

Baseline responses of Alphitobius diaperinus (Coleoptera: Tenebrionidae) to spinosad, and susceptibility of broiler populations in eastern and southern Australia

Spinosad was proposed as a potential chemical for control of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in Australian broiler houses after the detection of strong cyfluthrin resistance in many beetle populations. In 2004-2006, spinosad susceptibility of 13 beetle populations from eastern and southern Australian broiler houses and a cyfluthrin/fenitrothion-resistant reference population was determined using topical application, and was compared with the susceptibility of an insecticide-susceptible reference population. Comparisons of dose-response curves and baseline data showed that all populations, including the insecticide-susceptible population, were roughly equivalent in their response to spinosad, indicating no preexisting spinosad resistance. Two field populations, including the resistant reference population, which had confirmed cyfluthrin/fenitrothion- resistance, showed no cross-resistance to spinosad. There was no significant correlation between beetle weight and LC99.9. A discriminating concentration of 3% spinosad was set to separate resistant and susceptible individuals. Considering the levels of spinosad resistance that have been recorded in other insect pests, the sustained future usefulness of spinosad as a broiler house treatment will rely on effective integrated beetle management programs combined with carefully planned chemical use strategies.

Field assessment of the resistance of timber products to termite (Isoptera) damage: Results can change with the amount of bait wood

Whether or not termites initiate damage to timber via the end grain may determine the need for spot-treating the exposed untreated cut ends of envelope-treated softwood framing material. Australian Coptotermes acinaciformis (Froggatt) were field-tested for their ability to initiate feeding via the end grain of timber (35 × 90 mm) treated with a repellent Tanalith® T envelope. Specimens of commercial radiata pine Pinus radiata D.Don framing timber (untreated) and slash pine Pinus elliottii Englem. (untreated and envelope-treated) were partially clad in fine stainless steel mesh. Clad and unclad specimens were exposed to C. acinaciformis near Townsville, North Queensland, Australia, for four months. Results showed that this species of termite can indeed damage timber via the end grain, including exposed untreated cut ends of envelope-treated material as demonstrated earlier for different populations of C. acinaciformis. Differences between the test conditions in field trials carried out at different times (where C. acinaciformis either did or did not damage timber via the end grain) are discussed. Clearly, outcomes from field studies with preservative-treated materials are dependent upon experimental conditions. Notably, the amount of bait wood (highly termite-susceptible timber substrate) offered in a given method can strongly influence the termite response. Further investigation is required to standardise this aspect of conditions in protocols for the assessment of wood preservatives.

Host recognition by a polyphagous lepidopteran (Helicoverpa armigera): Primary host plants, host produced volatiles and neurosensory stimulation

An important question in the host-finding behaviour of a polyphagous insect is whether the insect recognizes a suite or template of chemicals that are common to many plants? To answer this question, headspace volatiles of a subset of commonly used host plants (pigeon pea, tobacco, cotton and bean) and nonhost plants (lantana and oleander) of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) are screened by gas chromatography (GC) linked to a mated female H. armigera electroantennograph (EAG). In the present study, pigeon pea is postulated to be a primary host plant of the insect, for comparison of the EAG responses across the test plants. EAG responses for pigeon pea volatiles are also compared between females of different physiological status (virgin and mated females) and the sexes. Eight electrophysiologically active compounds in pigeon pea headspace are identified in relatively high concentrations using GC linked to mass spectrometry (GC-MS). These comprised three green leaf volatiles [(2E)-hexenal, (3Z)-hexenylacetate and (3Z)-hexenyl-2-methylbutyrate] and five monoterpenes (α-pinene, β-myrcene, limonene, E-β-ocimene and linalool). Other tested host plants have a smaller subset of these electrophysiologically active compounds and even the nonhost plants contain some of these compounds, all at relatively lower concentrations than pigeon pea. The physiological status or sex of the moths has no effect on the responses for these identified compounds. The present study demonstrates how some host plants can be primary targets for moths that are searching for hosts whereas the other host plants are incidental or secondary targets.

New tools for management of phosphine resistance

Phosphine is the primary fumigant used to protect the majority of the world' s grain and a variety of other stored commodities from insect pests. Phosphine is playing an increasingly important role in the protection of commodities for two primary reasons. Firstly, use of the alternative fumigant, methyl bromide, has been sharply curtailed and is tightly regulated due to its role in ozone depletion, and secondly, consumers are becoming increasingly intolerant of contact pesticides. Niche alternatives to phosphine exist, but they suffer from a range of factors that limit their use, including: 1) Limited commercial adoption due to expense or slow mode of action; 2) Poor efficacy due to low toxicity, rapid sorption, limited volatility or high density; 3) Public health concerns due to toxicity to handlers or nearby residents, as well as risk of explosion; 4) Poor consumer acceptance due to toxic residues or smell. These same factors limit the prospects of quickly identifying and deploying a new fumigant. Given that resistance toward phosphine is increasing among insect pests, improved monitoring and management of resistance is a priority. Knowledge of the mode of action of phosphine as well as the mechanisms of resistance may also greatly reduce the effort and expense of identifying synergists or novel replacement compounds.

Gene interactions constrain the course of evolution of phosphine resistance in the lesser grain borer, Rhyzopertha dominica

Phosphine, a widely used fumigant for the protection of stored grain from insect pests, kills organisms indirectly by inducing oxidative stress. High levels of heritable resistance to phosphine in the insect pest of stored grain, Rhyzopertha dominica have been detected in Asia, Australia and South America. In order to understand the evolution of phosphine resistance and to isolate the responsible genes, we have undertaken genetic linkage analysis of fully sensitive (QRD14), moderately resistant (QRD369) and highly resistant (QRD569) strains of R. dominica collected in Australia. We previously determined that two loci, rph1 and rph2, confer high-level resistance on strain QRD569, which was collected in 1997. We have now confirmed that rph1 is responsible for the moderate resistance of strain QRD369, which was collected in 1990, and is shared with a highly resistant strain from the same geographical region, QRD569. In contrast, rph2 by itself confers only very weak resistance, either as a heterozygote or as a homozygote and was not discovered in the field until weak resistance (probably due to rph1) had become ubiquitous. Thus, high-level resistance against phosphine has evolved via stepwise acquisition of resistance alleles, first at rph1 and thereafter at rph2. The semi-dominance of rph2 together with the synergistic interaction between rph1 and rph2 would have led to rapid selection for homozygosity. A lack of visible fitness cost associated with alleles at either locus suggests that the resistance phenotype will persist in the field.

Structure and ultrastructure of the silk glands and spinneret of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

This study provides comprehensive documentation of silk production in the pest moth Helicoverpa armigera from gland secretion to extrusion of silk thread. The structure of the silk glands, accessory structures and extrusion apparatus are reported. The general schema of the paired silk glands follows that found for Lepidoptera. Morphology of the duct, silk press, muscle attachments and spigot are presented as a three-dimensional reconstruction and the cuticular crescent-shaped profile of the silk press is demonstrated in both open and closed forms with attendant muscle blocks, allowing advances in our knowledge of how the silk press functions to regulate the extrusion of silk. Growth of the spigot across instars is documented showing a distinctive developmental pattern for this extrusion device. Its shape and structure are related to use and load-bearing activity.