Antixenotic resistance of subterranean clover cotyledons to redlegged earth mite, Halotydeus destructor

The short-term response of redlegged earth mite, Halotydeus destructor (Tucker) (Acarina: Penthaleidae) to cotyledons of different varieties of subterranean clover (Trifolium subterraneum subsp. subterraneum L.) was assessed by means of paired choice tests, and no-choice tests. H. destructor had lower numbers and fed less on detached cotyledons of resistant than susceptible varieties, yielding a correlation between the numbers of mites and feeding damage to the cotyledons during a three hour test period. For a number of resistant and susceptible varieties, there was a negative correlation between cotyledon deterrence in the three hour choice test and feeding damage to seedling after a two week period. Since the response of the mites to different subclover varieties occurred within three hours, it is concluded that the resistance is based on antixenosis.No evidence was obtained for antifeedant activity in organic solvent extracts from the variety DGI007 (resistant) in comparison with those from the variety Dalkeith (susceptible). Water soluble compounds from DGI007 cotyledons were preferred by mites, in feeding tests in terms of numbers, over those from Dalkeith (susceptible). Squeezed sap from the cotyledons of both varieties showed the same effects on mites as 5% glucose and were more phagostimulatory than water extracts. Mechanically damaged cotyledons of Dalkeith and DGI007 attracted more mites than the undamaged counterparts. The toughness of cotyledons in 17 varieties of T. subterraneum subsp. subterraneum was measured with a manual penetrometer. Results showed a negative correlation between toughness values and mite feeding damage scores (r²=0.752) for all varieties except S3615D (resistant). This implies a likely involvement of epidermal toughness as a contributor in the antixenotic resistance of these varieties. Other mechanisms may be involved in the resistance of S3615D.     

Predictions of summer diapause in the redlegged earth mite, Halotydeus destructor (Acari: Penthaleidae), in Australia

A prediction for the onset of a summer diapause in the eggs of the redlegged earth mite, Halotydeus destructor, was developed for Australia. In this species diapause eggs pass the summer in the cadavers of adult female mites. Adult female mites were collected for several weeks from pastures in spring at 18 sites in south-western Australia and dissected to determine the timing of the production of diapause eggs. Some sites were sampled for several years between 1990 and 1997. A model was developed to predict the time for onset of diapause. The week at which 90% of eggs were in diapause was predicted best by daylength (80.1% of the variability), then by duration of the long-term plant growing season (10.4%, of variability), leaving 9.5% due to other factors. A single chemical spray in spring 2 weeks before the production of 90% diapause eggs resulted in 99% fewer mites present in autumn 7-8 months later at three sites. The timing of the spring spray was the factor leading to successful control. This model was tested at 17 sites across the whole geographical distribution of the redlegged earth mite in Australia between 1998 and 2001. The observed week of 90% diapause was within 1 week of the predicted week on 81% of occasions, and 2 weeks earlier on 15% of occasions. A database was created for the predicted date of onset of 90% diapause for the whole distribution of the redlegged earth mite in southern Australia on a 10 km(2) grid. Australian farmers are using this for timing a spring spray to control mites in the following autumn.     

Assays for the effects of volatile compounds from artificially damaged cotyledons of subterranean clover on the redlegged earth mite, Halotydeus destructor

The effects on the redlegged earth mite (Halotydeus destructor) (Acarina: Penthaleidae) of volatile compounds released from artificially damaged cotyledons of subterranean clover (Trifolium subterraneum L.), a host plant, were investigated with different assays. Mites were repelled by the volatile compounds in flask tube and in trimmed cotyledon disc tests. No differences could be shown between different tissue amounts and between resistant and susceptible varieties. When a membrane sachet was used containing either 2-(E)-hexenal, a compound produced by damaged subclover cotyledons, or the total volatile compounds collected from damaged cotyledons of Dalkeith (susceptible) admixed with 1% glucose, mites gathered to low but were deterred by high concentrations of the compounds. Volatile compounds collected from the resistant variety DGI007 were more deterrent than those from the susceptible Dalkeith. Membrane sachets containing 30 p.p.m. of 2-(E)-hexenal and 1% glucose were less preferred than cotyledons of Dalkeith (susceptible) but were not different from DGI007 (resistant). By increasing the glucose concentration to 5%, the membrane sachets with 30 p.p.m. of 2-(E)-hexenal were preferred to cotyledons of either variety. The results indicate that the membrane feeding technique provides a sensitive way of assaying volatile compounds from damaged subclover cotyledons against the mite.     

Genetic structure of Halotydeus destructor and Penthaleus major populations in Victoria (Acari: Penthaleidae)

The redlegged earth mite (Halotydeus destructor) and the blue oat mite (Penthaleus major) are major pests of pastures and crops in southern Australia. Reproductive modes, migration rates and levels of differentiation between populations were investigated using allozyme electrophoresis. Collections were made throughout Victoria and a sample was also obtained from Western Australia. Three enzyme loci were polymorphic in H. destructor (Mdh-1, Mdh-2 and Idh). Genotype frequencies of these loci did not differ between phenotypic males and females, providing no evidence for haplodiploidy. Allele frequencies were in Hardy-Weinberg equilibrium, indicating that H. destructor is diploid and sexual. This was confirmed via crosses between males and females. Allele frequencies differed between Victorian sites, although F statistics indicated little differentiation over all loci. A sample from Western Australia did not differ in allele frequencies from the Victorian sites. Four polymorphic loci were found in P. major (Mdh-1, Mdh-2, Idh and Gpi). Only a few multilocus genotypes occurred in a sample, indicating that P. major is parthenogenic. No male P. major were found in this study. A number of colour morphs were also identified and a genetic association between genital plate colour and clonal type was found in one population of P. major. Two different body colour morphs were associated with different clonal types.     

Cold storage of Halotydeus destructor (Acari: Penthaleidae) for use in experiments

Survival of medium sized nymphal stages of redlegged earth mite Halotydeus destructor (Tucker) (mainly tritonymphs and deutonymphs) stored under low temperature (1.5°C) in sealed plastic boxes remained more than 50% after 12 days of storage, with some mites surviving for up to eight weeks. Adding fresh subclover leaves into the storage box increased the survival rate of mites from 12% to 28%, 19 days after the storage started. Mites stored for two weeks at low temperatures showed feeding activity in a screening experiment similar to mites collected directly from the field. This indicated that cold storage of redlegged earth mite can be used to build up mite numbers for large screening experiments, or to extend the period of availability of mites collected from the field. However, their reproductive ability was greatly reduced after three weeks at low temperature. Thus, care should be taken when using mites for experiments measuring reproduction. The implications of low temperatures for reducing field populations of mites in midwinter are also discussed.     

An improved method for rearing Halotydeus destructor (Acari: Penthaleidae) in the laboratory

We developed an improved method for rearing Halotydeus destructor in the laboratory. Mite numbers increased rapidly over the summer of 1992–1993 when fed on vetch, Vicia sativa cv Blanchefleur; in contrast mite numbers were low and fell over the second half of the summer of 1990–91 when fed on subterranean clover, Trifolium subterraneum cv Junee. On V. sativa at a fluctuating temperature from 11 to 18°C 200 mites produced a mean of 1308 and 1455 progeny per transfer in 2 years, a 6–7-fold rate of increase per transfer. The mean transfer time was 35 days and using the interval between transfers as a measure of generation time, the mites completed six to seven generations with no evidence for development of diapause eggs from August until April. There was considerable variability in the numbers of mites produced per pot, associated in part with the presence of Verticillium sp. fungus in 35% of the pots during transfers. Improvement in H. destructor rearing resulted from the use of V. sativa cv Blanchefleur as a food source, maintenance of high humidity and adequate ventilation within cages, and the transfer of mites to fresh food sources at early nymphal stages, which reduced the spread of fungal infections.     

Effect of alphacypermethrin and bifenthrin on the survival of five acarine predators of Halotydeus destructor (Acari: Penthaleidae)

The synthetic pyrethroids, alphacypermethrin and bifenthrin, were evaluated in the laboratory and field for their impact on the survival of five acarine predators of the pasture pest, Halotydeus destructor (Tucker). In laboratory bioassays, alphacypermethrin at the proposed field rate (5 g a.i. ha^-1) for H. destructor and the scarab beetle pest, Aphodius tasmaniae Hope caused medium-high mortality to Walzia australica Womersley (Anystidae), Bdellodes affinis Atyeo (Bdellidae), Balaustium murorum (Hermann) (Erythraeidae) and Parasitus fimetorum (Berlese) (Parasitidae) but was not toxic to Cyta latirostris (Hermann) (Bdellidae). In the field, populations of W. australica and B. affinis treated with this rate were significantly reduced but recovered after 8 weeks. Bifenthrin at 5 g a.i. ha^-1 killed 96–100% of W. australica, B. affinis, B. murorum and P. fimetorum in laboratory bioassays and significantly reduced field populations of W. australica and B. affinis for 17 weeks. In bioassays both products at 2.5 g a.i. ha^-1 permitted 10–30% survival of B. affinis and B. murorum and alphacypermethrin permitted 59% survival of W. australica. A 2.5 g a.i. ha^-1 rate of alphacypermethrin is recommended for control of H. destructor in pasture when A. tasmaniae is not present, thus minimizing the impact of chemical control on the survival of some predatory mite species.     

Diapause and implications for control of Penthaleus species and Halotydeus destructor (Acari: Penthaleidae) in southeastern Australia

Blue oat mites, Penthaleus spp., and redlegged earth mites, Halotydeus destructor (Tucker) are major winter pests of a variety of crops and pastures. In southern Australia earth mites exhibit a facultative egg diapause to survive unfavorable summer conditions. The initiation of diapause egg production in earth mites was investigated using field and shade-house experiments. Species differed in their timing of diapause. H. destructor mainly produced diapausing eggs towards the end of the active mite season in spring, although small numbers were also produced in winter. In contrast, Penthaleus major (Dugés) produced diapause eggs almost immediately after emergence in autumn and continued producing these eggs throughout the season. Penthaleus falcatus (Qin and Halliday) also produced diapause eggs in early winter, although the first appearance of these eggs was slightly later in the season than for P. major. The diapause response of an undescribed species was also somewhat later than in P. major and P. falcatus, but earlier than in H. destructor. Electrophoresis of P. major samples indicated that clones of this parthenogenic species may differ in their timing of diapause egg production, providing another potential selective factor contributing to the maintenance of clonal diversity within this group. The results highlight the importance of determining species composition when devising control strategies for earth mite outbreaks.     

Biology, ecology and control of the Penthaleus species complex (Acari: Penthaleidae)

Blue oat mites, Penthaleus spp. (Acari: Penthaleidae), are major agricultural pests in southern Australia and other parts of the world, attacking various pasture, vegetable and crop plants. Management of these mites has been complicated by the recent discovery of three cryptic pest species of Penthaleus, whereas prior research had assumed a single species. The taxonomy, population genetics, ecology, biology and control of the Penthaleus spp. complex are reviewed. Adult Penthaleus have a dark blue-black body approximately 1 mm in length, and eight red-orange legs. Within Australia, they are winter pests completing two or three generations a season, depending on conditions. The summer is passed as diapausing eggs, when long-distance dispersal is thought to occur. The Penthaleus spp. reproduce by thelytokous parthenogenesis, with populations comprising clones that differ ecologically. The three pest Penthaleus spp. differ markedly in their distributions, plant hosts, timing of diapause egg production and response to pesticides, highlighting the need to develop control strategies that consider each species separately. Chemicals are the main weapons used in current control programs, however research continues into alternative more sustainable management options. Host plant resistance, crop rotations, conservation of natural enemies, and improved timing of pesticide application would improve the management of these pests. The most cost-effective and environmentally acceptable means of control will result from the integration of these practices combined with the development of a simple field-based kit to distinguish the different mite species.     

Newly isolated bacterial strains belonging to Bacillaceae (Bacillus sp.) and Micrococcaceae accelerate death of the honey bee mite, Varroa destructor (V. jacobsoni), in laboratory assays

Newly isolated bacterial strains belonging to Bacillaceae (Bacillus sp.), Micrococcaceae and three unidentified strains were tested for their pathogenicity against the mite, Varroa destructor. The Bacillus sp. strain and two of the strains belonging to the Micrococcaceae family significantly decreased the time for 50% mortality of the mite population (up to 57%) and hence may be potential control agents. In in vitro bioassay whole cells, extracellular broth and cellular extract of the Bacillus sp. strain effectively killed the mites, suggesting that both endotoxins and exotoxins contributed to the killing.     

Predicting the distribution of the crested tinamous, Eudromia spp. (Aves, Tinamiformes)

A bioclimatic analysis of the crested tinamous was conducted to explore climatic factors underpinning the distribution of both Eudromia elegans and E. formosa and to evaluate its potential application in paleontological studies. The study utilized records throughout the entire known range of Eudromia spp. in southern South America. Relationships between 20 environmental parameters and the presence of Eudromia species were established, mapping and characterizing their spatial distribution in a geographic information system using BIOCLIM and MAXENT algorithms. The MAXENT prediction map shows a more homogeneous pattern while BIOCLIM showed a patchier pattern. The models applied here generated maps that adjust to the well-known previous distributions of both species. Nevertheless, for Eudromia elegans, the distribution predicted by MAXENT includes areas where it is actually considered absent, and the BIOCLIM prediction does not include some areas where it is presumed present. Eudromia formosa were found in warmer and wetter sites than E. elegans. Low precipitation areas were identified as suitable for Eudromia elegans. Strong differences between the climatic profiles for both Eudromia spp distributions occurred, with the precipitation the most important influence. E. formosa tolerates the highest maximum temperatures, whereas E. elegans supports the lowest temperatures.     

Feeding Life Style of Redlegged Earth Mite, Halotydeus Destructor (Acari: Penthaleidae), in pastures and the role of broad-leafed weeds

Redlegged earth mite, Halotydeus destructor, is a major pest of pastures and crops in Australia, and also feeds on lower plants on the soil surface. Feeding behaviour is reviewed, and the role of Arctotheca calendula, capeweed, in determining occurrence and abundance in pastures is investigated. Mites fed more and produced more progeny on Trifolium subterraneum (subclover) than on capeweed in non-choice experiments. In pastures with mixed species three times more mites were feeding on subclover than on capeweed foliage. However, twice as many mites were found on the soil surface under pasture patches consisting mainly of capeweed than of subclover. Patches were selected on the basis of pasture height, irrespective of plant composition. Twice as many mites were found in populations under patches of tall than short pasture. The daytime relative humidity was higher in tall than short patches, and the temperature slightly lower, making tall patches a more favourable niche for these mites. Ninety per cent of H. destructor were on the soil surface, while 10% were feeding on the upper canopy of pasture. The proportion of each population that was feeding was greater on subclover than capeweed, and subclover was a more suitable food. In Australia H. destructor occurs in regions with mixed pasture species, grown in rotation with grain crops. The ability of H. destructor to utilize foliage of a range of plant species of differing suitability for food, while living mostly on the soil surface in niches favourable for survival, has enabled it to become very abundant.     

Effectiveness of three Glomus species in protecting pepper (Capsicum annuum L.) against verticillium wilt

The objective of this work was to study the influence of three Glomus species—Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe, Glomus intraradices (Schenck and Smith) and Glomus deserticola (Trappe, Bloss, and Menge)—on the development of Verticillium-induced wilt in Capsicum annuum cv. Piquillo. Results showed that the effectiveness of arbuscular mycorrhizal fungi (AMF) as biocontrol agents varied among different Glomus species. In pepper colonized by G. intraradices the severity of the disease was even higher than that observed in non-mycorrhizal plants in terms of plant growth and pepper yield. On the other hand, the high effectiveness exhibited by G. mosseae in improving plant growth and the early beginning of the reproductive stage in these plants was not associated with great plant protection and high pepper yield in diseased plants. Only plants associated with G. deserticola had greater yield than non-mycorrhizal ones despite the lower P fertilization applied to the mycorrhizal treatment and this fact was observed in both healthy and diseased plants. It is suggested that the higher specific phosphorus uptake in Verticillium-inoculated plants associated with G. deserticola could contribute to diminish the deleterious effect of pathogen on yield. On the other hand, the possible influence of endogenous phenolics in roots on the tolerance or resistance of pepper against wilt induced by Verticillium dahliae remains unclear.     

Chaff piles of harvester ant (Messor spp.) nests in a desert ecosystem

Summary The plant species composition of the chaff piles of three species of harvester ant (Messor spp.) and the contribution of the chaff to the organic pool were studied from August 1985 to July 1987. There were distinct differences in the plant species composition of the chaff of the three species. We attribute this to the different diets of the three species, which reflect the relative sizes of their individuals and their foraging strategies. The amount of chaff accumulated varies greatly between the three species (Messor rugossus: 127–196 g · ha^–1 · y^–1;Messor ebeninus: 2823–4437 g · ha^–1 · y^–1;Messor arenarius: 2165–2535 g · ha^–1 · y^–1), although the number of nests per hectare is virtually the same. We found that the amount of chaff is related to the rate of activity and the size of the individuals of each of the three ant species. The total chaff accumulated during the study period was 19.2 kg · ha^–1, which is an important contribution to the organic matter in the soil in the Negev desert ecosystem.     

Recent advances in the biocontrol of Orobanche (broomrape) species

Parasitic broomrapes (Orobanche spp.) are majoruncontrolled weeds in the Mediterranean regions of Europe and the NearEast causing major losses to vegetable, grain legume, and sunflowercrops. Selective herbicides alone cannot provide persistent, season-longcontrol of these parasites, and much methyl bromide is used for theircontrol, where affordable. Thus they are excellent targets forbiocontrol. The recent progress by the COST 816 Orobancheworking group in this area is reviewed herein. Natural infestation bythe fly Phytomyza orobanchia of seed capsules of Orobanchecrenata parasitising faba bean halved Orobanche seedproduction while inundative releases of adults reduced it to 5%of viable seeds. The fungi Fusarium arthrosporioides E4a andF. oxysporum E1d, as well as strains of bacteria were isolatedfrom diseased, juvenile, Orobanche flower stalks. They arepathogenic to O. aegyptiaca, O. crenata and O. ramosaon most vegetable crops. A F. oxysporum f. sp.orthoceras was specifically pathogenic to O. cumana onsunflowers. All were used in various experiments with a modicum ofsuccess. Methods were developed to formulate isolated mycelia, whichcould eventually allow the use of transgenic hypervirulent pathogens inasporogenic (deletion) mutants (as a failsafe against spread).Mycotoxins were also isolated from different Fusarium and otherfungal species that kill Orobanche, and are being consideredfor direct use, or to augment other strategies. All threeFusarium spp. used have been transformed with gusand/or gfp genes allowing tracing their movement in theenvironment, and opening the way to future transformations tohypervirulence.     

Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers

An efficient micropropagation protocol produced large number of plants of the three elite banana (Musa spp.) cultivars Robusta (AAA), Giant Governor (AAA) and Martaman (AAB) from shoot tip meristem. The genetic relationships and fidelity among the cultivars and micropropagated plants as assessed by random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers, revealed three somaclonal variants from Robusta and three from Giant Governor. A total of 5330 RAPD and 2741 ISSR fragments were generated with 21 RAPD and 12 ISSR primers in micropropagated plants. The percentage of polymorphic loci by RAPD and ISSR were found to be 1.75, 5.08 in Robusta and 0.83, 5.0 in Giant Governor respectively. Among the two marker systems used, ISSR fingerprinting detected more polymorphism than RAPD in Robusta and Giant Governor with most of the primers showing similar fingerprinting profile, whereas Martaman revealed complete genetic stability.     

Review of the genusBembras Cuvier, 1829 (Scorpaeniformes: Bembridae) with description of three new species collected from Australia and Indonesia

The bembrid genusBembras Cuvier is reviewed. Five species,B. japonica Cuvier,B. adenensis Imamura & Knapp and three undescribed species, were assigned to the genus. Type species of the genus,Bembras japonica is redescribed on the basis of 36 specimens including the holotype, and three new species,B. macrolepis, B. longipinnis andB. megacephala, previously misidentified asB. japonicus, are also described on the basis of specimens collected from Australia and Indonesia.Bembras macrolepis differs from its congeners by having large body scales, a long pectoral fin with 17–19 rays and a dark blotch on slightly upper portion to middle of margin, 14–15 anal-fin rays, small head and orbit, and caudal fin with a broad vertical dark band near posterior margin.Bembras longipinnis is distinguished from other members of the genus by having a slightly long pectoral fin with 17–19 rays and lacking a small black blotch near tip of upper rays, caudal fin with a large dark spot most intense in lower lobe, 1–2 gill rakers on upper gill arch, 13–14 anal-fin rays, slightly elong ated head and small orbit.Bembras megacephala is characterized by the following combination of characters: caudal fin with several irregular narrow vertical dark bands, small orbit, pectoral fin with 19–20 rays and lacking a small black blotch near tip of upper rays, head elongate, 2–4 gill rakers on upper gill arch, 15 anal-fin rays and small body scales. A key separating the five species ofBembras is given.     

Distribution and Seasonal Abundance of Trematode Parasites (Trematoda: Allocreadiidae: <Emphasis Type="Italic">Crepidostomum</Emphasis> spp.) in burrowing-mayfly nymphs (Ephemeroptera: Ephemeridae: <Emphasis Type="Italic">Hexagenia</Emphasis> spp.) from Connecting Rivers of the Laurentian Great Lakes

Burrowing-mayfly nymphs such as Hexagenia spp. have been used extensively in North America and Europe as a biomonitoring tool to indicate mesotrophic water quality, yet infestation by associated parasites has not been well documented. We performed laboratory analysis of archived samples of Hexagenia spp. nymphs collected in 1985 and 1986 to provide base-line data on the distribution (1985) and seasonal infestation (1986) of the trematode parasite Crepidostomum spp. in Hexagenia spp. nymphs in connecting rivers between Lakes Superior and Erie of the Laurentian Great Lakes. In May and June 1985, frequency of occurrence of metacercariae was widely distributed throughout the connecting rivers (63% of 203 stations with nymphs), except in areas where nymph densities were relatively low (i.e.,≤69 nymphs/m²). Distribution was probably underestimated in the present study because of low probability (mean = 31%, range = 0–57%) of detecting infestation in a small number of collected nymphs ( ≤10) at nymph densities ≤69/m². In 1986, seasonal infestation between April and October occurred in 3.3% (627) of 18696 nymphs. Overall prevalence, mean intensity, and mean abundance of parasites at one station in the St. Marys River indicate parasite transmission occurred between June and September. This period of transmission is dependent on the life-cycle of the parasite. In addition, the life-cycle of Hexagenia spp. determines which annual cohort of nymphs is infested and therefore, the duration of infestation. Although, no impacts of infestation on Hexagenia spp. nymphs were observed in the present study, infestation intensities were high enough (≥25 metacercariae per nymph) at one station in the St. Marys River to potentially cause tissue damage in a high proportion (53%) of infested nymphs.     

Influence of nitrogen fertilisation on the population of diazotrophic bacteria Herbaspirillum spp. and Acetobacter diazotrophicus in sugar cane (Saccharum spp.)

We report studies on the possible effects of fertilisation with high level of N (300 kg of N ha^-1) on the occurrence and numbers of the diazotrophic bacteria Herbaspirillum spp. and Acetobacter diazotrophicusin sugar cane plants. In the sugar cane genotype SP79-2312, the N fertilised plants generally showed higher concentrations of this element. These same plants also had lower numbers of A. diazotrophicus, while the population of Herbaspirillum spp. was not affected by N application. These differences in the concentration of N and the numbers of A. diazotrophicus due to N application were not shown in the variety SP70-1143. The numbers of A. diazotrophicus were also shown to be influenced by the harvest time, becoming reduced in the harvests that coincided with dry periods of the year.     

Fungi associated with storage rots of cocoyams (Colocasia spp.) in Nsukka,Nigeria

Cocoyam (Colocasia spp.) corms and cormels showing spoilage symptoms were collected from many stores in Nsukka locality and examined for rot and associated fungal pathogens. Aspergillus niger, Botryodiplodia theobromae, Corticium rolfsii, Geotrichum candidum, Fusarium oxysporum, and F. solani were recovered from rotten cocoyams. The representative isolates of these species caused cocoyam rot in pathogenicity tests. The rot due to A. niger, B. theobromae and C. rolfsii was extensive resulting in complete maceration of cocoyam tissue. Potassium sorbate (0.1 mg/ml) protected cocoyams from fungal rot with the exception of C. rolfsii.