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.     

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.     

The pest status and distribution of three cryptic blue oat mite species (Penthaleus spp.) and redlegged earth mite (Halotydeus destructor) in southeastern Australia

Earth mites are pests of crops and pastures in southeastern Australia.Recent studies show differences between earth mite species in their mode ofreproduction, preferred hosts and pesticide tolerance. This paper examines thedistribution and pest status of each species. The southeastern Australiandistribution for each species is mapped, incorporating new data from easternNewSouth Wales, South Australia and Tasmania. A new population of an undescribedspecies previously identified from northwestern Victoria was found in northernNew South Wales. CLIMEX was used to identify climatic factors limiting thedistribution of P. major and P.falcatus, the most broadly distributed species. This analysissuggests tolerance to heat and desiccation limits the inland distribution ofthese two species. A three-year survey of agricultural outbreaks indicates thatall Penthaleus species are major agricultural pestsalthough their pest status on crop types appears to differ. All speciescontributed to chemical control failures. However P.falcatus, previously identified in laboratory tests as havingincreased tolerance to pesticides, was the most common species associated withcontrol failures. A laboratory experiment indicated that mites are sometimespests on crops on which they cannot persist for a generation. Results arediscussed with respect to management of these agricultural pests.     

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.     

Understanding niche shifts: using current and historical data to model the invasive redlegged earth mite,Halotydeus destructor

Aim Niche conservatism is key to understanding species responses to environmental stress such as climate change or arriving in new geographical space such as biological invasion. Halotydeus destructor is an important agricultural pest in Australia and has been the focus of extensive surveys that suggest this species has undergone a niche shift to expand its invasive range inland to hotter and drier environments. We employ modern correlative modelling methods to examine niche conservatism in H. destructor and highlight ecological differences between historical and current distributions. Location Australia and South Africa. Methods We compile comprehensive distribution data sets for H. destructor, representing the native range in South Africa, its invasive range in Australia in the 1960s (40 yr post‐introduction) and its current range in Australia. Using MAXENT, we build correlative models and reciprocally project them between South Africa and Australia and investigate range expansion with models constructed for historical and current data sets. We use several recently developed model exploration tools to examine the climate similarity between native and invasive ranges and subsequently examine climatic variables that limit distributions. Results The invasive niche of H. destructor in Australia transgresses the native niche in South Africa, and the species has expanded in Australia beyond what is predicted from the native distribution. Our models support the notion that H. destructor has undergone a more recent range shift into hotter and drier inland areas of Australia since establishing a stable distribution in the 1960s. Main conclusions Our use of historical and current data highlights that invasion is an ongoing dynamic process and demonstrates that once a species has reached an established range, it may still expand at a later stage. We also show that model exploration tools help understand factors influencing the range of invasive species. The models generate hypotheses about adaptive shifts in H. destructor.     

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.     

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.     

Effects of seed phosphorus concentration on the emergence and growth of subterranean clover (Trifolium subterraneum)

Subterranean clover seed (Trifolium subterraneum cv. Dalkeith) with phosphorus concentrations of 0.75% (high P seed) and 0.48% (low P seed) and of uniform size (2.0–2.4 mm diameter) was used to measure the effect of seed P concentrations on seedling emergence and growth.Seedling emergence numbers were 35% greater for the high P seed, and this effect was independent of external P supply. High P seed also emerged more quickly than low P seed.Leaf emergence was faster and shoot dry weight was greater for seedlings grown from high compared with low P seed, but only when external P supply was deficient for plant growth. Phosphorus concentrations in the shoots of two-week old seedlings were 32–51% higher for high P seed, although by four weeks plants grown from high and low P seed had similar concentrations of P in their shoots. We suggest that establishing pastures using high P seed would improve both early and late season pasture production.     

Comparison of the growth rates of dinitrogen fixing subterranean clover swards with those assimilating nitrate ions

Summary Subterranean clover plants were grown as swards (about 2000 plants/m²) under controlled conditions with N provided either by N₂-fixation (NO ₃ ^– withheld) or by assimilation of NO ₃ ^– (NO ₃ ^– supplied). Crop growth rates were measured by dry matter sampling over periods of up to 70 days at PPFD values of 400–1000 mole quanta/m²/s. When NO ₃ ^– was supplied from sowing the swards grew more rapidly than when the swards were not supplied with NO ₃ ^– and plants had to establish an N₂-fixing apparatus. When inter-plant competition was reduced within the sward, a difference in growth rate in favour of NO ₃ ^– -fed plants continued for at least 50 days. When however, a closed canopy was allowed to form, the NO ₃ ^– -fed swards had more dry weight than the N₂-fed swards at the time of canopy closure but thereafter the two swards grew at similar rates at light flux densities of above about 800 mole quanta/m²/s. At light flux densities of about 400 mole quanta/m²/s N₂-fed swards had a growth rate 70–80% of that of NO ₃ ^– -fed plants. NO ₃ ^– -fed plants had a higher organic N content than did N₂-fed plants under all conditions.     

Effect of acaricide resistance on reproductive ability of the honey bee mite Varroa destructor

The reproduction of pyrethroid-resistant Varroa destructor mite, a brood parasite of honey bees, was observed in Weslaco, Texas, and the results compared with known susceptible mite populations from other studies. Seven Apis mellifera colonies that had mite populations resistant to the acaricide Apistan^™ were used. Pyrethroid-resistance was confirmed when only 17% rather than 90% of mites confined in dishes containing Apistan^™ died after 12 h of exposure. The average number of eggs laid by resistant mites invading worker and drone cells was 4.4 and 5.4 respectively. This is similar to the number of eggs laid by susceptible mites in worker (4.4–4.8) or drone (4.7–5.5) cells. Also the average number of fertilised V. destructor female mites produced by resistant mites in worker (1.0) and drone (2.1) cells were similar to the number produced by susceptible mites in worker (0.9) and drone (1.9–2.2) cells. In addition, no major differences between the resistant and susceptible mite populations were observed in either worker or drone cells when six different reproductive categories and offspring mortality rates were compared. Therefore, it appears that there is little or no reproductive fitness cost associated with pyrethroid resistance in V. destructor in Texas. This revised version was published online in July 2006 with corrections to the Cover Date.     

In vitro regeneration of commercial cultivars of subterranean clover

Regeneration of subterranean clover (Trifolium subterraneum L.) was achieved by both shoot organogenesis and somatic embryogenesis. Shoots derived via organogenesis were initiated from the hypocotyls of mature imbibed seed. The hypocotyl, including the emerging radicle, was sliced longitudinally into two halves and cultured on shoot induction medium. After 30 days, adventitious shoots were formed from the hypocotyl region while the radicle showed no development. Shoots were then subcultured onto shoot multiplication medium and finally onto a root initiation medium. Histological studies revealed that shoots arose de novo and did not originate from pre-existing meristems. In the second regeneration protocol, shoot apical meristems from young seedlings were induced to form callus. Following four to six weeks culture in the dark, somatic embryos appeared spontaneously on the calli. A majority of embryos had a well-defined root pole, two cotyledonary lobes, and were capable of germination, albeit at a low frequency. Regenerated plants obtained from both protocols appeared phenotypically normal.     

The effect of sulfur supply on the root characteristics of subterranean clover and annual ryegrass

Summary The root characteristics and sulfur absorption of Trikkala sub-clover and Wimmera ryegrass were compared in a pot experiment in which plants were grown at two levels of sulfur supply (0 and 64 mgS/pot) for a period of 40 days after emergence.Ryegrass roots had a greater weight and root weight ratio, but a lower sulfur content than sub-clover roots at both levels of sulfur supply. Ryegrass roots were longer, had a greater length per unit weight, longer root hairs, and hence a larger volume and surface area than sub-clover roots. However, when the sulfur content in the whole plant was related to the root parameters, sub-clover absorbed a far greater amount of sulfur, irrespective of whether the content was expressed on the basis of the weight, length, volume or surface area of the roots.Sulfur application had relatively little effect on the morphology of roots of either species. However, whilst sulfur application increased the weight of both shoots and roots, it decreased the root weight ratio for sub-clover but not for ryegrass. Increasing sulfur supply reduced the total length of sub-clover roots but tended to increase the length of ryegrass roots. For both species, the length per unit weight of roots decreased, and the average root diameter increased, with increasing sulfur supply. Sulfur application decreased the volume and surface area of sub-clover roots, but increased these parameters in ryegrass roots.     

Effects of phosphorus application and mycorrhizal inoculation on root characteristics of subterranean clover and ryegrass in relation to phosphorus uptake

The effects of phosphorus (P) application and mycorrhizal inoculation on the root characteristics of subterranean clover and ryegrass were examined. Phosphorus application increased total root length, root surface area and root volume of both plant species. In contrast, mycorrhizal infection only affected the root characteristics of subterranean clover. Ryegrass took up more P than non-mycorrhizal subterranean clover at all levels of application. However, mycorrhizal infection only increased P uptake by subterranean clover and there was no difference in P uptake between ryegrass and mycorrhizal subterranean clover at low levels of P application. When the P uptake was expressed on the basis of any of the root characteristics, subterranean clover were superior to ryegrass suggesting that the greater uptake of P by ryegrass is not due to a higher efficiency in absorption of P from soil solution, but rather to a large root system.     

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.     

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.     

Auxin-stimulated somatic embryogenesis from immature cotyledons of white clover

Cotyledons from immature embryos of white clover (Trifolium repens L.) cv. Osceola were exposed to 2,4-D or NAA to induce somatic embryogenesis. NAA at 10 or 20 mg 1^–1 was very inefficient at stimulating embryogenesis, while concentrations of 30 or 40 mg 1^–1 resulted in death of the explant tissue. Continuous exposure of cotyledons to 40 mg 1^–1 2,4-D resulted in somatic embryos which were arrested at the globular stage, or which underwent cycles of secondary embryogenesis, never proceeding beyond the globular stage. A 10 day exposure time to 2,4-D at the same concentration led to formation of somatic embryos, most of which had poorly developed cotyledons. Almost 10% of the somatic embryos converted into plants following transfer to medium devoid of growth regulators. Attempts to improve morphology of somatic embryos by using shorter exposure times to 2,4-D at 40 mg 1^–1, or by maintaining the 10 day exposure time while varying the concentration of 2,4-D, were not successful. Plants were obtained from all parents evaluated, although at different frequencies.     

Prolific direct plant regeneration from cotyledons of white clover

Summary A facile procedure has been developed to regenerate white clover (Trifolium repens L.) plants, rapidly and directly from cotyledon explants of 3 day old seedlings. Scanning electron microscopy and histological sectioning demonstrated that shoot meristems developed from individual epidermal cells on the adaxial surface of the cotyledonary stalk, proximal to the site of excision. Initial cell divisions occurred after 2 days of culture and regenerated plants were transferred to soil within 6–8 weeks. Regenerated plants were normal, flowered and set seed. The highest shoot regeneration frequency (an average of 20 shoots per cotyledon) was obtained using an MS based medium containing 1.0 mg 1^-1 6-benzylaminopurine and 0.05 mg 1^-1 -napthaleneacetic acid. A similar regeneration frequency was obtained from cotyledon explants taken from eight different white clover cultivars.Abbreviations BAP 6-benzylaminopurine - NAA -napthaleneacetic acid - MS Murashige and Skoog medium     

Effectiveness of the BT mite trap for detecting the storage mite pests, Acarus siro, Lepidoglyphus destructor and Tyrophagus longior

Traps have been used extensively to provide early warning of hidden pest infestations. To date, however, there is only one type of trap on the market in the U.K. for storage mites, namely the BT mite trap, or monitor. Laboratory studies have shown that under the test conditions (20 °C, 65% RH) the BT trap is effective at detecting mites for at least 10 days for all three species tested: Lepidoglyphus destructor, Tyrophagus longior and Acarus siro. Further tests showed that all three species reached a trap at a distance of approximately 80 cm in a 24 h period. In experiments using 100 mites of each species, and regardless of either temperature (15 or 20 °C) or relative humidity (65 or 80% RH), the most abundant species in the traps was T. longior, followed by A. siro then L. destructor. Trap catches were highest at 20 °C and 65% RH. Temperature had a greater effect on mite numbers than humidity. Tests using different densities of each mite species showed that the number of L. destructor found in/on the trap was significantly reduced when either of the other two species was dominant. It would appear that there is an interaction between L. destructor and the other two mite species which affects relative numbers found within the trap.The British Crowns right to retain a non-exclusive, royalty-free licence in and to any copyright is acknowledged.This revised version was published online in May 2005 with a corrected cover date.     

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.