Pollen on-twine for food provisioning and oviposition of predatory mites in protected crops

The establishment of predatory mites on protected crops is affected by the availability of shelter and alternative food. We suggest that a pollen covered twine, coined here “pollen on-twine”, may provide these necessities when attached to the plant. We evaluated the effect of twine types and two pollen species on the establishment of Euseius scutalis (Athias-Henriot) and Amblyseius swirskii Athias-Henriot on pre-flowering pepper plants in a growth chamber. For both phytoseiids, rayon jute twine was found more beneficial while corn and oak pollen did not differ in their effect. Though populations of both predators were best promoted when twine and pollen were applied separately, E. scutalis population increased by more than tenfold and A. swirskii doubled when plants were applied with pollen on-twine. We propose that after further refining, pollen on-twine can serve as a feasible solution for predatory mite establishment on protected crops.     

Preselection of predatory mites to improve year-round biological control of western flower thrips in greenhouse crops

In spring and summer, two groups of natural enemies are successfully used for biological control of western flower thrips,Frankliniella occidentalis (Pergande) in greenhouses: phytoseiid mites (Amblyseius cucumeris (Oudemans) and, to a lesser extent,A. barkeri (Hughes)) and anthocorid bugs (Orius spp.). During winter, however, these predators often fail to control the pest. One likely cause for failure is the predators' tendency to enter diapause under short day conditions. In addition, eggs of predatory mites are generally susceptible to low humidity conditions, which often arise in greenhouses when outside temperatures drop below zero, or at bright, hot days in summer. In search for a thrips predator that is not hampered by these conditions, five subtropical phytoseiid species were selected which were known to feed on thrips:A. hibisci (Chant),A. degenerans Berlese,A. limonicus s.s. Garman and McGregor,A. scutalis (Athias-Henriot) andA. tularensis (Congdon). These species were compared toA. cucumeris andA. barkeri, with respect to the following features: (1) predation and oviposition rate with youngF. occidentalis larvae as prey, (2) oviposition rate on a diet of sweet pepper pollen, (3) drought tolerance spectrum of eggs, and (4) incidence of reproductive diapause under short day conditions. The results showed thatA. limonicus exhibited the highest predation and oviposition rates on a diet of thrips larvae. Moreover,A. limonicus females showed total absence of diapause under the conditions tested. A major disadvantage of this species was, however, that its eggs were most sensitive to low air humidity conditions. Least sensitive to low air humidity were eggs ofA. degenerans andA. hibisci. Females ofA. degenerans andA. hibisci also showed total absence of diapause, and intermediate rates of predation and oviposition, on both thrips larvae and pollen. In conclusion, we argue thatA. degenerans andA. hibisci are the most promising candidates for biological control ofF. occidentalis under conditions of low humidity and short day length. The success of these candidates remains to be shown in greenhouse experiments.     

Rearing, release and establishment of imported predatory mites to control citrus rust mite in Israel

Citrus rust mite, Phyllocoptruta oleivora (Ashmead), is a major pest of citrus in Israel. Five species of predatory mites: Amblyseius herbicolus Chant, Euseius victoriensis (Womersley), Euseius elinae (Schicha), Typhlodromus rickeri Chant and Euseius stipulatus (Athias-Henriot), were imported to improve the biological control of the pest. Mite rearing and sampling methods were developed and improved, and the predators were released. Recovery in some citrus orchards was recorded, but only E. victoriensis became established in the north of Israel.     

A novel approach to the use of genetically modified herbicide tolerant crops for environmental benefit

The proposed introduction of genetically modified herbicide tolerant (GMHT) crops, with claims of improved weed control, has prompted fears about possible environmental impacts of their widespread adoption, particularly on arable weeds, insects and associated farmland birds. In response to this, we have developed a novel weed-management system for GMHT sugar beet, based on band spraying, which exploits the flexibility offered by the broad-spectrum partner herbicides. Here, we show the results from two series of field experiments which, taken together, demonstrate that, by using this system, crops can be managed for enhanced weed and insect biomass without compromising yield, thus potentially offering food and shelter to farmland birds and other wildlife. These results could be applicable widely to other row crops, and indicate that creative use of GMHT technology could be a powerful tool for developing more sustainable farming systems in the future.     

A novel cytomedical vehicle capable of protecting cells against complement

We have developed "Cytomedicine," which consists of functional cells entrapped in semipermeable polymer, and previously reported that APA microcapsules could protect the entrapped cells from injury by cellular immune system. However, microencapsulated cells were not protected from humoral immune system. Here, we developed a novel APA microcapsule, in which APA microbeads (APA(Ba) microbeads) were modified to contain a barium alginate hydrogel within their centers in an attempt to make it more difficult for antibody and complement to permeate the microcapsules. The permeability of APA(Ba) microbeads was clearly less than that of APA microcapsules, presumably due to the presence of barium alginate hydrogel. Cells encapsulated within APA(Ba) microbeads were protected against treatment with xenogeneic anti-serum. Furthermore, murine pancreatic beta-cells encapsulated in APA(Ba) microbeads remained viable and continued to secrete insulin in response to glucose. Therefore, APA(Ba) microbeads may be a useful carrier for developing anti-complement device for cytomedical therapy.     

Toxicity of organophosphorus systemic pesticides to predator mites and prey

Three systemic OP insecticides, Di-Syston^®, Meta-Systox-R^® and Systox^® were added at six concentrations (0.002–0.032% active ingredient) to nutrient solutions in which bean plants were cultured. On all concentrations the materials were less toxic to the two-spotted spider mite,Tetranychus urticae Koch feeding on the plant than toNeoseiulus fallacis (Garman) preying on theT. urticae. Of the three, Di-Syston was the least toxic to both mite species. WhenT. urticae were cultured on detached bean leaves, placed on cellucotton in nutrient solutions containing the same pesticide concentrations, their eggs were also toxic toN. fallacis. Longevity studies in conjunction with³²P labelled foliage showed thatN. fallacis are incapable of feeding directly on bean leaves.

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Zusammenfassung Drei systemische Phosphorinsektizide, Di-Syston^®, Meta-Systox^® und Systox^®, wurden, in sechs Konzentrationen (0,002–0,032% Wirkstoff) zu Nährstofflösungen hinzugefügt, in denen Bohnenpflanzen kultiviert wurden. Für die SpinnmilbeTetranychus urticae, die an den Bohnenblättern saugte, waren die drei Insektizide in sämtlichen Konzentrationen weniger toxisch als für die RaubmilbeNeoseiulus fallacis, die sich vonT. urticae ernährte. Es ergab sich demnach keine ökologische Selektivität. Von den drei geprüften Mitteln war Di-Syston für die beiden Milben am wenigsten toxisch. Wenn manT. urticae auf abgetrennten Bohnenblättern, die sich auf Cellucotton in Nährstofflösungen mit den genannten Insektizidkonzentrationen befanden, züchtete, wurden ihre Eier fürN. fallacis ebenfalls toxisch. Lebensdauer-Versuche mit Blättern, die³²P enthielten, zeigten, daßN. fallacis nicht fähig ist, sich direkt von Bohnenblättern zu ernähren.

     

A novel method for increasing the expression level of recombinant proteins

Expression of recombinant proteins is an important step towards elucidating the functions of many genes discovered through genomic sequencing projects. It is also critical for validating gene targets and for developing effective therapies for many diseases. Here we describe a novel method to express recombinant proteins that are extremely difficult to produce otherwise. The increased protein expression level is achieved by using a fusion partner, MTB32-C, which is the carboxyl terminal fragment of the Mycobacterium tuberculosis antigen, MTB32 (Rv0125). By fusing MTB32-C to the N-termini of target genes, we have demonstrated significant enhancement of recombinant protein expression level in Escherichia coli. The inclusion of a 6xHis tag and the 128-amino acid of MTB32-C will add 13.5 kDa to the fusion molecule. Comparison of the mRNA levels of the fusion and non-fusion proteins indicated that the increased fusion protein expression may be regulated at translational or post-translational steps. There are many potential applications for the generated fusion proteins. For example, MTB32-C fusion proteins have been used successfully as immunogens to generate both polyclonal and monoclonal antibodies. These antibodies have been used to characterize cellular localization of the proteins and to validate gene targets at protein level. In addition, these antibodies may be useful in diagnostic and therapeutic applications for many diseases. If desired, the MTB32-C portion in the fusion protein can be removed after protein expression, making it possible to study protein structure and function as well as to screen for potential drugs. Thus, this novel fusion expression system has become a powerful tool for many applications.     

Studies of the impact ofEuseius species (Acari: Phytoseiidae) on citrus mites using predator exclusion and predator release experiments

To create plots with differential levels ofEuseius, malathion sprays, highly toxic to phytoseiids but of low toxicity to citrus red mite,Panonychus citri (CRM), and citrus thrips,Scirtothrips citri, were applied to blocks of Valencia oranges for three seasons (1984, 1985 and 1986) and Eureka lemons for one season (1985). Releases ofEuseius spp. were made in plots within the treated blocks. In 1984, CRM populations were highest in the treated/no release plot, lowest in the untreated and intermediate in the treated/Euseius stipulatus-release plot. In 1985, CRM populations were high in all plots in the oranges (including theE. tularensis release) except theE. stipulatus-release plot, and in all plots in the lemons except the untreated.Euseius releases in the lemons had no effect, apparently because they were followed by very hot weather. In 1986, CRM was very low in all plots.Euseius stipulatus became the dominant phytoseiid species even in theE. tularensis-release plots after a few weeks, indicating that this species colonized more readily and spread to other trees more rapidly than didE. tularensis. The results support the hypothesis thatE. stipulatus is an important mortality factor contributing to CRM control at low levels.Euseius stipulatus also may have a negative impact on the citrus rust mite,Phyllocoptruta oleivora. Citrus thrips populations were low during the entire study period and did not differ significantly between plots.     

The relationship between dietary specialism and availability of food and water on cannibalistic interactions among predatory mites in protected crops

The interplay between dietary specialism, the tolerance of food and water stress and level of cannibalism is likely to be important in determining the outcome of biological control using inundative releases of multiple natural enemies, such as phytoseiid mites in protected crops. The dietary specialist, Phytoseiulus persimilis, with a short immature development time (4-5 days) when plentiful food was available had a low ability to survive without food (5 days), even with access to water. The dietary generalists, Neoseiulus californicus, N. cucumeris and lphiseius degenerans, had longer immature development times (by up to 2 days) than P. persimilis. Survival ability differed amongst the generalist species when they were starved but provided with constant access to water. Both N. californicus and N. cucumeris survived the longest (8-10 days) and I. degenerans survived the shortest period (4 days). No negative intra-specific interaction between immatures was observed with P. persimilis when food was available and in the absence of food this species tended to starve rather than act cannibalistically. Both N. californicus and N. cucumeris showed a low degree of cannibalism between immatures, either when food was available, or when starved but given access to water. Even when food was available survival of I. degenerans fell by 30% in 4 days and remained at 60-70% for 3 further days; survival continued to decline rapidly when they were starved but provided with water. This indicates that immatures of I. degenerans could either feed on dead conspecifics or that they were capable of a degree of cannibalism. Adult females of P. persimilis did not feed on conspecific eggs even when deprived of food but provided with water. Adult female N. californicus and N. cucumeris did feed on conspecific eggs but at a low level (<1 egg per day), which occurred only after 48 h starvation. Although egg cannibalism occurred more consistently with adult female I. degenerans than with other mite species it was at a low level (<1 egg per day). If the tendency to cannibalism, not just of eggs but with more susceptible life stages such as larvae, is reduced when water is available freely this could be important in determining the interactions that occur under natural conditions.     

Selection history and epistatic interactions impact dynamics of adaptation to novel environmental stresses

In rapidly changing environments, selection history may impact the dynamics of adaptation. Mutations selected in one environment may result in pleiotropic fitness trade-offs in subsequent novel environments, slowing the rates of adaptation. Epistatic interactions between mutations selected in sequential stressful environments may slow or accelerate subsequent rates of adaptation, depending on the nature of that interaction. We explored the dynamics of adaptation during sequential exposure to herbicides with different modes of action in Chlamydomonas reinhardtii. Evolution of resistance to two of the herbicides was largely independent of selection history. For carbetamide, previous adaptation to other herbicide modes of action positively impacted the likelihood of adaptation to this herbicide. Furthermore, while adaptation to all individual herbicides was associated with pleiotropic fitness costs in stress-free environments, we observed that accumulation of resistance mechanisms was accompanied by a reduction in overall fitness costs. We suggest that antagonistic epistasis may be a driving mechanism that enables populations to more readily adapt in novel environments. These findings highlight the potential for sequences of xenobiotics to facilitate the rapid evolution of multiple-drug and -pesticide resistance, as well as the potential for epistatic interactions between adaptive mutations to facilitate evolutionary rescue in rapidly changing environments.     

Combining plant- and soil-dwelling predatory mites to optimise biological control of thrips

The efficiency of a natural enemy combination compared to a single species release for the control of western flower thrips (WFT) Frankliniella occidentalis (Pergande) on cucumber plants was investigated. Since a large part of F occidentalis seems to enter the soil passage, a joint release of the plant-inhabiting predatory mite Amblyseius cucumeris (Oudemans) that feeds on thrips first-instar larvae and the soil-dwelling predatory mite Hypoaspis aculeifer (Canestrini) that preys on thrips pupae in the ground might offer a promising approach for a holistic control strategy. Therefore, two sets of experiments were conducted in cooperation with a commercial vegetable grower where the plants in plots were infested with a defined number of larval and adult F occidentalis. Two species of natural enemies were released either synchronously or solely, and their efficacy was compared to control plots devoid of antagonists. In both experiments, the predatory mites were released twice with a density of 46 A. cucumeris/m2, and 207 H. aculeifer/m2 (low-density) in the first experiment and 528 H. aculeifer/m2 (high-density) in the second one. Population growth of all arthropod species on the plants and in the soil was quantified at regular intervals and included all soil-dwelling mites and alternative preys present in the substrate. The results showed that H. aculeifer alone had a significant impact on thrips population development only when released at high-densities, but competence was lower compared to the other antagonist treatments. The impact of A. cucumeris alone and A. cucumeris & H. aculeifer combined was similar. Thus, the pooled exploitation of natural enemies did not boost thrips control compared to the single species application of A. cucumeris (non-additive effect), which could be explained by resource competition between both predatory mite species. Species number and population size in the soil of the experimental plots both showed a high variability, a possible consequence of their interaction with released soil-dwelling predatory H. aculeifer mites. The impact of resource competition and presence of alternative preys on thrips biological control is exhaustively discussed. From our study, we can extract the subsequent conclusions: (1) the combined use of H. aculeifer and A. cucumeris cannot increase thrips control on cucumber compared to the release of A. cucumeris alone, but the overall reliability of thrips biological control might be enhanced, (2) the availability of alternative preys seemed to affect the thrips predation rate of H. aculeifer, and (3) the impact of naturally occurring soil predatory mites on the control of WFT seemed to be partial.     

A novel method for nutritional studies of phytophagous mites, using vacuated leaf disks

Because many phytophagous mites often form serious pests in agricultural crops, there have been many investigations on the effect of the nutritional condition of the host plant on the development of phytophagous mite populations. Detached leaf cultures or leaf disks have been commonly used for such studies. However, the disadvantage of these methods is that nutrient levels can only be determined after chemical analyses of the leaves. We now describe a method in which air is pumped out of a closed container which holds a nutrient solution and the leaf disk. The nutrient solution is then pushed into the leaf disk. Using this method, the effect of all sorts of nutrients on phytophagous mites can be analysed. We used the method for studies on the effect of amino acids on the fecundity of the pink tea mite.     

一种捕食螨饲养观察装置

【目的】为在室内防止捕食螨逃逸,设计一种防止逃逸行为的饲养观察装置。【方法】对比研究了传统隔水式饲养装置和改进型装置在防止逃逸行为中的效果,及改进型装置对捕食螨发育及繁殖能力有无影响。【结果】改进型装置通过添加瓶塞、棉线、盖玻片组成密闭小室,可有效防止捕食螨的逃逸,逃逸率仅为3.3%。另外,使用该装置后的捕食螨生长发育及繁殖等能力与隔水式饲养法相比未受到显著影响。【结论】本文设计的饲养观察装置可在捕食螨生物学、生态学及农药抗性等基础研究中应用。     

Evaluation of predatory mites and Acramite for control of twospotted spider mites in strawberries in north central Florida

Greenhouse and field experiments were conducted from 2003 to 2005 to determine the effectiveness of two predatory mite species, Phytoseiulus persimilis Athias-Henriot and Neoseiulus californicus (McGregor), and a reduced-risk miticide, Acramite 50 WP (bifenazate), for control of twospotted spider mite, Tetranychus urticae Koch, in strawberries (Fragaria x ananassa Duchesne). In greenhouse tests, three treatments consisting of releases of P. persimilis, N. californicus, and an untreated control were evaluated. Both species of predatory mites significantly reduced twospotted spider mite numbers below those found in the control during the first 3 wk of evaluation. However, during week 4, twospotted spider mite numbers on the plants treated with P. persimilis increased and did not differ significantly from the control. Field studies used releases of P. persimilis and N. californicus, applications of Acramite, and untreated control plots. Both N. californicus and P. persimilis significantly reduced populations of twospotted spider mite below numbers recorded in the control plots. During the 2003-2004 field season P. persimilis took longer than N. californicus to bring the twospotted spider mite population under control (< 10 mites per leaflet). Acramite was effective in reducing twospotted spider mite populations below 10 mites per leaflet during the 2003-2004 field season but not during the 2004-2005 field season, possibly because of a late application. These findings indicate that N. californicus releases and properly timed Acramite applications are promising options for twospotted spider mite control in strawberries for growers in north Florida and other areas of the southeast.     

Susceptibility to deltamethrin in the predatory mites Neoseiulus californicus and Phytoseiulus macropilis (Acari: Phytoseiidae) populations in protected ornamental crops in Brazil

Knowledge of inter and intra-specific variation in the susceptibility of natural enemies to pesticides could help to better design integrated pest management strategies. The objective of this research was to evaluate the susceptibility to deltamethrin in populations of the predatory mites Neoseiulus californicus (McGregor) and Phytoseiulus macropilis (Banks) populations collected from protected ornamental crops in Brazil. The susceptibility to deltamethrin was characterized against immature and adult stages of both species. The impact of this insecticide was also measured by estimating the intrinsic rate of increase (r _i). The immature and adult stages of N. californicus were approximately 3,600 and 3,000-fold more tolerant to deltamethrin than those of P. macropilis. However, high variability in the susceptibility to this insecticide was detected among P. macropilis populations, with resistance ratios of up to 3,500-fold. The selection of deltamethrin-resistant strains of P. macropilis could be exploited in applied biological control programs.     

A novel method for increasing bacterial cellular yields of a fimbrial subunit polypeptide

Abstract An in-depth understanding of the genetic organisation of the K88 adhesion fimbriae determinant has been used to construct novel recombinant plasmids which direct the expression of high levels of K88 fimbriae suitable for use in vaccine preparations when harboured by Escherichia coli K12. This was achieved by placing the fimbrial subunit polypeptide cistron under the control of a powerful E. coli promoter while leaving the expression of the other cistrons encoded within the K88 determinant under the control of a separate promoter. This methodology could be used as a general approach to construct strains expressing high levels of other bacterial fimbriae.     

Disease resistance in rice and the role of molecular breeding in protecting rice crops against diseases

Rice diseases (bacterial, fungal, or viral) threaten food productivity. Host resistance is the most efficient, environmentally friendly method to cope with such diverse pathogens. Quantitative resistance conferred by quantitative trait loci (QTLs) is a valuable resource for rice disease resistance improvement. Although QTLs confer partial but durable resistance to many pathogen species in different crop plants, the molecular mechanisms of quantitative disease resistance remain mostly unknown. Quantitative resistance and non-host resistance are types of broad-spectrum resistance, which are mediated by resistance (R) genes. Because R genes activate different resistance pathways, investigating the genetic spectrum of resistance may lead to minimal losses from harmful diseases. Genome studies can reveal interactions between different genes and their pathways and provide insight into gene functions. Protein–protein interaction (proteomics) studies using molecular and bioinformatics tools may further enlighten our understanding of resistance phenomena.     

A novel protecting strategy for internucleosidic phosphate and phosphorothioate groups

The utility of 2-(N-isopropyl-N-anisoylamino)ethyl group for protection of internucleosidic phosphate linkages in oligonucleotide synthesis was studied. The group demonstrated high coupling yields, favorable deprotection kinetics and a high hydrolytic stability of phosphoramidite building blocks. The mechanism of deprotection was established using a model phosphate triester.