Occidentalis Females Research Articles

Characterization of Frankliniella occidentalis and Frankliniella bispinosa (Thysanoptera: Thripidae) Injury to Strawberry.

Frankliniella flower thrips are pests of numerous fruit and vegetable crops as they feed and reproduce in the flowers and fruits. The invasive Frankliniella occidentalis (Pergande) from the Southwestern United States, an economic pest in Florida since 2005, and the native Frankliniella bispinosa Morgan are both found in Florida strawberries. The objective of this research was to characterize injury to strawberry fruit from these species. Densities of 0, 2, 4, 8, 16, or 32 adult females of each species were caged on a strawberry plant with one flower. The cages were removed after 2, 5, or 10 d, and the number of surviving adults and the number of larvae determined. Injury was quantified on the mature fruit 21 d after thrips were introduced onto the experimental plant. Initial densities as low as two adult F. occidentalis females per flower decreased fruit set and increased cat-facing on the fruits that developed. There were no significant effects on fruit set and cat-facing by any initial density of F. bispinosa. Feeding by thrips on the small, green fruit affected the size and shape of the mature fruit: the diameters were decreased as a linear function of increased feeding by F. bispinosa, and the weights were decreased as a linear function of increased feeding by F. occidentalis. Overall, results showed that F. occidentalis was more damaging to strawberry than F. bispinosa. The differences in pest status between these common flower thrips species in Florida presents challenges to management programs.

Rosmarinus officinialis L. (Lamiales: Lamiaceae), a Promising Repellent Plant for Thrips Management.

A number of thrips species are among the most significant agricultural pests globally. Use of repellent intercrop plants is one of the key components in plant-based 'push-pull' strategies to manage pest populations. In this study, the behavioral responses of three thrips species, Frankliniella occidentalis (Pergande), Frankliniella intonsa (Trybom), and Thrips palmi Karny (Thysanoptera: Thripidae) to Rosmarinus officinalis were investigated in Y-tube olfactometer bioassays and cage experiments. In addition, the major volatile compounds from rosemary were identified and the effect of the individual compounds on thrips behavior was evaluated. Females and males of the three thrips species were significantly repelled by the volatiles from cut rosemary leaves. The presence of rosemary plants significantly reduced settlement of females of the three thrips species and eggs laid by F. occidentalis females on target host plants. In total, 47 compounds were identified in the volatiles collected from the cut leaves of rosemary plants. The responses of the three thrips species to 10 major volatile compounds showed significant differences. However, α-pinene, the most abundant volatile, was repellent to F. occidentalis and F. intonsa. Eucalyptol, the second most abundant volatile, showed significant repellent activity to all the three thrips species. Our findings showed that rosemary is a promising repellent plant against the three thrips pests we tested, which could be a good candidate for 'push' plants in plant-based 'push-pull' strategies. The identified volatile compounds that accounted for the repellent activity could be developed as repellents for sustainable thrips management.

Fumigant effect of essential oils on mortality and fertility of thrips Frankliniella occidentalis Perg.

The western flower thrips (Frankliniella occidentalis Perg.) is one of the most economically important insect pests of greenhouse plants. Plant protection against this pest is based predominantly on synthetic insecticides; however, this form of protection poses problems in terms of thrip resistance to the active substances, along with health risks associated with insecticide residues on the treated plants. Therefore, new active substances need to be sought. Essential oils could be a new, appropriate, and safe alternative for greenhouse culture protection. As greenhouses are enclosed areas, fumigation application of EOs is possible. This paper presents acute toxicity results for 15 commercial EOs applied by fumigation, as well as the effect of sublethal concentrations on fertility of F. occidentalis females. The most efficient EOs were obtained from Mentha pulegium and Thymus mastichina, with LC50(90) estimated as 3.1(3.8) and 3.6 (4.6) mg L-1 air, respectively. As found for the very first time, sublethal concentrations of EOs could result in a significant reduction in the fertility of surviving T. occidentalis females. Among the tested EOs, the EO from Nepeta cataria provided the highest inhibition of fertility, with EC50(90) estimated as 0.18 (0.36) mg L-1 air. Chemical composition of the most efficient EOs and possible applications of the results in practice are discussed. In conclusion, in light of the newly determined facts, EOs can be recommended as active substances for botanical insecticides to be applied against Thysanopteran pests by fumigation.

Laboratory and field investigation on the orientation of Frankliniella occidentalis (Thysanoptera: Thripidae) to more suitable host plants driven by volatiles and component analysis of volatiles.

Differences in population densities of Frankliniella occidentalis among different plant species suggest a preference for particular hosts. Host plant volatiles are often involved in insect fitness. However, few studies have explored the interaction between flower volatiles and fitness. The host fitness of F. occidentalis for different flowers was assessed through field investigation, and the olfactory preference of thrips for flower volatiles was tested in the laboratory. In the field study, 18 flower host plants were classified at four fitness levels by F. occidentalis. Olfactory tests showed that female F. occidentalis had significantly different preferences for different plant odours among the volatiles of the four tested flowers (each representing the four fitness levels), whereas male F. occidentalis did not. The relative response of F. occidentalis females to flower volatiles (Rosa rugosa > Dianthus caryophyllus > Gerbera jamesonii > Pelargonium hortorum) was consistent with the field performance of F. occidentalis. In total, 23, 29, 16 and 26 components were identified in the volatile profiles of R. rugosa, D. caryophyllus, G. jamesonii and P. hortorum, respectively. 3,5-Dimethoxytoluene (24.94%), nonanal (30.42%), (E)-3-penten-2-one (52.31%) and zingiberene (29.88%) were the single most abundant components of the volatiles of R. rugosa, D. caryophyllus, G. jamesonii and P. hortorum, respectively. Volatiles are important in attracting F. occidentalis to suitable hosts, and differences in the types and concentrations of volatile components among flowers may directly influence the olfactory responses and field performance of thrips. Potential attractants could be developed for integrated pest management programmes against this pest. © 2018 Society of Chemical Industry.

Continuous exposure to the deterrents cis-jasmone and methyl jasmonate does not alter the behavioural responses of Frankliniella occidentalis.

Behavioural responses of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), a generalist, cell sap-feeding insect species with piercing-sucking mouthparts, after continuous exposure to two deterrent secondary plant compounds are investigated. We compared in choice assays on bean leaf discs, the settling, feeding, and oviposition preferences of F. occidentalis females that had no experience with the two fatty acid derivatives methyl jasmonate and cis-jasmone before testing (naïve thrips) vs. females that had been exposed to the deterrent compounds before testing (experienced thrips). The thrips were exposed to the deterrents at low or high concentrations for varied time periods and subsequently tested on bean leaf discs treated with the respective deterrent at either a low or a high concentration. Frankliniella occidentalis females avoided settling on the deterrent-treated bean leaf discs for an observation period of 6 h, independent of their previous experience. Our results demonstrate that feeding and oviposition deterrence of the jasmonates to the thrips were not altered by continuous exposure of the thrips to the jasmonates. Habituation was not induced, neither by exposure to the low concentration of the deterrents nor by exposure to the high concentration. These results indicate that the risk of habituation to two volatile deterrent compounds after repeated exposure is not evident in F. occidentalis. This makes the two compounds potential candidates to be integrated in pest management strategies.

Cloning and Functional Characterization of Two BTB Genes in the Predatory Mite Metaseiulus occidentalis.

Proteins containing the BTB (Bric-à-brac, tramtrack, and Broad Complex) domain typically share low sequence similarities and are involved in a wide range of cellular functions. We previously identified two putative and closely related BTB genes, BTB1 and BTB2, in the genome of the predatory mite Metaseiulus occidentalis. In the current study, full-length BTB1 and BTB2 cDNAs were cloned and sequenced. BTB1 and BTB2 encode proteins of 380 and 401 amino acids, respectively. BTB1 and BTB2 proteins each contain an N-terminal BTB domain and no other identifiable domains. Thus, they belong to a large category of BTB-domain proteins that are widely distributed in eukaryotes, yet with largely unknown function(s). BTB1 and BTB2 gene knockdowns in M. occidentalis females using RNAi reduced their fecundity by approximately 40% and 73%, respectively, whereas knockdown had no impact on their survival or the development of their offspring. These findings suggest these two proteins may be involved in processes related to egg production in this predatory mite, expanding the list of functions attributed to these diverse proteins.

Asymmetrical sexual isolation but no postmating isolation between the closely related species Drosophila suboccidentalis and Drosophila occidentalis.

BackgroundDuring the speciation process several types of isolating barriers can arise that limit gene flow between diverging populations. Studying recently isolated species can inform our understanding of how and when these barriers arise, and which barriers may be most important to limiting gene flow. Here we focus on Drosophila suboccidentalis and D. occidentalis, which are closely related mushroom-feeding species that inhabit western North America and are not known to overlap in geographic range. We investigate patterns of reproductive isolation between these species, including premating, postmating prezygotic, and postzygotic barriers to gene flow.ResultsUsing flies that originate from a single population of each species, we find that the strength of premating sexual isolation between these species is asymmetric: while D. occidentalis females mate with D. suboccidentalis males at a reduced but moderate rate, D. suboccidentalis females discriminate strongly against mating with D. occidentalis males. Female hybrids will mate at high rates with males of either species, indicating that this discrimination has a recessive genetic basis. Hybrid males are accepted by females of both species. We do not find evidence for postmating prezygotic or postzygotic isolating barriers, as females use the sperm of heterospecific males and both male and female hybrids are fully fertile.ConclusionsPremating isolation is substantial but incomplete, and appears to be the primary form of reproductive isolation between these species. If these species do hybridize, the lack of postzygotic barriers may allow for gene flow between them. Given that these species are recently diverged and are not known to be sympatric, the level of premating isolation is relatively strong given the lack of intrinsic postzygotic isolation. Further work is necessary to characterize the geographic and genetic variation in reproductive isolating barriers, as well as to determine the factors that drive reproductive isolation and the consequences that isolating barriers as well as geographic isolation have had on patterns of gene flow between these species.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-015-0328-y) contains supplementary material, which is available to authorized users.

Clathrin heavy chain is important for viability, oviposition, embryogenesis and, possibly, systemic RNAi response in the predatory mite Metaseiulus occidentalis.

Clathrin heavy chain has been shown to be important for viability, embryogenesis, and RNA interference (RNAi) in arthropods such as Drosophila melanogaster. However, the functional roles of clathrin heavy chain in chelicerate arthropods, such as the predatory mite Metaseiulus occidentalis, remain unknown. We previously showed that dsRNA ingestion, followed by feeding on spider mites, induced systemic and robust RNAi in M. occidentalis females. In the current study, we performed a loss-of-function analysis of the clathrin heavy chain gene in M. occidentalis using RNAi. We showed that ingestion of clathrin heavy chain dsRNA by M. occidentalis females resulted in gene knockdown and reduced longevity. In addition, clathrin heavy chain dsRNA treatment almost completely abolished oviposition by M. occidentalis females and the few eggs produced did not hatch. Finally, we demonstrated that clathrin heavy chain gene knockdown in M. occidentalis females significantly reduced a subsequent RNAi response induced by ingestion of cathepsin L dsRNA. The last finding suggests that clathrin heavy chain may be involved in systemic RNAi responses mediated by orally delivered dsRNAs in M. occidentalis.

Role of silk webbing in the biology of Hemicheyletia wellsina (Acari: Cheyletidae)

Hemicheyletia wellsina is a predator of several families of arthropod plant pests. Hemicheyletia wellsina females produce silk webbing that covers their egg clutches, but the function of the webbing is unknown. Experiments indicate the webbing can protect the eggs from predation, resulting in approximately a 71% increase in egg hatch compared to when silk was removed in the presence of the phytoseiid predator Metaseiulus occidentalis. Within 2–3 days after larvae hatch, they appear to no longer need protection, as 18 of 20 H. wellsina larvae captured and fed on the much larger adult M. occidentalis females. The silk also may regulate the relative humidity around the eggs.

Oral delivery of double-stranded RNA induces prolonged and systemic gene knockdown in Metaseiulus occidentalis only after feeding on Tetranychus urticae

Metaseiulus (=Typhlodromus or Galendromus) occidentalis is an important biological control agent. Functional genomic studies on this predator have been hampered by the lack of reverse genetic tools such as RNA interference (RNAi). In the current study, we evaluated possible RNAi responses in M. occidentalis females by feeding double-stranded RNA (dsRNA) of RpL11, RpS2, RpL8, or Pros26.4 genes in 20% sucrose solution. Females needed to subsequently feed on two-spotted spider mites (Tetranychus urticae) to elicit a nearly complete loss of egg production. The corresponding gene knockdown was robust, long-term, and was observed in the very few eggs produced (systemic or parental RNAi). Interestingly, dsRNA-mediated gene knockdown could not be induced if these predators were provided only the sucrose diet after ingesting dsRNAs; T. urticae had to be provided to elicit the RNAi response. However, the spider mite diet was not needed for sustaining the dsRNA-mediated gene knockdown once it commenced. Oral delivery of dsRNA will be a valuable tool for efficient genome-wide functional screens in this important predatory mite.

Evaluation of proxies for quality of Metaseiulus occidentalis (Acari: Phytoseiidae) reared on different stages of Tetranychus urticae (Acari: Tetranychidae)

Experiments were conducted to determine if two different methods for assessing quality of the predatory mite Metaseiulus (=Galendromus or Typhlodromus) occidentalis (Nesbitt) (Acari: Phytoseiidae) could be used instead of the more-lengthy and tedious life-history assessment method. First, an experiment was conducted to confirm it was possible to develop females of confirmed different quality by rearing immatures on two diets (eggs only or actives only of Tetranychus urticae Koch (Acari: Tetranychidae). Metaseiulus occidentalis females had a higher total fecundity (15.6 vs. 8.5 eggs female−1), a higher daily fecundity (2.2 vs. 1.4 eggs female−1 day−1), a higher survival rate (100% vs. 56.7%) over 7days of evaluation, and a higher rate of developmental success of their progeny (96.4% vs. 69.5%) when fed eggs only rather than actives only, respectively. The results confirmed adult females of known different quality could be produced for evaluating potential quality-control assessment methods. Two potential quality-control metrics were investigated using these females: survival time of females when held without food and water, and size (length and width) of their dorsal shields. Starvation time and body size were influenced significantly by both diet type and mating status, but the differences obtained were insufficient to use as proxies for evaluating the quality of the M. occidentalis females of known different quality.

Cardinium is associated with reproductive incompatibility in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae)

Cardinium, a bacterium from the Bacteroidetes group, is associated with reproductive manipulations such as cytoplasmic incompatibility, parthenogenesis, and feminization in some arthropod species. We, and others, have shown that Cardinium, but not Wolbachia, is an endosymbiont in some populations of Metaseiulus occidentalis, a phytoseiid mite that is an important predator of spider mite pests of agricultural crops. However, the precise biological effects that Cardinium may have on M. occidentalis remain unclear. In this study we show, in two sets of crosses between different colonies of Cardinium-free (C−) M. occidentalis females and Cardinium-containing (C+) males, that fecundity was reduced in parental females, F1 progeny survival rates were reduced, and fewer female progeny were produced when compared to the reciprocal and control crosses. There were no differences in these attributes in the reciprocal and two control crosses. Cardinium was transmitted maternally but there was no observed paternal transmission. Finally, Cardinium did not cause asexual (thelytoky) reproduction in M. occidentalis. Thus, Cardinium is associated with nonreciprocal reproductive incompatibility in M. occidentalis and our results support the hypothesis that Cardinium is a reproductive parasite in this agriculturally important predator. Cardinium may therefore affect the evolution and ecology of M. occidentalis and biological control efforts using this mite.

Fruit fly larval trail acts as a cue in the host location process of the pupal parasitoid Coptera occidentalis

The pupal parasitoid Coptera occidentalis Muesebeck (Hymenoptera: Diapriidae) was reared in the 1970s in California for the biological control of the Walnut husk fly, Rhagoletis completa Cresson (Diptera: Tephritidae) and later introduced into Europe to be used as a biological control agent against fruit flies. Some behavioral aspects relating to the host location process were investigated in the laboratory in order to improve the chance of success in possible biological control programmes. The searching behavior of the female parasitoid was examined in an open arena in order to evaluate the role of larval trails or rotten-fruit trails (walnut) in the host location process. The response of C. occidentalis to four Tephritid hosts was evaluated: the natural host R. completa, the factitious host Ceratitis capitata (Wiedemann) and two fruit flies of economic importance, Rhagoletis cerasi Linnaeus and Bactrocera oleae (Rossi). In order to assess the response of the parasitoid, a number of behavioral parameters were considered, such as: arrestment behavior, resident time, walking distance, linear and angular speed. The larval trails of R. completa, as well as rotten-fruit liquid trails, were clearly detected by C. occidentalis females, even though the species has been reared for more than 80 generations on C. capitata. In addition, bioassays performed with C. capitata larvae showed a good behavioral response of parasitoid females to the larval trails. Finally, R. cerasi and B. oleae larval trails were barely detected in comparison to the traces left by R. completa and C. capitata larvae. The results are discussed in the light of the possible use of C. occidentalis in biological control programmes.

Extended starvation reduced and eliminated Wolbachia, but not Cardinium, from Metaseiulus occidentalis females (Acari: Phytoseiidae): A need to reassess Wolbachia’s status in this predatory mite?

The presence of Wolbachia and Cardinium bacteria has been documented in many arthropod species, including the predatory mite Metaseiulus (=Typhlodromus or Galendomus) occidentalis (Nesbitt) (Acari: Phytoseiidae). We show that Tetranychus urticae, the prey of Metaseiulus occidentalis, contains Wolbachia and no detectable Cardinium using quantitative PCR (qPCR). Starvation for 72h at 22°C eliminated most, if not all, Wolbachia in M. occidentalis adult females from 7 laboratory colonies. Refeeding of M. occidentalis with T. urticae after starvation for 72h restored the amounts of Wolbachia in M. occidentalis to those of prestarvation levels, suggesting that Wolbachia detected in M. occidentalis starved for shorter periods of time in current, and some previous, studies likely came from T. urticae. Furthermore, eggs from all M. occidentalis colonies examined were free of Wolbachia if they were surface-decontaminated with 0.3% sodium hypochlorite before DNA extraction. Cardinium was present in 6 of 14 laboratory colonies of M. occidentalis. Starvation for 3, 24, 48, and 72h had no effect on the amounts of Cardinium in adult females from the Cardinium-positive colonies. Eggs from these colonies were positive for Cardinium but contained less than 1% of the titers found in adult females. The data suggest that Cardinium, but not Wolbachia, is an endosymbiont in certain populations of M. occidentalis. In light of our current findings, we recommend specific practices for the identification of potential symbionts in predatory arthropod species using the PCR.

Variation within and betweenFrankliniellaThrips Species in Host Plant Utilization

Anthophilous flower thrips in the genus Frankliniella (Thysanoptera: Thripidae) exploit ephemeral plant resources and therefore must be capable of successfully locating appropriate hosts on a repeated basis, yet little is known of interspecific and intraspecific variation in responses to host plant type and nutritional quality. Field trials were conducted over two seasons to determine if the abundance of males and females of three common Frankliniella species, F. occidentalis (Pergande), F. tritici (Fitch) and F. bispinosa (Morgan), their larvae, and a key predator, Orius insidiosus (Say) (Hemiptera: Anthocoridae) were affected by host plant type and plant nutritional quality. Two host plants, pepper, Capsicum annuum L. (Solanales: Solanaceae) and tomato, Solanum lycopersicum L. that vary in suitability for these species were examined, and their nutritional quality was manipulated by applying three levels of nitrogen fertilization (101 kg/ha, 202 kg/ha, 404 kg/ha). F. occidentalis females were more abundant in pepper than in tomato, but males did not show a differential response. Both sexes of F. tritici and F. bispinosa were more abundant in tomato than in pepper. Larval thrips were more abundant in pepper than in tomato. Likewise, O. insidiosus females and nymphs were more abundant in pepper than in tomato. Only F. occidentalis females showed a distinct response to nitrogen fertilization, with abundance increasing with fertilization. These results show that host plant utilization patterns vary among Frankliniella spp. and should not be generalized from results of the intensively studied F. occidentalis. Given the different pest status of these species and their differential abundance in pepper and tomato, it is critical that scouting programs include species identifications for proper management.

Effects of transplants and extracts of thoracic nerve cord–ganglia on gonad maturation of penaeoid shrimp

It has been established recently that interspecific and intraspecific thoracic ganglia transplants from Penaeidae are gradually absorbed by the host without activating an encapsulation mechanism. Therefore, this research was designed to evaluate the thoracic ganglia extracts and implants from maturing Trachypenaeus byrdi (Burkenroad), Xiphopenaeus riveti (Bouvier) and Penaeus (Litopenaeus) occidentalis (Streets) females as potential inducers of sexual maturation in Penaeus (Litopenaeus) stylirostris (Stimpson), Penaeus (Litopenaeus) vannamei (Boone) and T. byrdi, from the Gulf of Nicoya, Costa Rica. Our findings suggest that interspecific and intraspecific thoracic ganglia extracts or implants from maturing penaeoid females are not capable of inducing a clear response in sexual maturation in males or females. Tissues were tested at increasing doses from 137, 386, 525 and 1500 μg g−1 body weight, without any positive response. It is proposed that a hypothetical hormone, vitellogenesis-stimulating hormone, from the thoracic ganglia, is under the strong negative control of eyestalks, by the gonad-inhibiting hormone in the subgenus Litopenaeus. Therefore, the use of thoracic ganglia extracts or implants would be ineffective when compared with injecting serotonin alone, as the present results seem to support.

Influence of salicylaldehyde and methyl salicylate on post‐landing behaviour of Frankliniella occidentalis Pergande

Abstract:This study investigates behavioural responses of adult western flower thrips (Frankliniella occidentalis Pergande; Thysan., Thripidae) females to direct contact with repellent phenylpropanoid plant compounds (salicylaldehyde and methyl salicylate) applied on bean and cucumber leaves. The residence time of F. occidentalis females until take off was significantly shorter on bean or cucumber leaf discs treated with salicylaldehyde at 1% concentration compared with control leaf discs. A methyl salicylate (1%) treatment of cucumber resulted in shorter time periods until thrips took off the treated leaf discs compared with the control leaf discs. In a choice experiment thrips avoided to settle on a 1% salicylaldehyde treatment of bean and cucumber leaf discs for a maximum of 3 h, on a 1% methyl salicylate treatment for a 5‐h period. Within a 24‐h period neither the egg‐laying nor the feeding activity of F. occidentalis was affected after salicylaldehyde application (0.1%, 1%) on bean or cucumber. In contrast, methyl salicylate (1%) applied on bean and cucumber significantly prevented thrips females from oviposition and reduced the percentage of damaged area caused by their feeding activity for 24 h. As olfactory repellent plant volatiles applied on crop plants may elicit diverse post‐landing responses of F. occidentalis, short‐ and long‐term effects should be considered when evaluating the factual applicability of secondary plant compounds in a successful thrips management strategy.

Microbial diversity in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae) and its prey, Tetranychus urticae (Acari: Tetranychidae)

Abstract Microorganisms associated with the predatory mite Metaseiulus (= Typhlodromus or Galendromus ) occidentalis (Nesbitt) and its prey, the two-spotted spider mite Tetranychus urticae (Koch), were assessed using a high-fidelity polymerase chain reaction (PCR) protocol and primers designed to identify Eubacteria, Archaeabacteria, iridoviruses, Helicosporidia, Cytophaga -like microorganisms, Wolbachia and its bacteriophage WO, fungi and yeast-like organisms. Sequences from four bacterial species related to Wolbachia (α-Proteobacteria), Cardinium , Bacteroidetes , and Enterobacter (γ-Proteobacteria) were obtained from M. occidentalis , and three sequences related to Wolbachia , Rickettsia , and Caulobacter (α-Proteobacteria) were obtained from T. urticae . No nucleotide differences were detected between the 16S rRNA, wspA or wspB Wolbachia sequences obtained from M. occidentalis and T. urticae , which suggest that horizontal transfer of Wolbachia could have occurred. Southern blot analyses of genomic DNA from both M. occidentalis and T. urticae using wspA probes were negative, indicating that this Wolbachia sequence is not integrated into the nuclear genome of either species. Two of the T. urticae colonies tested contained the WO bacteriophage, but none of the six M. occidentalis populations were infected. New M. occidentalis -specific forward and reverse 16S rRNA primers based on the Wolbachia , Cardinium , Bacteroidetes , and Enterobacter sequences obtained were designed and used to amplify PCR products from each of two laboratory and four field-collected samples of M. occidentalis females and eggs, indicating that these infections are widespread. Likewise, species-specific primers for T. urticae were designed for the Wolbachia , Rickettsia , and Caulobacter sequences obtained and used to evaluate T. urticae from strawberries, wine grapes, hops, almonds, and cherries from California, Washington, and Florida; all were positive for Wolbachia and Caulobacter but two of the six were negative for Rickettsia . None of the M. occidentalis colonies tested were positive for the microsporidium Oligosporidium occidentalis , which previously had been associated with a pathogenic condition in some of our laboratory colonies. The Gainesville colonies of M. occidentalis and T. urticae were negative for iridovirus, Archaeabacteria, fungi, Helicosporidia, and yeast-like organisms. So far, Wolbachia is the only symbiont that is shared by this predator and its prey.

Effects of fungicide residues on the survival, fecundity, and predation of the mites Tetranychus urticae (Acari: Tetranychidae) and Galendromus occidentalis (Acari: Phytoseiidae).

Representative fungicides from three or four families used for management of powdery mildew and other diseases in tree fruits were evaluated for their effects on a common spider mite and predator mite species, respectively. A modified Munger cell technique was effective in measuring the response of phytophagous and predaceous mites to fungicide residues on detached leaves in the laboratory. Demethylation-inhibiting (DMI) (imidazole [triflumazole] and triazole [myclobutanil]) and strobilurin (trifloxystrobin) fungicides were not toxic to female Tetranychus urticae Koch and Galendromus occidentalis (Nesbitt), and no sublethal effects were found on fecundity and predation rate after 3-5-d exposure to residues. Benomyl, a benzimidazole fungicide, increased adult mortality and reduced fecundity for both mite species; however, it did not alter the predation rate of G. occidentalis females on T. urticae eggs and larvae. Female G. occidentalis that survived the lethal effects of benomyl and the comparison acaricide pyridaben were unimpaired in predation. Our results for benomyl substantiate those of earlier studies and provide evidence for nontoxic effects of DMI and strobilurin fungicides on mites. We propose that DMI and strobilurin fungicides are a good fit for integrated mite management programs due to conservation of phytoseiid predatory mites.

Extrachromosomal Plasmid Dna Transmission and Amplification in Metaseiulus occidentalis (Acari: Phytoseiidae) Transformants Generated by Maternal Microinjection

The plasmid pJKP2 (12.5 kb) was injected into Metaseiulus occidentalis females using a maternal microinjection procedure (Presnail and Hoy 1992). DNA transmission to their progenies was followed by the polymerase chain reaction (PCR) for several generations. After selecting and retaining individuals that tested positive by the PCR for 5 generations, individual females were used to initiate 12 separate transgenic lines (T1-T12). Southern blot analysis performed 10 generations after injection indicated that in at least 8 lines the plasmid DNA was transmitted extrachromosomally, and in 5 of these lines the extrachromosomal plasmid DNA was highly amplified with an estimated 10 million copies per female. The longevity, fecundity, immature mortality, and sex ratio of these transgenic lines were not different from the wild-type control under laboratory conditions. However, after 20 generations, few transformants tested positive by the PCR and Southern blot analysis indicated no distinct banding pattern. Results from Southern blot analysis and the PCR indicated the extrachromosomal plasmid DNA was completely lost after 30 generations, suggesting a fitness cost did occur. Maternal microinjection delivers DNA efficiently to M. occidentalis and could be used for testing selectable markers or to deliver transposable-element vectors to obtain high rates of chromosomal integrations.