Incidence of eriophyid mites (Acariformes: Eriophyidae) and predatory mites (Parasitiformes: Phytoseiidae) in Florida citrus orchards under three different pest management programs.

The abundance and diversity of eriophyid and phytoseiid mites in south and central Florida were assessed in six citrus orchards under three different pest management systems, conventional, organic, and untreated. Tree canopy, ground cover, and leaf litter were sampled every two months in two groves for each of the three pest management systems from April 2019 to February 2021. The citrus rust mite, Phyllocoptruta oleivora (Ashmead) represented 95 to 99% of the rust mites sampled in each grove except in one untreated orchard where it accounted for 45% of the samples (n = 938 total P. oleivora mounted specimens). The pink citrus rust mite, Aculops pelekassi (Keifer) was present in organic and untreated orchards at 5% and 28%, respectively, but absent from conventional orchards (n = 134 total A. pelekassi mounted specimens). Twenty-nine species of phytoseiid mites were identified from 1778 specimens. Thirteen species were present in the canopy, fifteen in the ground cover, and eighteen in the leaf litter with some common species among these habitats. In the tree canopy, Typhlodromalus peregrinus (39%), Euseius spp. (25%), and Iphiseiodes quadripilis (19%) were the dominant species. Typhlodromalus peregrinus (43%), Typhlodromips dentilis (25%), and Proprioseiopsis mexicanus (13%) were the major species in the ground cover. Species richness was lower in organic orchards (3.0) compared to conventional and untreated orchards (5.0 and 4.7, respectively). In the leaf litter, Amblyseius curiosus (26%), Proprioseiopsis carolinianus (15%), Chelaseius floridanus (14%), and Amblyseius tamatavensis (12%) were the most common species. Shannon index was significantly higher in conventional orchards (1.45) compared to organic and untreated orchards (1.02 and 1.05, respectively). Evenness was also higher in conventional orchards (0.86) compared to organic and untreated (0.72 and 0.68, respectively). Finding of several phytoseiids in abundance across pest management programs suggest the need for identifying their role in pest suppression particularly mites.

Optimal dsRNA Concentration for RNA Interference in Asian Citrus Psyllid.

The Asian citrus psyllid (ACP) is a citrus pest and insect vector of "Candidatus Liberibacter asiaticus", the causal agent of citrus greening disease. Double-stranded RNA (dsRNA) biopesticides that trigger RNA interference (RNAi) offer an alternative to traditional insecticides. Standardized laboratory screening of dsRNA requires establishing the minimal effective concentration(s) that result in effective RNAi "penetrance" and trigger RNAi, resulting in one or more measurable phenotypes, herein, significant gene knockdown and the potential for mortality. In this study, knockdown was evaluated for a range of dsRNA concentrations of three ACP candidate genes, clathrin heavy chain (CHC), vacuolar ATPase subunit A (vATPase-A), and sucrose non-fermenting protein 7 (Snf7). Gene knockdown was quantified for ACP teneral adults and 3rd instar nymphs allowed a 48 h ingestion-access period (IAP) on 10, 50,100, 200, and 500 ng/µL dsRNA dissolved in 20% sucrose followed by a 5-day post-IAP on orange jasmine shoots. Significant gene knockdown (p < 0.05) in ACP third instar nymphs and adults ranged from 12-34% and 18-39%, 5 days post-IAP on dsRNA at 10-500 and 100-500 ng/µL, respectively. The threshold concentration beyond which no significant gene knockdown and adult mortality was observed post-48 h IAP and 10-day IAP, respectively, was determined as 200 ng/µL, a concentration indicative of optimal RNAi penetrance.

Species assemblage, abundance, and distribution of Phytoseiid mites (Parasitiformes: Phytoseiidae) in Citrus Under Protective Screen.

Citrus Under Protective Screen (CUPS) production system excludes the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, vector of Huanglongbing (HLB), and large predators. However, small pests and beneficial arthropods such as predatory mites may enter the CUPS. Predatory mites from the family Phytoseiidae attack several economically important pests, especially phytophagous mites common in CUPS. The species assemblage, abundance, and distribution of phytoseiid mites in CUPS and open-air (control) were investigated in Ray Ruby grapefruit from 2018 to 2020. Foliar sprays of chemicals for pest suppression were used in both systems and evaluated for effect on predatory mite populations. In the 2 production systems, a significant peak in the phytoseiid mite population was observed in February 2019 and 2020. This increase coincided with the blooming period of grapefruit, which likely provided pollen as an alternative food. A total of 2,234 specimens of 11 species were identified. Amblyseius tamatavensis Blommers and Typhlodromalus peregrinus (Muma) were dominant in both systems. Amblyseius tamatavensis was slightly more abundant in CUPS (50% of total specimens) than T. peregrinus (47%), while T. peregrinus was prevalent (82%) in open-air compared with A. tamatavensis (16%). Shannon index and evenness were significantly higher in CUPS, but species richness was similar. Chlorpyrifos with abamectin and oil, cyantraniliprole with spirodiclofen, copper, and oil as well as pyridaben alone reduced phytoseiid populations between 87% and 96%. Fenbutatin oxide mixed with copper reduced phytoseiids by 48% in open-air only. Our results demonstrate phytoseiid survival, reproduction, and contribution to biological control in CUPS, despite high temperature and humidity.

UV-LASER adjuvant-surfactant-facilitated delivery of mobile dsRNA to tomato plant vasculature and evidence of biological activity by gene knockdown in the potato psyllid.

BACKGROUND: Double-stranded RNA (dsRNA) biopesticides are of interest for the abatement of insect vectors of pathogenic bacteria such as 'Candidatus Liberibacter', which infects both its psyllid and plant hosts. Silencing of genes essential for psyllids, or for Liberibacter, is anticipated to lead to mortality or impeded bacterial multiplication. Foliar delivery is preferred for biopesticide application; however, the cuticle impedes dsRNA penetration into the vasculature. Here, conditions were established for wounding tomato leaves using ultraviolet light amplification by stimulated emissions of radiation (UV-LASER) to promote dsRNA penetration into leaves and vasculature. RESULTS: UV-LASER treatment with application of select adjuvants/surfactants resulted in vascular delivery of 100-, 300- and 600-bp dsRNAs that, in general, were correlated with size. The 100-bp dsRNA required no pretreatment, whereas 300- and 600-bp dsRNAs entered the vasculature after UV-LASER treatment only and UV-LASER adjuvant/surfactant treatment, respectively. Of six adjuvant/surfactants evaluated, plant-derived oil combined with an anionic organosilicon compound performed most optimally. Localization of dsRNAs in the tomato vasculature was documented using fluorometry and fluorescence confocal microscopy. The biological activity of in planta-delivered dsRNA (200-250 bp) was determined by feeding third-instar psyllids on tomato leaves post UV-LASER adjuvant/surfactant treatment, with or without psyllid cdc42- and gelsolin dsRNAs. Gene knockdown was quantified by quantitative, real-time polymerase chain reaction with reverse transcription (RT-qPCR) amplification. At 10 days post the ingestion-access period, knockdown of cdc42 and gelsolin expression was 61% and 56%, respectively, indicating that the dsRNAs delivered to the tomato vasculature were mobile and biologically active. CONCLUSION: Results indicated that UV-LASER adjuvant/surfactant treatments facilitated the delivery of mobile, biologically active dsRNA molecules to the plant vasculature. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Feeding behavior and hormoligosis associated with imidacloprid resistance in Asian citrus psyllid, Diaphorina citri.

Imidacloprid is a neonicotinoid insecticide used for managing the Asian citrus psyllid, Diaphorina citri Kuwayama, which serves as vector of phytopathogens causing citrus greening. However, development of resistance to neonicotinoids among populations of D. citri has coincided with occasional control failures in the field. The objectives of this research were to (1) survey current levels of imidacloprid resistance in Florida citrus; (2) compare feeding behavior between imidacloprid-resistant and susceptible D. citri using electrical penetration graph recordings, and (3) investigate the possible amplification of insecticide hormoligosis associated with resistance. Field surveys confirmed that the susceptibility of D. citri populations to imidacloprid has decreased in commercial Florida citrus groves compared with a laboratory-susceptible population. Following 12 generations of selection, resistance to imidacloprid increased by 438 fold compared with the susceptible strain. Imidacloprid-susceptible D. citri feeding on citrus exhibited significantly more bouts associated with intercellular pathway (C), phloem penetration (D), phloem salivation (E1), and nonprobing (Np) activities than imidacloprid-resistant counterparts. However, there were no differences observed in the frequency or duration of phloem ingestion or xylem feeding between susceptible and resistant D. citri. There was no statistical difference in fecundity between resistant and susceptible strains. However, the fecundity of imidacloprid-susceptible female D. citri treated with a sublethal concentration of imidacloprid (LC25 ) increased significantly compared with controls, while such hormoligosis was less pronounced among imidacloprid-resistant psyllids. Our results suggest that imidacloprid-resistant psyllids may cease feeding sooner than susceptible counterparts following sublethal exposure to this insecticide, indicative of a behavioral resistance mechanism.

Wild lime psyllid Leuronota fagarae Burckhardt (Hemiptera: Psylloidea) picorna-like virus full genome annotation and classification.

Picorna-like viruses of the order Picornavirales are a poorly defined group of positive-sense, single-stranded RNA viruses that include numerous pathogens known to infect plants, animals, and insects. A new picorna-like viral species was isolated from the wild lime psyllid (WLP), Leuronota fagarae, in the state of Florida, USA, and labelled: Leuronota fagarae picorna-like virus isolate FL (LfPLV-FL). The virus was found to have homology to a picorna-like virus identified in the Asian Citrus Psyllid (ACP), Diaphorina citri, collected in the state of Florida. Computational analysis of RNA extracts from WLP adult heads identified a 10,006-nucleotide sequence encoding a 2,942 amino acid polyprotein with similar functional domain structure to polyproteins of both Dicistroviridae and Iflaviridae. Sequence comparisons of nucleic acid and amino acid translations of the conserved RNA-dependent RNA polymerase, along with the entire N-terminal nonstructural coding region, provided insight into an evolutionary relationship of LfPLV-FL to insect-infecting iflaviruses. Viruses belonging to the family Iflaviridae encode a polyprotein of around 3000 amino acids in length that is processed post-translationally to produce components necessary for replication. The classification of a novel picorna-like virus in L. fagarae, with evolutionary characteristics similar to picorna-like viruses infecting Bactericera cockerelli and D. citri, provides an opportunity to examine virus host specificity, as well as identify critical components of the virus' genome required for successful transmission, infection, and replication. This bioinformatic classification allows for further insight into a novel virus species, and aids in the research of a closely related virus of the invasive psyllid, D. citri, a major pest of Floridian citriculture. The potential use of viral pathogens as expression vectors to manage the spread D. citri is an area that requires additional research; however, it may bring forth an effective control strategy to reduce the transmission of Candidatus Liberibacter asiaticus (CLas), the causative agent of Huanglongbing (HLB).

Integrated Water, Nutrient, and Pesticide Management of Huanglongbing-Affected Sweet Oranges on Florida Sandy Soils-A Review.

Citrus greening (huanglonbing (HLB)) disease has drastically reduced citrus fruit production in Florida over the last two decades. Scientists have developed sustainable nutrient management practices to live with the disease and continue fruit production. Best pesticide management practices have been devised to reduce the spread of HLB by Asian citrus psyllid (ACP). However, soil application of excessive nutrients and the use of soil drench application of pesticides to huanglongbing-infected citrus groves have been a serious environmental concern since the recent development of resistance to some pesticides. It is important to understand the consequences of applying pesticides and nutrients beyond the recommended application rates with an inappropriate method for citrus growth and development. Alternatively, foliar sprays of some nutrients proved effective to meet plants' requirements, and foliar sprays of effective insecticide products could provide an adequate mode of action for group rotation to minimize insecticide resistance by ACP and other pests. Sustainability in citrus production systems should include best management practices that improve pesticide and nutrient efficiency by including the total maximum daily load exiting the grove to reduce pesticide and nutrient exports into waterbodies.

Annotation of putative circadian rhythm-associated genes in Diaphorina citri (Hemiptera: Liviidae).

The circadian rhythm involves multiple genes that generate an internal molecular clock, allowing organisms to anticipate environmental conditions produced by the Earth's rotation on its axis. Here, we present the results of the manual curation of 27 genes that are associated with circadian rhythm in the genome of Diaphorina citri, the Asian citrus psyllid. This insect is the vector for the bacterial pathogen Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus greening disease (Huanglongbing). This disease severely affects citrus industries and has drastically decreased crop yields worldwide. Based on cry1 and cry2 identified in the psyllid genome, D. citri likely possesses a circadian model similar to the lepidopteran butterfly, Danaus plexippus. Manual annotation will improve the quality of circadian rhythm gene models, allowing the future development of molecular therapeutics, such as RNA interference or antisense technologies, to target these genes to disrupt the psyllid biology.

Predation of Florida red scale Chrysomphalus aonidum (L.) by the metallic blue ladybeetle Curinus coeruleus (Mulsant).

Florida red scale, Chrysomphalus aonidum (L.) is a species of armored scales, which attacks citrus crops. Biological control, particularly ladybeetles, are critical for its management in citrus production systems. We evaluated predation of C. aonidum by adult and larvae (3rd-4th instar) of the metallic blue ladybeetle, Curinus coeruleus. C. coeruleus were tested against C. aonidum in three separate treatments on Valencia orange leaves in Petri-dish arenas (1) individuals with armor intact, (2) individuals with armor removed, and (3) mix of individuals with armor intact or removed, to determine if armor inflicts distraction to predator consumption of scale and impact. Within 24 h of exposure to C. aonidum, adult beetle consumption rate averaged 64-68% in the two treatments containing all or half of the individuals with armor, compared to 100% in the treatment with armor removed. The consumption rate in the former two treatments increased to 83-89% within 72 h. Larval consumption of C. aonidum with armor intact was 25% at 24 h and 41% at 72 h, compared to 92-100% in the treatment with armor removed. The adults resulting from the larvae developed on C. aonidum with the armor intact or between the diets of armor intact or removed consumed more scales without armor than with armor. Significant consumption of C. aonidum by adults and larvae of C. coeruleus indicates that it is an efficient predator of this pest species. These novel findings suggest that this predator could be useful for suppressing C. aonidum populations in citrus production systems, particularly in habitats where both species are established such as Florida.

Crude Extracts and Alkaloids Derived from Ipomoea-Periglandula Symbiotic Association Cause Mortality of Asian Citrus Psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae).

Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important economic pest of citrus crops because it vectors the causal pathogen of huanglongbing (HLB; aka citrus greening). Population suppression of D. citri with insecticides has been disproportionally relied on for HLB management and a greater diversity of more sustainable tools is needed. Periglandula spp. is a fungal endosymbiont (family Clavicipitaceae) that forms a mutualistic relationship with members of plants in family Convolvulaceae. This association results in the production of ergot alkaloids that were previously documented as having psyllicidal properties. We investigated the mortality and behavior of D. citri exposed to crude extracts from morning glories in the plant family Convolvulaceae, as well as synthetic ergot alkaloids. Nymphs and adults were exposed to the crude plant extracts from Periglandula positive species of Convolvulaceae, as well as five synthetic ergot alkaloids. Treatments were prepared by exposing clippings of citrus to 100 ng/µL of crude extract from Periglandula-positive species of Ipomoea (I. imperati, I. leptophylla, I. pandurata and I. tricolor), and Turbina corymbosa, and from one Periglandula-negative species (I. alba) (100 ng/µL). Mortality of adult and nymphal D. citri was significantly higher than the control after exposure to extracts from I. tricolor and I. imperati. The synthetic ergot alkaloids, lysergol (10-100 ng/µL), ergonovine maleate (100 ng/µL), agroclavine (10-100 ng/µL), and ergosine (10-100 ng/µL) increased mortality of D. citri nymphs, while ergosine (100 ng/µL) and agroclavine (100 ng/µL) increased mortality of adults compared to water controls. Fewer D. citri adults settled on plants treated with crude extracts or synthetic ergot alkaloids than on water controls at 48 h after release. D. citri that fed on citrus leaves treated with 10 ng/µL solution of crude extract from the Periglandula-positive species Ipomoea (I. imperati, I. leptophylla, I. pandurata, I. tricolor), and Turbina corymbosa excreted significantly less honeydew compared with a negative water control and extract from Periglandula-negative species (I. alba). Our results indicate that crude extracts and ergot alkaloids exhibit toxic and sub-lethal effects on D. citri that could be useful for management of this pest.

Field Efficacy of Cordyceps javanica, White Oil and Spinetoram for the Management of the Asian Citrus Psyllid, Diaphorina citri.

Citrus greening disease is devastating the citrus industry in Florida, and the conventional synthetic pesticide applications used to control the vector, the Asian citrus psyllid (AsCP), Diaphorina citri, are rapidly becoming unsustainable. Various laboratory experiments indicate that the entomopathogenic fungus Cordyceps javanica, alone and in combination with horticultural oils, may offer a more sustainable strategy for the management of AsCP. Field studies conducted in 2018 and 2019 in mature citrus indicated that C. javanica alone, C. javanica mixed with white oil, and the chemical standard spinetoram mixed with white oil significantly suppressed AsCP adult populations by 61-83% up to 14 days after treatment in 2018, although colony-forming units of C. javanica were still present on the leaves 21 days after treatment (DAT). Only spinetoram + oil significantly suppressed AsCP, by 100%, up to 7 DAT in 2019. Natural enemies of AsCP, including lady beetles, lacewing larvae and the parasitoid Tamarixia radiata, were observed in the fungal treatments and the untreated control. The AsCP suppression by C. javanica and its compatibility with beneficial organisms suggest the potential use of this entomopathogenic fungus in citrus-integrated pest management.

Re-description of seven predatory mite species of family Phytoseiidae (Acari: Mesostigmata) sourced from Florida citrus groves.

Predatory mites in the family Phytoseiidae (Acari: Mesostigmata) are of great importance as biological control agents of pest mites and other arthropods. Correct identification of species is crucial to implement effective biological control of target pests. Here, we provide re-descriptions of seven phytoseiid mite species collected from citrus orchards in Florida. The several important morphological features including dorsal setae lengths, dorsal solenostomes, shape of calyx of spermatheca, chelicera dentition, measurements, and shape of macrosetae on legs currently used to discriminate phytoseiid species were missing in the original descriptions and re-descriptions of these species. Additionally, we observed the presence of a previously unnoted taxonomically important character on Proprioseius meridionalis Chant. Therefore, the re-description was essential for further diagnosis of this species. Accordingly, the validity of the presence/absence of this structure as a diagnostic character to separate species groups in the genus Proprioseius should be re-considered. Furthermore, Typhlodromalus peregrinus, a species for which a series of morphological variations are reported in previous descriptions, is re-described and illustrated from Clermont, Florida, a location very close (10 km) to its type location (Minneola), and the leaves of type host citrus. The macrosetae StIV was knobbed apically in all our specimens of T. peregrinus indicating invalidity of sharp-pointed or knobbed StIV to separate this species from a closely related species, T. aripo De Leon. These re-descriptions and species are important to utilizing authentic and promising candidates for biological control.

A Multimodal Attract-and-Kill Device for the Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae).

Phytophagous insects, including Asian citrus psyllids (Diaphorina citri Kuwayama), use multiple sensory modalities (vision, olfaction, and gustation,) to locate and accept host plants. We explored incorporation of several sensory cues into a multi-modal attract-and-kill device (AK device) using a three-dimensional shape to increase visibility, as well as elements of color, attractant, phagostimulant, UV reflectant, and toxicant. Attraction of adult D. citri to the device was mediated by a combination of a highly reflective yellow cylinder, a UV reflectant compound (magnesium oxide), and an odorant blend as a short-range attractant. The device surface was coated with a slow-release wax matrix (SPLAT™) augmented with a phagostimulant consisting of a 3-component blend (formic acid, acetic acid, and para-cymene) and an insecticide (ß-cyfluthrin). Psyllids landing on the device attempted to feed from the wax matrix, became intoxicated, died, and fell from the device. The device remained fully active over a period of 12 weeks partly because dead psyllids or nontargets did not adhere to the surface as occurs on adhesive yellow sticky cards, the industry standard. Laboratory and field assays showed that the device attracted and killed significantly more adult D. citri than ordinary yellow sticky cards. This device or a future iteration based on the design elements of this device is expected to contribute to sustainable and environmentally appropriate management of D. citri by exploiting the psyllid's innate behavioral responses to visual, olfactory, and gustatory stimuli.

Effectiveness of the Brown Lacewing, Sympherobius barberi Banks as a Biological Control Agent of the Asian Citrus Psyllid Diaphorina citri Kuwayama.

The Asian citrus psyllid (ACP) Diaphorina citri Kuwayama is an economically important pest of citrus because it vectors the causal pathogens of huanglongbing (HLB) or citrus greening disease. Biological control is an important component of citrus pest management but requires consistent strengthening of its impact on pest complex. The brown lacewing Sympherobius barberi Banks is a known predator of several insect pests from Asia, Europe, and America. However, there is not much information about its effectiveness against D. citri. We evaluated S. barberi against the D. citri and frozen eggs of the Mediterranean flour moth Ephestia kuehniella, the latter is a common diet used for rearing predators in laboratories. Adult S. barberi successfully fed on D. citri eggs and nymphs under both light and dark conditions. Diaphorina citri was also suitable for the development and reproduction of S. barberi except for slightly prolonged larval development compared with E. kuehniella diet. The egg hatch from the total number of eggs laid on D. citri and E. kuehniella diets averaged 65% and 52%, respectively. Females laid 64% eggs on dimpled white paper compared to 36% combined on plain paper and leaves of citrus, orange jasmine, eggplant and cantaloupe. Sympherobius barberi released at densities of 2-6 adults against eggs and nymphs of D. citri on infested orange jasmine plants in the cages provided a reduction of 43-81% in the number of provided eggs or nymphs. In the field tests on D. citri infested citrus trees, reduction averaged 35% in five cohorts in which developing colonies of 28-32 nymphs were provided to one S. barberi per cage. Findings suggest the significant potential of S. barberi as a predator of D. citri and to contribute to reducing huanglongbing.

Citrus Production Under Screen as a Strategy to Protect Grapefruit Trees From Huanglongbing Disease.

Citrus production under enclosed structures can exclude the Asian citrus psyllid (ACP, Diaphorina citri) and eliminate the negative effects of citrus greening or huanglongbing (HLB) disease caused by Candidatus Liberibacter asiaticus to the grapefruit (Citrus paradisi) fresh fruit industry. Physically impeding the insect vector from accessing trees is a logical method to have disease-free groves. Our objectives were to assess the ability of enclosed screenhouses to exclude the ACP, stop HLB inoculation and dissemination, and improve fruit yield of in-ground and container-grown 6-year-old "Ray Ruby" grapefruit at super-high planting densities relative to open-air trees. We built a large structure to allow commercial-scale trials and tested two production systems (screenhouse and open-air), two planting systems (in-ground and potted), and two rootstocks ("Sour Orange" [Citrus × aurantium] and "US-897" [Citrus reticulata × Poncirus trifoliate]). The experimental design was a randomized complete block design split-split-plot with four replications. Four passively ventilated 1,080-m2 completely enclosed screenhouses were constructed using a 50-mesh monofilament high-density polyethylene screen. The main support for each enclosed, covered structure consisted of pressure-treated, wooden utility poles. Trees were planted in Sept/2013 on a density of 1,957 trees/ha. Irrigation was performed on-demand using two 7.6-LPH drip emitters per tree, and fertigation was applied three times/week using 15N-2.6P-22.4K water-soluble fertilizer at 180 kg N/ha. Psyllids were monitored using sticky cards and detected inside the screenhouses post-Hurricane Irma, which damaged the screen structures in Sept/2017, leaving openings until repairs were completed in Apr/2018. Screen aging and a tropical storm in April/2019 caused another major screen opening fixed in Oct/2019. Despite the weather-related damages to the screens, only trees cultivated in open-air tested positive for Candidatus Liberibacter asiaticus after 6 years. There was fast disease progression for all outside treatments, with 100% infection. Covered, in-ground trees exhibited the highest trunk diameter and canopy volume (P < 0.0001). Trees grown inside screenhouses exhibited higher fruit yield than outside trees, with the highest yield observed for in-ground trees on "US-897" (51,081 kg/ha) (P < 0.0001). Several open-air treatments particularly in containers did not produce any fruit. On the other hand, potted grapefruit trees cultivated inside the enclosures had the highest soluble solids content (P < 0.001). The screenhouses provided disease exclusion, increased fruit yield, and fruit quality, representing an alternative for growers interested in producing high-quality fruit for the fresh market. Production cost and economic viability still need to be evaluated for large-scale implementation.

Demographic analysis of fenpyroximate and thiacloprid exposed predatory mite Amblyseius swirskii (Acari: Phytoseiidae).

Knowledge of the impact of pesticides on predators is crucial for developing integrated pest management (IPM) programs. Amblyseius swirskii (Acari: Phytoseiidae) is a predatory mite used to control several species of pest including Tetranychus urticae (Acari: Tetranychidae) and arthropods. T. urticae is a major pest of multiple greenhouse-grown and field crops including apples in Iran. Lethal and sublethal effects of fenpyroximate and thiacloprid were investigated on A. swirskii, using these chemicals separately at recommended rates or in combination at reduced rates. Recommended tested rates of both pesticides negatively influenced the biological parameters of A. swirskii such as the net reproductive rate (R0) and the intrinsic rate of increase (r). However, the combined treatment of the two pesticides at their reduced rates was less hazardous to A. swirskii. Our findings indicate that the combined use of these chemicals may be compatible with IPM programs utilizing A. swirskii as biological control tool against phytophagous mites and other pests. However, semifield and field studies to investigate the effects of reduced rate treatments of fenpyroximate and thiacloprid alone and in combination on T. urticae and A. swirskii are required for developing IPM programs.

Host Plant-Mediated Interactions Between 'Candidatus Liberibacter asiaticus' and Its Vector Diaphorina citri Kuwayama (Hemiptera: Liviidae).

The Diaphorina citri Kuwayama (Hemiptera: Liviidae) also known as Asian citrus psyllid transmits 'Candidatus Liberibacter asiaticus' (CLas) associated with the citrus Huanglongbing (HLB). Minimizing Asian citrus psyllid populations is one of the most important methods for HLB control. The development and survival of Asian citrus psyllid were evaluated on CLas+ and CLas- plants of Citrus reticulata Blanco. cv. Mashuiju (Sapindales: Rutaceae) or Citrus sunki Hort. ex Tanaka (Sapindales: Rutaceae) to assess the effects of CLas infection on vector populations. The development times were unaffected between CLas+ and CLas- plants for eggs and first to third instar nymphs but decreased for fourth instars, fifth instars, and total nymphal life on CLas+ plants. The survival rates of eggs and first and second instar nymphs were significantly lower, while those of third to fifth instar nymphs were significantly higher on CLas+ plants. However, overall nymphal survival did not differ between CLas+ and CLas- plants. The pre-oviposition period of Asian citrus psyllid on CLas+ plants decreased, while oviposition period increased, resulting in significantly higher fecundity and population trend index compared with those on CLas- plants, illustrating an overall fitness benefit of CLas infection to Asian citrus psyllid. On the other hand, the longevity and resistance to starvation and lower temperatures of adults from CLas+ plants were significantly lower than those from CLas- plants suggesting that there may be a fitness cost for Asian citrus psyllid on CLas+ plants. The development time of eggs and nymphs and adult longevity were always shorter on C. reticulata regardless of infection status. This knowledge of host plant, pathogen, and vector interactions is useful for developing Asian citrus psyllid-HLB management programs.

Physiological Effects of Citrus Leafminer Phyllocnistis citrella (Lepidoptera: Gracillariidae) Larval Feeding on Photosynthetic and Gaseous Exchange Rates in Citrus.

Feeding damage by arthropods exposes plants to pathogens and interferes with plant physiological processes. Feeding by the citrus leafminer (CLM), Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), larva exposes leaf cuticle to the causal pathogens of the citrus canker disease. We investigated the impact of larval feeding on photosynthetic rate (Pn), CO2 exchange rate (Ci), and H2O exchange rate (Wi) in eight citrus cultivars [Citrus mandarins (Kinnow, Seedless Kinnow, Feutrell's early), Citrus sinensis (Succari, Salustiana), Citrus tangerines (Fairchild), Citrus limon (China lemon), and Citrus paradisi Macfad (Grapefruit)] under natural conditions in comparison with control plants protected from larval feeding. The effects on gas exchange parameters were tested by allowing the single first instar larvae per leaf to feed till pupation at the rate of three leaves per plant and 10 plants per cultivar. A consistent reduction in leaf Pn, Ci, and Wi rates with larval development through 19 d was observed in all cultivars except Grapefruit in which all three parameters decreased through 13 d and then increased consistently through 19 d. Reductions in Pn, Ci, and Wi were significantly correlated. All three parameters of CLM-infested plants were reduced significantly compared to their levels in control plants except Pn in Kinnow, China lemon, and Grapefruit; Ci in Grapefruit; and Wi in Succari. Pn was reduced most in Fairchild, whereas Ci and Wi were reduced most in China lemon. The consequences of these physiological alterations in regions with high CLM populations could be even more intense and negatively impact plant health, tolerance to pests and diseases, and yield.

Localization and dynamics of Wolbachia infection in Asian citrus psyllid Diaphorina citri, the insect vector of the causal pathogens of Huanglongbing.

Wolbachia is a group of intracellular bacteria that infect a wide range of arthropods including the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. This insect is the vector of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of Huanglongbing or citrus greening disease. Here, we investigated the localization pattern and infection dynamics of Wolbachia in different developmental stages of ACP. Results revealed that all developmental stages of ACP including egg, 1st-5th instar nymphs, and adults of both gender were infected with Wolbachia. FISH visualization of an ACP egg showed that Wolbachia moved from the egg stalk of newly laid eggs to a randomly distributed pattern throughout the egg prior to hatching. The infection rate varied between nymphal instars. The titers of Wolbachia in fourth and fifth instar nymphs were significantly higher than those in the first and second instar nymphs. Wolbachia were scattered in all nymphal stages, but with highest intensity in the U-shaped bacteriome located in the abdomen of the nymph. Wolbachia was confined to two symmetrical organizations in the abdomen of newly emerged female and male adults. The potential mechanisms of Wolbachia infection dynamics are discussed.

Imidacloprid soil movement under micro-sprinkler irrigation and soil-drench applications to control Asian citrus psyllid (ACP) and citrus leafminer (CLM).

Imidacloprid (IM) is used to control the Asian Citrus Psyllid (ACP) and citrus leafminer (CLM), which are related to the spread of huanglongbing (HLB or citrus greening) and citrus canker diseases, respectively. In Florida citrus, imidacloprid is mainly soil-drenched around the trees for proper root uptake and translocation into plant canopy to impact ACP and CLM. The objective of this study was to determine the effect of imidacloprid rate, and irrigate amount on concentration of imidacloprid in the soil following drench application to citrus trees in three age classes. The plots were established at the Southwest Florida Research and Education Center, Immokalee, using a randomized complete-block design for three age classes of trees: one-year-old trees (B1), three to five-year-old trees (B2), and eight-year-old trees (B3). The treatments were a combination of two rates each of imidacloprid (1D, 2D) and micro-sprinkling irrigation (1I, 2I). Imidacloprid and bromide (Br-) used as tracer were applied simultaneously. Soil moisture and concentrations of imidacloprid and Br were monitored using soil cores from hand held augers. Soil moisture content (θV) did not differ under two irrigation rates at any given observation day or depth, except following heavy rainfall events. Br- was lost from the observation depths (0-45 cm) about two weeks after soil-drench. Contrarily, imidacloprid persisted for a much longer time (4-8 weeks) at all soil depths, regardless of treatment combinations. The higher retardation of imidacloprid was related to the predominantly unsaturated conditions of the soil (which in turn reduced soil hydraulic conductivities by orders of magnitude), the imidacloprid sorption on soil organic matter, and the citrus root uptake. Findings of this study are important for citrus growers coping with the citrus greening and citrus canker diseases because they suggest that imidacloprid soil drenches can still be an effective control measure of ACP and CLM, and the potential for imidacloprid leaching to groundwater is minimal.