Sucking insects and their predators on tree canopies of a monocultural stand of Caryocar brasiliense / Insetos sugadores e seus predadores na copa de Caryocar brasiliense em sistema de monocultura

Abstract Caryocar brasiliense Camb. (Malpighiales: Caryocaraceae) trees are widely distributed throughout the Cerrado ecosystem. The fruits of C. brasiliense trees are used by humans for food and as the main income source in many communities. C. brasiliense conservation is seriously threatened due to habitat loss caused by the land-use change. Sucking insects constitute an important ecological driver that potentially impact C. brasiliense survival in degraded environments. In addition, insects sampling methodologies for application in studies related to the conservation of C. brasiliense are poorly developed. In this study, sucking insects (Hemiptera) and their predators were recorded in three vertical strata of Caryocar brasiliense canopies. The distribution of sucking species showed vertical stratification along the canopy structure of C. brasiliense. The basal part of the canopy had the highest numbers of sucking insects Aphis gossypii (Glover 1877) (Hemiptera: Aphididae) and Bemisia tabaci (Genn. 1889) (Hemiptera: Aleyrodidae), and their predators Chrysoperla sp. (Neuroptera: Chrysopidae), spiders (Araneae), and Zelus armillatus (Lep. & Servi., 1825) (Hemiptera: Reduviidae). Predators' distribution follows the resource availability and preferred C. brasiliense tree parts with a higher abundance of prey.

Resumo Caryocar brasiliense Camb. (Malpighiales: Caryocaraceae) é amplamente distribuído por todo o ecossistema de cerrado. Os frutos de C. brasiliense são utilizados na alimentação humana e constitui uma importante fonte de renda para muitas comunidades. A perda de habitat provocada pelas mudanças de uso da terra coloca em risco a conservação de C. brasiliense. Insetos sugadores constituem um importante fator ecológico que, potencialmente, afeta o fitness de C. brasiliense em ambientes degradados. Além disso, as metodologias de amostragem de insetos para aplicação em estudos relacionados à conservação de C. brasiliense são pouco desenvolvidas. Neste estudo, o número de insetos sugadores (Hemiptera) e seus predadores foram avaliados em três estratos verticais do dossel de C. brasiliense. A distribuição das espécies sugadoras apresentou estratificação vertical ao longo da estrutura do dossel. O estrato basal do dossel apresentou o maior número de insetos sugadores Aphis gossypii (Glover 1877) (Hemiptera: Aphididae) e Bemisia tabaci (Genn. 1889) (Hemiptera: Aleyrodidae), e seus predadores Chrysoperla sp. (Neuroptera: Chrysopidae), aranhas (Araneae) e Zelus armillatus (Lep. & Servi., 1825) (Hemiptera: Reduviidae). Os predadores distribuíram-se de acordo com a disponibilidade de recursos, ocorrendo em maior número nas partes do dossel com maior abundância de suas presas.

Sucking insects and their predators on tree canopies of a monocultural stand of Caryocar brasiliense

Abstract Caryocar brasiliense Camb. (Malpighiales: Caryocaraceae) trees are widely distributed throughout the Cerrado ecosystem. The fruits of C. brasiliense trees are used by humans for food and as the main income source in many communities. C. brasiliense conservation is seriously threatened due to habitat loss caused by the land-use change. Sucking insects constitute an important ecological driver that potentially impact C. brasiliense survival in degraded environments. In addition, insects sampling methodologies for application in studies related to the conservation of C. brasiliense are poorly developed. In this study, sucking insects (Hemiptera) and their predators were recorded in three vertical strata of Caryocar brasiliense canopies. The distribution of sucking species showed vertical stratification along the canopy structure of C. brasiliense. The basal part of the canopy had the highest numbers of sucking insects Aphis gossypii (Glover 1877) (Hemiptera: Aphididae) and Bemisia tabaci (Genn. 1889) (Hemiptera: Aleyrodidae), and their predators Chrysoperla sp. (Neuroptera: Chrysopidae), spiders (Araneae), and Zelus armillatus (Lep. & Servi., 1825) (Hemiptera: Reduviidae). Predators' distribution follows the resource availability and preferred C. brasiliense tree parts with a higher abundance of prey.

Resumo Caryocar brasiliense Camb. (Malpighiales: Caryocaraceae) é amplamente distribuído por todo o ecossistema de cerrado. Os frutos de C. brasiliense são utilizados na alimentação humana e constitui uma importante fonte de renda para muitas comunidades. A perda de habitat provocada pelas mudanças de uso da terra coloca em risco a conservação de C. brasiliense. Insetos sugadores constituem um importante fator ecológico que, potencialmente, afeta o fitness de C. brasiliense em ambientes degradados. Além disso, as metodologias de amostragem de insetos para aplicação em estudos relacionados à conservação de C. brasiliense são pouco desenvolvidas. Neste estudo, o número de insetos sugadores (Hemiptera) e seus predadores foram avaliados em três estratos verticais do dossel de C. brasiliense. A distribuição das espécies sugadoras apresentou estratificação vertical ao longo da estrutura do dossel. O estrato basal do dossel apresentou o maior número de insetos sugadores Aphis gossypii (Glover 1877) (Hemiptera: Aphididae) e Bemisia tabaci (Genn. 1889) (Hemiptera: Aleyrodidae), e seus predadores Chrysoperla sp. (Neuroptera: Chrysopidae), aranhas (Araneae) e Zelus armillatus (Lep. & Servi., 1825) (Hemiptera: Reduviidae). Os predadores distribuíram-se de acordo com a disponibilidade de recursos, ocorrendo em maior número nas partes do dossel com maior abundância de suas presas.

A Novel Insect Overwintering Strategy: The Case of Mealybugs.

Insects have limited ability to regulate their body temperature and have thus required a range of strategies to withstand thermally stressful environments. Under unfavorable winter conditions, insects often take refuge under the soil surface to survive. Here, the mealybug insect family was selected for the study. Field experiments were performed in fruit orchards in eastern Spain. We used specifically designed floor sampling methods combined with fruit tree canopy pheromone traps. We found that in temperate climates, the large majority of the mealybugs move from the tree canopy to the roots during the winter, where they turn into belowground root-feeding herbivores to survive and continue underground the reproductive process. Within the rhizosphere, mealybugs complete at least one generation before emerging on the soil surface. The preferred area to overwinter is within 1 m in diameter around the fruit tree trunk, where more than 12,000 mealybug flying males per square meter can emerge every spring. This overwintering pattern has not previously been reported for any other group of insects showing cold avoidance behavior. These findings have implications at the winter ecology level but also at the agronomical level since treatments to control mealybug pests are, until now, only based on the fruit trees' canopy.

Nuances of Responses to Two Sources of Grapevine Leafroll Disease on Pinot Noir Grown in the Field for 17 Years.

Grapevine leafroll disease (GLD) is one of the most economically damaging virus diseases in grapevine, with grapevine leafroll-associated virus 1 (GLRaV-1) and grapevine leafroll-associated virus 3 (GLRaV-3) as the main contributors. This study complements a previously published transcriptomic analysis and compared the impact of two different forms of GLD to a symptomless control treatment: a mildly symptomatic form infected with GLRaV-1 and a severe form with exceptionally early leafroll symptoms (up to six weeks before veraison) infected with GLRaV-1 and GLRaV-3. Vine physiology and fruit composition in 17-year-old Pinot noir vines were measured and a gradient of vigor, yield, and berry quality (sugar content and berry weight) was observed between treatments. Virome composition, confirmed by individual RT-PCR, was compared with biological indexing. Three divergent viromes were recovered, containing between four to seven viruses and two viroids. They included the first detection of grapevine asteroid mosaic-associated virus in Switzerland. This virus did not cause obvious symptoms on the indicators used in biological indexing. Moreover, the presence of grapevine virus B (GVB) did not cause the expected corky bark symptoms on the indicators, thus underlining the important limitations of the biological indexing. Transmission of GLRaV-3 alone or in combination with GVB by Planococcus comstocki mealybug did not reproduce the strong symptoms observed on the donor plant infected with a severe form of GLD. This result raises questions about the contribution of each virus to the symptomatology of the plant.

Transmission of Areca Palm Velarivirus 1 by Mealybugs Causes Yellow Leaf Disease in Betel Palm (Areca catechu).

Yellow leaf disease (YLD) is the most destructive disease of betel palm (Areca catechu). A strong association between YLD and areca palm velarivirus 1 (APV1) has been observed. However, the causal relationship between APV1 and disease, and the transmission mode, warrant further investigation. This work showed that APV1 was transmitted by both Ferrisia virgata and Pseudococcus cryptus mealybugs and caused YLD symptoms in betel palm seedlings; therefore, we demonstrate that APV1 is a causal agent of YLD. APV1 was detected in the stylets, foreguts, midguts, and hindguts of the vectors via both immunocapture reverse transcription PCR and immunofluorescence assays. APV1 was not transmitted transovarially from viruliferous female F. virgata to their progeny. In summary, the transmission of APV1 by F. virgata may occur in a noncirculative, semipersistent manner. This study fills important gaps in our knowledge of velarivirus transmission, which is critical for developing YLD management practices.

Mealybug (Hemiptera: Pseudococcidae) Species Associated with Cacao Mild Mosaic Virus and Evidence of Virus Acquisition.

Theobroma cacao is affected by viruses on every continent where the crop is cultivated, with the most well-known ones belonging to the Badnavirus genus. One of these, cacao mild mosaic virus (CaMMV), is present in the Americas, and is transmitted by several species of Pseudococcidae (mealybugs). To determine which species are associated with virus-affected cacao plants in North America, and to assess their potential as vectors, mealybugs (n = 166) were collected from infected trees in Florida, and identified using COI, ITS2, and 28S markers. The species present were Pseudococcus jackbeardsleyi (38%; n = 63), Maconellicoccus hirsutus (34.3%; n = 57), Pseudococcus comstocki (15.7%; n = 26), and Ferrisia virgata (12%; n = 20). Virus acquisition was assessed by testing mealybug DNA (0.8 ng) using a nested PCR that amplified a 500 bp fragment of the movement protein-coat protein region of CaMMV. Virus sequences were obtained from 34.6 to 43.1% of the insects tested; however, acquisition did not differ among species, X2 (3, N = 166) = 0.56, p < 0.91. This study identified two new mealybug species, P. jackbeardsleyi and M. hirsutus, as potential vectors of CaMMV. This information is essential for understanding the infection cycle of CaMMV and developing effective management strategies.

Exploring Grower-sourced Data to Understand Spatiotemporal Trends in the Occurrence of a Vector, Pseudococcus maritimus (Hemiptera: Pseudococcidae) and Improve Grapevine Leafroll Disease Management.

Grapevine leafroll disease is a significant concern in the wine grape industry, as it spreads rapidly and contributes to economically significant reductions in yield and grape quality. Our objective was to utilize 5 yr of grower-sourced data from Napa (California, USA) to improve local and regional disease management efforts. Specifically, we applied a spatially integrated multivariate clustering technique to improve understanding of spatiotemporal trends in Pseudococcus maritimus (Ehrhorn) male populations-the primary vector in the region. We also implemented generalized linear mixed models to evaluate the effects of two key practices, insecticide sprays and roguing, on disease incidence. Results show P. maritimus has a biannual flight pattern in the study area, with the first flight peaking in early May and the second between early August and early September. Clusters of P. maritimus flight data fall largely within the vineyard footprints of individual growers, but also showed clear neighborhood effects. We found that when disease incidence within a block is <1%, consistent monitoring and removal of diseased vines is required to contain within-block spread. As within-block disease incidence grows to 1-20%, both insecticide applications and roguing are effective practices to reduce spread. At incidence levels >20%, roguing is a critical practice. Our results emphasize the importance of individual management efforts, but also the value of programs that engage the wider neighboring community and highlight the power of community data collection to guide decision-making.

Retention and Transmission of Grapevine Leafroll-Associated Virus 3 by Pseudococcus calceolariae.

Grapevine leafroll-associated virus 3 (GLRaV-3), an economically significant pathogen of grapevines, is transmitted by Pseudococcus calceolariae, a mealybug commonly found in New Zealand vineyards. To help inform alternative GLRaV-3 control strategies, this study evaluated the three-way interaction between the mealybug, its plant host and the virus. The retention and transmission of GLRaV-3 by P. calceolariae after access to non-Vitis host plants (and a non-GLRaV-3 host) White clover (Trifolium repens L. cv. "Grasslands Huia white clover"), Crimson clover (T. incarnatum), and Nicotiana benthamiana (an alternative GLRaV-3 host) was investigated. For all experiments, P. calceolariae first instars with a 4 or 6 days acquisition access period on GLRaV-3-positive grapevine leaves were used. GLRaV-3 was detected in mealybugs up to 16 days on non-Vitis plant hosts but not after 20 days. GLRaV-3 was retained by second instars (n = 8/45) and exuviae (molted skin, n = 6/6) following a 4 days acquisition period on infected grapevines leaves and an 11 days feeding on non-Vitis plant hosts. Furthermore, GLRaV-3 was transmitted to grapevine (40-60%) by P. calceolariae second instars after access to white clover for up to 11 days; 90% transmission to grapevine was achieved when no alternative host feeding was provided. The 16 days retention period is the longest observed in mealybug vectoring of GLRaV-3. The results suggest that an alternative strategy of using ground-cover plants as a disrupter of virus transmission may be effective if mealybugs settle and continue to feed on them for 20 or more days.

Use of Sugar Dispensers to Disrupt Ant Attendance and Improve Biological Control of Mealybugs in Vineyard.

Planococcus ficus (Signoret) and Pseudococcus comstocki (Kuwana) (Hemiptera: Pseudococcidae) are economically important pests occurring in vineyards, causing severe economic losses for growers and compromising bunch production. The partial effectiveness of insecticides used in controlling mealybug infestations as well as their high impact on the environment and on human health have led to the research of alternative and sustainable control methods, including biological control. Several natural enemies are reported to be effective against mealybugs, but their activity may be hindered by tending ants. These social insects are known to exhibit a mutualistic relationship with mealybugs, resulting in extremely aggressive behavior against beneficial insects. Consequently, this study explored a method to mitigate ant attendance by means of sugar dispensers in order to improve ecosystem services, as well as decrease mealybug infestation in vineyards. Field trials were carried out in four commercial vineyards of Northern Italy infested by mealybugs, in which Anagyrus vladimiri Triapitsyn (Hymenoptera: Encyrtidae) and Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) were released as biological control agents. Our results showed that sugar dispensers reduced ant activity and mealybug infestation, leading to a significant enhancement of ecosystem services. The technique showed a great potential in boosting biological control against mealybugs in field conditions, though the field application seemed to be labour intensive and needs to be replicated for a multi-year evaluation.

Non-Target Effects of dsRNA Molecules in Hemipteran Insects.

Insect pest control by RNA interference (RNAi)-mediated gene expression knockdown can be undermined by many factors, including small sequence differences between double-stranded RNA (dsRNA) and the target gene. It can also be compromised by effects that are independent of the dsRNA sequence on non-target organisms (known as sequence-non-specific effects). This study investigated the species-specificity of RNAi in plant sap-feeding hemipteran pests. We first demonstrated sequence-non-specific suppression of aphid feeding by dsRNA at dietary concentrations ≥0.5 µg µL-1. Then we quantified the expression of NUC (nuclease) genes in insects administered homologous dsRNA (with perfect sequence identity to the target species) or heterologous dsRNA (generated against a related gene of non-identical sequence in a different insect species). For the aphids Acyrthosiphon pisum and Myzus persicae, significantly reduced NUC expression was obtained with the homologous but not heterologous dsRNA at 0.2 µg µL-1, despite high dsNUC sequence identity. Follow-up experiments demonstrated significantly reduced expression of NUC genes in the whitefly Bemisia tabaci and mealybug Planococcus maritimus administered homologous dsNUCs, but not heterologous aphid dsNUCs. Our demonstration of inefficient expression knockdown by heterologous dsRNA in these insects suggests that maximal dsRNA sequence identity is required for RNAi targeting of related pest species, and that heterologous dsRNAs at appropriate concentrations may not be a major risk to non-target sap-feeding hemipterans.

Pseudococcidae (Hemiptera: Coccomorpha) in Uruguay: morphological identification and molecular characterization, with descriptions of two new species.

Mealybugs (Hemiptera: Coccomorpha: Pseudococcidae) are important pests in fruit production in Uruguay; however, very little is known about the species involved. A survey of mealybugs associated especially with fruit crops (apple, citrus, figs, grapes, pears, quince and strawberry), and other crops like vegetables and sugar cane, ornamentals and weeds was performed between 2017 and 2019 in Uruguay, using integrated taxonomy (morphology and DNA analyses) for their identification. A total of 19 mealybug species were identified. The most common species were Planococcus ficus (Signoret), Pseudococcus scatoterrae Granara de Willink and Pseudococcus viburni (Signoret) on fruits, and Phenacoccus madeirensis Green, Phenacoccus peruvianus Granara de Willink and Planococcus citri (Risso) on ornamental plants, all of them causing damage to their hosts. This study presents nine new species records for Uruguay, besides the description of two new species. An identification key to the mealybugs in Uruguay is provided.

Evaluation of RNA Interference for Control of the Grape Mealybug Pseudococcus maritimus (Hemiptera: Pseudococcidae).

The grape mealybug Pseudococcus maritimus (Ehrhorn, 1900) (Hemiptera: Pseudococcidae) is a significant pest of grapevines (Vitis spp.) and a vector of disease-causing grape viruses, linked to its feeding on phloem sap. The management of this pest is constrained by the lack of naturally occurring resistance traits in Vitis. Here, we obtained proof of concept that RNA interference (RNAi) using double-stranded RNA (dsRNA) molecules against essential genes for phloem sap feeding can depress insect survival. The genes of interest code for an aquaporin (AQP) and a sucrase (SUC) that are required for osmoregulation in related phloem sap-feeding hemipteran insects (aphids and whiteflies). In parallel, we investigated the grape mealybug genes coding non-specific nucleases (NUC), which reduce RNAi efficacy by degrading administered dsRNA. Homologs of AQP and SUC with experimentally validated function in aphids, together with NUC, were identified in the published transcriptome of the citrus mealybug Planococcus citri by phylogenetic analysis, and sequences of the candidate genes were obtained for Ps. maritimus by PCR with degenerate primers. Using this first sequence information for Ps. maritimus, dsRNA was prepared and administered to the insects via an artificial diet. The treatment comprising dsRNA against AQP, SUC and NUC significantly increased insect mortality over three days, relative to dsRNA-free controls. The dsRNA constructs for AQP and NUC were predicted, from sequence analysis to have some activity against other mealybugs, but none of the three dsRNA constructs have predicted activity against aphids. This study provides the basis to develop in planta RNAi strategies against Ps. maritimus and other mealybug pests of grapevines.

Revisiting an Aspergillus flavus Strain Isolated from an Egyptian Sugarcane Field in 1930.

The aflatoxin type B and G producer Aspergillus novoparasiticus was described in 2012 and was firstly reported from sputum, hospital air (Brazil), and soil (Colombia). Later, several survey studies reported the occurrence of this species in different foods and other agricultural commodities from several countries worldwide. This short communication reports on an old fungal strain (CBS 108.30), isolated from Pseudococcus sacchari (grey sugarcane mealybug) from an Egyptian sugarcane field in (or before) 1930. This strain was initially identified as Aspergillus flavus; however, using the latest taxonomy schemes, the strain is, in fact, A. novoparasiticus. These data and previous reports indicate that A. novoparasiticus is strongly associated with sugarcane, and pre-harvest biocontrol approaches with non-toxigenic A. novoparasiticus strains are likely to be more successful than those using non-toxigenic A. flavus strains. Further studies on the association between A. novoparasiticus and Pseudococcus sacchari might shed light on the distribution (and aflatoxin contamination) of this species in sugarcane. Additionally, the interaction between A. novoparasiticus, Pseudococcus sacchari, and sugarcane crop under different scenarios of climate change will be critical in order to get more insight into the host-pathogen interaction and host resistance and propose appropriate prevention strategies to decrease mycotoxin contamination and crop loss due to A. novoparasiticus attack.

Comparison of the Efficacy and Phytotoxicity of Phosphine and Ethyl Formate for Controlling Pseudococcus longispinus (Hemiptera: Pseudococcidae) and Pseudococcus orchidicola on Imported Foliage Nursery Plants.

The fumigation activity of phosphine (PH3) and ethyl formate (EF) and their phytotoxicity to 13 imported foliage nursery plant species were evaluated. The lethal concentration and time (LCT99) values of the PH3 indicated that the susceptibility of the nymphs (3.95 and <0.45 mg·h/liter, respectively) was higher than that of the adults (5.29 and 3.66 mg·h/liter, respectively) of two mealybugs [Pseudococcus longispinus (Targioni-Tozzetti) and P. orchidicola Takahashi]. The highest concentration reduction rate of PH3 and EF on the 13 foliage nursery plants in the 12-liter desiccator was 41.5% for Heteropanax fragrans and 71.7% for Schefflera arboricola, respectively, which indicates that PH3 has a lower sorption rate than EF. The phytotoxicities of PH3-treated foliage nursery plants did not significantly differ from those of the nontreated plants, but EF caused phytotoxicity in 11 foliage nursery plants a week after treatment. When the exposure time of PH3 increased to 24 h, the adults and nymphs of both mealybug species showed 100% mortality in the 0.5 m3 fumigation chamber. In the 10 m3 fumigation container used in the field, there was 100% mortality of both mealybugs after treatment with 2 g/m3 PH3 for 24 h at 16°C. These results indicate that EF is not a suitable mealybug fumigant due to its high sorption and phytotoxicity to foliage nursery plants, despite fumigation activity against the two species. However, PH3 seems to be suitable for mealybug fumigation in foliage nursery plants and can be used as a substitute for methyl bromide.

Biparental mealybugs may be more promiscuous than we thought.

Knowledge on the reproductive biology of target insect pest is essential for the effective implementation of pheromone-based pest management tactics. In mealybugs, the second largest family of scale insects, the existence of female multiple mating was recently suggested. In this study, we aimed at testing how general is this behavior in mealybugs, by investigating polygyny and polyandry in two cosmopolitan pest mealybugs, Planococcus citri and Pseudococcus calceolariae. Males of these species were able to mate an average of 11.9 and 13.3 females, respectively, during their lifespan. The number of fertilized females per male decreased with male age/mating history for both mealybugs. We found no differences in female fecundity and fertility, when fertilized by males with different mating history. When we used male age as a proxy of mating history, we observed a significant negative effect on female fecundity. The females of both species remained receptive after first copula and eventually mated multiple times. The percentage of remated females of P. citri decreased linearly with time since first copula, with some maintaining receptivity up to 24 h. Males showed no preference between virgin and mated females, in static-air olfactometer tests. We found no benefit of female multiple mating in relation to fecundity. In biparental mealybugs, the mating system of males is possibly scramble competition polygyny; and that of females is possibly polyandry, with female receptivity restricted to a relatively short period. We discuss the practical implications of the results for pest management.

DNA barcoding identification of Pseudococcidae (Hemiptera: Coccoidea) using the mitochondrial COI gene.

DNA barcoding is a recently developed technique for species-level identification that involves the use of short, standard DNA sequences as species labels. It is an effective complement to traditional taxonomic classification based on morphology. At present, research and applications involving the DNA barcoding of the Pseudococcidae are focused primarily on the cytochrome coxidase subunit I (COI) gene, but there is not yet a consensus on the preferred gene region for barcoding. The purpose of this study was to explore the effectiveness of identification of Pseudococcidae beetles using DNA barcoding technology. The COI gene sequences of 97 samples from 21 species of Asemini were analysed, followed by evaluation of the ability to identify species using a tree-building method and distance evaluation. The COI sequences (500 bp) exhibited distinct distributions of intra-specific and inter-specific variation and a significant barcoding gap. The success rate of identification was 97.84%. These results demonstrate the feasibility of using this segment of COI to identify most species of Pseudococcidae.

Physiological Response of Orchids to Mealybugs (Hemiptera: Pseudococcidae) Infestation.

The harmfulness of mealybugs resulting from sucking plant sap, secreting honeydew, and transmitting plant viruses can give them the status of serious pests. This study documents the influence of Pseudococcus maritimus (Ehrhorn) and Pseudococcus longispinus (Targioni Tozzetti) infestation on alterations in selected physiological parameters of Phalaenopsis x hybridum 'Innocence'. The condition of the cytoplasmic membranes was expressed as the value of thiobarbituric acid reactive substances. We have determined changes in the activities of catalase and guaiacol peroxidase and measured the following chlorophyll fluorescence parameters: maximum quantum yield of photosystem II (Fv/Fm), effective quantum yield (Y), photochemical quenching (qP), and nonphotochemical quenching (qN). The strongest physiological response of orchids was recorded in the initial period of mealybugs infestation. Prolonged insect feeding suppressed lipid peroxidation, peroxidase and catalase activity, as well as photosynthesis photochemistry. The pattern of changes was dependent on mealybug species. This indicated the complexity of the processes responsible for plant tolerance. Data generated in this study have provided a better understanding of the impact of two mealybug species infestation on Phalaenopsis and should be useful in developing pest management strategies.

Interactions Within Susceptible Hosts Drive Establishment of Genetically Distinct Variants of an Insect-Borne Pathogen.

Coinfections are common, leading to pathogen interactions during transmission and establishment in a host. However, few studies have tested the relative strengths of pathogen interactions in vectors and hosts that determine the outcome of infection. We tested interactions between two genetically distinct variants of the mealybug-transmitted Grapevine leafroll-associated virus 3. The transmission efficiency of each variant in single variant inoculations by two vector species was determined. The effects of vector species, a coinfected source, and simultaneous inoculation from multiple hosts to one host on variant establishment were examined. Within-vector interactions could have a role in transmission from hosts containing mixed infections, but not when vectors were moved from separate singly infected source plants to a single recipient plant. The invasive Planococcus ficus (Signoret) was a more efficient vector than Pseudococcus viburni (Signoret). Transmission efficiency of the two variants did not differ in single variant inoculations. Overall infections were the same whether from singly or coinfected source plants. In mixed inoculations, establishment of one variant was reduced. Mixed inoculations from two singly infected source plants resulted in fewer mixed infections than expected by chance. Therefore, the observed outcome was determined subsequent to host inoculation rather than in the vector. The outcome may be due to resource competition between pathogens. Alternatively apparent competition may be responsible; the pathogens' differential ability to overcome host defenses and colonize the host may determine the final outcome of new infections. Detailed knowledge of interactions between pathogens during transmission and establishment could improve understanding and management of disease spread.

Monitoring Pseudococcus calceolariae (Hemiptera: Pseudococcidae) in Fruit Crops Using Pheromone-Baited Traps.

The citrophilus mealybug, Pseudococcus calceolariae (Maskell), is an important pest of fruit crops in many regions of the world. Recently, its sex pheromone has been identified and synthesized. We carried out field experiments with the goal of developing monitoring protocols for P. calceolariae using pheromone-baited traps. Traps checked hourly for 24 hours showed a distinct diel pattern of male flight, between 18:00 and 21:00 h. The presence of unnatural stereoisomers did not affect trap captures, with isomeric mixtures capturing similar amounts of males as the biological active isomer. Dose of isomeric mixture pheromone (0-100 µg) had a nonlinear effect on male captures, with 10, 30, and 50 µg capturing similar amounts. The effective range of pheromone traps was determined by placing traps at different distances (15, 40, and 80 m) from an infested blueberry field, loaded with 0, 1 and 25 µg of the pheromone. For all distances, 25 µg dose captured more males, and was highly attractive up to 40 m. There was a significant effect of lure age on male captures (0-150 d), with similar amount of males captured up to 90-day-old lure, and lower captures in the 150-day-old lure compared with fresh ones. We found significant positive correlations between P. calceolariae males caught in pheromone traps with female abundance and fruit infestation at harvest. Our results show the usefulness of P. calceolariae pheromones for monitoring at field level and provide information for the design of monitoring protocols.

A survey of mealybugs infesting South-Brazilian wine vineyards

ABSTRACTMealybugs (Hemiptera: Pseudococcidae) are important pests of the grapevine Vitis spp. and are responsible for direct and indirect damage to production. The main mealybug species present in wine grapevine (Vitis vinifera L.) in Southern Brazil were identified and their incidence evaluated. Bunch-samples (n = 50) from 131 vineyards located in the Serra Gaúcha Region (RS) of Brazil were analyzed at harvest, and the occurrence of mealybugs in the roots was evaluated at the time of eradication of plants for replanting. Mealybugs were reared in laboratory until adulthood for species determination. The species Dysmicoccus brevipes (Cockerell, 1983), Dysmicoccus sp., Planococcus citri (Risso, 1813), Pl. minor(Maskell, 1897), Pseudococcus viburni (Signoret, 1875) and Pseudococcus sp. were identified in bunches. Dysmicoccus sp., D. umbambae Granara de Willink, 2009, Pl. citri and Pseudococcus sp. were found in the roots. Pl. citri (31.4%) and Dysmicoccus sp. (22.7%) were the most common species found in wine grape bunches in the Serra Gaúcha Region.