Isolation and characterization of two entomopathogenic fungi attackingDiaphorina citri (Homoptera, Psylloidea) in Indonesia

In an attempt to suppress the propagation of citrus greening disease in Indonesia, we explored pathogens ofDiaphorina citri which vectors the disease. At two orange orchards, manyD. citri adults were found to be dead and covered with fungal mycelia. Two fungi,Paecilomyces fumosoroseus andHirsutella citriformis, were consistently isolated from the infected insects. Molecular phylogeny of their 18S rDNA sequences showed that they belong to the ascomycetous clade of the Clavicipitales/Hypocreales, which embraces many entomopathogenic fungi. When healthy adults ofD. citri were inoculated with conidia of theP. fumosoroseus, the insects died within 6 d.     

Diaphorina citri populations carrying the bacterial plant pathogen Candidatus Liberibacter asiaticus are more susceptible to infection by entomopathogenic fungi than bacteria‐free populations

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Delivery of dsRNA through topical feeding for RNA interference in the citrus sap piercing‐sucking hemipteran,Diaphorina citri

RNA interference (RNAi) is a powerful means to study functional genomics in insects. The delivery of dsRNA is a challenging step in the development of RNAi assay. Here, we describe a new delivery method to increase the effectiveness of RNAi in the Asian citrus psyllid Diaphorina citri. Bromophenol blue droplets were topically applied to fifth instar nymphs and adults on the ventral side of the thorax between the three pairs of legs. In addition to video recordings that showed sucking of the bromophenol blue by the stylets, dissected guts turned blue indicating that the uptake was through feeding. Thus, we called the method topical feeding. We targeted the abnormal wing disc gene (awd), also called nucleoside diphosphate kinase (NDPK), as a reporter gene to prove the uptake of dsRNA via this method of delivery. Our results showed that dsRNA‐awd caused reduction of awd expression and nymph mortality. Survival and lifespan of adults emerged from treated nymphs and treated adults were affected. Silencing awd caused wing malformation in the adults emerged from treated nymphs. Topical feeding as a delivery of dsRNA is highly efficient for both nymphs and adults. The described method could be used to increase the efficiency of RNAi in D. citri and other sap piercing‐sucking hemipterans.     

Behavioral Response of <Emphasis Type="Italic">Tamarixia radiata</Emphasis> (Waterston) (Hymenoptera: Eulophidae) to Volatiles Emanating from <Emphasis Type="Italic">Diaphorina citri</Emphasis> Kuwayama (Hemiptera: Psyllidae) and Citrus

Tamarixia radiata Waterston (Hymenoptera: Eulophidae) is an effective idiobiont ectoparasitoid of the psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae), vector of the huanglongbing (HLB). We examined the olfactory responses of T. radiata to volatiles emanating from D. citri or plant volatiles using a custom designed T-maze olfactometer and open arena bioassays. We also examined the behavioral response of male and female T. radiata to conspecifics of the opposite sex to determine whether olfactory signals mediate mate location. T. radiata adults exhibited a sexually dimorphic response to volatiles emanating from D. citri and citrus. Female T. radiata responded positively to the odors emanating from D. citri nymphs in both olfactometer and open arena bioassays. However, female wasps showed no response to odors emanating from D. citri adults, D. citri honey dew secretions, intact citrus or orange jasmine leaves. Odors emanating from D. citri damaged citrus were not attractive to T. radiata females but stimulated attraction of wasps to D. citri on damaged plants. T. radiata females were not attracted to D. citri immatures when they were presented as visual cues. Male T. radiata did not show attraction to D. citri nymphs or other putative odors that were attractive to female T. radiata. In olfactometer bioassays, more male T. radiata responded to the odor of female conspecifics than blank controls in the absence of associated citrus host plant volatiles. Odors emanating from female T. radiata were not attractive to male T. radiata. Male or female T. radiata were not attracted to the odors emanating from same sex conspecifics. Both male and female T. radiata adults showed positive phototactic behavior. Collectively, our results provide behavioral evidence that: 1) female T. radiata use volatiles emanating from D. citri nymphs to locate its host and: 2) female T. radiata release a volatile pheromone that attracts male conspecifics.     

Feeding behavior of Diaphorina citri and its transmission of ‘Candidatus Liberibacter asiaticus’ to citrus

Huanglongbing (HLB), also known as citrus greening, is currently the most destructive disease of citrus, responsible for huge economic losses in the world's major citrus production areas. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), transmits ‘Candidatus Liberibacter asiaticus’ (Clas), the pathogen responsible to cause HLB. Understanding of vector, pathogen, and host plant interactions is important for the management of this vector‐disease complex. We used the direct‐current electrical penetration graph (DC‐EPG) system to evaluate feeding behavior of Clas‐infected D. citri adults, and their potential to transmit the pathogen to healthy citrus, Citrus reticulata Blanco cv. Sunki (Rutaceae), following a 24‐h inoculation access period. Plants were tested for the presence of Clas by qPCR 6 months after inoculation. Findings suggest that inoculation was associated with salivation into the phloem sieve elements (waveform E1). The minimum feeding time for successful transmission by a single adult was 88.8 min, with a minimum E1 duration of 5.1 min. Regression analysis indicated a significant relationship between E1 duration and transmission efficiency. The adults successful in transmitting Clas to healthy citrus were able to penetrate and feed in the phloem much earlier than those which did not transmit. The minimum duration of E1 for a female was shorter than that of a male, but transmission was higher. However, durations of other EPG parameters were not significantly different between male and female. Feeding by single Clas‐infected D. citri adults on 6‐month‐old plants (Sunki) resulted in 23% HLB‐positive plants 6 months after inoculation. Multiple nymphs or adults could transmit the pathogen more efficiently than individual adults in the field, and further enhance the severity of the disease. Effective tactics are warranted to control D. citri and disrupt transmission of Clas.     

Susceptibility of <Emphasis Type="Italic">Diaphorina citri</Emphasis> (Hemiptera: Liviidae) and Its Parasitoid <Emphasis Type="Italic">Tamarixia radiata</Emphasis> (Hymenoptera: Eulophidae) to Entomopathogenic Fungi under Laboratory Conditions

Diaphorina citri (Kuwayama) is a global pest of citrus that transmits the bacteria associated with the disease, Huanglongbing. Entomopathogenic fungi and the parasitoid Tamarixia radiata (Waterston) are important biological control agents of this pest and likely to interact in D. citri populations. As a basis for interaction studies, we determined the susceptibility of nymphs and adults of D. citri and adults of the parasitoid T. radiata to six fungal isolates from the species Beauveria bassiana s.l. (Bals.-Criv.) Vuill. (isolates B1 and B3), Metarhizium anisopliae s.s. (Metsch.) (Ma129 and Ma65) and Isaria fumosorosea Wize (I2 and Pae). We conducted experiments evaluating infection levels in all three insect groups following inoculation with a series of conidial concentrations (1 × 10⁴–1 × 10⁸ conidia mL^?1). Results showed that D. citri nymphs and T. radiata were more susceptible to fungal isolates than D. citri adults. Overall, B. bassiana and M. anisopliae isolates caused the greatest infection compared with I. fumosorosea isolates in all three groups of insects. Isolates B1 (B. bassiana) and Ma129 (M. anisopliae) infected a greater proportion of adults and nymphs of D. citri, respectively. Both isolates of B. bassiana caused greater infection in T. radiata compared with isolates of the other fungal species. We propose that isolates B1 and Ma129 are the strongest candidates for control of D. citri. Our results represent the first report of entomopathogenic fungi infecting T. radiata, and the basis for future studies to design a biological control programme that uses both agents more efficiently against D. citri populations.     

RNA interference of carboxyesterases causes nymph mortality in the Asian citrus psyllid,Diaphorina citri

Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an important pest of citrus. In addition, D. citri is the vector of Huanglongbing, a destructive disease in citrus, also known as citrus greening disease caused by Candidatus Liberibacter asiaticus. Huanglongbing causes huge losses for citrus industries. Insecticide application for D. citri is the major strategy to prevent disease spread. The heavy use of insecticides causes development of insecticide resistance. We used RNA interference (RNAi) to silence genes implicated in pesticide resistance in order to increase the susceptibility. The activity of dsRNA to reduce the expression of carboxyesterases including esterases FE4 (EstFE4) and acetylcholinesterases (AChe) in D. citri was investigated. The dsRNA was applied topically to the fourth and fifth instars of nymphs. We targeted several EstFE4 and AChe genes using dsRNA against a consensus sequence for each of them. Five concentrations (25, 50, 75, 100, 125 ng/μl) from both dsRNAs were used. The treatments with the dsRNA caused concentration dependent nymph mortality. The highest gene expression levels of both AChe and EstFE4 were found in the fourth and fifth nymphal instars. Gene expression analysis showed that AChe genes were downregulated in emerged adults from dsRNA‐AChe‐treated nymphs compared to controls. However, EstFE4 genes were not affected. In the same manner, treatment with dsRNA‐EstFE4 reduced expression level of EstFE4 genes in emerged adults from treated nymphs, but did not affect the expression of AChe genes. In the era of environmentally friendly control strategies, RNAi is a new promising venue to reduce pesticide applications.     

An assessment of interspecific competition between two introduced parasitoids of Diaphorina citri (Hemiptera: Liviidae) on caged citrus plants

Two parasitoids attacking nymphs of Asian citrus psyllid,Diaphorina citri Kuwayama (Hemiptera:Liviidae),Tamarixia radiata (Waterston)(Hymenoptera:Eulophi- dae)and Diaphoreneyrtus aligarhensis (Shafee,Alam &Agarwal)(Hymenoptera:Encyrtidae)are being released in California,USA in a classical biological control program. To evaluate the effect of multiple parasitoid species on D.citri mortality,we conducted mesocosm experiments under controlled conditions using a complete block design with 6 treatments (D.cirri nymphs exposed to:no parasitoids;D.aligarhensis or T.radiata alone;D.aligarhensis or T.radiata released first (by 48 h);and both species released simultaneously).Parasitism of D.citri nymphs by T.radiata exceeded 60%and was unchanged when D.aligarhensis were present.Parasitism by D.aligarhensis was greatest when T. radiata was absent (-28%)and was reduced in all treatments with T.radiata present (<3%).D.cirri mortality and parasitoid-related mortality olD.citri was consistent across parasitoid treatments.Laboratory results suggest that competition between D.aligarhensis and T.radiata is asymmetric and favors T.radiata.It may be difficult for D.aligarhensis to contribute significantly to D.cirri biological control where T.radiata is present.However, results reported here suggest that competition between T.radiata and D.aligarhensis is not likely to reduce parasitism by T.radiata or reduce parasitoid-induced mortality of D.citri.     

In vitro egg hatching of Diaphorina citri,the vector of Huanglongbing

Citrus greening (Huanglongbing, HLB) is a destructive disease associated with the uncultivable, phloem-limited, gram-negative bacteria Candidatus Liberibacter spp., which affects citrus crops and other Rutaceae species. HLB is rapidly transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). We developed an artificial medium on which D. citri eggs can hatch and first-instar nymphs can feed. The medium could be used to study insect physiology and screen molecules that may interfere with egg hatching and nymph development.     

Comparison of aerial conidia and blastospores from two entomopathogenic fungi against Diaphorina citri (Hemiptera: Liviidae) under laboratory and greenhouse conditions

This study compared the insecticidal activity of liquid culture-produced blastospores and solid substrate-produced aerial conidia of Beauveria bassiana GHA and Isaria fumosorosea ARSEF3581 strains against Diaphorina citri adults. Insects exposed to 10⁷ propagules/ml in a spray residue contact leaf bioassay died within 6 days at 25°C, with no significant differences between fungal treatments. At higher concentrations (10⁸ propagules/ml), I. fumosorosea conidia killed psyllids faster compared to its blastospore formulation, i.e. 4 versus 5 days, respectively. In greenhouse tests, the same treatments applied to infested citrus plants (2?×?10⁶ spores/ml) all significantly reduced the number of nymphs compared with the untreated controls over 3 weeks; however, only I. fumosorosea blastospores significantly reduced the number of F1 adult psyllids when compared with controls. Similar results were observed in the follow-up greenhouse test, where I. fumosorosea blastospores were the most effective treatment overall, reducing D. citri populations by about 60% after 21 days; by contrast, imidacloprid killed almost 100% of psyllids within a week in both tests. Fewer psyllids exhibited mycosis in the greenhouse (i.e. ≈20 versus?≥?87% in the laboratory). This is the first report comparing both conidial and blastospore formulations of B. bassiana and I. fumosorosea for the control of a psyllid pest. Field testing is required to determine how successful different spore formulations might be under various environmental conditions.     

Predation and functional response of the multi-coloured Asian ladybeetle Harmonia axyridis on the adult Asian citrus psyllid Diaphorina citri

Diaphorina citri (Hemiptera: Liviidae) is a vector for the citrus Huanglongbing greening disease, which can reduce crop yields. Harmonia axyridis (Coleoptera: Coccinellidae) is known to prey on D. citri nymphs; we investigated the predation capacity and functional response of H. axyridis on adult D. citri. H. axyridis larvae exhibited Holling’s type II functional response to different densities of adult D. citri. For second, third and fourth instar H. axyridis larvae, the successful attack rates were 0.75?±?0.22, 1.06?±?0.11, and 1.04?±?0.26, respectively; the handling times were 18.08?±?5.37, 0.45?±?0.07, and 0.24?±?0.07?h, respectively; and the estimated maximum predation rates were 4, 54, and 102, respectively. Fourth instar larvae had the lowest handling time. Adult H. axyridis consumed more D. citri adults maintained at 15–20°C than at 30–35°C, with an optimal temperature of 17.28°C. More adults were consumed during the day than at night, with peak consumption between 12:00 and 14:00?h, not differing significantly differ between predator genders. Adult H. axyridis reared on the aphid Acyrthosiphon pisum were tested for D. citri predation; these H. axyridis had successful attack rates, handling times, and estimated maximal predation rates of 0.66?±?0.18, 0.82?±?0.26?h, and 29, respectively. Conversely, those fed and tested on D. citri had corresponding rates of 0.99?±?0.17, 0.38?±?0.12?h, and 63, respectively. Significant H. axyridis predation on adult D. citri indicated its enhanced role in suppressing both nymph and adult D. citri.     

Selection of Bacillus thuringiensis strains in citrus and their pathogenicity to Diaphorina citri (Hemiptera: Liviidae) nymphs

Bacillus thuringiensis (Bt) toxins are effective in controlling insect pests either through the spraying of products or when expressed in transgenic crops. The discovery of endophytic Bt strains opened new perspectives for studies aimed at the control of sap‐sucking insects, such as the Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae), a vector of “Candidatus Liberibacter spp.,” associated with citrus huanglongbing (HLB). In this study, translocation of endophytic Bt strains in citrus seedlings inoculated with Bt suspension delivered by soil‐drench, and their systemic pathogenicity to D. citri nymphs were investigated. The pathogenicity of three wild‐type Bt strains against D. citri third‐instar nymphs was demonstrated. Among the 10 recombinant strains tested (each of them harboring a single cry or cyt gene), 3 can be highlighted, causing 42%–77% and 66%–90% nymphal mortality at 2 and 5 d after inoculation, respectively. The isolation of Bt cells from young citrus shoots and dead nymphs, and PCR performed with specific primers, confirmed the involvement of the Bt strains in the psyllid mortality. This is the first report showing the translocation of Bt strains from citrus seedling roots to shoots and their potential to control D. citri nymphs that fed on these soil‐drench inoculated seedlings. The Bt strains that caused the highest mortality rates have the potential to be used as bioinsecticides to control D. citri and the identified genes can be used for the production of transgenic Bt citrus.     

Effect of host deprivation on the foraging behavior of the Asian citrus psyllid parasitoid Tamarixia radiata: observations from the laboratory and the field

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a significant citrus pest and the parasitoid Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) has been released in various citrus‐producing areas in classical biological control programs targeting D. citri. We investigated the effect of host deprivation on the foraging behavior and patch utilization by T. radiata. In the laboratory, females deprived of hosts for 3 days tended to leave patches of 12 nymphs without parasitizing hosts during the ca. 30 min they spent in the patch before leaving. Moreover, half of these females failed to host feed, and those that did host feed, on average, needed more than 15 min to complete feeding. Conversely, non‐host‐deprived females parasitized on average three nymphs before leaving patches without host feeding during the ca. 39 min they spent in the patch. These laboratory observations were compared to mass‐reared female T. radiata that were released onto colonies of D. citri nymphs infesting citrus in the field. Release vials were provisioned with honey and these females had no opportunities to host feed over the 1‐ to 2‐day containment period prior to release. When introduced onto D. citri colonies, 68% T. radiata females abandoned D. citri patches prior to probing hosts, in part, because Argentine ants, Linepithema humile (Mayr) (Hymenoptera: Formicidae), tending colonies disturbed searching parasitoids. These results from laboratory and field studies are discussed in the context of classical biological control, with the aim of understanding how to manipulate host availability and ant activity so establishment rates and impact of T. radiata can be improved.     

Enhanced proliferation and efficient transmission of Candidatus Liberibacter asiaticus by adult Diaphorina citri after acquisition feeding in the nymphal stage

We carried out a quantitative detection of Candidatus Liberibacter asiaticus, the bacterium associated with the disease of huanglongbing, in the vector psyllid Diaphorina citri by using a TaqMan real‐time PCR assay. The concentration of the bacterium was monitored at 5‐day intervals for a period of 20 days after psyllids were exposed as fifth instar nymphs or adults to a Ca. L. asiaticus‐infected plant for an acquisition access period of 24 h. When adults fed on Ca. L. asiaticus‐infected plant, the concentration of the bacterium did not increase significantly and the pathogen was not transmitted to any citrus seedlings. In contrast, when psyllids fed on infected plant as nymphs, the concentration of the pathogen significantly increased by 25‐, 360‐ and 130‐fold from the initial acquisition day to 10, 15 and 20 days, respectively. Additionally, the pathogen was successfully transmitted to 67% of citrus seedlings by emerging adults. Our data suggested that multiplication of the bacterium into the psyllids is essential for an efficient transmission and show that it is difficult for adults to transmit the pathogen unless they acquire it as nymphs.     

Predation of the apefly,Spalgis epius (Lepidoptera: Lycaenidae) on citrus mealybug,Planococcus citri (Hemiptera: Pseudococcidae)

The citrus mealybug, Planococcus citri (Risso) (Hemiptera: Pseudococcidae) is a serious pest of economically important crops worldwide. The apefly, Spalgis epius (Westwood) (Lepidoptera: Lycaenidae) is a potential predator of various species of mealybugs. Earlier investigation on its daily preying capacity and preference for prey stages on P. citri is incomplete. Hence, a study was conducted to find out the daily prey consumption ability and preference for prey stages by different larval instars of S. epius reared on P. citri in the laboratory. Through the 8-day developmental period with four larval instars, the daily prey consumption of S. epius increased from the first to the seventh day and decreased on the eighth day prior to the prepupal stage. Generally, there was a significant difference in the prey consumption on different days. When the prey stages were offered separately, the first to fourth instar larva of S. epius consumed, respectively, a mean of 199.6, 722.6, 1908.8, and 4625.6 eggs or 21.5, 77.0, 168.5, and 670.5 nymphs or 3.2, 7.2, 16.0, and 35.1 adults of P. citri. When an S. epius larva was fed on P. citri eggs, nymphs and adults separately, it consumed a mean of 7456.7 eggs, 937.6 nymphs, or 62.3 adults during its entire development. When the prey stages were offered all together, a single S. epius larva consumed 2618.4 eggs, 170.4 nymphs, and 39.7 adults of P. citri throughout its entire development. The study revealed that S. epius is a voracious predator of P. citri and thus could be utilized as a major biological control agent.     

Prey preference and consumption capacity of Nephus arcuatus (Coleoptera: Coccinellidae): the influence of prey stage,prey size and feeding experience

This study examined the feeding habits of Nephus arcuatus Kapur (Coleoptera: Coccinellidae), an important predator of mealybugs in south-western Iran. The consumption capacity of male and female N. arcuatus adults was determined by their feeding on eggs, first-instar nymphs, and adult females of two destructive mealybugs, Nipaecoccus viridis (Newstead) and Planococcus citri Risso, over a 24-h period. N arcuatus consumed significantly more first-instar nymphs than eggs, and more eggs than female adults of both prey species. In addition, we also studied the developmental stage prey preference of adults reared on either N. viridis or P. citri and found that the prey preference of females did not change with the developmental stage of the mealybug. Meanwhile, the males reared on either N. viridis or P. citri showed a significant preference for the first-instar nymphs of P. citri over first-instar nymphs of N. viridis, while a preference for the eggs or adult females of these two mealybugs was not observed. This selection of first-instar nymphs by males was not tied to its previous feeding experience. Our findings suggest that prey stage, prey size and previous feeding experience had no effect on the prey selection of N. arcuatus, making it a good candidate for the biological control of mealybugs.     

Susceptibility of Asian citrus psyllid, <Emphasis Type="Italic">Diaphorina citri</Emphasis> (Hemiptera: Liviidae), to the insecticide afidopyropen: a new and potent modulator of insect transient receptor potential channels

A relatively new insecticide chemistry for controlling sucking insects, afidopyropen, was investigated for toxicity against Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). We evaluated the mortality of D. citri eggs, nymphs, and adults treated with afidopyropen using both laboratory-reared and field populations. We also quantified the effects of sublethal doses of afidopyropen on D. citri feeding, host choice selection, and fecundity. For laboratory susceptible adults, the contact LC₅₀, topical application LD₅₀, and leaf dip LC₅₀ were 2.13, 2.00, and 3.08 ng/µL, respectively. For adults collected from a commercially managed citrus grove in Florida, the contact LC₅₀, topical application LD₅₀, and leaf dip LC₅₀ were 1.37, 1.92, and 4.89 ng/µL, respectively. Egg hatch was significantly reduced following exposure to afidopyropen at 100 ng/µL. Furthermore, afidopyropen reduced D. citri nymph survival and adult emergence at concentrations ranging between 0.01 and 100 ng/µL. Diaphorina citri adult feeding decreased on citrus leaves treated with afidopyropen in a concentration-dependent manner as measured indirectly by honeydew excretion, and appeared almost completely inhibited after treatment with 10 and 100 ng/µL solutions of afidopyropen. In choice tests, significantly fewer D. citri adults settled on afidopyropen-treated plants than on control plants at 24, 48, and 72 h after release, with no differences in settling between males and females. Afidopyropen reduced the fecundity of D. citri in a concentration-dependent manner. Collectively, the results suggest that afidopyropen could contribute to the integrated management of D. citri and may therefore be useful in rotational programs to improve resistance management.     

The effects of conspecific and heterospecific interactions on foraging and oviposition behaviours of two parasitoids of Diaphorina citri

Direct observation was used to construct activity budgets for Diaphorencyrtus aligarhensis (Shafee, Alam & Agarwal) (Hymenoptera: Encyrtidae) and Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae), foraging for nymphs of Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Two populations of D. aligarhensis were used in experiments: a biparental population from Pakistan and a uniparental population from Taiwan. Parasitoids were provisioned with 20 D. citri nymphs, second through fifth instar; female parasitoids from all three populations foraged alone, in conspecific pairs, or in heterospecific pairs. Walking occupied approximately 40% of the activity budget of D. aligarhensis and ovipositor probing occupied 10–20%. In contrast, probing and walking each accounted for approximately 30% of the activity budget of T. radiata. The presence of conspecific females affected the activity budget of D. aligarhensis females, such that the focal female was observed walking less often and probing more frequently. Heterospecific females did not affect D. aligarhensis activity. These results suggest mutual interference may occur between D. aligarhensis females foraging on the same host patch. The behaviour of T. radiata was not significantly affected by the presence of conspecific or heterospecific competitors. These results are discussed with respect to the release of multiple species of natural enemies in the classical biological control programme targeting D. citri in California.     

Sex‐specific responses of Asian citrus psyllid to volatiles of conspecific and host‐plant origin

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the primary vector of Candidatus Liberibacter spp. bacteria that cause citrus greening, a disease of worldwide importance. Olfactometry was employed to test responses of D. citri to odours from intact citrus plants (Mexican lime, Citrus aurantifolia, sour orange, Citrus aurantium, Marsh grapefruit, Citrus paradisi and Valencia orange, Citrus sinensis), citrus plants previously infested with D. citri, and odours of conspecifics including nymphs, adult insects of same and opposite sex, and their products (honeydew), both alone and in combination. In contrast to other studies, psyllids of both sexes were attracted to volatiles of undamaged Mexican lime leaves, whereas undamaged grapefruit attracted only females, and leaves of Valencia and sour orange did not attract either sex. All four plant species attracted female psyllids when previously infested, but only Mexican lime and sour orange‐attracted males. Thus, Citrus species appear to vary in the production of both constituitive and induced volatiles that attract adult psyllids. Volatiles emitted by nymphs did not attract either sex, but psyllid honeydew was attractive to males, likely due to female pheromone residues. Males oriented to the odour of females, whereas the reverse was not true, and neither males nor females oriented to same‐sex volatiles. The addition of conspecific cues (adults, nymphs or honeydew) did not increase female attraction to previously infested leaves, but male response was increased by the presence of adults and honeydew, regardless of plant species. Thus, female psyllids appear to orient more strongly to volatiles of plant origin, whereas males respond more strongly to cues emanating from females and conspecific excretions. These results suggest that female psyllids drive the initial colonization of host plants, whereas males orient to females and infested plants. Identification of the specific volatiles involved may permit their use in monitoring and management of this pest.     

Relative importance of natural enemies and abiotic factors as sources of regulation of mealybugs (Hemiptera: Pseudococcidae) in Brazilian coffee plantations

In the present study, we determined the critical stages and the key factors of mortality for Planococcus citri (Hemiptera: Pseudococcidae) in Brazilian coffee plantations using a life table to understand the role of natural biological control on its population. Predators, parasitoids, rainfall, sunlight, physiological disturbances and fungal diseases were collectively responsible for 98.79% in the total mortality of P. citri. Predators belonging to the Chrysopidae, Syrphidae, Dolichopodidae and Coccinellidae families were the most important mortality factors in the early developmental stages of P. citri (i.e. eggs and 1st and 2nd instar nymphs), whereas predators belonging to the Coccinellidae and Chrysopidae families were the most important mortality factors for the last instars (i.e. 3rd instars and adults) for P. citri. The generalist predators Harmonia axyridis, Chrysoperla genanigra and Chrysoperla externa were the key mortality factors for P. citri. The third nymph stage was considered the critical life stage (i.e. the life stage that most influences population size). Our results show that generalist predators and climatic factors are important sources of natural mortality of P. citri governing the population dynamics of this pest in the field.