Electrophysiological and behavioral responses of Tamarixia radiata (Hymenoptera: Eulophidae) to volatiles of nymphal Diaphorina citri (Hemiptera: Liviidae).

Huanglongbing (HLB), a devastating citrus disease caused by Candidatus Liberibacter asiaticus, is efficiently vectored by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Tamarixia radiata (Waterston) plays a crucial role as an ectoparasitoid, preying on D. citri nymphs. By collecting and identifying headspace volatiles from fifth instar nymphs of D. citri using a gas chromatograph-mass spectrometer (GC-MS), we obtained a collection of 9 volatile compounds. These compounds were subsequently chosen to investigate the electrophysiological and behavioral responses of female T. radiata. At a concentration of 10 µg/µl, 9 compounds were compared with cis-3-hexen-1-ol (control), resulting in trans-2-nonenal inducing the highest relative electroantennogram (EAG) value, followed by hexanal, heptanal, n-heptadecane, tetradecanal, n-tetradecane, n-pentadecane, 1-tetradecanol, and 1-dodecanol. The top 5 EAG responses of female T. radiata to these compounds were further investigated through EAG dose-response experiments. The results showed positive dose-responses as concentrations increased from 0.01 to 10 µg/µl. In Y-tube olfactometer bioassays, female T. radiata exhibited a preference for specific compounds. They were significantly attracted to tetradecanal at a concentration of 10 µg/µl and trans-2-nonenal at 0.01 µg/µl, while no significant attraction was observed toward hexanal, heptanal, or n-heptadecane. Our report is the first to demonstrate that volatiles produced by D. citri nymphs attract T. radiata, which suggests that this parasitoid may utilize nymph volatiles to locate its host.

Transcriptome data reveal beneficial effects of Rickettsia (Rickettsiales: Rickettsiaceae) on Bemisia tabaci(Hemiptera: Aleyrodidae) through nutritional factors and defense mechanisms.

Whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a destructive insect pest of many crops. Rickettsia infection in different cryptic species of B. tabaci has been observed worldwide. Understanding the interactions between these 2 organisms is critical to developing Rickettsia-based strategies to control B. tabaci and thereby reduce the transmission of related vector-borne viruses. In this study, we investigated the effects of Rickettsia infection on the biological characteristics of the Middle East Asia Minor 1 (MEAM1) strain of B. tabaci through biological analysis of infected and uninfected individuals. The results of this study suggest that Rickettsia may confer fitness benefits. These benefits include increased fertility, improved survival rates, accelerated development, and resulted in female bias. We also investigated the transcriptomics impact of Rickettsia infection on B. tabaci by performing a comparative RNA-seq analysis of nymphs and adult females, both with and without the infection. Our analysis revealed 218 significant differentially expressed genes (DEGs) in infected nymphs compared to uninfected ones and 748 significant DEGs in infected female adults compared to their uninfected whiteflies. Pathway analysis further revealed that Rickettsia can affect many important metabolic pathways in whiteflies. The results suggest that Rickettsia plays an essential role in energy metabolism, and nutrient synthesis in the B. tabaci MEAM1, and depends on metabolites obtained from the host to ensure its survival. Overall, our findings suggest that Rickettsia has beneficial effects on B. tabaci and offered insights into the potential molecular mechanisms governing the interactions between Rickettsia and B. tabaci MEAM1.

Transcriptomic response of citrus psyllid salivary glands to the infection of citrus Huanglongbing pathogen.

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the key vector insect transmitting the Candidatus Liberibacter asiaticus (CLas) bacterium that causes the devastating citrus greening disease (Huanglongbing, HLB) worldwide. The D. citri salivary glands (SG) exhibit an important barrier against the transmission of HLB pathogen. However, knowledge on the molecular mechanism of SG defence against CLas infection is still limited. In the present study, we compared the SG transcriptomic response of CLas-free and CLas-infected D. citri using an illumine paired-end RNA sequencing. In total of 861 differentially expressed genes (DEGs) in the SG upon CLas infection, including 202 upregulated DEGs and 659 downregulated DEGs were identified. Functional annotation analysis showed that most of the DEGs were associated with cellular processes, metabolic processes, and the immune response. Gene ontology and Kyoto Encyclopaedia of Genes and Genomes enrichment analyses revealed that these DEGs were enriched in pathways involving carbohydrate metabolism, amino acid metabolism, the immune system, the digestive system, the lysosome, and endocytosis. A total of 16 DEGs were randomly selected to further validate the accuracy of RNA-Seq dataset by reverse-transcription quantitative polymerase chain reaction. This study provides substantial transcriptomic information regarding the SG of D. citri in response to CLas infection, which may shed light on the molecular interaction between D. citri and CLas, and provides new ideas for the prevention and control of citrus psyllid.

Rickettsia transmission from whitefly to plants benefits herbivore insects but is detrimental to fungal and viral pathogens.

Bacterial endosymbionts play important roles in the life histories of herbivorous insects by impacting their development, survival, reproduction, and stress tolerance. How endosymbionts may affect the interactions between plants and insect herbivores is still largely unclear. Here, we show that endosymbiotic Rickettsia belli can provide mutual benefits also outside of their hosts when the sap-sucking whitefly Bemisia tabaci transmits them to plants. This transmission facilitates the spread of Rickettsia but is shown to also enhance the performance of the whitefly and co-infesting caterpillars. In contrast, Rickettsia infection enhanced plant resistance to several pathogens. Inside the plants, Rickettsia triggers the expression of salicylic acid-related genes and the two pathogen-resistance genes TGA 2.1 and VRP, whereas they repressed genes of the jasmonic acid pathway. Performance experiments using wild type and mutant tomato plants confirmed that Rickettsia enhances the plants' suitability for insect herbivores but makes them more resistant to fungal and viral pathogens. Our results imply that endosymbiotic Rickettsia of phloem-feeding insects affects plant defenses in a manner that facilitates their spread and transmission. This novel insight into how insects can exploit endosymbionts to manipulate plant defenses also opens possibilities to interfere with their ability to do so as a crop protection strategy. IMPORTANCE: Most insects are associated with symbiotic bacteria in nature. These symbionts play important roles in the life histories of herbivorous insects by impacting their development, survival, reproduction as well as stress tolerance. Rickettsia is one important symbiont to the agricultural pest whitefly Bemisia tabaci. Here, for the first time, we revealed that the persistence of Rickettsia symbionts in tomato leaves significantly changed the defense pattern of tomato plants. These changes benefit both sap-feeding and leaf-chewing herbivore insects, such as increasing the fecundity of whitefly adults, enhancing the growth and development of the noctuid Spodoptera litura, but reducing the pathogenicity of Verticillium fungi and TYLCV virus to tomato plants distinctively. Our study unraveled a new horizon for the multiple interaction theories among plant-insect-bacterial symbionts.

Distribution and accumulation of Cadmium in different trophic levels affecting Serangium japonicum, the predatory beetle of whitefly Bemisia tabaci, biologically, physiologically and genetically: An experimental study.

Cadmium (Cd) is a heavy metal that is of great concern in agroecosystems due to its toxicity to plants, herbivores, carnivores, and human beings. The current study evaluated the allocation and bioaccumulation of Cd from soil to cotton plants, cotton plants to herbivore pests, and herbivorous pests to a natural enemy predator. When soil was spiked with 100 mg/kg Cd, results demonstrated that cotton roots accumulated more Cd than the stems and leaves. The bioaccumulation of Cd was less in 4th instar larvae, pupa, and adults of Serangium japonicum than in Bemisia tabaci adults. The bioaccumulation in S. japonicum elongated the immature development period and reduced adult longevity, oviposition days, fertility, and total pre-oviposition duration. The net reproduction of S. japonicum was also reduced, as was female mature weight and feeding potential; as a result, Cd exposure could reduce the future population size compared to uncontaminated populations. There was decreased activity of the antioxidant enzymes (SOD, CAT, and POD) and energy-conserving lipids (glycogen, triglyceride, and total cholesterol) in Cd-contaminated S. japonicum compared to controls. The detoxifying enzyme activity of GST and P450 increased while AChE activity did not change. The qRT-PCR research showed that SOD1, CAT, POD, glycogen, and triglyceride gene expression was higher than in controls, whereas detoxification gene expression did not change. Our results indicate that Cd exposure has a physiological trade-off between its adverse effects on life history traits and elevated detoxification and antioxidation of S. japonicum, which could result from gene expression alteration. Further studies are needed to assess whether Cd exposure causes irreversible DNA damage in S. japonicum.

Identification and transcriptional profiling of UV-A-responsive genes in Bemisia tabaci.

Ultraviolet-A (UV-A) radiation directly impacts the growth and spread of Bemisia tabaci. However, the mechanistic pathways of this phenomenon remain unknown. We analyzed B. tabaci transcriptome data after exposure to UV-A radiation for 6 h. The 453 genes were identified whose expression were significantly altered in response to the stress induced by UV-A irradiation. Forty genes were up-regulated, while 413 genes were down-regulated. Enrichment analysis using GO, KEGG, and Genomes databases revealed that the DEGs play key roles in antioxidation and detoxification, protein turnover, metabolic, developmental processes, and immunological response. Among the gene families involved in detoxification, shock, and development, down-regulated DEGs in transcriptional factor gene families were significantly greater than those up-regulated DEGs. Our findings demonstrated that exposure to UV-A stress can suppress immunity and affect the growth and biological parameters of B. tabaci by altering gene regulation. These results suggest a potential utility of UV-A stress in managing B. tabaci under greenhouse conditions.

Population changes of Bemisia tabaci (Hemiptera: Aleyrodidae) on different colored poinsettia leaves with different trichome densities and chemical compositions.

The whitefly, Bemisia tabaci, is a destructive and invasive pest of many horticultural plants including poinsettia (Euphorbia pulcherrima). Outbreaks of B. tabaci cause serious damage by direct feeding on phloem sap, and spreading 100+ plant viruses to crops. Bemisia tabaci were observed more frequently on green than red poinsettia leaves, and the factors responsible for this are unknown. Here, we investigated the development rate, survivorship, fecundity of B. tabaci feeding on green versus red leaves, as well as the leaves' volatiles, trichome density, anthocyanin content, soluble sugars, and free amino acids. Compared to red leaves, B. tabaci on green leaves showed increased fecundity, a higher female sex ratio, and survival rate. The green color alone was more attractive to B. tabaci than red. Red leaves of poinsettia contained more phenol, and panaginsene in their volatiles. Alpha-copaene and caryophyllene were more abundant in the volatiles of poinsettia green leaves. Leaf trichome density, soluble sugars and free amino acids were higher in green than red leaves of poinsettia, anthocyanin was lower in green than red leaves. Overall, green leaves of poinsettia were more susceptible and attractive to B. tabaci. The morphological and chemical variation between red and green leaves also differed; further investigation may reveal how these traits affect B. tabaci's responses.

The Dynamic Distribution of Wolbachia and Rickettsia in AsiaII1 Bemisia tabaci.

Wolbachia and Rickettsia are bacterial endosymbionts that can induce a number of reproductive abnormalities in their arthropod hosts. We screened and established the co-infection of Wolbachia and Rickettsia in Bemisia tabaci and compared the spatial and temporal distribution of Wolbachia and Rickettsia in eggs (3-120 h after spawning), nymphs, and adults of B. tabaci by qPCR quantification and fluorescent in situ hybridization (FISH). The results show that the titer of Wolbachia and Rickettsia in the 3-120 h old eggs showed a "w" patterned fluctuation, while the titers of Wolbachia and Rickettsia had a "descending-ascending descending-ascending" change process. The titers of Rickettsia and Wolbachia nymphal and the adult life stages of Asia II1 B. tabaci generally increased with the development of whiteflies. However, the location of Wolbachia and Rickettsia in the egg changed from egg stalk to egg base, and then from egg base to egg posterior, and finally back to the middle of the egg. These results will provide basic information on the quantity and localization of Wolbachia and Rickettsia within different life stages of B. tabaci. These findings help to understand the dynamics of the vertical transmission of symbiotic bacteria.

The phylogeny and distribution of Wolbachia in two pathogen vector insects, Asian citrus psyllid and Longan psyllid.

Background: Wolbachia is the most abundant bacterial endosymbiont among insects. It can play a prominent role in the development, reproduction and immunity of its given insect host. To date, Wolbachia presence is well studied within aphids, whiteflies and planthoppers, but relatively few studies have investigated its presence in psyllids. Methods: Here, the infection status of Wolbachia in five species of psyllid, including Asian citrus psyllid Diaphorina citri and longan psyllid Cornegenapsylla sinica was investigated. The phylogenetic relationships of different Wolbachia lines and their infection density and patterns in D. citri and C. sinica from different countries was also examined. Results: The infection rates of Wolbachia in D. citri and C. sinica were both 100%, and their sequencing types are ST173 and ST532 respectively. Phylogenetic analysis revealed that the Wolbachia lines in D. citri and C. sinica both belong to the Con subgroup of Wolbachia supergroup B. In addition, Wolbachia displayed a scattered localization pattern in the 5th instar nymphs and in the reproductive organs of both D. citri and C. sinica but differed in other tissues; it was highest in the midgut, lowest in the salivary glands and medium in both the testes and ovaries. Conclusion: Our findings assist in further understanding the coevolution of Wolbachia and its psyllid hosts. Given that Wolbachia could play an important role in insect pest control and pathogen transmission inhibition, our findings may also provide new insights for development of control strategies for D. citri and C. sinica.

Blue light stimulates light stress and phototactic behavior when received in the brain of Diaphorina citri.

Blue light with a wavelength of 400-470 nm is the composition of the visible light. However, in recent years, blue light contributed the most significance to light pollution due to the artificial light at night. Previously, we have demonstrated that the Asian citrus psyllid (ACP), Diaphorina citri, an important pest in citrus production, has significant positive phototaxis with a light-emitting diode light of 400 nm. In this study, ACP with positive phototactic behavior to 400 nm light (PH) and non-phototactic behavior to 400 nm light (NP) were collected, individually. Transcriptome dynamics of head tissues of PH and NP groups were captured by using RNA-sequencing technology, respectively. Forty-three to 46 million clean reads with high-quality values were obtained, and 1773 differential expressed genes (DEGs) were detected. Compared with the NP group, there were 841 up-regulated DEGs and 932 down-regulated DEGs in the PH group. Eight pathways were significantly enriched in the PH group in the KEGG database, while 43 up-regulated pathways and 25 down-regulated pathways were significantly enriched in the PH group in the GO database. The DGE approach was reliable validated by real time quantitative PCR. Results indicated that the blue light acted as an abiotic stress causing physiological and biochemical responses such as oxidative stress, protein denaturation, inflammation and tumor development in ACPs. Additionally, the light was absorbed by photoreceptors of ACPs, and converted into electrical signal to regulate neuromodulation. This study provides basic information for understanding the molecular mechanisms of ACP in response to blue light and provides a reference for further studies to elucidate phototactic behavior.

Silencing of V-ATPase-E gene causes midgut apoptosis of Diaphorina citri and affects its acquisition of Huanglongbing pathogen.

The Asian citrus psyllid, Diaphorina citri Kuwayama, is among the most important pests of citrus. It is the main vector of the Huanglongbing (HLB) pathogen Candidatus Liberibacter asiaticus (CLas), which causes severe losses in citrus crops. Control of D. citri is therefore of paramount importance to reduce the spread of HLB. In this regard, using RNA interference (RNAi) to silence target genes is a useful strategy to control psyllids. In this study, using RNAi, we examined the biological functions of the V-ATPase subunit E (V-ATP-E) gene of D. citri, including its effect on acquisition of CLas. The amino acid sequence of V-ATP-E from D. citri had high homology with proteins from other insects. V-ATP-E was expressed at all D. citri life stages analyzed, and the expression level in mature adults was higher than that of teneral adults. Silencing of V-ATP-E resulted in a significant increase in mortality, reduced body weight, and induced cell apoptosis of the D. citri midgut. The reduced expression of V-ATP-E was indicated to inhibit CLas passing through the midgut and into the hemolymph, leading to a majority of CLas being confined to the midgut. In addition, double-stranded RNA of D. citri V-ATP-E was safe to non-target parasitic wasps. These results suggest that V-ATP-E is an effective RNAi target that can be used in D. citri control to block CLas infection.

Rickettsia Infection Benefits Its Whitefly Hosts by Manipulating Their Nutrition and Defense.

Endosymbionts play an essential role in the biology, physiology and immunity of insects. Many insects, including the whitefly Bemisia tabaci, are infected with the facultative endosymbiont Rickettsia. However, the mutualism between Rickettsia and its whitefly host remains unclear. This study investigated the biological and physiological benefits of Rickettsia infection to B. tabaci. Results revealed that infection of Rickettsia increased the fertility, the survival rate from nymph to adult and the number of female whiteflies. In addition, this facilitation caused a significant reduction in nymphal developmental duration but did not affect percentage rate of egg hatching. Rickettsia infected B. tabaci had significantly higher glycogen, soluble sugar and trehalose contents than Rickettsia negative B. tabaci individuals. Rickettsia also improved the immunity of its whitefly hosts. Rickettsia infested B. tabaci had lower mortality rates and higher semi-lethal concentrations (LC50) when exposed to the fungus Akanthomyces attenuatus and the insecticides imidacloprid and spirotetramat. The percentage of parasitism by Encarsia formosa was also reduced by Rickettsia infection. Overall, Rickettsia infection benefits B. tabaci by improving the nutritional composition of its host, and also protects B. tabaci by enhancing its resistance towards insecticides (imidacloprid and spirotetramat), entomopathogenic fungi (A. attenuatus) and its main parasitoid (E. formosa); all of which could significantly impact on current management strategies.

Physiological and Molecular Response Modifications by Ultraviolet-C Radiation in Plutella xylostella and Its Compatibility with Cordyceps fumosorosea.

Ultraviolet-C (UV-C) radiation significantly impacts living organisms. UV-C radiation can also be used as a pest management tool. Therefore, this study was designed to investigate the effect of UV-C radiation on the physiology and gene expression level of Plutella xylostella, a destructive vegetable pest. Results showed that, after exposure to UV-C radiation for 3, 6, 12, and 24 h, the activity of SOD (superoxide dismutase) and CAT (catalase) of P. xylostella increased, while the activity of PPO (polyphenol oxidase), POD (peroxidase), AChE (acetylcholinesterase), CarE (carboxylesterase), and ACP (acid phosphatase) decreased with increased exposure time. Correlation coefficient analyses indicated that the activity of CAT correlated positively, while PPO and CarE correlated negatively, with exposure time. Gene regulation analysis via qRT-PCR confirmed a significant increase in regulation in CAT, CarE, and PPO-related genes. We also investigated the effect of UV-C exposure on the virulence of Cordyceps fumosorosea against P. xylostella. Here, results indicated that when the fungal treatment was applied to larvae before UV-C radiation, the virulence of C. fumosorosea was significantly reduced. However, this decline in virulence of C. fumosorosea due to UV-C exposure remained only for one generation, and no effect was observed on secondary infection. On the other hand, when larvae were exposed to UV-C radiation before fungal application, the mortality rate significantly increased as the exposure time to UV-C radiation increased. From the current study, it could be concluded that UV-C exposure suppressed the immunity to P. xylostella, which later enhanced the virulence of entomopathogenic fungi. Moreover, the study also suggested that UV irradiation is an effective pest management tool that could be incorporated into pest management strategies, which could help reduce pesticide application, be economically beneficial for the farmer, and be environmentally safe.

Parasitoid-mediated horizontal transmission of Rickettsia between whiteflies.

Intracellular bacterial endosymbionts of arthropods are mainly transmitted vertically from mother to offspring, but phylogenetically distant insect hosts often harbor identical endosymbionts, indicating that horizontal transmission from one species to another occurs in nature. Here, we investigated the parasitoid Encarsia formosa-mediated horizontal transmission of the endosymbiont Rickettsia between different populations of whitefly Bemisia tabaci MEAM1. Rickettsia was successfully transmitted from the positive MEAM1 nymphs (R +) into E. formosa and retained at least for 48 h in E. formosa adults. Fluorescence in situ hybridization (FISH) visualization results revealed that the ovipositors, mouthparts, and digestive tract of parasitoid adults get contaminated with Rickettsia. Random non-lethal probing of Rickettisia-negative (R- ) MEAM1 nymphs by these Rickettsia-carrying E. formosa resulted in newly infected MEAM1 nymphs, and the vertical transmission of Rickettsia within the recipient females can remain at least up to F3 generation. Further phylogenetic analyses revealed that Rickettsia had high fidelity during the horizontal transmission in whiteflies and parasitoids. Our findings may help to explain why Rickettsia bacteria are so abundant in arthropods and suggest that, in some insect species that shared the same parasitoids, Rickettsia may be maintained in populations by horizontal transmission.

Impact of Huanglongbing Pathogen Infection on the Amino Acid Composition in Both Citrus Plants and the Asian Citrus Psyllid.

The Asian citrus psyllid (ACP) Diaphorina citri is the main vector of the pathogen Candidatus Liberibacter asiaticus (CLas), which is the causal agent of citrus Huanglongbing disease. Feeding by both ACP nymphs and adults on host plants allows them to obtain nutrition. Therefore, the nutritional content within the plant phloem is of much importance for the development and reproduction of ACP. The infection by pathogenic microbiomes may affect the amino acid contents of their host plants and then indirectly affect the biology of sap-feeding insects. In this study, we investigated the amino acid contents and their proportions in both CLas-infected and CLas-free citrus plants, ACP adults, and also in honeydew produced by ACP nymphs. Results showed that infection by CLas had a large impact on the amino acid species and proportion in all the tested target plants, ACP adults, and in the honeydew of ACP nymphs. The content of total amino acids in CLas-infected citrus was much higher than that of CLas-free citrus. However, CLas infection significantly reduced the proportion of essential amino acids (EAAs) in these plants. When feeding on CLas-infected citrus plants, ACP adults absorbed less total amino acids than those adults feeding on healthy plants, but the proportion of EAAs was significantly higher when they fed on CLas-infected citrus plants. The proportion of EAAs also significantly increased in the honeydew secreted by ACP nymphs that fed on CLas-infected citrus plants. However, EAA detection in the honeydew of ACP nymphs indicated that the utilization rate of EAAs by CLas positive ACP nymphs was reduced. Our study has revealed that CLas infection significantly affects the contents, proportion, and utilization efficiency of different amino acids in citrus plants, ACP adults, and nymphs, leading to a developmental pattern of ACP that is more conducive to CLas transmission.

Parasitoid vectors a plant pathogen, potentially diminishing the benefits it confers as a biological control agent.

Huanglongbing (HLB) is a destructive disease of citrus primarily transmitted by the Asian citrus psyllid (ACP). Biocontrol of ACP is an environmentally sustainable alternative to chemicals. However, the risk of parasitoid rational application in ACP biocontrol has never been evaluated. Here we show, the dominant parasitoid of ACP, Tamarixia radiata, can acquire the HLB pathogen Candidatus Liberibacter asiaticus (CLas) and transmit it horizontally when probing ACP nymphs. If these ACP nymphs survive the probing, develop to adults and move to healthy plants, CLas can be transmitted to citrus leaves during feeding. We illustrate the formerly unrecognized risk that a parasitoid can potentially serve as a phoretic vector of the pathogen transmitted by its host, thus potentially diminishing some of the benefits it confers via biocontrol. Our findings present a significant caution to the strategy of using parasitoids in orchards with different infection status of insect-vectored pathogens.

Lethal and sublethal effects of emamectin benzoate on life-table and physiological parameters of citrus red mite, Panonychus citri.

The citrus red mite (Panonychus citri) is a challenge to manage in citrus orchards due to resistance against several pesticides. There is a necessity therefore to find new pesticides for effective control of P. citri. This study was designed to evaluate the lethal and sublethal effects of emamectin benzoate against P. citri. The results showed that the LC50 of emamectin benzoate to adults of P. citri was 0.35 (0.26-0.43) mg a.i. L-1 and the LC90 was 1.44 (1.16-1.96) mg a.i. L-1. The sublethal concentration exposures (LC10 and LC30) had a significant negative impact on the larval, protonymph, and deutonymph developmental periods. Male longevity was much lower in LC30 treatments than in the controls. Although female longevity was unaffected, the fecundity (eggs per female) was decreased in the sublethal concentration treatments. Results revealed that the adult pre-oviposition period (APOP) and total pre-oviposition period (TPOP) were increased. Other growth parameters r, λ, and R0 decreased, whereas mean generation time (T) increased due to pesticide exposure. The survival rate (Sxj), age-specific fecundity and net maternity, life expectancy (Exj), and reproduction (Vxj) was reduced by LC10 and LC30 exposure. An increase in malondialdehyde (MDA) contents with increasing emamectin benzoate concentration demonstrates that emamectin benzoate induces oxidative stress in P. citri. The activity of antioxidant enzymes (superoxide dismutase, SOD and catalase, CAT) was decreased due to LC30 and LC10 treatments compared to the control. Detoxification enzyme activity (cytochrome P450, glutathione-S-transferases, GST and acetylcholinesterase, AChE) was increased in treated mites compared to the control. This study demonstrates that emamectin benzoate has both a lethal effect on citrus red mite and sublethal effects on its biology and physiology. It is, therefore, potentially an effective pesticide for management of P. citri.

Phototoxicity of Ultraviolet-A against the Whitefly Bemisia tabaci and Its Compatibility with an Entomopathogenic Fungus and Whitefly Parasitoid.

Ultraviolet (UV) radiation significantly affects insect life and, as a result, has been widely used to control different invertebrate pests. The current results demonstrate that when Bemisia tabaci first instar nymphs are exposed to UV-A light for 12, 24, 48, and 72 h, their developmental and biological parameters are negatively affected by UV-A exposure; the effect increased with an increase in exposure time. We hypothesized that UV-A light is compatible with other biological control agents. Results showed that when the entomopathogenic fungus Cordyceps fumosorosea was applied to third instar nymphs of B. tabaci previously exposed to UV-A light, the LC50 was 3.4% lower after 72 h of exposure to UV-A light compared to the control. However, when the fungus was exposed to UV-A light, its virulence decreased with an increase in UV-A exposure time. The parasitism rate of Encarsia formosa against 24 h UV-A-exposed third instar nymphs of B. tabaci increased while the adult emergence from parasitized nymphs was not affected after UV-A light exposure. Parasitism rate was significantly reduced however following E. formosa exposure to UV-A light; but again, adult emergence was not affected from parasitized nymphs. The percentage mortality of E. formosa increased with increasing exposure time to UV-A light. The enzyme activity of SOD, CAT, GST, and AChE and the energy reserve contents were negatively affected due to UV-A exposure. Collectively, this study has demonstrated that UV-A light significantly suppresses the immune system of B. tabaci and that UV-A light is compatible with other biological control agents if it is applied separately from the biological agent.

Rickettsia increases its infection and spread in whitefly populations by manipulating the defense patterns of the host plant.

The whitefly Bemisia tabaci is a destructive agricultural pest that frequently harbors various species of secondary symbionts including Rickettsia. Previous studies have revealed that the infection of Rickettsia can improve whitefly performance on food plants; however, to date, no evidence has shown, if, and how, Rickettsia manipulates the plant-insect interactions. In the current study, the effects of Rickettsia persistence on the induced plant defenses and the consequent performance of whitefly B. tabaci were investigated. Results revealed that Rickettsia can be transmitted into plants via whitefly feeding and remain alive within the cotton plants for at least 2 weeks. The different expression genes of cotton plants were mostly concentrated in the phytohormone signaling pathways, the marker genes of jasmonic-acid signaling pathway (AOC, AOS, LOX, MYC2) were significantly downregulated, while the marker genes of the salicylic-acid signaling pathway (WRKY70, PR-1) were upregulated. Biological experiments revealed that the fecundity of Rickettsia negative B. tabaci significantly increased when they fed on Rickettsia-persistent cotton plants. Taken together, we provide experimental evidence that the persistence of Rickettsia and its induced defense responses in cotton plants can increase the fitness of whitefly and, by this, Rickettsia may increase its infection and spread within its whitefly host.

Antagonistic interaction between male-killing and cytoplasmic incompatibility induced by Cardinium and Wolbachia in the whitefly, Bemisia tabaci.

Cardinium and Wolbachia are maternally inherited bacterial symbionts of arthropods that can manipulate host reproduction by increasing the fitness of infected females. Here, we report that Cardinium and Wolbachia coinfection induced male-killing and cytoplasmic incompatibility (CI) when they coexisted in a cryptic species of whitefly, Bemisia tabaci Asia II7. Cardinium and Wolbachia symbionts were either singly or simultaneously localized in the bacteriocytes placed in the abdomen of B. tabaci nymphs and adults. Cardinium-Wolbachia coinfection induced male-killing and resulted in a higher female sex ratio in the intraspecific amphigenetic progeny of Asia II7 ICWH and ICWL lines; interestingly, male-killing induction was enhanced with increased Cardinium titer. Moreover, single infection of Wolbachia induced partial CI in the Asia II7 IW line and resulted in reduced fecundity, higher embryonic mortality, and lower female sex ratio. The uninfected Asia II7 IU line had significantly higher fecundity, lower embryonic and nymphal mortalities, and a lower level of CI than both the Wolbachia-infected Asia II7 IW line and the Cardinium-Wolbachia-coinfected Asia II7 ICWH line. Our findings indicate that Cardinium-Wolbachia coinfection induced male-killing, which may have had antagonistic effects on Wolbachia-induced CI in the Asia II7 whiteflies. For the first time, our study revealed that B. tabaci Asia II7 reproduction is co-manipulated by Cardinium and Wolbachia endosymbionts.