Assessment of Variation in Feeding Behavior by Color Morph in the Asian citrus Psyllid (Diaphorini Citri Kuwayama)

The Asian citrus psyllid (ACP) Diaphorina citri (Hemiptera:Liviidae), vector of huanglongbing disease, displays a high degree of color polyphenism. In the adult stage, ACP exhibits abdominal colors that can be separated into three color groupings: blue-green, grey-brown and orange-yellow. Color morphology has been shown to influence important and energetically costly psyllid life traits including reproduction, dispersion, immune defense and resistance to insecticides. Despite this, it remains unclear how color morphology is correlated with feeding behavior. Understanding variation in feeding behavior of the ACP color morphs is critical to better understanding how ACP populations utilize host-plants and to assess potential risk for transmission of the causal agent of huanglongbing disease. We compared the feeding behavior of the three ACP color morphs by using electropenetrography (EPG). We did not detect differences in the feeding behavior activities at phloem or xylem tissues when comparing the three-color morphs. Furthermore, there were no differences in feeding behavioral parameters before reaching phloem or xylem tissues. Our results suggest energy requirements are similar between color morphs and feeding behavior parameters associated with CLas transmission are potentially similar between color morphs.

Monitoring Population Phenology of Asian Citrus Psyllid Using Deep Learning

Asian citrus psyllid, Diaphorina citri Kuwayama (Liviidae: Hemiptera) is a menacing and notorious pest of citrus plants. It vectors a phloem vessel-dwelling bacterium Candidatus Liberibacter asiaticus, which is a causative pathogen of the serious citrus disease known as Huanglongbing. Huanglongbing disease is a major bottleneck in the export of citrus fruits from Pakistan. It is being responsible for huge citrus economic losses globally. In the current study, several prediction models were developed based on regression algorithms of machine learning to monitor different phenological stages of Asian citrus psyllid to predict its population about different abiotic variables (average maximum temperature, average minimum temperature, average weekly temperature, average weekly relative humidity, and average weekly rainfall) and biotic variable (host plant phenological patterns) in citrus-growing regions of Pakistan. The pest prediction models can be used for proper applications of pesticides only when needed for reducing the environmental and cost impacts of pesticides. Pearson’s correlation analysis was performed to find the relationship between different predictor (abiotic and biotic) variables and pest infestation rate on citrus plants. Multiple linear regression, random forest regressor, and deep neural network approaches were compared to predict population dynamics of Asian citrus psyllid. In comparison with other regression techniques, a deep neural network-based prediction model resulted in the least root mean squared error values while predicting egg, nymph, and adult populations.

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.

The Influence of Temperature and Host Gender on Bacterial Communities in the Asian Citrus Psyllid

The Asian citrus psyllid, D. citri Kuwayama is the primary vector for Candidatus Liberibacter asiaticus (CLas), which causes a destructive disease in citrus plants. Bacterial symbionts are important determinants of insect physiology, and they can be impacted by many external factors. Temperature is an important abiotic factor affecting insect physiology, and it is also known that differences in symbiont proportions may vary in different insect genders. To date, it is unclear how the symbionts of D. citri are affected by temperature and gender. This study used high-throughput sequencing of 16S ribosomal RNA amplicons to determine how temperature and gender affect the bacterial communities present in D. citri. We identified 27 amplicon sequence variants (ASVs) belonging to 10 orders, seven classes, and five phyla. The dominant phylum was Proteobacteria (99.93%). Other phyla, including Firmicutes, Bacteroidota, Deinococcota, Cyanobacteria, and Actinobacteriota, were less abundant (<0.1%). Profftella (71.77–81.59%) and Wolbachia (18.39–28.22%) were the predominant taxa in all samples. Under high-temperature treatment, Profftella was more common in females, while Wolbachia had a higher abundance in males. In males, Profftella was more abundant under low-temperature treatments than under high-temperature treatments. In contrast, Wolbachia showed a higher abundance under high-temperature treatments than under low-temperature treatments. An RT-qPCR (quantitative real-time PCR) approach confirmed the results obtained with high-throughput DNA sequencing. Our results provide a basis for understanding the co-adaptation of D. citri and its symbionts to environmental temperature stress.

Controlling Citrus Huanglongbing: Green Sustainable Development Route Is the Future

Huanglongbing (HLB) is the most severe bacterial disease of citrus crops caused by Candidatus Liberibacter spp. It causes a reduction in fruit yield, poor fruit quality, and even plants death. Due to the lack of effective medicine, HLB is also called citrus “AIDS.” Currently, it is essential for the prevention and control of HLB to use antibiotics and pesticides while reducing the spread of HLB by cultivating pathogen-free seedlings, removing disease trees, and killing Asian citrus psyllid (ACP). New compounds [e.g., antimicrobial peptides (AMPs) and nanoemulsions] with higher effectiveness and less toxicity were also found and they have made significant achievements. However, further evaluation is required before these new antimicrobial agents can be used commercially. In this review, we mainly introduced the current strategies from the aspects of physical, chemical, and biological and discussed their environmental impacts. We also proposed a green and ecological strategy for controlling HLB basing on the existing methods and previous research results.

Individual Protective Covers (IPCs) for Young Tree Protection from the HLB Vector, the Asian Citrus Psyllid

Psyllid exclusion is the most effective strategy to keep citrus trees free from HLB. Individual protective covers (IPCs) are a novel strategy based on psyllid exclusion of individual trees using a protective mesh bag. This new 3-page publication summarizes the knowledge we have accrued from our 3-year-long field studies using the IPC system and provides recommendations that can help with effective use of this technology. By F. Alferez, U. Albrecht, S. Gaire, O. Batuman, J. Qureshi, and M. Zekri.https://edis.ifas.ufl.edu/hs1425

Silencing of Aquaporin Homologue Accumulates Uric Acid and Decreases the Lifespan of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae)

The Asian citrus psyllid, Diaphorina citri Kuwayama is devastating the citrus industry worldwide. It transmits ‘Candidatus Liberibacter asiaticus’, the pathogen of Huanglongbing in citrus. RNA interference is an excellent tool for functional genomics and for screening target genes for pest control. Herein, we silenced the aquaporin (AQP) gene (DcAQP) homologue in D. citri to study its functionality and whether it could be a good target for a control strategy. AQP is an integral membrane channel protein that aids in the rapid flux of water and other small solutes that move across the lipid membrane. In Hemiptera, it is well established that AQP plays important roles in adjusting to physiological challenges including (1) regulating osmotic stress between the gut lumen and hemolymph after imbibing large quantities of a low nitrogen, sugar-rich liquid diet; (2) avoiding or preventing dehydration and desiccation; and (3) surviving at elevated temperatures. The dsRNA-DcAQP was applied twice to nymphs of the 4th and 5th instars through a soaking technique. Silencing AQP caused a significant increase in nymph mortality. Emerged adults showed malformations and a shorter lifespan. Silencing DcAQP provoked alterations in some metabolites and increased the uric acid content in emerged adults. DcAQP could be a useful target to control D. citri.