Abstract The fruit fly (Diptera: Tephritidae) species, Ceratitis capitata , Ceratitis cosyra , Ceratitis rosa , Ceratitis quilicii , and Bactrocera dorsalis are of economic importance in South Africa. These agricultural pests cause extensive damage to a range of commercially produced fruit, primarily for export. These pests are of phytosanitary significance, and their presence in fruit-producing regions in South Africa has led to restrictions in export trade of fresh produce. Accurate identification of these flies, particularly at immature stages intercepted in fruit consignments originating from South Africa, is essential but remains an ongoing challenge. A rapid and accurate identification assay to differentiate these five species is needed for inspection and pest surveillance. High throughput sequencing data were generated for each of the five fruit fly species, and five sets of species-specific primers were designed for use in a multiplex PCR. Each primer set amplifies an amplicon of a different size for each species allowing for accurate identification. PCR sensitivity tests demonstrate that the limit of detection for this assay is 10 ng and 4 ng of DNA when extracted from larvae and adult specimens, respectively. The assay developed can be applied in fruit inspection and survey activities within the country and at ports of entry. 

/pdf/a-multiplex-pcr-assay-for-the-identification-of-fruit-flies-7ugnson7.pdf

A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa

Abstract. Tarno H, Octavia E, Himawan T, Setiawan Y. 2022. Detection of fruit flies (Diptera: Tephritidae) Using Cue-Lure and Methyl Eugenol in Steiner Traps in Depok City and Bogor District, West Java, Indonesia. Biodiversitas 23: 4202-4208. Fruit flies (Diptera: Tephritidae) are one of the most economically important insect pests worldwide. Two types of attractants are recommended for monitoring and controlling fruit flies: methyl eugenol (ME) and cue-lure (CL). The objective of this study was to investigate the species richness and abundance of fruit flies (Diptera: Tephritidae) using methyl eugenol (ME) and cue-lure (CL) in Depok City and Bogor District, West Java, Indonesia. The study was conducted in three sites of fruit orchards in Bogor District i.e., Malay apple (Syzygium malaccense), Lemon (Citrus limon), Papaya (Carica papaya) and two sites of fruit orchards in Depok District i.e., carambola (Averrhoa carambola) and guavas (Psidium guajava). Male fruit flies were trapped using Steiner traps baited with methyl eugenol (ME) and cue-lure (CL). There were 11 fruit fly species collected from this study i.e., Bactrocera albistrigata, B. carambolae, B. dorsalis, B. neocognata, B. umbrosa, B. verbascifoliae, Dacus conopsoides, D. longicornis, Zeugodacus calumniata, Z. caudatus, and Z. cucurbitae. In this study, B. was , D. conopsoides, D. longicornis, and Z. caudatus were only collected in Steiner traps baited with CL and B. dorsalis and B. umbrosa also only responded to ME. During sampling periods, 20,217 male adult fruit flies were collected. The highest relative abundance attracted to ME was B. carambolae and CL was Z. cucurbitae.  In this study, B. albistrigata, B. calumniate, D. longicornis, Z. caudatus and Z. cucurbitae were significantly more attracted to CL than ME. Whereas, B. carambolae, B. dorsalis, B. umbrosa, and B. verbascifoliae were significantly more attracted to ME than CL. Our study showed also did not find invasive species in Depok City and Bogor District, West Java, Indonesia. Our study can be used as a method for monitoring and controlling fruit flies, namely Steiner trap with CL for Z. cucurbitae and Steiner trap with ME for B. carambolae, B. dorsalis and B. umbrosa in Depok City and Bogor District, West Java, Indonesia.  

Diversity and abundance of fruit flies (Diptera: Tephritidae) on different host plants in Depok and Bogor Districts, West Java, Indonesia

The quarantine pests, Anastrepha grandis (Macquart) and Zeugodacus cucurbitae (Coquillett), are economically important for the fruit fly–free area in Brazilian semiarid, an area in which they are not yet present. Therefore, the objective of this study was to assess the probability of establishment of A. grandis and Z. cucurbitae based on the estimate number of generations in different climatic regions of the Brazilian semiarid. For scenarios of future projections, it was estimated the number of generations with increase of 1°C (low radiative forcing scenario) and 4°C (high radiative forcing scenario) in the air temperature. Finally, we also estimate the quarantine period to eliminate the invading population of cucurbit fruit flies in Brazilian semiarid. For this, the average historical air temperature of 32 semiarid municipalities was used and the biology data of fruit flies (thermal threshold of development and thermal constant) were used. The fruit flies are able to present several generations per year in Brazilian semiarid. Anastrepha grandis can present from 7.99 (Sergipe) to 9.66 (Piaui) generations. The melon fly Z. cucurbitae may present from 31.25 (Sergipe) to 40.66 (Piaui) generations. The estimation of species multiplication is accentuated in any season, with greater amplitude in spring and summer. The municipalities of Piaui, Ceara, and Rio Grande do Norte presented the highest estimates of fruit fly generations. The increase of air temperature in a future scenario may favor the quarantine pests, A. grandis and Z. cucurbitae, in Brazilian semiarid. In conclusion, the species A. grandis and Z. cucurbitae can be established in Brazilian semiarid, with particular concern for the fruit fly–free area.

Establishment Probability of Anastrepha grandis and Zeugodacus cucurbitae (Diptera: Tephritidae) in Brazilian Semiarid Based on Thermal Requirements.

Sweet potato virus disease (SPVD), caused by synergistic infection of Sweet potato chlorotic stunt virus (SPCSV) and Sweet potato feathery mottle virus (SPFMV), is responsible for substantial yield loss all over the world. However, there are currently no approved treatments for this severe disease. The crucial role played by RNase III of SPCSV (CSR3) as RNA silencing suppressor during the viruses' synergistic interaction in sweetpotato makes it an ideal drug target for developing antiviral treatment. In this study, high-throughput screening (HTS) of small molecular libraries targeting CSR3 was initiated by a virtual screen using Glide-docking, allowing the selection of 6,400 compounds out of 136,353. We subsequently developed and carried out a kinetic-based HTS using fluorescence resonance energy transfer technology that isolated 112 compounds. These compounds were validated with dose-response assays including the kinetic-based HTS and binding affinity assays using surface plasmon resonance and microscale thermophoresis. Finally, the interference of the selected compounds with viral accumulation was verified in planta In summary, we identified five compounds belonging to two structural classes that inhibited CSR3 activity and reduced viral accumulation in plants. These results provide the foundation for developing antiviral agents targeting CSR3 to provide new strategies for controlling sweetpotato virus diseases.Significance statementWe report here a high-throughput inhibitor identification that targets a severe sweetpotato virus disease caused by co-infection with two viruses (SPCSV and SPFMV). The disease is responsible for up to 90% yield loss. Specifically, we targeted the RNase III enzyme encoded by SPCSV, which plays an important role in suppressing the RNA silencing defense system of sweetpotato plants. Based on virtual screening, laboratory assays, and confirmation in planta, we identified five compounds that could be used to develop antiviral drugs to combat the most severe sweetpotato virus disease.

https://journals.asm.org/doi/pdf/10.1128/JVI.00107-21

In Vitro Identification and In Vivo Confirmation of Inhibitors for Sweet Potato Chlorotic Stunt Virus RNA Silencing Suppressor, a Viral RNase III

Abstract Climate change and biological invasions of insect pests are interlinked global concerns that drive shifts in the distribution of invasive insects. The peach fruit fly, Bactrocera zonata Saunders, is one of the most economically important Tephritidae species that attack several host plants and causes serious damage in Asia and Africa. Currently, B. zonata is absent from many countries and regions but has a risk of invasion. Therefore, it is crucial to investigate the impact of climate change on the global potential distribution of B. zonata . In this study, we used MaxEnt and CLIMEX models to estimate the risk area for B. zonata under near current and future climate conditions. The MaxEnt and CLIMEX results showed that the south of North and Central America was suitable for B. zonata . The European countries were slightly suitable for B. zonata . In Asia, the highly suitable regions of B. zonata included Saudi Arabia, United Arab Emirates, Oman, Iran, Pakistan, India, Nepal, Bangladesh, Bhutan, Myanmar, Thailand, Vietnam, and Laos. Moreover, China, Philippines, Indonesia, and Japan showed highly climate suitability for B. zonata . The climate suitability of B. zonata was increasingly high in the projection under climate change. The result of the two models showed that the climatic suitability for B. zonata will increase under climate change in China. Taken together, these predictive results support the quarantine of B. zonata for high-risk countries and provide in-depth information on how climatic changes may affect its possible geographic range. 

Estimation of the potential geographical distribution of invasive peach fruit fly under climate change by integrated ecological niche models

To control plant viruses, viral RNA silencing suppressors are important drug targets due to their key roles in interfering antiviral RNA silencing. Here we have presented a strategy, combining virtual and experimental screening, to discover the inhibitors of viral suppressor. By docking 157 026 compounds from a natural product library into P19 model, eighteen candidate compounds were selected. Candidates VS2, VS12, VS14 and VS15 displayed strong binding ability to P19 in the surface plasmon resonance imaging assay with KD values of 136.2, 111.6, 81.2 and 124.5 nM, respectively. Then the inhibition activities of these inhibitors on the association between P19 and siRNA were also affirmed by electrophoretic mobility shift assay. Moreover, the antiviral effects on plants showed that compounds VS14 and VS15 both exhibited antiviral activities against Tomato bushy stunt virus (TBSV) in vivo with inhibition rates of 32.35% and 16.61% in 11 dpi, respectively. This strategy would be a powerful tool for the discovery of novel antiviral agents and provide new insights into the control of plant viruses.

/pdf/discovery-of-novel-inhibitors-of-rna-silencing-suppressor-5csexxkf4d.pdf

Discovery of novel inhibitors of RNA silencing suppressor P19 based on virtual screening

The genera Anastrepha, Bactrocera, Ceratitis, Dacus and Rhagoletis in the family Tephritidae order Diptera are economically important, worldwide distributed and cause damage to a large number of commercially produced fruits and vegetables. China had regulated these five genera as quarantine pests, including the species Carpomya vesuviana. An accurate molecular method not depending on morphology able to detect all the quarantine fruit flies simultaneously is required for quarantine monitoring. This study contributes a comparative analysis of 146 mitochondrial genomes of Diptera species and found variable sites at the mt DNA cox2 gene only conserved in economically important fruit flies species. Degenerate primers (TephFdeg/TephR) were designed specific for the economically important fruit flies. A 603 bp fragment was amplified after testing each of the 40 selected representative species belonging to each economically important Tephritid genera, no diagnostic fragments were detected/amplified in any of the other Tephritidae and Diptera species examined. PCR sensitivity assays demonstrated the limit of detection of targeted DNA was 0.1 ng/μl. This work contributes an innovative approach for detecting all reported economically important fruit flies in a single-step PCR specific for reported fruit fly species of quarantine concern in China.

/pdf/a-conserved-motif-within-cox-2-allows-broad-detection-of-h0w6gmyabm.pdf

A conserved motif within cox 2 allows broad detection of economically important fruit flies (Diptera: Tephritidae)

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Discovery of novel inhibitors of RNA silencing suppressor P19 based on virtual screening

A conserved motif within cox 2 allows broad detection of economically important fruit flies (Diptera: Tephritidae)