Bemisia Tabaci Research Articles

Functional analysis of a NPC1 gene from the whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).

Niemann-Pick C (NPC) disease is a neurodegenerative disorder related to cellular sterol trafficking and mutation of NPC1 gene is the main cause for this disease. The function of NPC1 have been reported in a few insects but rarely studied in hemipterans. In the present study, we investigate the function of NPC1 in a hemipteran pest, the whitefly Bemisia tabaci. It was found that B. tabaci had only one NPC1 homolog (BtNPC1), in contrast to two homologs in many other insects. BtNPC1 was ubiquitously expressed at all developmental stages and body parts of whiteflies, with the highest level in adult abdomen, and the expression of BtNPC1 was induced by cholesterol feeding. To further investigate the function of BtNPC1, leaf-mediated RNA interference experiments were carried out. Results showed that knockdown of BtNPC1 led to reduced survival of whiteflies, as well as reduced fecundity. Moreover, knockdown of BtNPC1 affected the development and metamorphosis of whitefly nymphs. Taken these together, we conclude that BtNPC1 played a crucial role in sterol-related biological processes of B. tabaci and might be used as an insecticide target for development of novel pest management approaches.

Akanthomyces dipterigenus acts as a potential biocontrol agent by causing mortality and inhibiting population growth of Bemisia tabaci

Bemisia tabaci is a worldwide pest, and has developed severe resistance to many insecticide. Therefore, it is great significance to find new substitute for chemical insecticide to control B. tabaci. In this study, selected efficient strains by comparing the virulence of Akanthomyces dipterigenus, Cordyceps javanica, Purpureocillium lilacinum, Penicillium camemberti, Metarhizium anisopliae, and Beauveria bassiana against 2nd-instar nymphs of B. tabaci and bioassays were conducted on various developmental stages of B. tabaci using highly pathogenic strain. Additionally, the effects of medium lethal concentration of high-efficiency strains on the biological parameters of B. tabaci were also studied. The bioassay showed that six entomopathogenic fungi have significant effects on the 2nd-instar nymphs of B. tabaci, the median lethal concentration (LC50) of A. dipterigenus against 2nd-instar nymphs of B. tabaci was only 6.54 × 103 (3.35 × 103–1.13 × 104) conidia mL−1, which was lower than that of the other five fungi, and the median lethal time (LT50) was the shortest. In addition, A. dipterigenus also had lethal effects on eggs, nymphs and adults of B. tabaci, among which the mortality rate of 2nd-instar nymphs was the highest. The fungal treatment affected the population parameters of B. tabaci, R0, r, and λ were significantly reduced, T was significantly prolonged, and had higher preadult mortality and shorter adult longevity, but had no significant effect on the total fecundity. These findings highlight the potential of using A. dipterigenus as a sustainable and eco-friendly alternative to chemical insecticides for controlling B. tabaci populations in agricultural settings.

Transcriptomic Analysis of Tobacco Plants in Response to Whitefly Infection.

The whitefly Bemisia tabaci is one of the most destructive pests worldwide, and causes tremendous economic losses. Tobacco Nicotiana tabacum serves as a model organism for studying fundamental biological processes and is severely damaged by whiteflies. Hitherto, our knowledge of how tobacco perceives and defends itself against whiteflies has been scare. In this study, we analyze the gene expression patterns of tobacco in response to whitefly infestation. A total of 244 and 2417 differentially expressed genes (DEGs) were identified at 12 h and 24 h post whitefly infestation, respectively. Enrichment analysis demonstrates that whitefly infestation activates plant defense at both time points, with genes involved in plant pattern recognition, transcription factors, and hormonal regulation significantly upregulated. Notably, defense genes are more intensely upregulated at 24 h post infestation than at 12 h, indicating an increased immunity induced by whitefly infestation. In contrast, genes associated with energy metabolism, carbohydrate metabolism, ribosomes, and photosynthesis are suppressed, suggesting impaired plant development. Taken together, our study provides comprehensive insights into how plants respond to phloem-feeding insects, and offers a theoretical basis for better research on plant-insect interactions.

Resistance in Soybean Genotypes to Whitefly <i>Bemisia Tabaci</i>

This study demonstrates the defensive responses of eight soybean genotypes based on plant metabolites to the whitefly Bemisia tabaci. At two sampling intervals (30 and 50 days after sowing) secondary metabolites viz., total phenols, o-dihydroxy phenols, flavonols and tannins and primary metabolites viz., total soluble sugars, reducing sugars and total soluble proteins have been estimated from uninfested and infested soybean plants (125 whitefly adults/plant). The genotypes revealed significant variation in secondary metabolites while primary metabolites decreased. The correlation between whitefly and metabolites showed that secondary metabolites were significantly negatively correlated incidence while primary metabolites were significantly positively correlated.

Genetics of begomovirus resistance in Cucurbita moschata Duchesne: A novel resistant source PVR-1343

Whitefly transmitted begomoviruses are the most destructive virus(es) that causes quantitative and qualitative losses to pumpkin (Cucurbita moschata) crop worldwide. In present study we have molecularly characterized three types of viral infections, which were caused by squash leaf curl China virus (SLCCNV), tomato leaf curl New Delhi virus (ToLCNDV), and their mixed infection (MI-Sq/To). Screening of diverse germplasm through whitefly (Bemisia tabaci mitotype Asia II) mediated artificial inoculation and PCR confirmed the 'PVR-1343' genotype as a resistance source against identified begomoviruses. To understand the genetic control of begomovirus resistant gene(s), segregating populations derived from crosses of 'PVR-1343' and four susceptible genotypes (MVSR-6711, Punjab Samrat, MBN-6242 and P-278) inoculated with SLCCNV, ToLCNDV and MI-Sq/To by whitefly in closed environment for three consecutive years (2018–2020). Disease scoring at 75 days after sowing rendered all F1 progenies as susceptible indicating recessive nature of resistance with no maternal or cytoplasmic effects. Chi-square analysis indicated a good fit of 9(S):3(MS):3(MR):1(R) ratio in F2 progenies of all crosses. Backcross of the F1’s to the resistant parent segregated into 1:1:1:1 ratio, while all progeny were susceptible in backcross of the F1’s with susceptible parent. Phenotypic segregation ratio of F2:3 confirmed the F2 genotypic ratio of 1:2:2:4:1:2:1:2:1. This substantiated that multiple begomovirus resistance in 'PVR-1343' is controlled by digenic recessive genes. Moreover, F2:3 progenies resistant to SLCCNV were also resistant to ToLCNDV and MI-Sq/To and vice versa, suggesting that common or tightly linked gene(s) might be involved in resistance to both viruses.

Pathogens are an important driving force for the rapid spread of symbionts in an insect host.

One of the biggest challenges for pathogens invading hosts is microbial symbionts but the role of pathogens in symbionts in nature is unknown. By tracking the dynamics of the entomopathogenic fungal Cordyceps javanica and symbionts in natural populations of the whitefly Bemisia tabaci from 2016 to 2021 across China, we reveal that Rickettsia, a newly invaded symbiont, is positively correlated with the pathogen in both frequency and density. We confirm that applying pathogen pressure can selectively drive Rickettsia to sudden fixation in whiteflies both in the laboratory and in the field. Furthermore, the driving force is elucidated by the Rickettsia-conferred suppression of pathogen infection quantity, proliferation and sporulation, acting as a potential barrier of onward transmission of the pathogen. These results show that pathogens are an important driving force for rapid shifts in host symbionts in the natural niche.

Post-Application Field Persistence and Efficacy of Cordyceps javanica against Bemisia tabaci.

Previously, Cordyceps javanica Wf GA17, a causing agent of whitefly epizootics in southern Georgia, demonstrated superior temperature tolerance and higher virulence against the whitefly Bemisia tabaci than commercial strains in the laboratory. The post-application persistence and efficacy of this fungus against B. tabaci were compared with that of the commercially available C. javanica Apopka97 strain over a two-year field study in cotton and vegetable crops. When blastospores of both strains were applied alone, whitefly populations were not effectively suppressed. Thus, JMS stylet oil was added to fungal treatments for enhancing efficacy and persistence. For 0-day samples, all fungal treatments caused similar but significant levels of immature mortality regardless of fungal strain, propagule form (conidia vs. blastospores), and application method (alone or mixed with JMS). In follow-up samplings, Wf GA17 blastospores + JMS achieved higher control levels than other treatments in some trials, but the efficacy did not last long. The JMS oil alone caused significant mortality and suppressed whiteflies. Over 90% of spores lost viability 24 h after treatment in all fungal treatments. Across evaluation times, there was no difference between the two fungal strains (conidia or blastospores, alone or combined with JMS), but conidia persisted better than blastospores for both strains. Overall, the field persistence and efficacy of C. javanica did not last long; therefore, improved delivery methods and formulations are needed for enhancement.

Leaf-mediated response of soybean genotypes to infestation by whitefly,Bemisia tabaci(Gennadius)

AbstractThe whitefly,Bemisia tabaci(Gennadius) is documented as a major pest on soybean. It was reported that whitefly response towards its hosts and their cultivars varies, and is mediated through various host-related factors. Considering the significance of leaf morphological characteristics in influencing the host–whitefly responses, the present investigation was conducted in screen-house conditions to study the prevailing variations in leaf morphological characteristics of soybean genotypes and their role in governing the adult whitefly attractiveness and oviposition preference. In the multiple-choice test, the whitefly population (eggs, nymphs and adults) was found to be lowest in moderately resistant genotypes (SL 1028 and SL 1074) compared to highly susceptible (DS 3105) and susceptible genotypes (SL 688, SL 958 and SL 1113). The foliar trichomes were characterized using scanning electron microscopy and leaf area, leaf lamina thickness and leaf shape data were acquired using standard procedures. To determine the factors involved in the resistance/susceptible responses towards whitefly, Pearson correlation was applied between the morphological characteristics and the whitefly population. The results show that the leaf area, trichome density, trichome length and trichome angle showed a significant positive correlation with the whitefly population, whereas leaf lamina thickness was negatively correlated. Thus, for developing whitefly-resistant germplasm, breeders should choose genotypes having narrow and thick leaves with sparse, short and flat trichomes.

First Report of Tomato Leaf Curl Cebu Virus Associated with Stachytarpheta jamaicensis Golden Yellow Mosaic Disease in Taiwan.

Stachytarpheta jamaicensis (L.) Vahl, also known as snack weed, is an exotic plant in Taiwan. In April 2021, severe golden yellow mosaic leaves (Fig. S1) were observed on S. jamaicensis plants in Taichung City, Taiwan. Samples from eight symptomatic and two asymptomatic plants were collected from the public flowerbed. Total DNA was extracted from each of the collected samples by using a modified CTAB method (Echevarría-Machado et al. 2005). PCR with Begomovirus degenerate primers (PAL1v1978/PAR1c715; Rojas et al. 1993) was conducted. The expected 1.5-kb fragment was amplified only from the 8 symptomatic samples. Two randomly selected amplicons were cloned on pCRII-TOPO TA vector (Invitrogen Co., San Diego, CA, USA) and sequenced with the ABI3730 automatic sequencer (Applied Biosystems, Hammonton, NJ, USA) at National Chung Hsing University (NCHU). After NCBI BLASTn analysis, the sequences were shown to be most closely related to tomato leaf curl Cebu virus (ToLCCeV) isolates (EU487042, EU487025, KU946997), with 92.4-92.5% nucleotide sequence identity by using the CLUSTAL W method of MegAlign program (DNASTAR, Inc., Madison, WI, USA). A ToLCCeV specific primer pair (FJJ2021-165 /166 5'-ACTTACAGGCCCATGTATCG-3' / 5'-GAATGGGTATCCGAGCACG-3') was designed to amplify and sequence the remaining half of viral DNA. The expected 1.6-kb amplicon was amplified only from the symptomatic samples. The full-length of DNA-A consisted of 2.7-kb nucleotides (ToLCCeV isolate stachy, ON525110 and ON525111) and contained six open reading frames (two in viral sense, V1 to V2 and four in the viral complementary sense, C1 to C4) and the conserved nonanucleotide motif (TAATATTAC). The full-length DNA-A of ToLCCeV stachy isolates shared 99.9% nucleotide identity to each other and 91.2-92.4% and 91.3-92.5% nucleotide identities to other ToLCCeV isolates (EU487042, EU487025, KU946997) available in NCBI GenBank. Besides, ToLCCeV is a monopartite begomovirus that harbors no DNA-B. Thus, there were no bands amplified from the degenerate primer pair for DNA-B (DNABLC2 / DNABLV2; Green et al. 2001). Furthermore, the infectious clone was constructed by using phi29 DNA polymerase (New England Biolabs, Ipswich, MA, USA) for rolling circle amplification (RCA). The RCA product was partially digested with ApaI (NEB) and ligated into the binary vector pCambia0380 (AF234290). The resulting recombinant vector was transformed into Agrobacterium tumefaciens C58. A. tumefaciens C58, containing the infectious ToLCCeV-Stachy DNA-A vector, was grown overnight in LB broth containing kanamycine (50 μg/ml) at 28°C. S. jamaicensis and Nicotiana benthamiana (Nb, four to six leaf stage) plants were agroinoculated to confirm the infectivity of the ToLCCeV clone. The leaf curling and blister symptoms were observed on the Nb systemic leaves 17-day post inoculation (dpi) and the golden yellow mosaic symptom noticed on S. jamaicensis systemic leaves 30-dpi. The presence of the viral DNA in the inoculated plants was confirmed by PCR using the specific primer pair of ToLCCeV. To the best of our knowledge, this is the first report of the monopartite begomovirus, ToLCCeV, associated with golden yellow mosaic disease in S. jamaicensis in Taiwan. The existence of ToLCCeV might severely impact the tomato and pepper industry because they are the natural hosts of ToLCCeV (Tsai et al. 2011) and ToLCCeV may be transmitted by the whitefly, Bemisia tabaci, in Taiwan (Ko et al. 2005).

Evaluation of Insecticides in the Management of Whiteflies (Bemisia tabaci Gennadius) and their Impacts on Yield of Eggplants

Evaluation of Insecticides in the Management of Whiteflies (Bemisia tabaci Gennadius) and their Impacts on Yield of Eggplants

Flavonoid production in tomato mediates both direct and indirect plant defences against whiteflies in tritrophic interactions.

The role of plant flavonoids in direct defences against chewing and sap-sucking herbivorous insects has been extensively characterized. However, little is known about flavonoid-mediated tritrophic interactions between plants, herbivorous insects and natural enemies. In this study, we investigated how flavonoids modulate plant-insect interactions in a tritrophic system involving near-isogenic lines (NILs) of cultivated tomato (Solanum lycopersicum) with high (line NIL-purple hypocotyl [PH]) and low (line NIL-green hypocotyl [GH]) flavonoid levels, with a generalist herbivore whitefly (Bemisia tabaci) and its predatory bug (Orius sauteri). By contrasting levels of tomato flavonoids (direct defence) while manipulating the presence of predators (indirect defence), we found that high production of flavonoids in tomato was associated with a higher inducibility of direct defences and a stronger plant resistance to whitefly infestation and stimulated the emissions of induced volatile organic compounds, thereby increasing the attractiveness of B. tabaci-infested plants to the predator O. sauteri. Furthermore, suppression of B. tabaci population growth and enhancement of plant growth were mediated directly by the high production of flavonoids and indirectly by the attraction of O. sauteri, and the combined effects were larger than each effect individually. Our results conclude that high flavonoid production in tomato enhances herbivore-induced direct and indirect defences to better defend against herbivores in tritrophic interactions. Thus, the development of transgenic plants may present an opportunity to utilize the beneficial role of flavonoids in integrated pest management, while simultaneously maintaining or improving resistance against other pests and pathogens. This article is protected by copyright. All rights reserved.

Occurrence of Tomato Leaf Curl New Delhi Virus in Cucurbit Plants in China.

In August 2022, melon (Cucumis melo), cucumber (Cucumis sativus) and luffa (Luffa aegyptiaca) plants showed virus-like symptoms characteristic of geminiviruses (yellowish green, mosaic patterns and severe curling of leaves, short internodes, and stunting) in 10 greenhouses in Fengxian district and 20 greenhoues in Jiading district of Shanghai, China. Fifty symptomatic leaf samples were randomly collected: 28 from melon, 17 from cucumber, and 5 from luffa. To investigate the etiology of the observed disease, total DNA and RNA was extracted via a DNA extraction kit (Tiangen, Not: DP350) and TRIzol reagent (Sigma-Aldrich, Not: T9424), respectively. Healthy melon plants grown in a growth chamber served as negative control. The DNA and RNA samples were screened for the presence of geminiviruses, Cucurbit chlorotic yellow virus (CCYV), Melon yellow spot virus (MYSV), Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV) and Cucumber green mottle mosaic virus (CGMMV) through PCR or RT‒PCR with geminiviruses (Deng et al. 1994), CCYV, MYSV, CMV, ZYMV and CGMMV (Zeng et al. 2011, 2019) primers. The PCR results showed that 28 melon leaves and 17 cucumber leaves were positive for geminivirus and CCYV, respectively, 5 luffa samples were infected with only geminivirus, and virus was not detected from the healthy plants. These results indicate that these two viruses are widely distributed throughout cucurbit crops in Shanghai, China. All the geminiviruses sequences (approximately 510 bp) were quite similar to each other and were most similar (99.4%) to the Tomato leaf curl New Delhi virus (ToLCNDV) sequence (GenBank Accession No. OP356207) (Li et al. 2022). To confirm the presence of geminiviruses, the segments of DNA-A and DNA-B were amplified by PCR with 4 ToLCNDV-specific primer sets (Mizutani et al. 2011) and sequenced from 10 samples (4 melon, 4 cucumber and 2 luffa). Both DNA-A and DNA-B of the ToLCNDV sequences and features were deposited in GenBank under the accession numbers OQ190939-OQ190948 (DNA-A, 2739 nt) and OQ190949-OQ190958 (DNA-B, 2693 nt). BLASTn analysis of Shanghai isolates of ToLCNDV (DNA-A and DNA-B) showed that the sequences shared nucleotide identities ranged from 99.3% to 100% among them and with values of more than 99.4% nucleotide identity with ToLCNDV isolates from tomato in China (OP356207 and OP356208) (Li et al. 2022). To confirm the virus infection, we have successfully constructed an infectious clone for 0823-1 isolate in the binary plasmid and inoculated melon with and without an infectious clone. The melon plants inoculated with ToLCNDV 15 dpi showed the high accumulation of the virus and displayed symptoms similar to viruses in greenhouse. Based on the complete sequences, results of the molecular phylogenetic analysis (Fig. 2) and infectious clone, these geminiviruses were identified as ToLCNDV. ToLCNDV has been reported to occur and spread by the whitefly (Bemisia tabaci) in many Asian countries (Sohrab et al. 2003; Sohrab et al. 2011; Aamir et al. 2020) and Europe (Juárez et al. 2014; Ruiz et al. 2015; Luigi et al. 2019). Large populations of whiteflies were also present in all our surveyed areas. However, to our knowledge, this is the first report of the occurrence of ToLCNDV in cucurbit plants in China. The presence of ToLCNDV and CCYV can cause severe losses in crop yields or even crop failure. In addition to TYLCV, ToLCNDV is another major geminivirus-induced disease threatening cucurbit and other vegetable production in China.

Role of etofenprox nanoformulation in suppression of the silver whitefly, Bemisia tabaci and its residue in eggplant fruits

Role of etofenprox nanoformulation in suppression of the silver whitefly, Bemisia tabaci and its residue in eggplant fruits

Effects of holy basil (Ocimum sanctum) on viral disease of chili (Capsicum annum L.) under mixed crop cultivation

Chili is one of the important commodities in Indonesia. Chili cultivation faces many obstacles, leading to a decrease in production, such as virus infections. Yellow leaf curl and curly top viruses are groups of viruses that are often found in chili fields. This research was conducted to investigate the effect of intercropping (basil-chili) in suppressing viral diseases, especially yellow leaf curl and curly top viruses. A Randomized block design was used for this research, including intercropping and monoculture. The parameters observed in this study included disease incidence and intensity, the number and weight of chilies. Symptoms of virus infection include leaf curl, yellowing, and stunting. The disease has been reported to be transmitted by insect vectors, such as Aphis gossypii and Bemisia tabaci. The results showed that the incidence and intensity of yellow leaf curl and curly top virus diseases in chili pepper and basil plants were not significantly different in each treatment. The number of healthy chilies and their weight were also not significantly different. Damaged chilies are fruits that have been attacked by fruit flies. Basil intercropped with chilies attracts fruit flies, which affects the quantity and weight of the harvested chilies. The population of B. tabaci was significantly different in each treatment, while A. gossypii did not show significant differences among the treatments. Intercropping basil and chili increased the population of A. gossypii, while the population of B. tabaci decreased in this treatment. These two insect pests are vectors for yellow leaf curl and curly top viruses.

Evaluation of the Stability of a 1,8-Cineole Nanoemulsion and Its Fumigant Toxicity Effect against the Pests Tetranychus urticae, Rhopalosiphum maidis and Bemisia tabaci.

Pest control is a main concern in agriculture. Indiscriminate application of synthetic pesticides has caused negative impacts leading to the rapid development of resistance in arthropod pests. Plant secondary metabolites have been proposed as a safer alternative to conventional pesticides. Monoterpenoids have reported bioactivities against important pests; however, due to their high volatility, low water solubility and chemical instability, the application of these compounds has been limited. Nanosystems represent a potential vehicle for the broad application of monoterpenoids. In this study, an 1,8-cineole nanoemulsion was prepared by the low energy method of phase inversion, characterization of droplet size distribution and polydispersity index (PDI) was carried out by dynamic light scattering and stability was evaluated by centrifugation and Turbiscan analysis. Fumigant bioactivity was evaluated against Tetranychus urticae, Rhopalosiphum maidis and Bemisia tabaci. A nanoemulsion with oil:surfactant:water ratio of 0.5:1:8.5 had a droplet size of 14.7 nm and PDI of 0.178. Formulation was stable after centrifugation and the Turbiscan analysis showed no particle migration and a delta backscattering of ±1%. Nanoemulsion exhibited around 50% more bioactivity as a fumigant on arthropods when compared to free monoterpenoid. These results suggest that nanoformulations can provide volatile compounds of protection against volatilization, improving their bioactivity.

A temperature-dependent phenology model for Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae).

The sweetpotato whitefly, Bemisia tabaci (Gennadius) Middle East-Asia Minor 1 (MEAM1), is widespread across tropical and subtropical regions, affecting hundreds of cultivated and wild plant species. Because the species transmits a variety of viruses, the whitefly has become one of the most economically significant insect pests in the world. Determining a pest's population growth potential as a function of temperature is critical for understanding a species population dynamics, predicting the potential range of the species and its associated diseases, and designing adaptive pest management strategies. The life history of B. tabaci MEAM1 was studied in life-table experiments at 7 constant temperatures ranging from 12 to 35 °C. Nonlinear equations were fitted to development, mortality, and reproduction data and compiled into an overall phenology rate-summation model using Insect Life Cycle Modeling (ILCYM) software, to simulate life-table parameters based on temperature. Life tables of B. tabaci MEAM1 observed at naturally variable temperature in La Molina, Lima, during different seasons, covering the entire temperature range of the species' predicted performance curve, were used to validate the model. Simulations predicted population growth within temperature between 13.9 and 33.4 °C, revealing a maximum finite rate of population increase (λ = 1.163), with a generation time of 33.3 days at 26.4 °C. Predicted species performance agreed well when compared against observed life tables and published data. The process-based physiological model presented here for B. tabaci MEAM1 should prove useful to predict the potential spatial distribution of the species based on temperature and to adjust pest control measures taking different population growth potentials due to prevailing temperature regimes into account.

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.

Comparative evolutionary analyses of eight whitefly Bemisia tabaci sensu lato genomes: cryptic species, agricultural pests and plant-virus vectors

BackgroundThe group of > 40 cryptic whitefly species called Bemisia tabaci sensu lato are amongst the world’s worst agricultural pests and plant-virus vectors. Outbreaks of B. tabaci s.l. and the associated plant-virus diseases continue to contribute to global food insecurity and social instability, particularly in sub-Saharan Africa and Asia. Published B. tabaci s.l. genomes have limited use for studying African cassava B. tabaci SSA1 species, due to the high genetic divergences between them. Genomic annotations presented here were performed using the ‘Ensembl gene annotation system’, to ensure that comparative analyses and conclusions reflect biological differences, as opposed to arising from different methodologies underpinning transcript model identification.ResultsWe present here six new B. tabaci s.l. genomes from Africa and Asia, and two re-annotated previously published genomes, to provide evolutionary insights into these globally distributed pests. Genome sizes ranged between 616—658 Mb and exhibited some of the highest coverage of transposable elements reported within Arthropoda. Many fewer total protein coding genes (PCG) were recovered compared to the previously published B. tabaci s.l. genomes and structural annotations generated via the uniform methodology strongly supported a repertoire of between 12.8—13.2 × 103 PCG. An integrative systematics approach incorporating phylogenomic analysis of nuclear and mitochondrial markers supported a monophyletic Aleyrodidae and the basal positioning of B. tabaci Uganda-1 to the sub-Saharan group of species. Reciprocal cross-mating data and the co-cladogenesis pattern of the primary obligate endosymbiont ‘Candidatus Portiera aleyrodidarum’ from 11 Bemisia genomes further supported the phylogenetic reconstruction to show that African cassava B. tabaci populations consist of just three biological species. We include comparative analyses of gene families related to detoxification, sugar metabolism, vector competency and evaluate the presence and function of horizontally transferred genes, essential for understanding the evolution and unique biology of constituent B. tabaci. s.l species.ConclusionsThese genomic resources have provided new and critical insights into the genetics underlying B. tabaci s.l. biology. They also provide a rich foundation for post-genomic research, including the selection of candidate gene-targets for innovative whitefly and virus-control strategies.

Development of a multiplex RT-RPA assay for simultaneous detection of three viruses in cucurbits.

Begomoviruses and criniviruses, vectored by whiteflies (Bemisia tabaci), are important threats to crops worldwide. In recent years, the spread of cucurbit leaf crumple virus (CuLCrV), cucurbit yellow stunting disorder virus (CYSDV) and cucurbit chlorotic yellows virus (CCYV) on cucurbit crops has been reported to cause devastating crop losses in many regions of the world. In this study, a multiplex recombinase polymerase amplification (RPA) assay, an isothermal technique for rapid and simultaneous detection of DNA and RNA viruses CuLCrV, CYSDV and CCYV was developed. Highly specific and sensitive multiplex RPA primers for the coat protein region of these viruses were created and evaluated. The sensitivity of the multiplex RPA assay was examined using serially diluted plasmid containing the target regions. The results demonstrated that multiplex RPA primers have high sensitivity with a detection limit of a single copy of the viruses. The multiplex RPA primers were specific to the target as indicated by testing against other begomoviruses, potyviruses and an ilarvirus, and no nonspecific amplifications were noted. The primers simultaneously detected mixed infection of CCYV, CYSDV and CuLCrV in watermelon and squash crude extracts. This study is the first report of a multiplex RPA assay for simultaneous detection of mixed infection of DNA and RNA plant viruses.

Precision and Accuracy of Field Versus Laboratory Bioassay Insecticide Efficacy for the Control of Immature Bemisia tabaci.

Ecotoxicological studies often result in reports on the limitation and sometime failures of biological assay data to predict field response to similar treatments. Nevertheless, it is widely accepted that controlled bioassays can better quantify the specific mortality response of a target pest species to a specific toxin. To quantify the relationship between whitefly bioassay and field response data, we evaluated a controlled laboratory bioassay and a concurrent cucurbit field trial method to assess insecticide efficacy for controlling the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). This was based on oviposition and nymphal development. We specifically tested the assumptions that a maximum dose bioassay would more precisely measure insecticide efficacy as compared with a comparable field spray test evaluation, and the response would be equal between the bioassay and the field as a measure of control accuracy for both adult oviposition and development of nymphal stages. To make a direct comparison, we tested the same whitefly population subsamples from 352 plots in eight cucurbit field experiments in Georgia, USA, in 2021 and 2022. The bioassays provide significantly precision for estimating proportional whitefly response. As expected, treatment-specific nonequivalence in immature whitefly counts between the bioassay and field, i.e., a lack of accuracy, only occurred with insecticides that were not highly toxic to all growth stages of whiteflies.