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BACKGROUND: Double‐stranded RNA (dsRNA) biopesticides are of interest for the abatement of insect vectors of pathogenic bacteria such as 'Candidatus Liberibacter', which infects both its psyllid and plant hosts. Silencing of genes essential for psyllids, or for Liberibacter, is anticipated to lead to mortality or impeded bacterial multiplication. Foliar delivery is preferred for biopesticide application; however, the cuticle impedes dsRNA penetration into the vasculature. Here, conditions were established for wounding tomato leaves using ultraviolet light amplification by stimulated emissions of radiation (UV‐LASER) to promote dsRNA penetration into leaves and vasculature. RESULTS: UV‐LASER treatment with application of select adjuvants/surfactants resulted in vascular delivery of 100‐, 300‐ and 600‐bp dsRNAs that, in general, were correlated with size. The 100‐bp dsRNA required no pretreatment, whereas 300‐ and 600‐bp dsRNAs entered the vasculature after UV‐LASER treatment only and UV‐LASER adjuvant/surfactant treatment, respectively. Of six adjuvant/surfactants evaluated, plant‐derived oil combined with an anionic organosilicon compound performed most optimally. Localization of dsRNAs in the tomato vasculature was documented using fluorometry and fluorescence confocal microscopy. The biological activity of in planta‐delivered dsRNA (200–250 bp) was determined by feeding third‐instar psyllids on tomato leaves post UV‐LASER adjuvant/surfactant treatment, with or without psyllid cdc42‐ and gelsolin dsRNAs. Gene knockdown was quantified by quantitative, real‐time polymerase chain reaction with reverse transcription (RT–qPCR) amplification. At 10 days post the ingestion‐access period, knockdown of cdc42 and gelsolin expression was 61% and 56%, respectively, indicating that the dsRNAs delivered to the tomato vasculature were mobile and biologically active. CONCLUSION: Results indicated that UV‐LASER adjuvant/surfactant treatments facilitated the delivery of mobile, biologically active dsRNA molecules to the plant vasculature. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND: RNA‐mediated interference (RNAi) has become a promising biopesticide technology with which to direct sequence‐specific gene knockdown of key targets in the potato psyllid (PoP) Bactericera cockerelli, resulting in significant mortality. In this study, three strategically selected target genes, ATF4, C7 and D24, essential for the biosynthesis and regulation of ecdysteroids, were evaluated for knockdown and mortality using oral delivery of individual, paired and all three double‐stranded RNAs (dsRNAs), in five replicated experiments. Knockdown was determined as the fold‐change in gene expression using a quantitative polymerase chain reaction. RESULTS: Knockdown of the D24 target, at 39%–45%, resulted in 51% PoP mortality by 10 days post‐ingestion (dpi) of dsRNA. Knockdown of C7, at 38%–61%, resulted in 53% mortality by 10 dpi, whereas dsD24 ingestion resulted in 65% mortality by 10 dpi when dsD24 and dsC7 were co‐delivered. Three phenotypes, INCOMEC, PREMEC and SWOLLEN, were observed at a frequency of 4%–12%, and are consistent with incomplete ecdysis in immature and/or adult PoP. Adult PoP exhibiting INCOMEC survived for several days but were unable to mate or fly, whereas SWOLLEN and PREMEC were lethal to the immature instars. Knockdown of ATF4 did not result in the mortality or malformations in immature and adult PoP. CONCLUSIONS: Compared with knockdown of individual D24 and C7 targets, significantly greater RNAi penetrance was achieved following delivery of combined dsRNAs. The highest knockdown that resulted in incomplete ecdysis and/or mortality was obtained for targets with predicted involvement in the same or interacting pathway(s). Knockdown of ATF4 was apparently "rescued" by uncharacterized compensatory gene(s) or effects. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

During 1991, in Brazil, the presence of the exotic Bemisia tabaci B mitotype was reported in São Paulo state. However, the duration from the time of initial introduction to population upsurges is not known. To investigate whether the 1991 B mitotype outbreaks in Brazil originated in São Paulo or from migrating populations from neighboring introduction sites, country‐wide field samples of B. tabaci archived from 1989–2005 collections were subjected to analysis of mitochondrial cytochrome oxidase I (mtCOI) and nuclear RNA‐binding protein 15 (RP‐15) sequences. The results of mtCOI sequence analysis identified all B. tabaci as the NAFME 8 haplotype of the B mitotype. Phylogenetic analyses of RP‐15 sequences revealed that the B mitotype was likely a hybrid between a B type parent related to a haplotype Ethiopian endemism (NAFME 1–3), and an unidentified parent from the North Africa‐Middle East (NAF‐ME) region. Results provide the first evidence that this widely invasive B mitotype has evolved from a previously undocumented hybridization event. Samples from Rio de Janeiro (1989) and Ceará state (1990), respectively, are the earliest known B mitotype records in Brazil. A simulated migration for the 1989 introduction predicted a dispersal rate of 200–500 km/year, indicating that the population was unlikely to have reached Ceará by 1990. Results implicated two independent introductions of the B mitotype in Brazil in 1989 and 1990, that together were predicted to have contributed to the complete invasion of Brazil in only 30 generations. [ABSTRACT FROM AUTHOR]

The Bemisia tabaci complex in Ecuador was studied with respect to phylogenetic relationships and eco-geographical distribution. Whitefly samples were collected from natural and agricultural environments in nine provinces of Ecuador (latitude, 2°N–5°S; longitude, 78°–81°W). Mitotypes were identified based on phylogenetic analysis of the 30-mtCOI-tRNAleuregion (832 bp) and corrected pairwise distance analysis. The distribution of mitotypes was modeled using MaxEnt, and their predicted niches were characterized according to environmental gradients. Four B. tabacimitotypes were identified, of which three are endemic, herein ECU1–3, and the other is the introduced B mitotype. Mitotypes ECU1 (44%), ECU2 (0.74%), and ECU3 (1.47%) grouped in the American Tropics (AMTROP) species and diverged by as much as 10%, which was higher than previous estimates for the AMTROP clade of 7–8.6%. Although haplotypes of ECU1 and ECU2 are known from the American Tropics, this is the first report of the ECU3 mitotype, which may possibly be restricted to southern Ecuador. The distribution of the three ECU-endemic mitotypes spanned the high-altitude niches of the western slope of the Andes, rich in microclimates with variable temperature and humidity conditions. The non-endemic B mitotype (47%) occurred only in the irrigated cropping systems located in hot and/or dry-tropical ecological niches. Of the endemic mitotypes, ECU1 occupied the most ecological niches. Among variables contributing to ECU1 and B mitotype niche range assignments, the most significant to influence ecological range was rainfall. The B. tabaciendemic to Ecuador were more diverse with respect to mtCOI-tRNAleu sequence than previously known, and occupied distinct microclimate niches suggestive of ecological resilience. [ABSTRACT FROM AUTHOR]

Endosymbionts associated with the whitefly Bemisia tabaci cryptic species are known to contribute to host fitness and environmental adaptation. The genetic diversity and population complexity were investigated for endosymbiont communities of B. tabaci occupying different micro‐environments in Pakistan. Mitotypes of B. tabaci were identified by comparative sequence analysis of the mitochondria cytochrome oxidase I (mtCOI) gene sequence. Whitefly mitotypes belonged to the Asia II‐1, ‐5, and ‐7 mitotypes of the Asia II major clade. The whitefly–endosymbiont communities were characterized based on 16S ribosomal RNA operational taxonomic unit (OTU) assignments, resulting in 43 OTUs. Most of the OTUs occurred in the Asia II‐1 and II‐7 mitotypes (r2 =.9, p <.005), while the Asia II‐5 microbiome was less complex. The microbiome OTU groups were mitotype‐specific, clustering with a basis in phylogeographical distribution and the corresponding ecological niche of their whitefly host, suggesting mitotype‐microbiome co‐adaptation. The primary endosymbiont Portiera was represented by a single, highly homologous OTU (0%–0.67% divergence). Two of six Arsenophonus OTUs were uniquely associated with Asia II‐5 and ‐7, and one occurred exclusively in Asia II‐1, two only in Asia II‐5, and one in both Asia II‐1 and ‐7. Four other secondary endosymbionts, Cardinium, Hemipteriphilus, Rickettsia, and Wolbachia OTUs, were found at ≤29% frequencies. The most prevalent Arsenophonus OTU was found in all three Asia II mitotypes (55% frequency), whereas the same strain of Cardinium and Wolbachia was found in both Asia II‐1 and ‐5, and a single Hemipteriphilus OTU occurred in Asia II‐1 and ‐7. This pattern is indicative of horizontal transfer, suggestive of a proximity between mitotypes sufficient for gene flow at overlapping mitotype ecological niches. [ABSTRACT FROM AUTHOR]