The Combination of Bacillus Thuringiensis and Its Engineered Strain Expressing dsRNA Increases the Toxicity against Plutella Xylostella

RNA interference (RNAi) has been developed and used as an emerging strategy for pest management. Here, an entomopathogen Bacillus thuringiensis (Bt) was used to express the dsRNA for the control of Plutella xylostella. A vector containing a 325-bp fragment of the conserved region of P. xylostella arginine kinase gene (PxAK) flanking in two ends with the promoter Pro3α was developed and transferred into Bt 8010 and BMB171, and consequently engineered Bt strains 8010AKi and BMB171AKi expressing dsRNA of PxAK were developed. The two engineered Bt strains were separately mixed with Bt 8010 in a series of ratios, and then fed to the P. xylostella larvae. We found that 8010:8010AKi of 9:1 and 8010:BMB171AKi of 7:3 caused a higher mortality than Bt 8010. PxAK expression levels in the individuals treated with the mixtures, 8010AKi and BMB171Aki, were lower than that in the control. The intrinsic rate of increase (r) and net reproductive rate (R0) of the population treated with 8010:8010AKi of 9:1 were lower than those of the population treated with Bt 8010 or 8010AKi. We developed a Bt-mediated insect RNAi for the control of P. xylostella and demonstrated a practical approach to integrating the entomopathogen with RNAi technique for the pest management.

The scuttle flies (Diptera: Phoridae) of Iran with the description of Mahabadphora aesthesphora as a new genus and species

Scuttle flies (Diptera: Phoridae) are mega-diverse and often synanthropic insects that play superb roles in various ecosystems. Identification of this group of insects is challenging due to their small size, morphological identification difficulties, niche diversity, and lack of taxonomic keys. To pave the way, an in-depth investigation was directed toward the scuttle flies in Iran using morphological and molecular data. A dichotomous key was also developed to identify the genus and species of the phorids reported in the country. The faunistic findings revealed the presence of about 22,000 (13,903 male and 8,097 female) phorid materials organized into 11 genera. Megaselia species (n = 13768), made up about 99% of the specimens studied. Moreover, 71 morphologically defined species belonging to nine genera were molecularly characterized using COI, 28S rRNA, and Arginine kinase datasets. Excluding four Megaselia Rondani, 1856 species, our results specified that morphologically delimited species were in agreement with the molecular analyses inferred from the COI/28S rRNA and COI/Arginine kinase sequences with genetic distances and phylogenetic trees. According to the results of the present study and previously published data, the Phoridae recorded for Iran are a total of 97 species that are ordered in 13 genera and three subfamilies, including Chonocephalinae, Metopininae and Phorinae. By comparing the known world phorid genera, a new monotypic genus of scuttle flies, Mahabadphora aesthesphora gen. nov., sp. nov., was identified based on its morphological and molecular characteristics and included in an updated key. Our results could comprehensively determine the taxonomic status of scuttle flies in Iran, scrutinize their phylogenetic structures and facilitate their identification.

Occupational Allergic Sensitization Among Workers Processing King Crab (Paralithodes camtschaticus) and Edible Crab (Cancer pagurus) in Norway and Identification of Novel Putative Allergenic Proteins

Introduction: Asthma and allergy occur frequently among seafood processing workers, with the highest prevalence seen in the crustacean processing industry. In this study we established for the first time the prevalence of allergic sensitization in the Norwegian king- and edible crab processing industry and characterized the IgE-reactive proteins.Materials and Methods: Two populations of crab processing workers participated; 119 king crab and 65 edible crab workers. The investigation included information on work tasks and health through a detailed questionnaire. Allergic sensitization was investigated by crab-specific IgE quantification and skin prick tests (SPT) to four in-house prepared crab extracts; raw meat, cooked meat, raw intestines and raw shell. Allergen-specific IgE binding patterns were analyzed by IgE immunoblotting to the four allergen extracts using worker serum samples. Total proteins in crab SPT extracts and immunoblot-based IgE binding proteins were identified by mass spectrometric analysis.Results: Positive SPTs were established in 17.5% of king- and 18.1% of edible crab workers, while elevated IgE to crab were demonstrated in 8.9% of king- and 12.2% of edible crab processing workers. There was no significant difference between the king and edible crab workers with respect to self-reported respiratory symptoms, elevated specific IgE to crab or SPT results. Individual workers exhibited differential IgE binding patterns to different crab extracts, with most frequent binding to tropomyosin and arginine kinase and two novel IgE binding proteins, hemocyanin and enolase, identified as king- and edible crab allergens.Conclusions: Occupational exposure to king- and edible crabs may frequently cause IgE mediated allergic sensitization. Future investigations addressing the diagnostic value of crab allergens including tropomyosin and arginine kinase and the less well-known IgE-binding proteins hemocyanin and enolase in a component-resolved diagnostic approach to crab allergy should be encouraged.

McsB forms a gated kinase chamber to mark aberrant bacterial proteins for degradation

In Gram-positive bacteria, the McsB protein arginine kinase is central to protein quality control, labelling aberrant molecules for degradation by the ClpCP protease. Despite its importance for stress response and pathogenicity, it is still elusive how the bacterial degradation labelling is regulated. Here, we delineate the mechanism how McsB targets aberrant proteins during stress conditions. Structural data reveal a self-compartmentalized kinase, in which the active sites are sequestered in a molecular cage. The 'closed' octamer interconverts with other oligomers in a phosphorylation-dependent manner and, contrary to these 'open' forms, preferentially labels unfolded proteins. In vivo data show that heat-shock triggers accumulation of higher-order oligomers, of which the octameric McsB is essential for surviving stress situations. The interconversion of open and closed oligomers represents a distinct regulatory mechanism of a degradation labeler, allowing the McsB kinase to adapt its potentially dangerous enzyme function to the needs of the bacterial cell.

A Protective dsRNA is Crucial for Optimum RNAi Gene Silencing in Chilo partellus

RNAi technology is currently employed as an alternate control measure for agricultural pests. However, the variability of RNAi efficiency in insect pests limits the extensive usage of this technology and demands identifying the best target gene for effective RNAi. Four different bacterially-expressed dsRNA and purified dsRNAs coated on artificial diet were fed to the larvae. The transcripts expression was analyzed at 5 days and 15 days post-exposure to various dsRNAs. In the larvae fed on bacterially-expressed dsRNA, knockdown percentages were 80 and 57% knockdown in Acetylcholinesterase transcript, 40 and 60% gene knockdown in Arginine kinase, 74 and 73% knockdown in Chymotrypsin, and 80 and 20% reduction in V-ATPase transcript expression. Overall, the mRNA knockdown percentages in the targeted genes were more pronounced at 5 days of exposure to bacterially-expressed crude dsRNA than 15 days of exposure. However, most purified dsRNAs rarely induce any significant knockdown except dsARG, which reduced the arginine kinase transcript by 40%. Our findings suggest that for optimum RNAi in C. partellus, the dsRNA must be protected from direct access with nucleases. © 2021 Friends Science Publishers