Non-Target Effects of dsRNA Molecules in Hemipteran Insects

Insect pest control by RNA interference (RNAi)-mediated gene expression knockdown can be undermined by many factors, including small sequence differences between double-stranded RNA (dsRNA) and the target gene. It can also be compromised by effects that are independent of the dsRNA sequence on non-target organisms (known as sequence-non-specific effects). This study investigated the species-specificity of RNAi in plant sap-feeding hemipteran pests. We first demonstrated sequence-non-specific suppression of aphid feeding by dsRNA at dietary concentrations ≥0.5 µg µL−1. Then we quantified the expression of NUC (nuclease) genes in insects administered homologous dsRNA (with perfect sequence identity to the target species) or heterologous dsRNA (generated against a related gene of non-identical sequence in a different insect species). For the aphids Acyrthosiphon pisum and Myzus persicae, significantly reduced NUC expression was obtained with the homologous but not heterologous dsRNA at 0.2 µg µL−1, despite high dsNUC sequence identity. Follow-up experiments demonstrated significantly reduced expression of NUC genes in the whitefly Bemisia tabaci and mealybug Planococcus maritimus administered homologous dsNUCs, but not heterologous aphid dsNUCs. Our demonstration of inefficient expression knockdown by heterologous dsRNA in these insects suggests that maximal dsRNA sequence identity is required for RNAi targeting of related pest species, and that heterologous dsRNAs at appropriate concentrations may not be a major risk to non-target sap-feeding hemipterans.

Eigenfunction Expansions of Ultradifferentiable Functions and Ultradistributions. III. Hilbert Spaces and Universality

In this paper we analyse the structure of the spaces of smooth type functions, generated by elements of arbitrary Hilbert spaces, as a continuation of the research in our papers (Dasgupta and Ruzhansky in Trans Am Math Soc 368(12):8481–8498, 2016) and (Dasgupta and Ruzhansky in Trans Am Math Soc Ser B 5:81–101, 2018). We prove that these spaces are perfect sequence spaces. As a consequence we describe the tensor structure of sequential mappings on the spaces of smooth type functions and characterise their adjoint mappings. As an application we prove the universality of the spaces of smooth type functions on compact manifolds without boundary.

Evaluation of genetic structure in European wheat cultivars and advanced breeding lines using high-density genotyping-by-sequencing approach

Background The genetic diversity and gene pool characteristics must be clarified for efficient genome-wide association studies, genomic selection, and hybrid breeding. The aim of this study was to evaluate the genetic structure of 509 wheat accessions representing registered varieties and advanced breeding lines via the high-density genotyping-by-sequencing approach. Results More than 30% of 13,499 SNP markers representing 2162 clusters were mapped to genes, whereas 22.50% of 26,369 silicoDArT markers overlapped with coding sequences and were linked in 3527 blocks. Regarding hexaploidy, perfect sequence matches following BLAST searches were not sufficient for the unequivocal mapping to unique loci. Moreover, allelic variations in homeologous loci interfered with heterozygosity calculations for some markers. Analyses of the major genetic changes over the last 27 years revealed the selection pressure on orthologs of the gibberellin biosynthesis-related GA2 gene and the senescence-associated SAG12 gene. A core collection representing the wheat population was generated for preserving germplasm and optimizing breeding programs. Conclusions Our results confirmed considerable differences among wheat subgenomes A, B and D, with D characterized by the lowest diversity but the highest LD. They revealed genomic regions that have been targeted by breeding.

A universal RT-qPCR assay for “One Health” detection of influenza A viruses

The mutual dependence of human and animal health is central to the One Health initiative as an integrated strategy for infectious disease control and management. A crucial element of the One Health includes preparation and response to influenza A virus (IAV) threats at the human-animal interface. The IAVs are characterized by extensive genetic variability, they circulate among different hosts and can establish host-specific lineages. The four main hosts are: avian, swine, human and equine, with occasional transmission to other mammalian species. The host diversity is mirrored in the range of the RT-qPCR assays for IAV detection. Different assays are recommended by the responsible health authorities for generic IAV detection in birds, swine or humans. In order to unify IAV monitoring in different hosts and apply the One Health approach, we developed a single RT-qPCR assay for universal detection of all IAVs of all subtypes, species origin and global distribution. The assay design was centred on a highly conserved region of the IAV matrix protein (MP)-segment identified by a comprehensive analysis of 99,353 sequences. The reaction parameters were effectively optimised with efficiency of 93–97% and LOD95% of approximately ten IAV templates per reaction. The assay showed high repeatability, reproducibility and robustness. The extensive in silico evaluation demonstrated high inclusivity, i.e. perfect sequence match in the primers and probe binding regions, established as 94.6% for swine, 98.2% for avian and 100% for human H3N2, pandemic H1N1, as well as other IAV strains, resulting in an overall predicted detection rate of 99% on the analysed dataset. The theoretical predictions were confirmed and extensively validated by collaboration between six veterinary or human diagnostic laboratories on a total of 1970 specimens, of which 1455 were clinical and included a diverse panel of IAV strains.

A universal RT-qPCR assay for “One Health” detection of influenza A viruses

The mutual dependence of human and animal health is central to the One Health initiative as an integrated strategy for infectious disease control and management. A crucial element of the One Health includes preparation and response to influenza A virus (IAV) threats at the human-animal interface. The IAVs are characterized by extensive genetic variability, they circulate among different hosts and establish host-specific lineages. The four main host reservoirs are: avian, swine, human and equine, with occasional transmission to other mammalian species. The host diversity is mirrored in the range of the RT-qPCR assays for IAV detection. Different assays are recommended by the responsible health authorities for generic IAV detection in birds, swine or humans. In order to unify IAV monitoring in different hosts and apply the One Health approach, we developed a single RT-qPCR assay for universal detection of all IAVs of all subtypes, species origin and global distribution. The assay design was centred on a highly conserved region of the IAV MP-segment identified by a comprehensive analysis of 99,353 sequences. The reaction parameters were effectively optimised with efficiency of 93-97% and LOD95% of approximately ten IAV templates per reaction. The assay showed high repeatability, reproducibility and robustness. The extensive in silico evaluation demonstrated high inclusivity, i.e. perfect sequence match in the primers and probe binding regions, established as 94.6% for swine, 98.2% for avian and 100% for human H3N2, pandemic H1N1, as well as other IAV strains, resulting in an overall predicted detection rate of 99% on the analysed dataset. The theoretical predictions were confirmed and extensively validated by collaboration between six veterinary or human diagnostic laboratories on a total of 1970 specimens, of which 1455 were clinical and included a diverse panel of IAV strains.

A putative miRNA in the spike gene of SARS-CoV-2 has perfect sequence identity to both the forward and reverse complementary strands of hsa-mir-8055 involved in T-cell response to antigen

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), also known as COVID-19, encodes for a spike protein that is responsible for both attachment and membrane fusion, thereby being critical in the pathogenicity of this virus. Here, I report that a putative miRNA localized in the spike gene of SARS-CoV-2 matches to the forward strand of hsa-miR-8055, a miRNA expressed during T-cell response to antigen, and also binds with perfect complementarity to its seed region

A Community Empowerment Model through Pesantren-Based Family Empowerment Post (POSDAYA)

Purpose: This paper aims to analyze A Community Empowerment Model through Pesantren-based Family Empowerment Post (Posdaya). Design/methodology/approach: The method used is composes a reality to be a story, describing a problem, event, phenomenon orderly, followed by analysis and interpretation to analyze the data in a perfect sequence. Findings: 3 hypotheses are rejected while the other 3 is accepted. Research limitations/implications: These components were: 1) Data collection; this was done through interviews. Besides, the data were also collected through a literature study. 2) Data Reduction, after the data were gathered, the researcher focused on removing unnecessary data and arrange the data to gain a conclusion. 3) Data display, this was done through sentences and structured story. 4) Conclusion drawing, this was done to gain full meaning from the processed data, creating a clear, complete synopsis. Practical implications: Results show that from the 3 hypotheses proposed accepted, 1) Pesantren-based Posdaya needs to improve its resources through training or organizational education in order to improve the quality of empowerment program and the continuity of Posdaya activities. 2) Pesantren-based Posdaya needs to strengthen its social capital by establishing stronger social relationships with stakeholders, thus opening accessibility for community empowerment, or family empowerment in particular. 3) Pesantren-based Posdaya should strengthen its organizational structure and workgroup as the program implementer, and continuously monitor the program. Originality/value: This paper is original.