Recent advances in the delivery and applications of nonviral CRISPR/Cas9 gene editing.

The CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 genome editing system has been a major technological breakthrough that has brought revolutionary changes to genome editing for therapeutic and diagnostic purposes and precision medicine. With the advent of the CRISPR/Cas9 system, one of the critical limiting factors has been the safe and efficient delivery of this system to cells or tissues of interest. Several approaches have been investigated to find delivery systems that can attain tissue-targeted delivery, lowering the chances of off-target editing. While viral vectors have shown promise for in vitro, in vivo and ex vivo delivery of CRISPR/Cas9, their further clinical applications have been restricted due to shortcomings including limited cargo packaging capacity, difficulties with large-scale production, immunogenicity and insertional mutagenesis. Rapid progress in nonviral delivery vectors, including the use of lipid, polymer, peptides, and inorganic nanoparticle-based delivery systems, has established nonviral delivery approaches as a viable alternative to viral vectors. This review will introduce the molecular mechanisms of the CRISPR/Cas9 gene editing system, current strategies for delivering CRISPR/Cas9-based tools, an overview of strategies for overcoming off-target genome editing, and approaches for improving genome targeting and tissue targeting. We will also highlight current developments and recent clinical trials for the delivery of CRISPR/Cas9. Finally, future directions for overcoming the limitations and adaptation of this technology for clinical trials will be discussed.

Evaluating the influence of nature connection and values on conservation attitudes at a tropical deforestation frontier.

Inner phenomena, such as personal motivations for pursuing sustainability, may be critical levers for improving conservation outcomes. Most conservation research and policies, however, focus on external phenomena (e.g., ecological change or economic processes). We explored the factors shaping 9 conservation attitudes toward forest and wildlife protection among colonist farmers around an Amazonian deforestation frontier. Our data comprised 241 face-to-face quantitative surveys, complemented with qualitative insights from open-ended questionnaire responses and opportunistic semistructured interviews. To account for the full spectrum of possible inner motivations, we employed measures of nature connection (indicating biospheric motivation) and personal values organized around the traditionalism (traditionalist through to high openness to change) and universalism dimensions (egoistic through to altruistic motivations). We used averaged beta-binomial generalized linear models to assess the role of external factors (socioeconomic, sociodemographic, and environmental) and personal (inner) motivations on the variation in attitudes. Each attitude was modeled separately. The relative importance of each predictor was judged by the proportion of models where it appeared as significant. Proconservation views were expressed by the majority (at least 65%) of the respondents in 7 out of the 9 attitude models. The most consistent predictors were emotional nature connection and personal values (significant in 4-6 out of 9 models), rather than external phenomena (significant in 0-5 models). However, the poorest farmers had lower scores on the agreement with prioritizing nature over development (𝛽 = -0.52, 95% CI: -0.96 to -0.07). Qualitative data also indicated that economic barriers hinder forest conservation on farms. These results suggest that biospheric, traditionalistic, and altruistic motivations promote people's proconservation attitudes, but nurturing these latent motivations is unlikely to improve conservation outcomes if material poverty remains unaddressed. Integrating the inner-outer perspective into conservation thinking and practical interventions could foster environmental stewardship and increase human well-being.

Evaluación de la influencia de la conexión y los valores naturales sobre las actitudes de conservación es una frontera tropical de deforestación Resumen Los fenómenos interiores, como los motivos personales para ser sustentables, pueden ser niveladores importantes para aumentar los resultados de conservación. Sin embargo, la mayor parte de la investigación y las políticas se enfocan en los fenómenos exteriores (cambios ecológicos o procesos económicos, por ejemplo). Exploramos los factores que moldean nueve actitudes de conservación relacionadas con la protección de los bosques y la fauna en un grupo de agricultores colonos cercanos a una frontera de deforestación en la Amazonía. Nuestros datos se obtuvieron de 241 encuestas presenciales cuantitativas complementadas con información cualitativa tomada de las respuestas en cuestionarios de preguntas abiertas y entrevistas oportunistas semiestructuradas. Para considerar el espectro completo de las posibles motivaciones interiores empleamos medidas de la conexión natural (lo que indica motivos relacionados a la biósfera) y valores personales organizados en torno a las dimensiones del tradicionalismo (del tradicionalista hasta una gran apertura al cambio) y el universalismo (del egoísta hasta los motivos altruistas). Usamos modelos lineales generalizados betabinomiales promediados para evaluar el papel que tienen los factores externos (socioeconómicos, sociodemográficos y ambientales) y los motivos personales (interiores) en la variación de las actitudes. Modelamos cada actitud por separado. Juzgamos la importancia relativa de cada predictor mediante la proporción de modelos en los que aparecían como significativos. La mayoría (al menos el 65%) de los respondientes expresó las opiniones en pro de la conservación en siete de los nueve modelos de actitud. Los predictores más uniformes fueron la conexión emocional natural y los valores personales (significativos en cuatro a seis de los nueve modelos), en lugar de los fenómenos externos (significativos en cero a cinco modelos). Sin embargo, los agricultores más pobres tuvieron un puntaje más bajo en cuanto a estar de acuerdo con la priorización de la naturaleza sobre el desarrollo (𝛽 = -0.52, 95% CI -0.96 a -0.07). Los datos cualitativos también indicaron que las barreras económicas impiden la conservación de los bosques en las fincas. Estos resultados sugieren que los motivos tradicionalistas, altruistas y aquellos relacionados con la biósfera promueven las actitudes en pro de la conservación de las personas, pero es poco probable que propiciar estos motivos latentes aumente los resultados de conservación si sigue sin solucionarse la pobreza material. La inclusión de las perspectivas internas y externas dentro del pensamiento de conservación y las intervenciones prácticas podría fomentar la administración ambiental e incrementar el bienestar humano.

Eight new species of Cycloneuroterus Melika & Tang gallwasps from Taiwan and mainland China (Hymenoptera: Cynipidae: Cynipini).

Eight new species of cynipid gallwasps, Cycloneuroterus abei Melika & Tang, C. ergei Tang & Melika, C. gilvus Melika & Tang, C. globosus Melika & Tang, C. jianwui Tang & Melika, C. lohsei Melika & Tang, C. tumiclavus Tang & Melika, C. uraianus Tang & Melika, from Taiwan and mainland China are described. Descriptions, diagnoses, biology, and host associations for the new species and a key to all Cycloneuroterus species are given. All taxa are supported by morphological and molecular data. Seven newly described species induce galls on Quercus subgenus Cyclobalanopsis, while one, C. uraianus, induces galls on Castanopsis. This is the first Cycloneuroterus species known to associate with Castanopsis.

Food for Pollinators: Quantifying the Nectar and Pollen Resources of Urban Flower Meadows.

Planted meadows are increasingly used to improve the biodiversity and aesthetic amenity value of urban areas. Although many 'pollinator-friendly' seed mixes are available, the floral resources these provide to flower-visiting insects, and how these change through time, are largely unknown. Such data are necessary to compare the resources provided by alternative meadow seed mixes to each other and to other flowering habitats. We used quantitative surveys of over 2 million flowers to estimate the nectar and pollen resources offered by two exemplar commercial seed mixes (one annual, one perennial) and associated weeds grown as 300m2 meadows across four UK cities, sampled at six time points between May and September 2013. Nectar sugar and pollen rewards per flower varied widely across 65 species surveyed, with native British weed species (including dandelion, Taraxacum agg.) contributing the top five nectar producers and two of the top ten pollen producers. Seed mix species yielding the highest rewards per flower included Leontodon hispidus, Centaurea cyanus and C. nigra for nectar, and Papaver rhoeas, Eschscholzia californica and Malva moschata for pollen. Perennial meadows produced up to 20x more nectar and up to 6x more pollen than annual meadows, which in turn produced far more than amenity grassland controls. Perennial meadows produced resources earlier in the year than annual meadows, but both seed mixes delivered very low resource levels early in the year and these were provided almost entirely by native weeds. Pollen volume per flower is well predicted statistically by floral morphology, and nectar sugar mass and pollen volume per unit area are correlated with flower counts, raising the possibility that resource levels can be estimated for species or habitats where they cannot be measured directly. Our approach does not incorporate resource quality information (for example, pollen protein or essential amino acid content), but can easily do so when suitable data exist. Our approach should inform the design of new seed mixes to ensure continuity in floral resource availability throughout the year, and to identify suitable species to fill resource gaps in established mixes.

New species of cynipid inquilines of the genus Saphonecrus (Hymenoptera: Cynipidae: Synergini) from the Eastern Palaearctic, with a re-appraisal of known species world-wide.

Fifteen new species of cynipid inquilines, Saphonecrus chinensis Tang & Schwéger, S. gilvus Melika & Schwéger, S. globosus Schwéger & Tang, S. leleyi Melika & Schwéger, S. lithocarpii Schwéger & Melika, S. longinuxi Schwéger & Melika, S. morii Schwéger & Tang, S. nantoui Tang, Schwéger & Melika, S. nichollsi Schwéger & Melika, S. pachylomai Schwéger, Tang & Melika, S. robustus Schwéger & Melika, S. saliciniai Melika, Tang & Schwéger, S. shanzhukui Melika & Tang, S. symbioticus Melika & Schwéger, and S. taitungi Schwéger, Tang & Melika, from the Eastern Palaearctic are described. Descriptions, diagnoses, biology, and host associations for the new species, and a key to Palaearctic Saphonecrus species are given. All new taxa form distinct units as demonstrated by the molecular phylogenetic analyses of Palaearctic Saphonecrus species. The status of some earlier described Saphonecrus species is discussed also. The Synergini genus Lithonecrus Nieves-Aldrey & Butterill, 2014 is synonymized with Lithosaphonecrus Tang, Melika & Bozsó, 2013. Three Saphonecrus species are transferred to Synergus: Synergus brevis (Weld) comb. nova, Synergus hupingshanensis (Liu, Yang & Zhu) comb. nova, and Synergus yukawai (Wachi, Ide & Abe) comb. nova. Synophrus vietnamensis Abe, Ide, Konishi & Ueno is transferred to Lithosaphonecrus: Lithosaphonecrus vietnamensis Abe, Ide, Konishi & Ueno), comb. nova. The current number of valid Saphonecrus species worldwide is 36.

Impacts of local adaptation of forest trees on associations with herbivorous insects: implications for adaptive forest management.

Disruption of species interactions is a key issue in climate change biology. Interactions involving forest trees may be particularly vulnerable due to evolutionary rate limitations imposed by long generation times. One mitigation strategy for such impacts is Climate matching - the augmentation of local native tree populations by input from nonlocal populations currently experiencing predicted future climates. This strategy is controversial because of potential cascading impacts on locally adapted animal communities. We explored these impacts using abundance data for local native gallwasp herbivores sampled from 20 provenances of sessile oak (Quercus petraea) planted in a common garden trial. We hypothesized that non-native provenances would show (i) declining growth performance with increasing distance between provenance origin and trial site, and (ii) phenological differences to local oaks that increased with latitudinal differences between origin and trial site. Under a local adaptation hypothesis, we predicted declining gallwasp abundance with increasing phenological mismatch between native and climate-matched trees. Both hypotheses for oaks were supported. Provenance explained significant variation in gallwasp abundance, but no gall type showed the relationship between abundance and phenological mismatch predicted by a local adaptation hypothesis. Our results show that climate matching would have complex and variable impacts on oak gall communities.

A new genus of oak gallwasp, Cyclocynips Melika, Tang & Sinclair (Hymenoptera: Cynipidae: Cynipini), with descriptions of two new species from Taiwan.

A new genus of cynipid oak gallwasp-Cyclocynips Melika, Tang, & Sinclair (Hymenoptera: Cynipidae: Cynipini), with two new species--C. uberis and C. tumorvirgae--reared from galls on oaks of the Quercus subgenus Cyclobalanopsis is described from Taiwan. Descriptions of asexual generation adults and their diagnostic characters are presented. The likelihood of yet undiscovered sexual generations and the evolution of host-plant associations in these species are discussed.

Reconstructing community assembly in time and space reveals enemy escape in a Western Palearctic insect community.

How geographically widespread biological communities assemble remains a major question in ecology. Do parallel population histories allow sustained interactions (such as host-parasite or plant-pollinator) among species, or do discordant histories necessarily interrupt them? Though few empirical data exist, these issues are central to our understanding of multispecies evolutionary dynamics. Here we use hierarchical approximate Bayesian analysis of DNA sequence data for 12 herbivores and 19 parasitoids to reconstruct the assembly of an insect community spanning the Western Palearctic and assess the support for alternative host tracking and ecological sorting hypotheses. We show that assembly occurred primarily by delayed host tracking from a shared eastern origin. Herbivores escaped their enemies for millennia before parasitoid pursuit restored initial associations, with generalist parasitoids no better able to track their hosts than specialists. In contrast, ecological sorting played only a minor role. Substantial turnover in host-parasitoid associations means that coevolution must have been diffuse, probably contributing to the parasitoid generalism seen in this and similar systems. Reintegration of parasitoids after host escape shows these communities to have been unsaturated throughout their history, arguing against major roles for parasitoid niche evolution or competition during community assembly.