Sarah N. Inwood

Honeybees, Apis mellifera, play a vital role as pollinators in global agricultural ecosystems. Nutrition, particularly dietary protein content, profoundly impacts honeybee health and reproduction. Yet, the molecular mechanisms connecting diet composition and gene expression in honeybee eggs remain underexplored. In this study, we investigate the intricate relationship between diet, gene expression, and honeybee egg development. Using RNA-seq analysis, we explore the effects of different protein-to-carbohydrate (P: C) ratios in honeybee diets on differential gene expression in the eggs laid by the queen and potential associated molecular responses. Our research identifies 1007 differentially expressed genes (DEGs) across various dietary conditions, highlighting the pivotal role of nutritional composition in shaping gene expression during egg development.Cluster analysis revealed two DEG profiles corresponding to low protein diets (LPD) and high protein diets (HPD). LPD conditions upregulate genes linked to protein catabolism, autophagy, and ubiquitin-mediated proteolysis, indicating potential cellular responses to nutritional stress. Conversely, HPD conditions upregulate genes related to RNA processing, spliceosome activity, and the MAPK signalling pathway, suggesting normal cellular development.Notably, the Hippo signalling pathway exhibits distinct gene regulation patterns under LPD and HPD conditions, potentially influencing cellular growth and differentiation in response to nutrient availability.Our findings underscore the critical role of nutrition in honeybee health and reproduction, providing insights into optimizing honeybee diets for colony health and resilience. As honeybee populations confront challenges from changing environmental conditions and resource availability, understanding these molecular responses is crucial for their effective management and conservation as essential pollinators. This study establishes a foundation for further investigations into the functional consequences of these molecular responses at the individual level and their broader implications for honeybee colony development and health.

Modified, agricultural landscapes are susceptible to damage by insect pests. Biological control of pests is typically successful once a control agent has established, but this depends on the agent’s capacity to co-evolve with the host. Theoretical studies have shown that different levels of genetic variation between the host and the control agent will lead to rapid evolution of resistance in the host. Although this has been reported in one instance, the underlying genetics have not been studied. To address this, we measured the genetic variation in New Zealand populations of the pasture pest, Argentine stem weevil (Listronotus bonariensis), which is controlled with declining effectiveness by a parasitoid wasp, Microctonus hyperodae. We constructed a draft reference genome of the weevil, collected samples from a geographical survey of 10 sites around New Zealand, and genotyped them using a modified genotyping-by-sequencing approach. New Zealand populations of Argentine stem weevil have high levels of heterozygosity and low population structure, consistent with a large effective population size and frequent gene flow. This implies that Argentine stem weevils were able to evolve more rapidly than their biocontrol agent, which reproduces asexually. These findings show that monitoring genetic diversity in biocontrol agents and their targets is critical for long-term success of biological control.

In the past few years genetic technologies springing from advances in DNA sequencing (so-called high-throughput sequencing), and/or from CRISPR/Cas9 gene editing, have been proposed as being useful in bioheritage research. The potential scope for the use of these genetic technologies in bioheritage is vast, including enabling the recovery of threatened species, engineering proxies of extinct species and genetically controlling pests. While these technologies are often complex, they provide new opportunities that may help support New Zealand's beleaguered flora and fauna, and thus warrant scientific examination. Here we discuss these genetic technologies, focussing on scientific benefits and risks of each. We also acknowledge the social, cultural, ethical and regulatory constraints on their use, with emphasis on the importance of partnership with tangata whenua to determine when, whether or how these technologies should be used in enhancing New Zealand's bioheritage. We hope this will provide source material to support future decision making around the use of new genetic technologies in bioheritage.

Species of the genus Trichoderma are ubiquitous in the environment and are widely used in agriculture, as biopesticides, and in the industry for the production of plant cell wall-degrading enzymes. Trichoderma represents an important genus of endophytes, and several Trichoderma species have become excellent models for the study of fungal biology and plant–microbe interactions; moreover, are exceptional biotechnological factories for the production of bioactive molecules useful in agriculture and medicine. Next-generation sequencing technology coupled with systematic construction of recombinant DNA molecules provides powerful tools that contribute to the functional analysis of Trichoderma genetics, thus allowing for a better understanding of the underlying factors determining its biology. Here, we present the creation of diverse vectors containing (i) promoter-specific vectors for Trichoderma, (ii) gene deletions (using hygromycin phosphotransferase as selection marker), (iii) protein localization (mCherry and eGFP, which were codon-optimized for Trichoderma), (iv) gene complementation (neomycin phosphotransferase) and (v) overexpression of encoding gene proteins fused to fluorescent markers, by using the Golden Gate cloning technology. Furthermore, we present the design and implementation of a binary vector for Agrobacterium-mediated transformation in Trichoderma to increase the homologous recombination rate and the generation of a novel selection marker based on carboxin resistance.