Eurasian spruce bark beetle Ips typographus, a natural part of forest ecosystems, is a major threat to Norway spruce forests during outbreaks. Olfaction plays a crucial role in the survival and range expansion of these beetles, amid forest disturbances and climate change. As the current management strategies are suboptimal for controlling outbreaks, the reverse chemical ecology approaches based on pheromone receptors offer promising alternatives. While the search for pheromone receptors is in progress, recently found chromosomal inversions indicates signs of adaptation in this species. Our attempts to characterise one of the highly expressed odorant receptors, ItypOR33, located in an inversion, led to the discovery of polymorphic variants distributed with similar frequency across 18 European populations. Deorphanizing ItypOR33 and its variant ItypOR33a using the Drosophila empty-neuron system (DeNS) revealed ItypOR33 tuned to amitinol, a heterospecific pheromone component in Ips spp., whereas its variant tuned to (S)-(-)-ipsenol, a conspecific pheromone component of I. typographus. The in silico approaches revealed the structural basis of variations by predicting putative ligand-binding sites, tunnels and ligand-receptor interactions. However, no sex-specific differences were found in the ItypOR33 expression, and its ligand amitinol elicited behavioural and electrophysiological responses. Reporting population-level functional olfactory polymorphisms for the first time in a non-model organism-bark beetles, provides key evidence for further exploring their survival and adaptation in forests. Additionally, these findings indicate potential long-term complexities of managing bark beetles in forests.
- Klíčová slova
- Drosophila empty‐neuron system, bark beetles, deorphanization, functional polymorphism, olfactory adaptation, pheromone receptor, population genomics,
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- časopisecké články MeSH
Fungal diseases threaten the forest ecosystem, impacting tree health, productivity, and biodiversity. Conventional approaches to combating diseases, such as biological control or fungicides, often reach limits regarding efficacy, resistance, non-target organisms, and environmental impact, enforcing alternative approaches. From an environmental and ecological standpoint, an RNA interference (RNAi) mediated double-stranded RNA (dsRNA)-based strategy can effectively manage forest fungal pathogens. The RNAi approach explicitly targets and suppresses gene expression through a conserved regulatory mechanism. Recently, it has evolved to be an effective tool in combating fungal diseases and promoting sustainable forest management approaches. RNAi bio-fungicides provide efficient and eco-friendly disease control alternatives using species-specific gene targeting, minimizing the off-target effects. With accessible data on fungal disease outbreaks, genomic resources, and effective delivery systems, RNAi-based biofungicides can be a promising tool for managing fungal pathogens in forests. However, concerns regarding the environmental fate of RNAi molecules and their potential impact on non-target organisms require an extensive investigation on a case-to-case basis. The current review critically evaluates the feasibility of RNAi bio-fungicides against forest pathogens by delving into the accessible delivery methods, environmental persistence, regulatory aspects, cost-effectiveness, community acceptance, and plausible future of RNAi-based forest protection products.
- Klíčová slova
- Plant fungal pathogen, RNA interference, forest disease management, host-induced gene silencing, host–pathogen interaction, spray-induced gene silencing,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
BACKGROUND: Climate change has recently boosted the severity and frequency of pine bark beetle attacks. The bacterial community associated with these beetles acts as "hidden players," enhancing their ability to infest and thrive on defense-rich pine trees. There is limited understanding of the environmental acquisition of these hidden players and their life stage-specific association with different pine-feeding bark beetles. There is inadequate knowledge on novel bacterial introduction to pine trees after the beetle infestation. Hence, we conducted the first comparative bacterial metabarcoding study revealing the bacterial communities in the pine trees before and after beetle feeding and in different life stages of two dominant pine-feeding bark beetles, namely Ips sexdentatus and Ips acuminatus. We also evaluated the bacterial association between wild and lab-bred beetles to measure the deviation due to inhabiting a controlled environment. RESULTS: Significant differences in bacterial amplicon sequence variance (ASVs) abundance existed among different life stages within and between the pine beetles. However, Pseudomonas, Serratia, Pseudoxanthomonas, Taibaiella, and Acinetobacter served as core bacteria. Interestingly, I. sexdentatus larvae correspond to significantly higher bacterial diversity and community richness and evenness compared to other developmental stages, while I. acuminatus adults displayed higher bacterial richness with no significant variation in the diversity and evenness between the life stages. Both wild and lab-bred I. sexdentatus beetles showed a prevalence of the bacterial family Pseudomonadaceae. In addition, wild I. sexdentatus showed dominance of Yersiniaceae, whereas Erwiniaceae was abundant in lab-bred beetles. Alternatively, Acidobacteriaceae, Corynebacteriaceae, and Microbacteriaceae were highly abundant bacterial families in lab-bred, whereas Chitinophagaceae and Microbacteriaceae were highly abundant in wild I. accuminatus. We validated the relative abundances of selected bacterial taxa estimated by metagenomic sequencing with quantitative PCR. CONCLUSION: Our study sheds new insights into bacterial associations in pine beetles under the influence of various drivers such as environment, host, and life stages. We documented that lab-breeding considerably influences beetle bacterial community assembly. Furthermore, beetle feeding alters bacteriome at the microhabitat level. Nevertheless, our study revisited pine-feeding bark beetle symbiosis under the influence of different drivers and revealed intriguing insight into bacterial community assembly, facilitating future functional studies.
- Klíčová slova
- Ips acuminatus, Ips sexdentatus, amplicon sequence variances (ASVs), core bacteriome, holobiont, microhabitat,
- Publikační typ
- časopisecké články MeSH
Plant defence mechanisms, including physical barriers like toughened bark and chemical defences like allelochemicals, are essential for protecting them against pests. Trees allocate non-structural carbohydrates (NSCs) to produce secondary metabolites like monoterpenes, which increase during biotic stress to fend off pests like the Eurasian spruce bark beetle, ESBB (Ips typographus). Despite these defences, the ESBB infests Norway spruce, causing significant ecological damage by exploiting weakened trees and using pheromones for aggregation. However, the mechanism of sensing and resistance towards host allelochemicals in ESBB is poorly understood. We hypothesised that the exposure of ESBB to spruce allelochemicals, especially monoterpenes, leads to an upsurge in the important detoxification genes like P450s, GSTs, UGTs, and transporters, and at the same time, genes responsible for development must be compromised. The current study demonstrates that exposure to monoterpenes like R-limonene and sabiene effectively elevated detoxification enzyme activities. The differential gene expression (DGE) analysis revealed 294 differentially expressed (DE) detoxification genes in response to R-limonene and 426 DE detoxification genes in response to sabiene treatments, with 209 common genes between the treatments. Amongst these, genes from the cytochrome P450 family 4 and 6 genes (CP4 and CP6), esterases, glutathione S-transferases family 1 (GSTT1), UDP-glucuronosyltransferase 2B genes (UDB), and glucose synthesis-related dehydrogenases were highly upregulated. We further validated 19 genes using RT-qPCR. Additionally, we observed similar high expression levels of detoxification genes across different monoterpene treatments, including myrcene and α-pinene, suggesting a conserved detoxification mechanism in ESBB, which demands further investigation. These findings highlight the potential for molecular target-based beetle management strategies targeting these key detoxification genes.
- Klíčová slova
- Norway spruce, RNA-seq, RT-qPCR, bark beetles, bioassay, detoxification, enzyme assay, monoterpenes,
- MeSH
- brouci * metabolismus genetika účinky léků MeSH
- kůra rostlin chemie metabolismus MeSH
- metabolická inaktivace * MeSH
- monoterpeny * metabolismus farmakologie MeSH
- regulace genové exprese u rostlin MeSH
- smrk * metabolismus genetika MeSH
- systém (enzymů) cytochromů P-450 metabolismus genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- monoterpeny * MeSH
- systém (enzymů) cytochromů P-450 MeSH
Anthropogenic activities and subsequent global climate change instigate drastic crop productivity and yield changes. These changes comprise a rise in the number and severity of plant stress factors, which can arise simultaneously or sequentially. When abiotic stress factors are combined, their impact on plants is more substantial than that of a singleton stress factor. One such impact is the alteration of redox cellular homeostasis, which, in turn, can regulate downstream stress-responsive gene expression and resistance response. The epigenetic regulation of gene expression in response to varied stress factors is an interesting phenomenon, which, conversely, can be stable and heritable. The epigenetic control in plants in response to abiotic stress combinations and their interactions with cellular redox alteration is an emerging field to commemorate crop yield management under climate change. The article highlights the integration of the redox signaling pathways and epigenetic regulations as pivotal components in the complex network of plant responses against multi-combinatorial stresses across time and space. This review aims to lay the foundation for developing novel approaches to mitigate the impact of environmental stresses on crop productivity, bridging the gap between theoretical understanding and practical solutions in the face of a changing climate and anthropogenic disturbances.
- Klíčová slova
- abiotic stress, anthropogenic disturbances, crop resilience, epigenetic regulation, histone modification, reactive oxygen species,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
The rhizosphere is the hotspot for microbial enzyme activities and contributes to carbon cycling. Precipitation is an important component of global climate change that can profoundly alter belowground microbial communities. However, the impact of precipitation on conifer rhizospheric microbial populations has not been investigated in detail. In the present study, using high-throughput amplicon sequencing, we investigated the impact of precipitation on the rhizospheric soil microbial communities in two Norway Spruce clonal seed orchards, Lipová Lhota (L-site) and Prenet (P-site). P-site has received nearly double the precipitation than L-site for the last three decades. P-site documented higher soil water content with a significantly higher abundance of Aluminium (Al), Iron (Fe), Phosphorous (P), and Sulphur (S) than L-site. Rhizospheric soil metabolite profiling revealed an increased abundance of acids, carbohydrates, fatty acids, and alcohols in P-site. There was variance in the relative abundance of distinct microbiomes between the sites. A higher abundance of Proteobacteria, Acidobacteriota, Ascomycota, and Mortiellomycota was observed in P-site receiving high precipitation, while Bacteroidota, Actinobacteria, Chloroflexi, Firmicutes, Gemmatimonadota, and Basidiomycota were prevalent in L-site. The higher clustering coefficient of the microbial network in P-site suggested that the microbial community structure is highly interconnected and tends to cluster closely. The current study unveils the impact of precipitation variations on the spruce rhizospheric microbial association and opens new avenues for understanding the impact of global change on conifer rizospheric microbial associations.
- Klíčová slova
- FUNGuild, Norway spruce, PICRUSt2, amplicon sequencing, microbial communities, network analysis, precipitation, rhizosphere, seed orchards, soil metabolites,
- MeSH
- Bacteria genetika klasifikace metabolismus MeSH
- déšť MeSH
- klimatické změny MeSH
- mikrobiota * genetika MeSH
- půda chemie MeSH
- půdní mikrobiologie * MeSH
- rhizosféra * MeSH
- semena rostlinná růst a vývoj mikrobiologie MeSH
- smrk * mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- půda MeSH
The northern bark beetle, Ips duplicatus, is an emerging economic pest, reportedly infesting various species of spruce (Picea spp.), pine (Pinus spp.), and larch (Larix spp.) in Central Europe. Recent climate changes and inconsistent forest management practices have led to the rapid spread of this species, leaving the current monitoring strategies inefficient. As understanding the molecular components of pheromone detection is key to developing novel control strategies, we generated antennal transcriptomes from males and females of this species and annotated the chemosensory proteins. We identified putative candidates for 69 odorant receptors (ORs), 50 ionotropic receptors (IRs), 25 gustatory receptors (GRs), 27 odorant-binding proteins (OBPs), including a tetramer-OBP, 9 chemosensory proteins (CSPs), and 6 sensory neuron membrane proteins (SNMPs). However, no sex-specific chemosensory genes were detected. The phylogenetic analysis revealed conserved orthology in bark beetle chemosensory proteins, especially with a major forest pest and co-habitant, Ips typographus. Recent large-scale functional studies in I. typographus chemoreceptors add greater significance to the orthologous sequences reported here. Nevertheless, identifying chemosensory genes in I. duplicatus is valuable to understanding the chemosensory system and its evolution in bark beetles (Coleoptera) and, generally, insects.
- Klíčová slova
- Coleoptera, Ips duplicatus, RNAseq, antennal transcriptome, bark beetles, chemosensory proteins, olfaction, orthologs, tetramer OBP,
- MeSH
- brouci * genetika metabolismus MeSH
- fylogeneze * MeSH
- hmyzí proteiny * genetika metabolismus MeSH
- receptory pachové * genetika metabolismus MeSH
- stanovení celkové genové exprese metody MeSH
- transkriptom * MeSH
- tykadla členovců * metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- hmyzí proteiny * MeSH
- odorant-binding protein MeSH Prohlížeč
- receptory pachové * MeSH
Plants and insects coevolved as an evolutionarily successful and enduring association. The molecular arms race led to evolutionary novelties regarding unique mechanisms of defence and detoxification in plants and insects. While insects adopt mechanisms to conquer host defence, trees develop well-orchestrated and species-specific defence strategies against insect herbivory. However, current knowledge on the molecular underpinnings of fine-tuned tree defence responses against different herbivore insects is still restricted. In the current study, using a multi-omics approach, we unveiled the defence response of Populus tremula against aphids (Chaitophorus populialbae) and spongy moths (Lymantria dispar) herbivory. Comparative differential gene expression (DGE) analyses revealed that around 272 and 1203 transcripts were differentially regulated in P. tremula after moth and aphid herbivory compared to uninfested controls. Interestingly, 5716 transcripts were differentially regulated in P. tremula between aphids and moth infestation. Further investigation showed that defence-related stress hormones and their lipid precursors, transcription factors, and signalling molecules were over-expressed, whereas the growth-related counterparts were suppressed in P. tremula after aphid and moth herbivory. Metabolomics analysis documented that around 37% of all significantly abundant metabolites were associated with biochemical pathways related to tree growth and defence. However, the metabolic profiles of aphid and moth-fed trees were quite distinct, indicating species-specific response optimization. After identifying the suitable reference genes in P. tremula, the omics data were further validated using RT-qPCR. Nevertheless, our findings documented species-specific fine-tuning of the defence response of P. tremula, showing conservation on resource allocation for defence overgrowth under aphid and moth herbivory. Such findings can be exploited to enhance our current understanding of molecular orchestration of tree responses against herbivory and aid in developing insect pest resistance P. tremula varieties.
- Klíčová slova
- Populus tremula, RT-qPCR, aphids, induced defence, reference gene analysis, spongy moth, transcriptome and metabolomics,
- MeSH
- býložravci * MeSH
- metabolom MeSH
- metabolomika metody MeSH
- mšice * fyziologie MeSH
- můry * fyziologie genetika MeSH
- Populus * genetika parazitologie metabolismus MeSH
- regulace genové exprese u rostlin * MeSH
- stanovení celkové genové exprese MeSH
- transkriptom * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Vacuolar-type (H+)-ATPase (vATPase) is a conserved multi-subunit eukaryotic enzyme composed of 14 subunits that form a functional complex consisting of an ATP-hydrolytic domain (V1) and a proton-translocation domain (V0). ATP hydrolysis and subsequent H+ translocation rely heavily on a fully assembled V1/V0 complex. Since vATPase is crucial for insect survival, it is a viable molecular target for pest control. However, detailed functional analyses of the 14 subunits and their suitability for pest control have not been fully explored in a single insect species. In this study, we identified 22 vATPase subunit transcripts that correspond to 13 subunits (A1, A2, B, C, D, E, F, G, H, a1, a2, c and d) in the white-backed planthopper (WBPH), Sogatella furcifera, a major hemipteran pest of rice. RNAi screens using microinjection and spray-based methods revealed that the SfVHA-F, SfVHA-a2 and SfVHA-c2 subunits are critical. Furthermore, star polymer (SPc) nanoparticles were utilized to conduct spray-induced and nanoparticle-delivered gene silencing (SI-NDGS) to evaluate the pest control efficacy of RNAi targeting the SfVHA-F, SfVHA-a2 and SfVHA-c2 transcripts. Target mRNA levels and vATPase enzymatic activity were both reduced. Honeydew excreta was likewise reduced in WBPH treated with dsRNAs targeting SfVHA-F, SfVHA-a2 and SfVHA-c2. To assess the environmental safety of the nanoparticle-wrapped dsRNAs, Cyrtorhinus lividipennis Reuter, a major natural enemy of planthoppers, was also sprayed with dsRNAs targeting SfVHA-F, SfVHA-a2 and SfVHA-c2. Post-spray effects of dsSfVHA-a2 and dsSfVHA-c2 on C. lividipennis were innocuous. This study identifies SfVHA-a2 and SfVHA-c2 as promising targets for biorational control of WBPH and lays the foundation for developing environment-friendly RNAi biopesticides.
- Klíčová slova
- Natural enemy, RNA interference, Spray-induced and nanoparticle-delivered gene silencing, Star polymer, White-backed planthopper, vATPase,
- MeSH
- adenosintrifosfát MeSH
- Hemiptera * MeSH
- Heteroptera * MeSH
- hodnocení rizik MeSH
- pesticidy * MeSH
- RNA interference MeSH
- rýže (rod) * genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- adenosintrifosfát MeSH
- pesticidy * MeSH
