Sex pheromone biosynthesis in the sugarcane borer Diatraea saccharalis: paving the way for biotechnological production.

BACKGROUND: The sugarcane borer Diatraea saccharalis (Lepidoptera) is a key pest on sugarcane and other grasses in the Americas. Biological control as well as insecticide treatments are used for pest management, but economic losses are still significant. The use of female sex pheromones for mating disruption or mass trapping in pest management could be established for this species, provided that economical production of pheromone is available. RESULTS: Combining in vivo labelling studies, differential expression analysis of transcriptome data and functional characterisation of insect genes in a yeast expression system, we reveal the biosynthetic pathway and identify the desaturase and reductase enzymes involved in the biosynthesis of the main pheromone component (9Z,11E)-hexadecadienal, and minor components hexadecanal, (9Z)-hexadecenal and (11Z)-hexadecenal. We next demonstrate heterologous production of the corresponding alcohols of the pheromone components, by expressing multiple steps of the biosynthetic pathway in yeast. CONCLUSION: Elucidation of the genetic basis of sex pheromone biosynthesis in D. saccharalis, and heterologous expression in yeast, paves the way for biotechnological production of the pheromone compounds needed for pheromone-based pest management of this species. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effects of long-term fertilization with contemporary Danish human urine, composted household waste and sewage sludge on soil nematode abundance and community structure.

It is desirable to recycle the urban waste products human urine, composted household waste and sewage sludge as fertilizers to agricultural fields. This could minimize the use of NPK fertilizer, improve soil structure and store carbon. However, waste products may contain heavy metals, persistent organic pollutants (POP) and plastics, and there are concerns that long-term build-up of these substances will cause unwanted effects on soil health. Nematodes are ubiquitous and numerous in soil ecosystems. Abundance and community structure of soil nematodes can be used as indicators of soil health, as some species are vulnerable to pollution. There are well-developed methods for detecting environmental changes based on nematode community structure. At the long-term CRUCIAL field experiment, where alternative fertilizer products have been applied since 2003, we measured effects of long-term fertilization with human urine, composted household waste and sewage sludge on soil properties (pH, soil organic matter and nitrogen availability), abundance of soil microorganisms (bacteria, fungi, small protozoa and ciliates) and nematode trophic groups compared to plots with unfertilized, NPK and cattle manure treatment. Sampling and assessments were done three times during a growth season. Further, we assessed the composition of nematode communities using metabarcoding. Treatments with a high input of organic matter (cattle manure, composted household waste and sewage sludge) had high abundances of bacteria and thus bacterial grazers (small protozoa, ciliates, and bacterial feeding nematodes). We found a significant correlation between nematode community structure and pH and organic matter. We calculated the nematode Maturity Index 2-5 (pollution indicator) based on metabarcoding data, which did not differ significantly between the treatments. We conclude that long-term fertilization with different types of contemporary Danish urban waste products affects both soil properties and abundance of soil organisms, the latter largely reflecting the organic matter input of the fertilizer treatments. We found no adverse effect on nematode communities that could indicate pollution-induced stress on nematofauna or decreased soil fertility.

Production of moth sex pheromones for pest control by yeast fermentation.

The use of insect sex pheromones is an alternative technology for pest control in agriculture and forestry, which, in contrast to insecticides, does not have adverse effects on human health or environment and is efficient also against insecticide-resistant insect populations. Due to the high cost of chemically synthesized pheromones, mating disruption applications are currently primarily targeting higher value crops, such as fruits. Here we demonstrate a biotechnological method for the production of (Z)-hexadec-11-en-1-ol and (Z)-tetradec-9-en-1-ol, using engineered yeast cell factories. These unsaturated fatty alcohols are pheromone components or the immediate precursors of pheromone components of several economically important moth pests. Biosynthetic pathways towards several pheromones or their precursors were reconstructed in the oleaginous yeast Yarrowia lipolytica, which was further metabolically engineered for improved pheromone biosynthesis by decreasing fatty alcohol degradation and downregulating storage lipid accumulation. The sex pheromone of the cotton bollworm Helicoverpa armigera was produced by oxidation of fermented fatty alcohols into corresponding aldehydes. The resulting yeast-derived pheromone was just as efficient and specific for trapping of H. armigera male moths in cotton fields in Greece as a conventionally produced synthetic pheromone mixture. We further demonstrated the production of (Z)-tetradec-9-en-1-yl acetate, the main pheromone component of the fall armyworm Spodoptera frugiperda. Taken together our work describes a biotech platform for the production of commercially relevant titres of moth pheromones for pest control via yeast fermentation.

A global database of soil nematode abundance and functional group composition.

As the most abundant animals on earth, nematodes are a dominant component of the soil community. They play critical roles in regulating biogeochemical cycles and vegetation dynamics within and across landscapes and are an indicator of soil biological activity. Here, we present a comprehensive global dataset of soil nematode abundance and functional group composition. This dataset includes 6,825 georeferenced soil samples from all continents and biomes. For geospatial mapping purposes these samples are aggregated into 1,933 unique 1-km pixels, each of which is linked to 73 global environmental covariate data layers. Altogether, this dataset can help to gain insight into the spatial distribution patterns of soil nematode abundance and community composition, and the environmental drivers shaping these patterns.

A single-host fermentation process for the production of flavor lactones from non-hydroxylated fatty acids.

Lactone flavors with fruity, milky, coconut, and other aromas are widely used in the food and fragrance industries. Lactones are produced by chemical synthesis or by biotransformation of plant-sourced hydroxy fatty acids. We established a novel method to produce flavor lactones from abundant non-hydroxylated fatty acids using yeast cell factories. Oleaginous yeast Yarrowia lipolytica was engineered to perform hydroxylation of fatty acids and chain-shortening via ß-oxidation to preferentially twelve or ten carbons. The strains could produce γ-dodecalactone from oleic acid and δ-decalactone from linoleic acid. Through metabolic engineering, the titer was improved 4-fold, and the final strain produced 282 mg/L γ-dodecalactone in a fed-batch bioreactor. The study paves the way for the production of lactones by fermentation of abundant fatty feedstocks.

Soil nematode abundance and functional group composition at a global scale.

Soil organisms are a crucial part of the terrestrial biosphere. Despite their importance for ecosystem functioning, few quantitative, spatially explicit models of the active belowground community currently exist. In particular, nematodes are the most abundant animals on Earth, filling all trophic levels in the soil food web. Here we use 6,759 georeferenced samples to generate a mechanistic understanding of the patterns of the global abundance of nematodes in the soil and the composition of their functional groups. The resulting maps show that 4.4 ± 0.64 × 1020 nematodes (with a total biomass of approximately 0.3 gigatonnes) inhabit surface soils across the world, with higher abundances in sub-Arctic regions (38% of total) than in temperate (24%) or tropical (21%) regions. Regional variations in these global trends also provide insights into local patterns of soil fertility and functioning. These high-resolution models provide the first steps towards representing soil ecological processes in global biogeochemical models and will enable the prediction of elemental cycling under current and future climate scenarios.

EasyCloneYALI: CRISPR/Cas9-Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica.

The oleaginous yeast Yarrowia lipolytica is an emerging host for production of fatty acid-derived chemicals. To enable rapid iterative metabolic engineering of this yeast, there is a need for well-characterized genetic parts and convenient and reliable methods for their incorporation into yeast. Here, the EasyCloneYALI genetic toolbox, which allows streamlined strain construction with high genome editing efficiencies in Y. lipolytica via the CRISPR/Cas9 technology is presented. The toolbox allows marker-free integration of gene expression vectors into characterized genome sites as well as marker-free deletion of genes with the help of CRISPR/Cas9. Genome editing efficiencies above 80% were achieved with transformation protocols using non-replicating DNA repair fragments (such as DNA oligos). Furthermore, the toolbox includes a set of integrative gene expression vectors with prototrophic markers conferring resistance to hygromycin and nourseothricin.

Long-term and realistic global change manipulations had low impact on diversity of soil biota in temperate heathland.

In a dry heathland ecosystem we manipulated temperature (warming), precipitation (drought) and atmospheric concentration of CO2 in a full-factorial experiment in order to investigate changes in below-ground biodiversity as a result of future climate change. We investigated the responses in community diversity of nematodes, enchytraeids, collembolans and oribatid mites at two and eight years of manipulations. We used a structural equation modelling (SEM) approach analyzing the three manipulations, soil moisture and temperature, and seven soil biological and chemical variables. The analysis revealed a persistent and positive effect of elevated CO2 on litter C:N ratio. After two years of treatment, the fungi to bacteria ratio was increased by warming, and the diversities within oribatid mites, collembolans and nematode groups were all affected by elevated CO2 mediated through increased litter C:N ratio. After eight years of treatment, however, the CO2-increased litter C:N ratio did not influence the diversity in any of the four fauna groups. The number of significant correlations between treatments, food source quality, and soil biota diversities was reduced from six to three after two and eight years, respectively. These results suggest a remarkable resilience within the soil biota against global climate change treatments in the long term.

Five year change in alcohol intake and risk of breast cancer and coronary heart disease among postmenopausal women: prospective cohort study.

OBJECTIVE: To test the hypothesis that postmenopausal women who increase their alcohol intake over a five year period have a higher risk of breast cancer and a lower risk of coronary heart disease compared with stable alcohol intake. DESIGN: Prospective cohort study. SETTING: Denmark, 1993-2012. PARTICIPANTS: 21 523 postmenopausal women who participated in the Diet, Cancer, and Health Study in two consecutive examinations in 1993-98 and 1999-2003. Information on alcohol intake was obtained from questionnaires completed by participants. MAIN OUTCOME MEASURES: Incidence of breast cancer, coronary heart disease, and all cause mortality during 11 years of follow-up. Information was obtained from the Danish Cancer Register, Danish Hospital Discharge Register, Danish Register of Causes of Death, and National Central Person Register. We estimated hazard ratios according to five year change in alcohol intake using Cox proportional hazards models. RESULTS: During the study, 1054, 1750, and 2080 cases of breast cancer, coronary heart disease, and mortality occurred, respectively. Analyses modelling five year change in alcohol intake with cubic splines showed that women who increased their alcohol intake over the five year period had a higher risk of breast cancer and a lower risk of coronary heart disease than women with a stable alcohol intake. For instance, women who increased their alcohol intake by seven or 14 drinks per week (corresponding to one or two drinks more per day) had hazard ratios of breast cancer of 1.13 (95% confidence interval 1.03 to 1.23) and 1.29 (1.07 to 1.55), respectively, compared to women with stable intake, and adjusted for age, education, body mass index, smoking, Mediterranean diet score, parity, number of births, and hormone replacement therapy. For coronary heart disease, corresponding hazard ratios were 0.89 (0.81 to 0.97) and 0.78 (0.64 to 0.95), respectively, adjusted for age, education, body mass index, Mediterranean diet score, smoking, physical activity, hypertension, elevated cholesterol, and diabetes. Results among women who reduced their alcohol intake over the five year period were not significantly associated with risk of breast cancer or coronary heart disease. Analyses of all cause mortality showed that women who increased their alcohol intake from a high intake (≥14 drinks per week) to an even higher intake had a higher mortality risk that women with a stable high intake. CONCLUSION: In this study of postmenopausal women over a five year period, results support the hypotheses that alcohol intake is associated with increased risk of breast cancer and decreased risk of coronary heart disease.

Defoliation reduces soil biota - and modifies stimulating effects of elevated CO2.

To understand the responses to external disturbance such as defoliation and possible feedback mechanisms at global change in terrestrial ecosystems, it is necessary to examine the extent and nature of effects on aboveground-belowground interactions. We studied a temperate heathland system subjected to experimental climate and atmospheric factors based on prognoses for year 2075 and further exposed to defoliation. By defoliating plants, we were able to study how global change modifies the interactions of the plant-soil system. Shoot production, root biomass, microbial biomass, and nematode abundance were assessed in the rhizosphere of manually defoliated patches of Deschampsia flexuosa in June in a full-factorial FACE experiment with the treatments: increased atmospheric CO 2, increased nighttime temperatures, summer droughts, and all of their combinations. We found a negative effect of defoliation on microbial biomass that was not apparently affected by global change. The negative effect of defoliation cascades through to soil nematodes as dependent on CO 2 and drought. At ambient CO 2, drought and defoliation each reduced nematodes. In contrast, at elevated CO 2, a combination of drought and defoliation was needed to reduce nematodes. We found positive effects of CO 2 on root density and microbial biomass. Defoliation affected soil biota negatively, whereas elevated CO 2 stimulated the plant-soil system. This effect seen in June is contrasted by the effects seen in September at the same site. Late season defoliation increased activity and biomass of soil biota and more so at elevated CO 2. Based on soil biota responses, plants defoliated in active growth therefore conserve resources, whereas defoliation after termination of growth results in release of resources. This result challenges the idea that plants via exudation of organic carbon stimulate their rhizosphere biota when in apparent need of nutrients for growth.

Are there links between responses of soil microbes and ecosystem functioning to elevated CO2, N deposition and warming? A global perspective.

In recent years, there has been an increase in research to understand how global changes' impacts on soil biota translate into altered ecosystem functioning. However, results vary between global change effects, soil taxa, and ecosystem processes studied, and a synthesis of relationships is lacking. Therefore, here we initiate such a synthesis to assess whether the effect size of global change drivers (elevated CO2, N deposition, and warming) on soil microbial abundance is related with the effect size of these drivers on ecosystem functioning (plant biomass, soil C cycle, and soil N cycle) using meta-analysis and structural equation modeling. For N deposition and warming, the global change effect size on soil microbes was positively associated with the global change effect size on ecosystem functioning, and these relationships were consistent across taxa and ecosystem processes. However, for elevated CO2, such links were more taxon and ecosystem process specific. For example, fungal abundance responses to elevated CO2 were positively correlated with those of plant biomass but negatively with those of the N cycle. Our results go beyond previous assessments of the sensitivity of soil microbes and ecosystem processes to global change, and demonstrate the existence of general links between the responses of soil microbial abundance and ecosystem functioning. Further we identify critical areas for future research, specifically altered precipitation, soil fauna, soil community composition, and litter decomposition, that are need to better quantify the ecosystem consequences of global change impacts on soil biodiversity.

Smoking and risk of liver cirrhosis: a population-based cohort study.

OBJECTIVE: Alcohol is the most acknowledged risk factor for liver cirrhosis. Smoking is rarely considered to be a cause of liver cirrhosis even though a few studies have suggested the opposite. The aim of this study was to assess the independent effect of smoking on alcoholic liver cirrhosis and liver cirrhosis in general. MATERIALS AND METHODS: The authors used data from the Copenhagen City Heart Study including a cohort of 9889 women and 8590 men from the Danish general population. Smoking and covariates were assessed at four consecutive examinations in 1976-78, 1981-83, 1991-94 and 2001-03. Updated measures were used in the analyses. Information on incident cases of alcoholic liver cirrhosis and liver cirrhosis was obtained from national hospital registries. Data were analyzed by means of Cox regression. RESULTS: A total of 225 cases of alcoholic liver cirrhosis and 431 cases of liver cirrhosis occurred during follow-up. The hazard ratios (HR) of alcoholic liver cirrhosis was 3.9, 95% confidence interval (CI): 1.6, 9.4 for women and 1.6, 95% CI: 0.9, 3.0 for men smoking >10 g of tobacco per day compared with never-smokers. For liver cirrhosis, corresponding HRs were 2.2, 95% CI: 1.4, 3.4 for women and 1.4, 95% CI: 0.9, 2.2 for men. The HRs were adjusted for age, alcohol intake, education and body mass index. CONCLUSIONS: Smoking was associated with an increased risk of liver cirrhosis independent of alcohol intake.

[New unifying diagnosis of functional diseases]. / Ny faelles diagnose for de funktionelle sygdomme.

Functional somatic symptoms are prevalent in all medical settings, but their management is hampered by an obsolete theoretical framework and inadequate classification systems. Epidemiological and neurobiological studies suggest that the functional somatic syndromes, e.g. fibromyalgia, chronic fatigue syndrome, irritable bowel syndrome and somatoform disorders belong to the same family of disorders. An empirically based diagnosis including different subtypes and severities is proposed as a unifying diagnostic construct: bodily distress syndrome. This construct provides a common language for functional disorders across medical specialties.