Vitamin E: An assistant for black soldier fly to reduce cadmium accumulation and toxicity.

Cadmium (Cd) is a toxic heavy metal associated with osteoporosis, liver, and kidney disease. The black soldier fly (BSF) Hermetia illucens may be exposed to Cd during the transformation of livestock manure. The BSF has a high tolerance to Cd. In the previous work of the laboratory, we found that vitamin E (VE) may play a role in the tolerance of BSF to Cd exposure. The main findings are as follows: The BSF larvae pretreated with exogenous VE had heavier body weight, lower content and toxicity of Cd under similar Cd exposure. Even in high Cd exposure at the concentrations of 300 and 700 mg/kg, the BSF larvae pretreated with exogenous VE at a concentration of 100 mg/kg still reduced the Cd toxicity to 85.33 % and 84.43 %, respectively. The best-fitting models showed that metallothionein (MT) content, oxidative damage (8-hydroxydeoxyguanosine content, malondialdehyde content), antioxidant power (total antioxidant power, peroxidase activity) had a great influence on content and toxicity of Cd bioaccumulated in the larvae. The degree of oxidative damage was reduced in the larvae with exogenous VE pretreatments. This variation can be explained by their changed MT content and increased antioxidant power because of exogenous VE. These results reveal the roles of VE in insects defense against Cd exposure and provide a new option for the prevention and therapy of damage caused by Cd exposure.

Specific functions of single pistil S-RNases in S-gene homozygous Pyrus germplasm.

Gametophytic self-incompatibility (SI) is regulated by S-allele recognition; that is, pollen in a style with the same S-genotype will undergo programmed cell death and stop growing so that it is unable to complete double fertilization, ultimately resulting in the SI response. S-RNase is the female determinant of SI in pear (Pyrus). In the Pyrus genome, there are two different S-RNase alleles at the S-locus, which generate two different S-RNase products in the pistil. The extracted S-glycoprotein is actually a protein complex. In this study, artificial self-pollination was conducted at the bud stage to overcome SI in 'Huanghua' (S1S2) pear. Seven plants homozygous for S1-RNase and four homozygous for S2-RNase were selected from the selfed progeny of 'Huanghua' by S-gene molecular identification biotechnology. We investigated the function of single S-RNases isolated from the pistils of S-gene homozygous Pyrus germplasm. The pollen of 'Huanghua' could smoothly pass through the style of the S-gene homozygous germplasm and complete fertilization. S-RNases were extracted from flower styles of different genotypes and used to treat different types of pollen. The S-RNase from 'Huanghua' completely inhibited the growth of S1S2, S1S1, and S2S2 pollen, while the S-RNase from homozygous germplasm allowed some S1S2 pollen and different single genotypes of pollen to continue growing. These results further validate the core events of SI including cytoskeleton depolymerization and programmed cell death. By iTRAQ-based proteomic analysis of style proteins, a total of 13 S-RNase-related proteins were identified. In summary, we have created reliable S-RNase gene homozygous germplasm, which will play a crucial role in further research on SI in pear and in the development of the pear industry.

Life cycle assessment and cost-benefit analysis of nature-based solutions for contaminated land remediation: A mini-review.

Nature-based solutions (NbS) have gained significant attention as a promising approach for remediating contaminated lands, offering multiple ecosystem services (ESs) benefits beyond pollution mitigation. However, the quantitative sustainability assessment of NbS remediation systems, particularly with regard to post-remediation impacts, remains limited. This mini-review aims to address the existing gaps in the assessment of NbS remediation systems by evaluating the limitations of life cycle assessment (LCA) and cost-benefit analysis (CBA) methodologies. A systematic literature search was conducted resulting in the review of 44 relevant studies published between 2006 and 2023. The review highlights an increasing trend in the coverage in the sustainability assessment literature of NbS remediation systems. Phytoextraction was identified as the main NbS mechanism employed in 65 % of the reviewed works, targeting contaminants such as heavy metals and hydrocarbons. However, the post-remediation aspects, including impacts on ESs and the end-of-life management of NbS biomass, were often neglected in the assessments with only a subset of studies partially exploring such aspects. The findings underscore the need for a comprehensive and integrated approach to assess the sustainability of NbS remediation systems, including the incorporation of economic factors, site-specific considerations, and post-remediation impacts. Addressing these gaps will enhance the understanding of NbS effectiveness and facilitate informed decision-making for contaminated land remediation.

Environmental, economic, and social sustainability assessment: A case of using contaminated tailings stabilized by waste-based geopolymer as road base.

Contaminated tailings, e.g., iron tailings, could be stabilized by low-carbon geopolymers for recycling as road base, but the sustainability has yet to be evaluated comprehensively. This study developed a sustainable framework from the life cycle perspective, consisting of quantitative indicators in environment, society, and economy, to assess five stabilization cases (i.e., M1, M2, C1, C2, and cement). Moreover, a modified AHP (Analytic Hierarchy Process)-CRITIC (CRiteria Importance Through Inter-criteria Correlation)-TOPSIS (Technique for Order of Preference by Similarity to Ideal Solution) model was applied to select the most sustainable stabilization method. The sustainability scores of four cases using geopolymers were higher than that of the cement case (0.22), with C2 (0.75), C1 (0.64), M1 (0.56), and M2 (0.54), respectively. Sensitivity analysis confirmed that the assessment results were relatively stable, especially when the subjective weight of the economy was not the highest (the cement counterpart had economic advantages). This study constituted a novel approach to complement the selection of sustainable stabilization cases, bridging the gap of focusing exclusively on green stabilization performance.

Coalbed Methane Enrichment Characteristics and Exploration Target Selection in the Zhuozishan Coalfield of the Western Ordos Basin, China.

The Zhuozishan coalfield at the western margin of the Ordos Basin is one of the main coal-mining areas in China, and recent explorations have revealed the great potential for coalbed methane (CBM) resources in its Carboniferous and Permian strata. In this paper, the controlling factors of CBM enrichment of the major coals are studied in this coalfield and the CBM resources are estimated based on the analysis of the coal petrology and compilation of literature data on the gas content. The result of the coal petrology analysis of 10 samples shows that the vitrinite content of No. 16 coal (71.9-77.3%) is higher than that of No. 9 coal (59.1-65.1%), and the inertinite content of No. 16 coal (18.9-23.5%) is lower than that of No. 9 coal (30.1-34.9%). The R o,max value of No. 16 coal (1.18-1.35%) is higher than that of No. 9 coal (1.04-1.13%), and both coals are of medium rank. Due to greater thickness, deeper burial depth, and better coal petrology characteristics, the No. 16 coal seam of the Taiyuan Formation is selected as the major coal seam for CBM resource estimation, which has a thickness of 1-6 m and a present-day burial depth of 200-1100 m. The gas content of this coal seam varies mostly between 4 and 10 m3/t. Positive correlation between the coal seam thickness as well as present-day burial depth and the gas content suggests that the thick and deeply buried coal seams are favorable for CBM preservation. The ash yield shows an insignificant negative correlation with the gas content, indicating that ash yield is not an important factor for CBM enrichment. The syncline hinges located below the thrust zones show higher gas content due to greater burial depths. In contrast, the anticline hinges at shallower depths tend to have lower gas contents. Based on the combined information about sedimentary environments, structural patterns, and hydrogeology, two CBM accumulation models are put forward in the study area that include syncline-hydraulic plugging below thrust nappe and fault-confined aquifer plugging. The volumetric method is used to estimate the CBM resources, and results indicate that the CBM resource in the whole coalfield is 428.78 × 108 m3, and the total resource abundance is 0.74 × 108 m3/km2. Two favorable areas for the CBM exploration are optimized based on the resource amount and resource abundance. One of the favorable areas is the Kabuqi area in the northern part of the coalfield, and another is the Baiyunwusu area in the central part of the coalfield. These two areas will be the CBM priority exploration areas at the western margin of the Ordos Basin.

Two low-toxic Klebsiella pneumoniae strains from gut of black soldier fly Hermetia illucens are multi-resistance to sulfonamides and cadmium.

In recent years, pollution of antibiotics and heavy metal has often been reported in organic wastes. Saprophytic insects have been recorded as biological control agents in organic waste management. During organic waste conversion, the intestinal bacteria of the saprophytic insects play an important role in digestion, physiology, immunity and prevention of pathogen colonization. Black soldier fly (BSF) Hermetia illucens has been widely used as saprophytic insects and showed tolerance to sulfonamides (SAs) and cadmium (Cd). Diversity and changes in gut microbiota of black soldier fly larvae (BSFL) were evaluated through 16S rRNA high-throughput sequencing, and a decrease in diversity of gut microbiota along with an increase in SAs stress was recorded. Major members identified were Actinomycetaceae, Enterobacteriaceae, and Enterococcaceae. And fourteen multi-resistance Klebsiella pneumoniae strains were isolated. Two strains BSFL7-B-5 (from middle midgut of 7-day BSFL) and BSFL11-C-1 (from posterior midgut of 11-day BSFL) were found to be low-toxic and multi-resistance. The adsorption rate of SAs in 5 mg/kg solutions by these two strains reached 65.2% and 61.6%, respectively. Adsorption rate of Cd in 20 mg/L solutions was 77.2% for BSFL7-B-5. The strain BSFL11-C-1 showed higher than 70% adsorption rates of Cd in 20, 30 and 40 mg/L solutions. This study revealed that the presence of multi-resistance bacterial strains in the gut of BSFL helped the larvae against SAs or Cd stress. After determining how and where they are used, selected BSFL gut bacterial strains might be utilized in managing SAs or Cd contamination at suitable concentrations in the future.

Identification of three metallothioneins in the black soldier fly and their functions in Cd accumulation and detoxification.

The black soldier fly (BSF) Hermetia illucens has a strong tolerance to cadmium stress. This helps to use BSF in entomoremediation of heavy metal pollution. Rich metallothionein (MT) proteins were thought to be important for some insects to endure the toxicity of heavy metal. We identified and characterized three MTs genes in BSF (BSFMTs), including BSFMT1, BSFMT2A, and BSFMT2B. Molecular modeling was used to predict metal binding sites. Phylogenetic analysis was used to identify gene families. Overexpression of the recombinant black soldier fly metallothioneins was found to confer Cd tolerance in Escherichia coli. Finally, functions of BSFMTs in BSF were explored through RNA interference (RNAi). RNAi results of BSFMT2B showed that the larval fresh weight decreased significantly, and the larvae mortality increased significantly. This study suggests that BSFMTs have important properties in Cd detoxification and tolerance in BSF. Further characterization analyses of physiological function about metallothioneins are necessary in BSF and other insects.

Materials Engineering in Perovskite for Optimized Oxygen Evolution Electrocatalysis in Alkaline Condition.

Developing robust and highly efficient electrocatalysts for oxygen evolution reaction (OER) is critical for renewable, secure, and emission-free energy technologies. Perovskite Ba0.5 Sr0.5 Co0.8 Fe0.2 O3-δ (BSCF) has emerged as a promising OER electrocatalyst with desirable intrinsic activity. Inspired by the factor that substituting in transition-metal sublattice of the perovskite can further optimize the OER activity, herein, nickel-substituted BSCF is adopted, that is, Ba0.5 Sr0.5 Co0.8- x Fe0.2 Nix O3-δ (x = 0.05, 0.1, 0.2, denoted as BSCFNx, x = 5, 10, 20, respectively), as efficient and stable OER catalysts in alkaline solution. The phase structure, microchemistry, oxygen vacancy, and electrochemical activity of such samples are well-investigated. Endowed with an overpotential of only 278 mV at 10 mA cm-2 and a Tafel slope of merely 47.98 mV dec-1 , BSCFN20 exhibits the optimum OER activity. When constructing a two-electrode cell with BSCFN20 as anode and Pt/C as cathode (BSCFN20||Pt/C) for water splitting, it only requires a voltage of 1.63 V to achieve 50 mA cm-2 , and the BSCFN20||Pt/C remains stable within 80 h at 10 mA cm-2 , superior to the state-of-the-art RuO2 ||Pt/C counterpart. This work provides a feasible strategy for designing stable and highly active perovskite electrocatalysts for future energy storage and conversion.

Diversity of metabolite accumulation patterns in inner and outer seed coats of pomegranate: exploring their relationship with genetic mechanisms of seed coat development.

The expanded outer seed coat and the rigid inner seed coat of pomegranate seeds, both affect the sensory qualities of the fruit and its acceptability to consumers. Pomegranate seeds are also an appealing model for the study of seed coat differentiation and development. We conducted nontarget metabolic profiling to detect metabolites that contribute to the morphological differentiation of the seed coats along with transcriptomic profiling to unravel the genetic mechanisms underlying this process. Comparisons of metabolites in the lignin biosynthetic pathway accumulating in seed coat layers at different developmental stages revealed that monolignols, including coniferyl alcohol and sinapyl alcohol, greatly accumulated in inner seed coats and monolignol glucosides greatly accumulated in outer seed coats. Strong expression of genes involved in monolignol biosynthesis and transport might explain the spatial patterns of biosynthesis and accumulation of these metabolites. Hemicellulose constituents and flavonoids in particular accumulated in the inner seed coat, and candidate genes that might be involved in their accumulation were also identified. Genes encoding transcription factors regulating monolignol, cellulose, and hemicellulose metabolism were chosen by coexpression analysis. These results provide insights into metabolic factors influencing seed coat differentiation and a reference for studying seed coat developmental biology and pomegranate genetic improvement.

Rapid Detection of Alternaria Species Involved in Pear Black Spot Using Loop-Mediated Isothermal Amplification.

Alternaria species are the most important fungal pathogens that attack various crops as well as fruit trees such as pear and cause black spot disease. Here, a loop-mediated isothermal amplification (LAMP) assay is developed for the detection of Alternaria species. A. alternata cytochrome b (cyt-b) gene was used to design two pairs of primers and amplified a 229-bp segment of Aacyt-b gene. The results showed that LAMP assay is faster and simpler than polymerase chain reaction (PCR). LAMP assay is highly sensitive method for the detection of about 1 pg of genomic DNA of A. alternata by using optimized concentration of MgCl2 (4 mM) in final LAMP reaction. In contrast, the limit of detection was 1 ng of target DNA via conventional PCR. Among the genomic DNA of 46 fungal species, only the tubes containing DNA of Alternaria spp. except A. porri, A. solani, and A. infectoria changed color from orange to yellowish green with SYBR Green I including the main pathogens of pear black spot. The yellowish green color was indicative of DNA amplification. Moreover, LAMP assay was used for testing infected tissues among 22 healthy and diseased pear tissues; the orange color changed to yellowish green for infected tissues only. Altogether, we conclude that cyt-b gene can be used for the detection of Alternaria spp. via LAMP assay, which is involved in pear black spot disease.

Reference gene selection for quantitative gene expression analysis in black soldier fly (Hermetia illucens).

Hermetia illucens is an important resource insect for the conversion of organic waste. Quantitative PCR (qPCR) is the primary tool of gene expression analysis and a core technology of molecular biology research. Reference genes are essential for qPCR analysis; however, a stability analysis of H. illucens reference genes has not yet been carried out. To find suitable reference genes for normalizing gene expression data, the stability of eight housekeeping genes (including ATP6V1A, RPL8, EF1, Tubulin, TBP, GAPDH, Actin and RP49) was investigated under both biotic (developmental stages, tissues and sex) and abiotic (heavy metals, food, antibiotics) conditions. Gene expression data were analysed by geNorm, NormFinder, BestKeeper, and ΔCt programs. A set of specific reference genes was recommended for each experimental condition using the results of RefFinder synthesis analysis. This study offers a solid foundation for further studies of the molecular biology of H. illucens.

Effect of sulfonamide pollution on the growth of manure management candidate Hermetia illucens.

Antibiotics are commonly used in livestock and poultry farming. Residual antibiotics in manure may lead to antibiotic pollution of soil, surface water, and groundwater through land application and run-off rainfall. The black soldier fly (BSF) Hermetia illucens is a good candidate for manure management. We evaluated the effect of sulfonamide pollution on the growth of H. illucens. Four treatments were considered with a sulfonamide content in the feed of 0 (control group), 0.1, 1, and 10 mg/kg. The control larvae were fed without sulfonamide. Survival and development status of the individuals were recorded daily. The weights of the fifth instar larvae, prepupae, and pupae were checked. Antioxidant enzyme activity was determined with the fifth instar larvae. The results showed that a low (0.1 and 1 mg/kg) concentration of sulfonamides had no effects on larval survival, pupation, and eclosion of BSFs. A high sulfonamide concentration of 10 mg/kg had a significant effect on the survival of larvae and pupae and on the body weight of larvae, prepupae and pupae. Peak of the cumulated pupation rate and eclosion rate in the sulfonamide treatment of 10 mg/kg was very low. Pupation and eclosion in this group peaked later than that of the control and low sulfonamide concentration treatment groups (0.1 mg/kg and 1 mg/kg). Larvae from the sulfonamides group showed lower antioxidase activities than that of the control. In sulfonamide groups, the activity of peroxidase and superoxide dismutase was reduced in a concentration-dependent manner. Sulfamonomethoxine, sulfamethoxazole, and sulfamethazine were not detected in the harvested prepupae. Only sulfadiazine was discovered in the sulfonamide treatments of 1 and 10 mg/kg. In conclusion, BSFs can tolerate certain concentrations of sulfonamide contamination.

The pomegranate (Punica granatum L.) genome and the genomics of punicalagin biosynthesis.

Pomegranate (Punica granatum L.) is a perennial fruit crop grown since ancient times that has been planted worldwide and is known for its functional metabolites, particularly punicalagins. We have sequenced and assembled the pomegranate genome with 328 Mb anchored into nine pseudo-chromosomes and annotated 29 229 gene models. A Myrtales lineage-specific whole-genome duplication event was detected that occurred in the common ancestor before the divergence of pomegranate and Eucalyptus. Repetitive sequences accounted for 46.1% of the assembled genome. We found that the integument development gene INNER NO OUTER (INO) was under positive selection and potentially contributed to the development of the fleshy outer layer of the seed coat, an edible part of pomegranate fruit. The genes encoding the enzymes for synthesis and degradation of lignin, hemicelluloses and cellulose were also differentially expressed between soft- and hard-seeded varieties, reflecting differences in their accumulation in cultivars differing in seed hardness. Candidate genes for punicalagin biosynthesis were identified and their expression patterns indicated that gallic acid synthesis in tissues could follow different biochemical pathways. The genome sequence of pomegranate provides a valuable resource for the dissection of many biological and biochemical traits and also provides important insights for the acceleration of breeding. Elucidation of the biochemical pathway(s) involved in punicalagin biosynthesis could assist breeding efforts to increase production of this bioactive compound.