Transcription factor shapes chromosomal conformation and regulates gene expression in bacterial adaptation.

Genomic mutations allow bacteria to adapt rapidly to adverse stress environments. The three-dimensional conformation of the genome may also play an important role in transcriptional regulation and environmental adaptation. Here, using chromosome conformation capture, we investigate the high-order architecture of the Zymomonas mobilis chromosome in response to genomic mutation and ambient stimuli (acetic acid and furfural, derived from lignocellulosic hydrolysate). We find that genomic mutation only influences the local chromosome contacts, whereas stress of acetic acid and furfural restrict the long-range contacts and significantly change the chromosome organization at domain scales. Further deciphering the domain feature unveils the important transcription factors, Ferric uptake regulator (Fur) proteins, which act as nucleoid-associated proteins to promote long-range (>200 kb) chromosomal communications and regulate the expression of genes involved in stress response. Our work suggests that ubiquitous transcription factors in prokaryotes mediate chromosome organization and regulate stress-resistance genes in bacterial adaptation.

Cellulosimicrobium composti sp. nov., a thermophilic bacterium isolated from compost.

A Gram-stain-positive, yellow-pigmented, non-motile actinobacterial strain, designated as BIT-GX5T, was isolated from a sesame husks compost collected in Beijing, PR China. This bacterium was found to be able to grow in the temperature range from 16 to 50 °C and had an optimal growth temperature at 45 °C. Its taxonomic position was analysed using a polyphasic approach. The 16S rRNA gene sequence (1482 bp) of strain BIT-GX5T was most similar to Cellulosimicrobium funkei ATCC BAA-886T (99.45%), Cellulosimicrobium cellulans LMG 16121T (99.17%) and Cellulosimicrobium marinum RS-7-4T (98.75%). The results of phylogenetic analyses, based on the 16S rRNA gene, concatenated sequences of five housekeeping genes (gyrB, rpoB, recA, atpD and trpB) and genome sequences, placed strain BIT-GX5T in a separate lineage among the genus Cellulosimicrobium within the family Promicromonosporaceae. The major polar lipids of strain BIT-GX5T were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminophospholipid and aminolipid. The major isoprenoid quinone was MK-9(H4), while the cell-wall sugars were galactose, rhamnose, glucose and mannose. The peptidoglycan type was A4α l-Lys-d-Ser-d-Asp. The major fatty acids were anteiso-C15:0 and iso-C15: 0, which were similar to other members in the genus Cellulosimicrobium. Results of in silico DNA-DNA hybridization and average nucleotide identity calculations plus physiological and biochemical tests exhibited the genotypic and phenotypic differentiation of strain BIT-GX5T from the other members of the genus Cellulosimicrobium. Therefore, strain BIT-GX5T is considered to represent a novel species within the genus Cellulosimicrobium, for which the name Cellulosimicrobium composti sp. nov. is proposed. The type strain is BIT-GX5T (= CGMCC 1.17687T = KCTC 49391T).

Intestinirhabdus alba gen. nov., sp. nov., a novel genus of the family Enterobacteriaceae, isolated from the gut of plastic-eating larvae of the Coleoptera insect Zophobas atratus.

A bacterial strain, BIT-B35T, was isolated from the gut of plastic-eating larvae of the Coleoptera insect Zophobas atratus. Its taxonomic position was determined by using a polyphasic approach. Cells were white-pigmented, Gram-stain-negative, motile short rods with terminal flagella. The 16S rRNA gene sequence (1411 bp) of strain BIT-B35T showed highest similarity (98.1%) to Escherichia fergusonii ATCC 35469T and Citrobacter koseri LMG 5519T. The results of phylogenetic analyses, based on the 16S rRNA gene, concatenated sequences of seven housekeeping genes (atpD, gyrB, infB, rpoB, pyrG, fusA and leuS) and genome sequences, placed strain BIT-B35T in a separate lineage among the family of Enterobacteriaceae. The major fatty acids were C16 : 0, C17 : 0 cyclo and C19 : 0 cyclo ω8c. The genomic DNA G+C content of strain BIT-B35T was 57.1 mol%. The chemotaxonomic data plus results of physiological and biochemical tests also distinguished strain BIT-B35T from members of other genera within the family Enterobacteriaceae. Therefore, strain BIT-B35T is considered to represent a novel species of a novel genus within the family Enterobacteriaceae, for which the name Intestinirhabdus alba gen. nov., sp. nov. is proposed. The type strain is BIT-B35T (=CGMCC 1.17042T=KCTC 72448T).

Myroides albus sp. nov., isolated from the gut of plastic-eating larvae of the coleopteran insect Zophobas atratus.

A bacterial strain, BIT-d1T, was isolated from the gut of plastic-eating larvae of the coleopteran insect Zophobas atratus. Its taxonomic position was analysed using a polyphasic approach. Cells were white-pigmented, Gram-stain-negative, non-motile, long rods without flagella. The 16S rRNA gene sequence (1401 bp) of strain BIT-d1T showed highest similarity (98.0%) to Myroides pelagicus SM1T and 96.6~92.6 % similarity to the other species of the genus Myroides. The results of phylogenetic analyses, based on the 16S rRNA gene, concatenated sequences of six housekeeping genes (gyrB, dnaK, tuf, murG, atpA and glyA) and genome sequences, placed strain BIT-d1T in a separate lineage among the genus Myroides, family Flavobacteriaceae. The major isoprenoid quinone was menaquinone-6 (MK-6) and the major fatty acids were C15 : 0 iso, C17 : 0 iso 3-OH and summed feature 9 (comprising iso-C17 : 1 ω9c and/or C16 : 0 10-methyl), which were similar to other members in the genus Myroides. In silico DNA-DNA hybridization and average nucleotide identity calculations plus physiological and biochemical tests exhibited the genotypic and phenotypic differentiation of strain BIT-d1T from the other members of the genus Myroides. Therefore, strain BIT-d1T is considered to represent a novel species within the genus Myroides, for which the name Myroides albus sp. nov is proposed. The type strain is BIT-d1T (=CGMCC 1.17043T=KCTC 72447T).

Mixta tenebrionis sp. nov., isolated from the gut of the plastic-eating mealworm Tenebrio molitor L.

A bacterial strain, BIT-26T, was isolated from the gut of plastic-eating mealworm Tenebrio molitor L. The taxonomic position of this new isolate was investigated by using a polyphasic approach. Cells of the strain were Gram-stain-negative, facultatively anaerobic, motile rods with peritrichous flagella. The 16S rRNA gene sequence (1412 bp) of strain BIT-26T showed the highest similarity (97.4 %) to Erwinia piriflorinigrans CFBP 5888T, followed by Citrobacter sedlakii NBRC 105722T (97.3 %), Mixta calida LMG 25383T (97.3 %), Cronobacter muytjensii ATCC 51329T (97.2 %) and Mixta theicola QC88-366 T (97.2 %). The results of phylogenetic analyses, based on the 16S rRNA gene and concatenated sequences of four housekeeping genes (atpD, gyrB, infB and rpoB), placed strain BIT-26T within the genus Mixta of the family Erwiniaceae. This affiliation was also supported by the chemotaxonomic data. Strain BIT-26T had similar predominant fatty acids, including C12 : 0, C14 : 0, C16 : 0, C17 : 0 cyclo and C19 : 0 cyclo ω8c, to species of the genus Mixta. In silico DNA-DNA hybridization and average nucleotide identity calculations plus physiological and biochemical tests allowed the genotypic and phenotypic differentiation of strain BIT-26T from other species of the genus Mixta with validly published names. Therefore, strain BIT-26T is considered to represent a novel species, for which the name Mixta tenebrionis sp. nov is proposed. The type strain is BIT-26T (=CGMCC 1.17041T=KCTC 72449T).

Photo-Oxidative Degradation Mitigated the Developmental Toxicity of Polyamide Microplastics to Zebrafish Larvae by Modulating Macrophage-Triggered Proinflammatory Responses and Apoptosis.

Microplastics (MPs) are ubiquitous in the environment and pose substantial threats to the water ecosystem. However, the impact of natural aging of MPs on their toxicity has rarely been considered. This study found that visible light irradiation with hydrogen peroxide at environmentally relevant concentration for 90 days significantly altered the physicochemical properties and mitigated the toxicity of polyamide (PA) fragments to infantile zebrafish. The size of PA particles was reduced from ∼8.13 to ∼6.37 µm, and nanoparticles were produced with a maximum yield of 5.03%. The end amino groups were volatilized, and abundant oxygen-containing groups (e.g., hydroxyl and carboxyl) and carbon-centered free radicals were generated, improving the hydrophilicity and colloidal stability of degraded MPs. Compared with pristine PA, the depuration of degraded MPs mediated by multixenobiotics resistance was much quicker, leading to markedly lower bioaccumulation in fish and weaker inhibition on musculoskeletal development. By integrating transcriptomics and transgenic zebrafish [Tg(lyz:EGFP)] tests, differences in macrophages-triggered proinflammatory effects, apoptosis via IL-17 signaling pathway, and antioxidant damages were identified as the underlying mechanisms for the attenuated toxicity of degraded MPs. This work highlights the importance of natural degradation on the toxicity of MPs, which has great implications for risk assessment of MPs.

Incubation behaviours of oviraptorosaur dinosaurs in relation to body size.

Most birds sit on their eggs during incubation, a behaviour that likely evolved among non-avian dinosaurs. Several 'brooding' specimens of smaller species of oviraptorosaurs and troodontids reveal these non-avian theropods sat on their eggs, although little is known of incubation behaviour in larger theropod species. Here we examine egg clutches over a large body size range of oviraptorosaurs in order to understand the potential effect of body size on incubation behaviour. Eggshell porosity indicates that the eggs of all oviraptorosaurs were exposed in the nest, similar to brooding birds. Although all oviraptorosaur clutches consist of radially arranged eggs in a ring configuration, clutch morphology varies in that the central opening is small or absent in the smallest species, becomes significantly larger in larger species, and occupies most of the nest area in giant species. Our results suggest that the smallest oviraptorosaurs probably sat directly on the eggs, whereas with increasing body size more weight was likely carried by the central opening, reducing or eliminating the load on the eggs and still potentially allowing for some contact during incubation in giant species. This adaptation, not seen in birds, appears to remove the body size constraints of incubation behaviour in giant oviraptorosaurs.

Biodegradation of vulcanized rubber by a gut bacterium from plastic-eating mealworms.

The disposal of vulcanized rubber waste is difficult due to the presence of three-dimensional crosslinking network structure. Here, we report that a bacterium Acinetobacter sp. BIT-H3, isolated from the gut of plastic-eating mealworm, can grow on and degrade vulcanized poly(cis-1,4-isoprene) rubber (vPR). Scanning electronic microscopy (SEM) shows that strain BIT-H3 can penetrate into the vPR and produce craters and cracks. The tensile strength and the crosslink density of vPR decreased by 53.2% and 29.3% after ten weeks' incubation, respectively. The results of Horikx analysis, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, and X-ray absorption near-edge structure (XANES) spectroscopy reveal that strain BIT-H3 can break down both sulfide bridges and double bonds of polymeric backbone within vPR. Sulfate and oligo(cis-1,4 isoprene) with terminal aldehyde and keto groups were identified as metabolic products released during vPR degradation. Through genomic and transcriptional analyses, five enzymes of dszA, dszC1, dszC2, Laccase2147, and Peroxidase1232 were found to be responsible for vPR degradation. Based on the chemical structure characterizations and molecular analyses, a vPR biodegradation pathway was proposed for strain BIT-H3. These findings pave a way for exploiting vulcanized rubber-degrading microorganisms from insect gut and contribute to establish a biodegradation method for vulcanized rubber waste disposal.

Photoaging enhanced the adverse effects of polyamide microplastics on the growth, intestinal health, and lipid absorption in developing zebrafish.

The safety of microplastics (MPs) and associated health effects has been one of the major concerns worldwide. However, the role of photoaging toward the risk of MPs in water ecosystems remains inconclusive yet. In this study, the size of polyamide (PA, ∼32.50 µm) MPs was obviously decreased after photoaging in water containing fulvic acid (FA) and humic acid (HA) (∼19.75 and âˆ¼24.30 µm, respectively). Nanoplastics were formed (4.65% and 2.03%, respectively) and hydrophilia and colloidal stability was improved due to the formation of oxygen-containing functional groups. FA-aged PA exhibited higher inhibition on body length and weight of developing zebrafish than HA-aged and pristine PA. Photoaged MPs in intestine were more difficult to be depurated by zebrafish, leading to the disappearance of intestinal folding, shedding of more enterocytes, and emaciation of intestinal microvilli. Dietary lipid digestion in larvae was inhibited by aged PA due to oxidative stress-triggered lipid peroxidation and inhibition of lipase activities and bile acids secretion. Exposure of photoaged MPs down-regulated genes (cd36, dgat1a, dgat2, mttp, etc.) associated with triglyceride resynthesis and transportation, resulting in lipid maladsorption and growth inhibition. Our findings highlight the potential negative effects of environmentally aged MPs on diet digestion and nutrient assimilation in fish.

Photolytic degradation elevated the toxicity of polylactic acid microplastics to developing zebrafish by triggering mitochondrial dysfunction and apoptosis.

Biodegradable plastics (BPs), as alternatives to conventional plastics, are increasingly consumed, but pose potential threats to aquatic ecosystems. In addition, the impact of natural aging on the toxicity of BPs is poorly understood. In this study, the photodegradation of polylactic acid (PLA, a typical BP) microplastics (MPs) under ultraviolet irradiation in water for 90 days was investigated, and the toxicities of virgin and degraded PLA to infantile zebrafish were compared. The results revealed that the size of MPs was reduced from ~25.56 to ~11.22 µm after degradation and nanoparticles were generated with a maximum yield of 7.13%. The formation of abundant oxygen-containing groups (i.e. CË­O and C-O-C) improved the hydrophilia and stability of MPs. Compared with pristine PLA, the efflux and detoxification of degraded PLA mediated by ABC transporters and P450 enzymes were slower, leading to higher bioaccumulation and skeletal development inhibition of zebrafish. Further, oxidative stress-triggered mitochondrial structural damage, depolarization, fission inhibition, and apoptosis were identified as crucial mechanisms underlying the elevated toxicity of PLA after degradation. These findings highlight the importance and necessity of considering natural degradation of BPs and related toxicity, which poses great implications for risk assessment and management of BPs.

Relatively low tooth replacement rate in a sauropod dinosaur from the Early Cretaceous Ruyang Basin of central China.

Tooth replacement rate is an important feature related to feeding mechanics and food choices for dinosaurs. However, only a few data points are available for sauropod dinosaurs, partially due to rarity of relevant fossil material. Four somphospondylan sauropod species have been recovered from the Lower Cretaceous Aptian-Albian Haoling Formation in the Ruyang Basin, Henan Province of central China, but no cranial material has been reported except for a single crown. Here we report the discovery of the rostral portion of a left dentary with replacement teeth in its first five alveoli. Comparative anatomical study shows the partial dentary can be assigned to a member of early diverging somphospondylans. The non-destructive tooth length-based approach to estimating tooth formation time and replacement rate is adopted here. The estimated tooth replacement rate is 76 days, faster than that of Brachiosaurus (83 days) and much lower than typical late diverging lithostrotian titanosaurians (20 days). Thus, this discovery adds an intermediate tooth replacement rate in the evolution of titanosauriform sauropods and supports the idea that evolution of tooth replacement rate is clade-specific. This discovery also provides more information to understand the Ruyang sauropod assemblage, which includes one of the most giant dinosaurs to have walked our Earth (Ruyangosaurus giganteus).

Biodegradation and mineralization of polystyrene by plastic-eating superworms Zophobas atratus.

Polystyrene (PS) is one of the major plastic debris accumulated in environment. Previously, we reported that mealworm (Tenebrio molitor) was capable of degrading and mineralizing Styrofoam (PS foam). This finding arouses our curiosity to explore whether more other insect species have the same capability as mealworms. Here, an insect larva, superworm (Zophobas atratus), was newly proven to be capable of eating, degrading and mineralizing PS. Superworms could live with Styrofoam as sole diet as well as those fed with a normal diet (bran) over a 28-day period. The average consumption rate of Styrofoam for each superworm was estimated at 0.58 mg/d that was 4 times more than that of mealworm. Analyses of frass, using gel permeation chromatography (GPC), solid-state 13C cross-polarization/magic angle spinning nuclear magnetic resonance (CP/MAS NMR) spectroscopy, and thermogravimetric interfaced with Fourier transform infrared (TG-FTIR) spectroscopy, demonstrated that the depolymerization of long-chain PS molecules and the formation of low molecular-weight products occurred in the larval gut. A respirometry test showed that up to 36.7% of the ingested Styrofoam carbon was converted into CO2 during a 16-day test period. The PS-degrading capability of superworm was inhibited by the antibiotic suppression of gut microbiota, indicating that gut microbiota contributed to PS degradation. This new finding extends the PS-degrading insects beyond the species within the Tenebrio genus and indicates that the gut microbiota of superworm would be a novel bioresource for pursuit of plastic-degrading enzymes.

Context-dependency of synthetic minimal promoters in driving gene expression: a case study.

Synthetic promoters are considered ideal candidates in driving robust gene expression. Most of the available synthetic promoters are minimal promoters, for which the upstream sequence of the 5' end of the core region is usually excluded. Although the upstream sequence has been shown to mediate transcription of natural promoters, its impact on synthetic promoters has not been widely studied. Here, a library of chromosomal DNA fragments is randomly fused with the 5' end of the J23119 synthetic promoter, and the transcriptional performance of the promoter is evaluated through ß-galactosidase assay, fluorescence intensity and chemical biosynthesis. Results show that changes in the upstream sequence can induce significant variation in the promoter strength of up to 5.8-fold. The effect is independent of the length of the insertions and the number of potential transcription factor binding sites. Several DNA fragments that are able to enhance the transcription of both the natural and the synthetic promoters are identified. This study indicates that the synthetic minimal promoters are susceptible to the surrounding sequence context. Therefore, the upstream sequence should be treated as an indispensable component in the design and application of synthetic promoters, or as an independent genetic part for the fine-tuning of gene expression.

[Microbial degradation of polyurethane plastics].

Plastic pollution has become a global environmental issue, making it necessary to explore the environmental disposal technology for plastic waste. Recently, we and other researchers have individually found microorganisms or enzymes from nature that can degrade synthetic plastic. These findings indicated that the capability of these microorganisms or enzymes to degrade plastic could be used for the disposal of plastic waste. Polyurethane (PUR) was one of the most used general plastic and its plastic waste occupied 30% of the total volume of different plastic waste. This review tried to provide a comprehensive summary of the researches on microbial degradation of PUR plastic in the past 70 years since its invention, and focused on the PUR-degrading fungi, bacteria, genes or enzymes, degradation products and the corresponding biological disposal technologies. We finally proposed the key scientific challenges on the development of high efficient biological disposal for PUR waste in the perspective researches.