Dioxygenase Gene Research Articles

Genomic and transcriptomic characterization revealed key adaptive mechanisms of Marinobacter hydrocarbonoclasticus NI9 for proliferation and degradation of jet fuel

Marinobacter hydrocarbonoclasticus NI9 has the ability to degrade short-chain alkanes (n-C6 to n-C8), which provides a competitive advantage over other microorganisms. Therefore, we sequenced NI9's genome and analyzed its transcriptome using RNAseq when growing in jet fuel. The genome (3.98 Mb) has at least two alkane monooxygenase genes that are differentially expressed with monooxygenase FH712_RS17105 being highly induced compared to monooxygenase FH712_RS06210. FH712_RS17105 is clustered in an operon with a long-chain-fatty-acid--CoA ligase, an aldehyde dehydrogenase family protein, and rubredoxin. The entire gene cluster was induced along with several dioxygenase and cytochrome P450 genes. Genome analyses further revealed aupA and aupB genes encoding proteins involved in the uptake of micelle-solubilized alkanes. M. hydrocarbonoclasticus NI9 cells achieved fuel tolerance through the activation of efflux pumps and blockage of the efflux pumps with an efflux pump inhibitor significantly reduced NI9 growth in fuel. We identified a fuel-inducible RND-efflux pump, which is an orthologue of the P. aeruginosa MexB. M. hydrocarbonoclasticus NI9 also induced ABC iron transport mechanisms suggesting that there is a high demand for iron during fuel degradation. The results collectively suggest that M. hydrocarbonoclasticus NI9 employ similar adaptive mechanisms to those in P. aeruginosa to proliferate in hydrocarbon-containing environments.

Aerobic Bioaugmentation to Decrease Polychlorinated Biphenyl (PCB) Emissions from Contaminated Sediments to Air.

We conducted experiments to determine whether bioaugmentation with aerobic, polychlorinated biphenyl (PCB)-degrading microorganisms can mitigate polychlorinated biphenyl (PCB) emissions from contaminated sediment to air. Paraburkholderia xenovorans strain LB400 was added to bioreactors containing PCB-contaminated site sediment. PCB mass in both the headspace and aqueous bioreactor compartments was measured using passive samplers over 35 days. Time-series measurements of all 209 PCB congeners revealed a 57% decrease in total PCB mass accumulated in the vapor phase of bioaugmented treatments relative to non-bioaugmented controls, on average. A comparative congener-specific analysis revealed preferential biodegradation of lower-chlorinated PCBs (LC-PCBs) by LB400. Release of the most abundant congener (PCB 4 [2,2′-dichlorobiphenyl]) decreased by over 90%. Simulations with a PCB reactive transport model closely aligned with experimental observations. We also evaluated the effect of the phytogenic biosurfactant, saponin, on PCB bioavailability and biodegradation by LB400. Time-series qPCR measurements of biphenyl dioxygenase (bphA) genes showed that saponin better maintained bphA abundance, compared to the saponin-free treatment. These findings indicate that an active population of bioaugmented, aerobic PCB-degrading microorganisms can effectively lower PCB emissions and may therefore contribute to minimizing PCB inhalation exposure in communities surrounding PCB-contaminated sites.

In-silico Profiling of Deleterious Non Synonymous SNPs of Homogentisate 1, 2 Dioxygenase (HGD) Gene for Early Diagnosis of “Alkaptonuria”

In-silico characterization and molecular modelling of a single amino acid substitution in HGD (Homogentisate 1,2dioxygenase) gene are mainly caused by the deficiency of enzyme Homogentisate 1,2dioxygenase (HGD). An enzyme HGD involved in the catabolism of amino acids such as tyrosine and phenylalanine. The objective of this study was to analyse non-synonymous SNPs from highly deleterious missense mutations which affect the protein function of HGD gene. Based on 3D structure different computational algorithms were performed to identify deleterious SNPs and assess the influence of mutation by using molecular dynamics simulations and molecular docking. Bioinformatics analysis like SIFT, PolyPhen 2.0, I mutant 3.0, PANTHER, SNPs and GO were performed to predict non deleterious ns-SNPs from missense mutations. Energy minimization was done by using GROMACS followed by RMSD calculations and free-energy values under SWISS-PDB viewer and PyMoL respectively. Later, Trajectory analysis was performed using computational tools like SRIDE, CONSURF, SPPIDER, PSIPRED, FLEXPRED for predicting the probably damaged ns-SNPs. Moreover, molecular docking was performed and identified highly deleterious probably damaging mutation. By operating 10 bioinformatics analysis, we obtained 5 mutations R53W, L61P, G121R, G361R and L430H which have an adverse effect on HGD gene. The results of the ConSurf analysis showed that all of these ns-SNPs are in the highly conserved positions and influence the structure of native proteins. L61P mutation had more effect on protein structure. Later, for future studies these mutations assists to develop an effective drug for the associated disease Alkaptonuria.

Sandaracinobacteroides saxicola sp. nov., a Zeaxanthin-Producing and Halo-Sensitive Bacterium Isolated from Fully Weathered Granitic Soil, and the Diversity of Its ARHDs

A yellow, Gram-stain-negative, aerobic, non-spore-forming, motile, and rod-shaped bacterial strain designated M6T was isolated from fully weathered granitic soil. The strain showing the highest 16S rRNA gene sequence similarity to M6T was Sandaracinobacteroides hominis SZY PN-1T (96.3%), the only species in the genus Sandaracinobacteroides. The average nucleotide identity and digital DNA-DNA hybridization value between these two strains were 72.6% and 18.0% respectively. Growth was inhibited by NaCl (≥0.1% (w/v)). Strain M6T contained C18:1ω7c (33.8%), C14:0 2-OH (16.6%), summed feature 3 (15.8%), and C16:0 (12.6%) as the major fatty acids. The polar lipids profile consisted of phosphatidylglycerol, phosphatidylethanolamine, an unidentified glycolipid, four unidentified phospholipids, and four unidentified lipids. The genome of strain M6T was 3.4 Mb with 67.7% GC content. Further genomic analysis revealed a biosynthetic gene cluster for zeaxanthin, the production of which was verified by a high-resolution mass spectrum. The existence of multiple genes for aromatic ring-hydroxylating dioxygenases implies the potential ability for organic pollution controlling. The morphological, physiological, chemotaxonomic, and phylogenetic analysis clearly distinguished this strain from its phylogenetic neighbors, thus strain M6T represents a novel species of the genus Sandaracinobacteroides, for which the name Sandaracinobacteroides saxicola sp. nov. is proposed. The type of strain is M6T (=CGMCC 1.19164T=NBRC 115420T).

Risk variants of obesity associated genes demonstrate BMI raising effect in a large cohort.

Obesity is highly polygenic disease where several genetic variants have been reportedly associated with obesity in different ethnicities of the world. In the current study, we identified the obesity risk or protective association and BMI raising effect of the minor allele of adiponectin, C1Q and collagen domain containing (ADIPOQ), cholesteryl ester transfer protein (CEPT), FTO alpha-ketoglutarate dependent dioxygenase (FTO), leptin (LEP), and leptin receptor (LEPR) genes in a large cohort stratified into four BMI-based body weight categories i.e., normal weight, lean, over-weight, and obese. Based on selected candidate genetic markers, the genotyping of all study subjects was performed by PCR assays, and genotypes and allele frequencies were calculated. The minor allele frequencies (MAFs) of all genetic markers were computed for total and BMI-based body weight categories and compared with MAFs of global and South Asian (SAS) populations. Genetic associations of variants with obesity risk were calculated and BMI raising effect per copy of the minor allele were estimated. The genetic variants with higher MAFs in obese BMI group were; rs2241766 (G = 0.43), rs17817449 (G = 0.54), rs9939609 (A = 0.51), rs1421085 (C = 0.53), rs1558902 (A = 0.63), and rs1137101 (G = 0.64) respectively. All these variants were significantly associated with obesity (OR = 1.03-4.42) and showed a high BMI raising effect (β = 0.239-0.31 Kg/m2) per copy of the risk allele. In contrast, the MAFs of three variants were higher in lean-normal BMI groups; rs3764261 A = 0.38, rs9941349 T = 0.43, and rs7799039 G = 0.40-0.43). These variants showed obesity protective associations (OR = 0.68-0.76), and a BMI lowering effect per copy of the protective allele (β = -0.103-0.155 Kg/m2). The rs3764261 variant also showed significant and positive association with lean body mass (OR = 2.38, CI = 1.30-4.34). Overall, we report six genetic variants of ADIPOQ, FTO and LEPR genes as obesity-risk markers and a CETP gene variant as lean mass/obesity protective marker in studied Pakistani cohort.

Paclobutrazol Ameliorates Low-Light-Induced Damage by Improving Photosynthesis, Antioxidant Defense System, and Regulating Hormone Levels in Tall Fescue.

Paclobutrazol (PBZ) is a plant-growth regulator (PGR) in the triazole family that enhances plant tolerance to environmental stresses. Low-light (LL) intensity is a critical factor adversely affecting the growth of tall fescue (Festuca arundinacea Schreb.). Therefore, in this study, tall fescue seedlings were treated with PBZ under control and LL conditions to investigate the effects of PBZ on enhancing LL stress resistance by regulating the growth, photosynthesis, oxidative defense, and hormone levels. Our results reveal that LL stress reduced the total biomass, chlorophyll (Chl) content, photosynthetic capacity, and photochemical efficiency of photosystem II (PSII) but increased the membrane lipid peroxidation level and reactive oxygen species (ROS) generation. However, the application of PBZ increased the photosynthetic pigment contents, net photosynthetic rate (Pn), maximum quantum yield of PSII photochemistry (Fv/Fm), ribulose-1,5-bisphosphate carboxylase (RuBisCO) activity, and starch content. In addition, PBZ treatment activated the antioxidant enzyme activities, antioxidants contents, ascorbate acid-glutathione (AsA-GSH) cycle, and related gene expression, lessening the ROS burst (H2O2 and O2∙−). However, the gibberellic acid (GA) anabolism was remarkably decreased by PBZ treatment under LL stress, downregulating the transcript levels of kaurene oxidase (KO), kaurenoic acid oxidase (KAO), and GA 20-oxidases (GA20ox). At the same time, PBZ treatment up-regulated 9-cis-epoxycarotenoid dioxygenase (NCED) gene expression, significantly increasing the endogenous abscisic acid (ABA) concentration under LL stress. Thus, our study revealed that PBZ improves the antioxidation and photosynthetic capacity, meanwhile increasing the ABA concentration and decreasing GA concentration, which ultimately enhances the LL stress tolerance in tall fescue.

Efficient degradation of hydroquinone by a metabolically engineered Pseudarthrobacter sulfonivorans strain.

Pseudarthrobacter sulfonivorans strain Ar51 can degrade crude oil and multi-substituted benzene compounds efficiently at low temperatures. However, it cannot degrade hydroquinone, which is a key intermediate in the degradation of several other compounds of environmental importance, such as 4-nitrophenol, g-hexachlorocyclohexane, 4-hydroxyacetophenone and 4-aminophenol. Here we co-expressed the two subunits of hydroquinone dioxygenase from Sphingomonas sp. strain TTNP3 with different promoters in the strain Ar51. The strain with 2 hdnO promoters exhibited the strongest hydroquinone catabolic activity. However, in the absence of antibiotic selection this ability to degrade hydroquinone was lost due to plasmid instability. Consequently, we constructed a hisD knockout strain, which was unable to synthesise histidine. By introducing the hisD gene onto the plasmid, the ability to degrade hydroquinone in the absence of antibiotic selection was stabilised. In addition, to make the strain more stable for industrial applications, we knocked out the recA gene and integrated the hydroquinone dioxygenase genes at this chromosomal locus. This strain exhibited the strongest activity in catabolizing hydroquinone, up to 470mg/L in 16h without antibiotic selection. In addition, this activity was shown to be stable when the strain has cultured in medium without antibiotic selection after 20 passages.

Characterization and Expression Analysis of Extradiol and Intradiol Dioxygenase of Phenol-Degrading Haloalkaliphilic Bacterial Isolates

Haloalkophilic bacteria have a potential advantage as a bioremediation organism of high oil-polluted and industrial wastewater. In the current study, Haloalkaliphilic isolates were obtained from Hamralake, Wadi EL-Natrun, Egypt. The phenotype script, biochemical characters, and sequence analysis of bacterial-16S rRNA were used to identify the bacterial isolates; Halomonas HA1 and Marinobacter HA2. These strains required high concentrations of NaCl to ensure bacterial growth, especially Halomonas HA1 strain. Notably, both isolates can degrade phenol at optimal pH values, between 8 and 9, with the ability to grow in pH levels up to 11, like what was seen in the Halomonas HA1 strain. Moreover, both isolates represent two different mechanistic pathways for phenol degradation. Halomonas HA1 exploits the 1,2 phenol meta-cleavage pathway, while Marinobacter HA2 uses the 2,3 ortho-cleavage pathway as indicated by universal primers for 1,2 and 2,3 CTD genes. Interestingly, Marinobacter HA2 isolate eliminated the added phenol within an incubation period of 72 h, while the Halomonas HA1 isolate invested 96 h in degrading 84% of the same amount of phenol. Phylogenetic analysis of these 1,2 CTD (catechol dioxygenase) sequences clearly showed an evolutionary relationship between 1,2 dioxygenases of both Halomonadaceae and Pseudomonadaceae. In comparison, 2,3 CTD of Marinobacter HA2 shared the main domains of the closely related species. Furthermore, semi-quantitative RT-PCR analysis proved the constitutive expression pattern of both dioxygenase genes. These findings provide new isolates of Halomonas sp. and Marinobacter sp. that can degrade phenol at high salt and pH conditions via two independent mechanisms.

Investigation of proteins' interaction network and the expression pattern of genes involved in the ABA biogenesis and antioxidant system under methanol spray in drought-stressed rapeseed.

Drought is one of the most critical abiotic stresses, which significantly impair rapeseed (Brassica napus L.) productivity. Several factors can regulate the stress response, including changes in gene expression in biological pathways, extensive protein interaction networks, and post-translational regulatory factors like microRNAs. External factors can also affect the intensity of the stress response. Therefore, this study investigated protein-protein interactions of some essential genes involved in abscisic acid (ABA) production, antioxidant system, and Krebs cycle. The expression of phyton synthase (PSY), 9-cis-epoxycarotenoid dioxygenase (NCED3), aldehyde oxidase (AAO3), thioredoxin reductase (NTRC), and glutathione reductase (GR) genes in two rapeseed genotypes, i.e., Hyola308 (drought-sensitive) and SLM046 (drought-tolerant) were evaluated using qRT-PCR technique under 72h of drought stress and methanol foliar application. In the SLM046 (tolerant) genotype, the expression levels of PYS, NCED, AAO3, and GR genes were increased after 8h of foliar application. The expression level of the NTR gene was increased 8 and 24h after stress and methanol treatment. In the Hyola308 genotype, PYS, AAO3, NTR, and GR genes' expression level was increased 8h after methanol foliar application, and the NCED gene was increased 24h after stress with methanol treatment. In general, methanol foliar application increased the expression levels of several genes. Particularly, the gene expression was considerably higher in the SLM046 genotype than in Hyola308. Bioinformatics prediction of microRNAs targeting PSY, NCED, GR, NTRC, and AAO3 genes was performed, and 38, 38, 13, 11, and 11 microRNAs were predicted for these genes, respectively. The study of effective microRNAs showed that sometimes more than one type of microRNA could affect the desired gene, and in some cases, a conserved family of microRNAs caused the main effect on gene expression. Overall, our results lay the foundation for functional characterization of these genes or gene-miRNA modules in regulating drought stress tolerance in rapeseed.

Identification of the carotenoid cleavage dioxygenase genes and functional analysis reveal DoCCD1 is potentially involved in beta-ionone formation in Dendrobium officinale.

The carotenoids are the most widely distributed secondary metabolites in plants and can be degraded by carotenoid cleavage dioxygenase (CCD) to form apocarotenoids including an important C13 compound beta-ionone. Volatile beta-ionone can confer the violet and woody fragrance to plant essential oils, flowers, fruits, and vegetables, which therefore has been used in various industries. Dendrobium officinale is a traditional medicinal plant. However, there was limited information on the key enzymes involved in the biosynthesis of beta-ionone in D. officinale. In the present study, beta-ionone was detected in stems and leaves of D. officinale and genome-wide identification and expression profiles of CCD genes were subsequently carried out. There were nine DoCCD members in D. officinale. According to the phylogenetic relationship, DoCCD proteins were classified into six subfamilies including CCD1, CCD4, CCD7, CCD8, nine-cis-epoxycarotenoid dioxygenase (NCED) and zaxinone synthase (ZAS). DoCCD genes showed distinctive expression profiles and DoCCD1 gene was abundantly expressed in eight tissues. Induced expression of DoCCD1 gene resulted in discoloration of Escerichia coli strains that can accumulate carotenoids. Analysis of Gas Chromatography/Mass Spectrometer showed that DoCCD1 enzyme can cleave lycopene to produce 6-methyl-5-hepten-2-one and pseudoionone and also catalyze beta-carotene to form beta-ionone. Expression of DoCCD1 gene in Nicotiana benthamiana leaf resulted in production of abundant beta-ionone. Overall, the present study first provides valuable information on the CCD gene family in D. officinale, function of DoCCD1 gene as well as production of beta-ionone through genetic modification.

Metabolic Profiling and Transcriptional Analysis of Carotenoid Accumulation in a Red-Fleshed Mutant of Pummelo (Citrus grandis).

Citrus grandis ‘Tomentosa’, commonly known as ‘Huajuhong’ pummelo (HJH), is used in traditional Chinese medicine and can moisten the lungs, resolve phlegm, and relieve coughs. A spontaneous bud mutant, named R-HJH, had a visually attractive phenotype with red albedo tissue and red juice sacs. In this study, the content and composition of carotenoids were investigated and compared between R-HJH and wild-type HJH using HPLC–MS analysis. The total carotenoids in the albedo tissue and juice sacs of R-HJH were 4.03- and 2.89-fold greater than those in HJH, respectively. The massive accumulation of carotenoids, including lycopene, β-carotene and phytoene, led to the attractive red color of R-HJH. However, the contents of flavones, coumarins and most volatile components (mainly D-limonene and γ-terpinene) were clearly reduced in R-HJH compared with wild-type HJH. To identify the molecular basis of carotenoid accumulation in R-HJH, RNA-Seq transcriptome sequencing was performed. Among 3948 differentially expressed genes (DEGs), the increased upstream synthesis genes (phytoene synthase gene, PSY) and decreased downstream genes (β-carotene hydroxylase gene, CHYB and carotenoid cleavage dioxygenase gene, CCD7) might be the key factors that account for the high level of carotenoids in R-HJH. These results will be beneficial for determining the molecular mechanism of carotenoid accumulation and metabolism in pummelo.

Co-option of a carotenoid cleavage dioxygenase gene (CCD4a) into the floral symmetry gene regulatory network contributes to the polymorphic floral shape-color combinations in Chrysanthemum sensu lato.

Morphological novelties, including formation of trait combinations, may result from de novo gene origination and/or co-option of existing genes into other developmental contexts. A variety of shape-color combinations of capitular florets occur in Chrysanthemum and its allies. We hypothesized that co-option of a carotenoid cleavage dioxygenase gene into the floral symmetry gene network would generate a white zygomorphic ray floret. We tested this hypothesis in an evolutionary context using species in Chrysanthemum sensu lato, a monophyletic group with diverse floral shape-color combinations, based on morphological investigation, interspecific crossing, molecular interaction and transgenic experiments. Our results showed that white color was significantly associated with floret zygomorphy. Specific expression of the carotenoid cleavage dioxygenase gene CCD4a in marginal florets resulted in white color. Crossing experiments between Chrysanthemum lavandulifolium and Ajania pacifica indicated that expression of CCD4a is trans-regulated. The floral symmetry regulator CYC2g can activate expression of CCD4a with a dependence on TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING (TCP) binding element 8 on the CCD4a promoter. Based on all experimental findings, we propose that gene co-option of carotenoid degradation into floral symmetry regulation, and the subsequent dysfunction or loss of either CYC2g or CCD4a, may have led to evolution of capitular shape-color patterning in Chrysanthemum sensu lato.

Mode of Action of a Novel Synthetic Auxin Herbicide Halauxifen-Methyl

Halauxifen-methyl is a new auxin herbicide developed by Corteva Agriscience (Wilmington, DE, USA). It has been suggested that ABF5 may be the target of halauxifen-methyl, as AFB5 mutants of Arabidopsis thaliana are resistant to halauxifen-methyl, which preferentially binds to AFB5. However, the mode of action of halauxifen-methyl has not yet been reported. Therefore, the aim of the present study was to reveal the mode of action of halauxifen-methyl by exploring its influence on indole-3-acetic acid (IAA) homeostasis and the biosynthesis of ethylene and Abscisic Acid (ABA) in Galium aparine. The results showed that halauxifen-methyl could disrupt the homeostasis of IAA and stimulate the overproduction of ethylene and ABA by inducing the overexpression of the 1-aminocyclopropane-1-carboxylate synthase (ACS) and 9-cis-epoxycarotenoid dioxygenase (NCED) genes involved in ethylene and ABA biosynthesis, finally leading to senescence and plant death.

Targeted metabolomics suggests a probable role of the FTO gene in the kynurenine pathway in prediabetes.

BackgroundGenome-wide association studies have identified the alpha-ketoglutarate dependent dioxygenase gene (FTO) as the first susceptibility gene of obesity. In the present study, we utilized targeted metabolomics in an attempt to further elucidate mechanisms underlying the action of the FTO gene.MethodsThis study was part of a health survey of employees of the Electricity Generating Authority of Thailand (n = 79, 10 female and 69 male). Targeted metabolomics was performed by liquid chromatography–mass spectrometry using Biocrates AbsoluteIDQ-p180 kit. Genotyping of FTO rs9939609 was performed by real-time PCR (TaqMan™ MGB probes).ResultsUsing OPLS-DA variable importance in projection (VIP), tryptophan was found to be among the metabolites with the 10 highest VIP scores. Pearson’s correlation analysis showed that kynurenine and tryptophan were positively correlated only in subjects with the rs9939609 A allele (n = 32, r = 0.56, p < 0.001) and the correlation coefficients were significantly higher in subjects having the A allele than in those without the A allele (p < 0.05). Moreover, the kynurenine/tryptophan ratio was significantly associated with the presence of the A allele, independently of body mass index and sex.ConclusionsThe FTO gene is likely to influences the conversion of tryptophan to kynurenine.

Electroporation-mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes.

This protocol describes a fast and effective method for isolating primary mouse hepatocytes followed by electroporation-mediated delivery of CRISPR-Cas9 as ribonucleoproteins (RNPs) and mRNA. Primary mouse hepatocytes were isolated using a three-step retrograde perfusion method resulting in high yields of up to 50 × 106 cells per liver and cell viability of >85%. This protocol provides detailed instructions for plating, staining, and culturing hepatocytes. The results indicate that electroporation provides a high transfection efficiency of 89%, as measured by the percentage of green fluorescent protein (GFP)-positive cells and modest cell viability of >35% in mouse hepatocytes. To demonstrate the utility of this approach, CRISPR-Cas9 targeting the hydroxyphenylpyruvate dioxygenase gene was electroporated into primary mouse hepatocytes as proof-of-principle gene editing to disrupt a therapeutic gene related to an inherited metabolic disease (IMD) of the liver. A higher on-target edit of 78% was observed for RNPs compared to 47% editing efficiency with mRNA. The functionality of hepatocytes was evaluated in vitro using an albumin assay that indicated that delivering CRISPR-Cas9 as RNPs and mRNA results in comparable cell viability in primary mouse hepatocytes. A promising application for this protocol is the generation of mouse models for human genetic diseases affecting the liver.

Biochar enhanced polycyclic aromatic hydrocarbons degradation in soil planted with ryegrass: Bacterial community and degradation gene expression mechanisms

Biochar and ryegrass have been used in the remediation of polycyclic aromatic hydrocarbons (PAHs)-contaminated soils; however, the effects of different biochar application levels on the dissipation of PAHs, bacterial communities, and PAH-ring hydroxylating dioxygenase (PAH-RHDα) genes in rhizosphere soil remain unclear. In this study, enzyme activity tests, real-time quantitative polymerase chain reaction (PCR), and high-throughput sequencing were performed to investigate the effects of different proportions of rape straw biochar (1%, 2%, and 4% (w/w)) on the degradation of PAHs, as well as the associated changes in the soil bacterial community and PAH-RHDα gene expression. The results revealed that biochar enhanced the rhizoremediation of PAH-contaminated soil and that 2% biochar-treated rhizosphere soil was the most effective in removing PAHs. Furthermore, urease activity, abundance and activity of total bacteria, and PAH-degrading bacteria were enhanced in soil that was amended with biochar and ryegrass. Additionally, the activity of 16S rDNA and PAH-RHDα gram-negative (GN) genes increased with increasing biochar dosage and had a positive correlation with the removal of PAHs. Biochar changed the rhizosphere soil bacterial composition and α-diversity, and promoted the growth of Pseudomonas and Zeaxanthinibacter. In addition, the relative abundance of Pseudomonas was positively correlated with PAH removal. These findings imply that rape straw biochar can enhance the rhizoremediation of PAH-contaminated soil by changing soil bacterial communities and stimulating the expression of PAH-RHDα GN genes. The 2% of rape straw biochar combined with ryegrass would be an effective method to remediate the PAH-contaminated soil.

Complete Genome Sequence Resource for Xanthomonas translucens pv. undulosa MAI5034, a Wheat Pathogen from Uruguay.

Complete Genome Sequence Resource for Xanthomonas translucens pv. undulosa MAI5034, a Wheat Pathogen from Uruguay.

Metabolism of Carotenoids and β-Ionone Are Mediated by Carotenogenic Genes and PpCCD4 Under Ultraviolet B Irradiation and During Fruit Ripening.

Carotenoids are essential pigments widely distributed in tissues and organs of higher plants, contributing to color, photosynthesis, photoprotection, nutrition, and flavor in plants. White- or yellow-fleshed colors in peach were determined by expression of carotenoids cleavage dioxygenase (PpCCD) genes, catalyzing the degradation of carotenoids. The cracked volatile apocarotenoids are the main contributors to peach aroma and flavor with low sensory threshold concentration. However, the detailed regulatory roles of carotenoids metabolism genes remained unclear under UV-B irradiation. In our study, metabolic balance between carotenoids and apocarotenoids was regulated by the expression of phytoene synthase (PSY), β-cyclase (LCY-B), ε-cyclase (LCY-E), and PpCCD4 under UV-B irradiation. The transcript levels of PpPSY, PpLCY-B, PpLCY-E, and PpCHY-B were elevated 2- to 10-fold compared with control, corresponding to a nearly 30% increase of carotenoids content after 6 h UV-B irradiation. Interestingly, the total carotenoids content decreased by nearly 60% after 48 h of storage, while UV-B delayed the decline of lutein and β-carotene. The transcript level of PpLCY-E increased 17.83-fold compared to control, partially slowing the decline rate of lutein under UV-B irradiation. In addition, the transcript level of PpCCD4 decreased to 30% of control after 48 h UV-B irradiation, in accordance with the dramatic reduction of apocarotenoid volatiles and the delayed decrease of β-carotene. Besides, β-ionone content was elevated by ethylene treatment, and accumulation dramatically accelerated at full ripeness. Taken together, UV-B radiation mediated the metabolic balance of carotenoid biosynthesis and catabolism by controlling the transcript levels of PpPSY, PpLCY-B, PpLCY-E, and PpCCD4 in peach, and the transcript level of PpCCD4 showed a positive relationship with the accumulation of β-ionone during the ripening process. However, the detailed catalytic activity of PpCCD4 with various carotenoid substrates needs to be studied further, and the key transcript factors involved in the regulation of metabolism between carotenoids and apocarotenoids need to be clarified.

L‐DOPA dioxygenases from diverse natural product pathways

L‐DOPA dioxygenase is part of a mini‐pathway to the synthon 3‐vinyl‐2,3‐pyrroline‐5‐carboxylic acid (VPCA) that is elaborated and embedded within the final product structures of lincomycin, anthramycin, sibiromycin, tomaymycin and hormaomycin. Using the VPCA mini‐pathway as a starting point, we searched sequence space to identify novel natural product pathways containing a VPCA synthon. From among these novel natural product pathways, representative L‐DOPA dioxygenase gene products from Streptomyces hygroscopicus subsp. jinggangensis and Nocardia arthriditis were studied as pure proteins for their stability and activity on L‐DOPA and related catechols in steady‐state assays for catechol cleavage. These results were analyzed in comparison to characterized L‐DOPA dioxygenases from Streptomyces lincolnensis and Streptomyces sclerotialus.

Ectopic expression of a rice triketone dioxygenase gene confers mesotrione tolerance in soybean.

BACKGROUNDHerbicide‐resistant weeds pose a challenge to agriculture and food production. New herbicide tolerance traits in crops will provide farmers with more options to effectively manage weeds. Mesotrione, a selective pre‐ and post‐emergent triketone herbicide used in corn production, controls broadleaf and some annual grass weeds via hydroxyphenylpyruvate dioxygenase (HPPD) inhibition. Recently, the rice HIS1 gene, responsible for native tolerance to the selective triketone herbicide benzobicyclon, was identified. Expression of HIS1 also confers a modest level of mesotrione resistance in rice. Here we report the use of the HIS1 gene to develop a mesotrione tolerance trait in soybean.RESULTSConventional soybean is highly sensitive to mesotrione. Ectopic expression of a codon‐optimized version of the rice HIS1 gene (TDO) in soybean confers a commercial level of mesotrione tolerance. In TDO transgenic soybean plants, mesotrione is rapidly and locally oxidized into noninhibitory metabolites in leaf tissues directly exposed to the herbicide. These metabolites are further converted into compounds similar to known classes of plant secondary metabolites. This rapid metabolism prevents movement of mesotrione from treated leaves into vulnerable emerging leaves. Minimizing the accumulation of the herbicide in vulnerable emerging leaves protects the function of HPPD and carotenoid biosynthesis more generally while providing tolerance to mesotrione.CONCLUSIONSMesotrione has a favorable environmental and toxicological profile. The TDO‐mediated soybean mesotrione tolerance trait described here provides farmers with a new option to effectively manage difficult‐to‐control weeds using familiar herbicide chemistry. This trait can also be adapted to other mesotrione‐sensitive crops (e.g. cotton) for effective weed management. © 2022 Bayer Crop Science. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.