Transcript Abundance Of Genes Research Articles

L-carnitine enhances developmental potential of bovine oocytes matured under high lipid concentrations in vitro

Maternal obesity elevates non-esterified fatty acids (NEFA) follicular concentrations. Bovine cumulus-oocyte complexes (COCs) matured in vitro under high NEFA have altered metabolism and reduced quality. Systemically, obesity promotes altered mitochondrial metabolism linked to L-carnitine insufficiency. We hypothesized that L-carnitine supplementation during IVM of bovine COCs in the presence of high NEFA would lessen the negative effects of exposure to excessive lipids on embryonic development and oxidative stress. COCs were collected from abattoir ovaries and matured in four groups: CON (control), LC (3 mM L-carnitine), HN (high NEFA: 200uM oleic, 150uM palmitic and 75uM stearic acid), and HNLC (HN and LC). Mature oocytes were assayed for aerobic and anaerobic metabolism utilizing oxygen and pH microsensors or fertilized in vitro (D0). Cleavage (D3) and blastocyst (D7, D8) rates were assessed. D3 embryos with ≥ 4 cells were stained for cytosolic and mitochondrial ROS. D8 blastocysts were assayed for gene transcript abundance of metabolic enzymes. Oocyte metabolism was not affected by IVM treatment. D3 formation of embryos with ≥ 4 cells were lower in LC or HN than CON or HNLC; blastocyst rates were greater for CON and lower for HN than LC and HNLC. D3 embryo mitochondrial and cytosolic ROS were reduced in HNLC when compared to other groups. IVM in HN altered blastocyst gene transcript abundance when compared to CON, but not LC or HNLC. In conclusion, supplementation with L-carnitine protects oocytes exposed to high NEFA during IVM and improves their developmental competence, suggesting that high lipid exposure may lead to L-carnitine insufficiency in bovine oocytes.

Alternative splicing regulates autophagy in response to environmental stresses in cucumber (Cucumis sativus)

Autophagy is important for cell survival during stress and nutrient recycle. However, the mechanisms involved in regulating autophagy in cucumber have not been determined. Here, we analyzed the transcript abundance of key autophagy genes in the conjugation pathway including ATG3, ATG4, ATG5, ATG7, ATG8, ATG10, ATG12 and ATG16 in response to nitrogen deficiency, pathogen infection and oxidative stress in Cucumis sativus. ATG8 protein abundance was analyzed using immunoblot analysis. Seven out of 13 autophagy genes studied have splice variances including ATG3, ATG4, ATG5, ATG8b, ATG8c, ATG8e and ATG10. All except ATG5 have changes in transcript variances abundance upon nitrogen starvation, indicating regulation via alternative splicing is pervasive in autophagy genes. This is the first report of alternative splicing in autophagy genes in cucumbers. However, transcription control is also important, especially in ATG4, and those without transcript variances. Moreover, different environment cues regulate autophagy genes through different mechanisms. The ATG8 protein tag was also regulated at the protein level through post-translational modification and blockage of degradation. This work shows that the regulation of autophagy in cucumber is complex and involves many mechanisms. Better understanding of autophagy regulation would thus help breeding plants that perform better even under environmental stresses.

Exogenous triiodothyronine application increases survivorship in larval lake sturgeon (Acipenser fulvescens): Molecular and endocrine regulation of appetite, growth, and stress axes

Lake sturgeon (Acipenser fulvescens) are critically endangered across their native Canadian range and thus conservation aquaculture is practiced to restock historically decimated wild populations. However, as in many aquaculture settings, episodes of mass mortality occur throughout larval development and are often linked to periods of dietary transition. Thyroid hormones play a critical role in larval development and metamorphosis, including the transition from endogenous to exogenous feeding, and thus have been utilized in aquaculture to promote hatch rate, growth, and survivability in multiple fish species. The objective of the present study was to determine whether exogenous application of the biologically active thyroid hormone, triiodothyronine (T3), could improve lake sturgeon growth and survival during early ontogeny in two distinct rearing environments; tumbling jars representing hatchery conditions or egg mats representing more natural conditions. Waterborne application of 3 ppm T3 during fertilization improved growth and survival of lake sturgeon during the first month of life, with greater effects observed in the egg mat environment. Concomitant increases in mRNA abundance of genes involved in appetite regulation (e.g., neuropeptide Y and orexin) and somatic growth (e.g., insulin-like growth factor 1) support accelerated growth of T3 exposed fish through multiple neuroendocrine axes. Following the transition to an exogenous diet, transcript abundance was significantly reduced in the T3 group when compared to controls, perhaps indicating negative feedback regulation. T3 application also caused transient reductions in whole body cortisol content and reduced transcript abundance of genes related to the stress axis later in development; however, no apparent negative consequences were observed as a result. Finally, oral dosing of 3 ppm T3 during an exogenous dietary transition in juvenile lake sturgeon at ∼1.5 months post fertilization significantly increased sturgeon mass, but not survival, indicating that exogenous T3 can contribute to the success of young-of-the-year sturgeon during dietary transitions at multiple life stages. Therefore, these data demonstrate that elevated survivorship and growth associated with exogenous T3 application during fertilization are associated with upregulation of hypothalamus-pituitary-thyroid/interrenal, gut-brain, and somatotropic axes. Overall, exogenous T3 application represents a cost-effective way to positively influence lake sturgeon growth and survival at key dietary transitions during early development. Further, this study adds to a growing body of literature demonstrating beneficial effects of thyroid hormone treatment in fish aquaculture.

Glucuronic acid: not just another brick in the cell wall.

This article is a Commentary on Debra‐Maceluch et al. (2023), 238: 297–312.

Expression profile of key genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin or fetal calf serum

Cumulus cells from cumulus-oocyte complexes (COC) matured in vitro in serum-free medium show high incidence of apoptosis and DNA double-strand breaks (DSB). This study aimed to characterize the transcript expression profile of selected genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin (BSA) or fetal calf serum (FCS). Briefly, bovine cumulus-oocyte complexes were in vitro matured with either, 0.4% BSA or 10% FCS for 3, 6, 12 or 24h. The total RNA of cumulus cells was used for real-time PCR analysis. Transcript abundance of XRCC6, XRCC5, DNAPK, GAAD45B, TP53BP1, RAD50, RAD52, ATM and BRCA2 target genes changed as the IVM proceeded (P<0.05). However, an interaction between protein source (FCS or BSA) and time was not detected (P≥0.05). Cumulus cells from COCs matured with BSA presented higher mRNA expression of two genes compared to FCS group: TP53BP1 at 6h and BRCA1 at 3, 6, 12 and 24h (P<0.05). In summary, our results showed for the first time the expression profile of the key genes involved in DSB repair mechanisms in cumulus cells obtained from bovine COCs matured with FCS or BSA. The higher mRNA expression of BRCA1 and TP53BP1 and lower mRNA expression of TNFAIP6 suggests an increase in apoptosis rate and DNA damage in cumulus cells cultured in BSA-supplemented medium and may explain, at least to some extent, the reduced developmental potential of bovine oocytes matured in serum-free medium.

Metabolomic Investigation of Major Depressive Disorder Identifies a Potentially Causal Association With Polyunsaturated Fatty Acids

BackgroundMetabolic differences have been reported between individuals with and without major depressive disorder (MDD), but their consistency and causal relevance have been unclear. MethodsWe conducted a metabolome-wide association study of MDD with 249 metabolomic measures available in the UK Biobank (n = 29,757). We then applied two-sample bidirectional Mendelian randomization and colocalization analysis to identify potentially causal relationships between each metabolite and MDD. ResultsA total of 191 metabolites tested were significantly associated with MDD (false discovery rate–corrected p < .05), which decreased to 129 after adjustment for likely confounders. Lower abundance of omega-3 fatty acid measures and a higher omega-6 to omega-3 ratio showed potentially causal effects on liability to MDD. There was no evidence of a causal effect of MDD on metabolite levels. Furthermore, genetic signals associated with docosahexaenoic acid colocalized with loci associated with MDD within the fatty acid desaturase gene cluster. Post hoc Mendelian randomization of gene-transcript abundance within the fatty acid desaturase cluster demonstrated a potentially causal association with MDD. In contrast, colocalization analysis did not suggest a single causal variant for both transcript abundance and MDD liability, but rather the likely existence of two variants in linkage disequilibrium with one another. ConclusionsOur findings suggest that decreased docosahexaenoic acid and increased omega-6 to omega-3 fatty acids ratio may be causally related to MDD. These findings provide further support for the causal involvement of fatty acids in MDD.

Comparison of an Antioxidant Source and Antioxidant Plus BCAA on Athletic Performance and Post Exercise Recovery of Horses.

Antioxidant supplementation decreases postexercise oxidative stress but could also decrease muscle protein synthesis. This study compared the effects of three diets: low antioxidant (control, CON), high antioxidant (AO), and branched-chain amino acid high antioxidant (BCAO) supplementation on postexercise protein synthesis and oxidative stress. We hypothesized that supplementing antioxidants with branched-chain amino acids(BCAA) would reduce oxidative stress without hindering muscle protein synthesis. Eighteen mixed-breed polo horses (11 mares and 7 geldings, with age range between 5 and 18 years, were on CON diet for 30 days (from day -45 until day 0) and then were assigned to one of the treatments after the first lactate threshold test (day 0, LT). LT were also conducted on days 15 and 30 of supplemenation. Oxidative stress was assessed by measuring blood glutathione peroxidase, superoxide dismutase, and malondialdehyde concentrations before 2 and 4 hours after each LT. Muscle biopsies were taken before and 4 hours after each LT and analyzed for gene expression of protein synthesis by RTqPCR. Data were analyzed by ANOVA and compared by least-square means. A reduction in oxidative stress occurred over time (P < .05), from day 0 to day 30. An up-regulation in the abundance of muscle protein mRNA transcripts was found for CD36, CPT1, PDK4, MYF5, and MYOG (P < .05) after all lactate threshold tests, without a treatment effect. A treatment-by-exercise effect was observed for MYOD1 (P=.0041). Transcript abundance was upregulated in AO samples post exercise compared to other treatments. MYF6 exhibited a time-by-treatment effect (P=.045), where abundance increased more in AO samples from day 0 to day 15 and 30 compared to other treatments. Transcript abundance for metabolic and myogenic genes was upregulated in post exercise muscle samples with no advantage from supplementation of antioxidants with branched-chain amino acids compared to antioxidants alone.

Identification of gene markers to predict proanthocyanidins biosynthesis in cell culture of Croton lechleri

Croton lechleri Müll. Arg. is a traditional medicinal tree growing in South America, which produces a deep red sap known as Dragon’s Blood Sap (DBS) used for its many therapeutic benefits. DBS bioactivity is mainly due to the high content of proanthocyanidins (PAs), important plant secondary metabolites involved in plant defence, but also utilised as antioxidant for human health. Since the process to directly obtain DBS from the C. lechleri tree is expensive, time-consuming and strongly dependent on the environmental conditions, the use of plant cell cultures for DBS production seems to represent a suitable alternative. The aptitude of plant cell cultures to synthesize specific secondary metabolites can be elicited by different types of abiotic stresses. In this study, the C. lechleri main genes putatively involved in the PAs biosynthetic pathway were identified and their expression was evaluated in both calli in solid medium and cell culture suspensions to identify the most suitable predictive gene markers for PAs biosynthesis. C. lechleri cell cultures were grown and elicited by N-starvation to induce the production of PAs and were then monitored by assessing the expression of previously identified gene markers, by quantifying the content of PAs and measuring few phenotypical parameters. The results show a new perspective on how PA gene transcript abundance could be used to predict the biosynthesis of useful secondary metabolites in plant cell cultures and to speed up the screening of putative elicitors.

Mechanisms Driving the Distribution and Activity of Mineralization and Nitrification in the Reservoir Riparian Zone.

The riparian zone ecosystems have greater energy flow and elemental cycling than adjacent terrestrial and aquatic ecosystems. Mineralization and nitrification are important initiating processes in the nitrogen cycle, but their distribution and activity under different environmental conditions in the riparian zone and the driving mechanisms are still not clear. We investigated the effects of environmental and microbial factors on mineralization and nitrification activities by analyzing the community of alkaline (apr) and neutral (npr) metallopeptidase, ammonia-oxidizing archaea (AOA), and bacteria (AOB) in soils and sediments under different land-use types in the riparian zone of Miyun Reservoir, as well as measuring potential nitrogen mineralization and ammonia oxidation rates (AOR). The results showed that the mineralization and nitrification activities of soils were greater than those of sediments. AOA and AOB dominate the ammonia oxidation activity of soil and sediment, respectively. NH4+ content was a key factor influencing the ecological niche differentiation between AOA and AOB. The high carbon and nitrogen content of the woodland significantly increased mineralization and nitrification activity. Microbial communities were significantly clustered in the woodland. The land-use type, not the flooding condition, determined the distribution of microbial community structure. The diversity of npr was significantly correlated with potential N mineralization rates, while the transcript abundance of AOA was significantly correlated with ammonia oxidation rates. Our study suggests that environmental changes regulate the distribution and activity of mineralization and nitrification processes in the reservoir riparian zone by affecting the transcript abundance, diversity and community structure of the microbial functional genes.

Comparative Transcriptomics Reveal Metabolic Rather than Genetic Control of Divergent Antioxidant Metabolism in the Primary Root Elongation Zone of Water-Stressed Cotton and Maize.

Under water stress, the primary root elongation zones of cotton and maize exhibit both conserved and divergent metabolic responses, including variations in sulfur and antioxidant metabolism. To explore the relative importance of metabolic and genetic controls of these responses for each species, and the extent to which responses are mediated by similar gene expression networks within the framework of ortholog groups, comparative transcriptomics analyses were conducted under conditions of equivalent tissue water stress. Ortholog analysis revealed that 86% of the transcriptome response to water stress was phylogenetically unrelated between cotton and maize. Elevated transcript abundances for genes involved in abscisic acid (ABA) biosynthesis and signaling, as well as key enzymes that enable osmotic adjustment, were conserved between the species. In contrast, antioxidant responses, at least with regard to glutathione metabolism and anti-oxidative enzymes, did not exhibit such a transcript abundance adaptive signature. In particular, previously characterized differential responses of the glutathione and sulfur metabolic pathways between cotton and maize were not evident in the transcriptomic responses. The findings indicate that the antioxidant response in both species results from a metabolic acclimation to water stress, and thus represents an example of water stress-related metabolic plasticity.

Combining time-resolved transcriptomics and proteomics data for Adverse Outcome Pathway refinement in ecotoxicology

Conventional Environmental Risk Assessment (ERA) of pesticide pollution is based on soil concentrations and apical endpoints, such as the reproduction of test organisms, but has traditionally disregarded information along the organismal response cascade leading to an adverse outcome. The Adverse Outcome Pathway (AOP) framework includes response information at any level of biological organization, providing opportunities to use intermediate responses as a predictive read-out for adverse outcomes instead. Transcriptomic and proteomic data can provide thousands of data points on the response to toxic exposure. Combining multiple omics data types is necessary for a comprehensive overview of the response cascade and, therefore, AOP development. However, it is unclear if transcript and protein responses are synchronized in time or time lagged. To understand if analysis of multi-omics data obtained at the same timepoint reveal one synchronized response cascade, we studied time-resolved shifts in gene transcript and protein abundance in the springtail Folsomia candida, a soil ecotoxicological model, after exposure to the neonicotinoid insecticide imidacloprid. We analyzed transcriptome and proteome data every 12 h up to 72 h after onset of exposure. The most pronounced shift in both transcript and protein abundances was observed after 48 h exposure. Moreover, cross-correlation analyses indicate that most genes displayed the highest correlation between transcript and protein abundances without a time-lag. This demonstrates that a combined analysis of transcriptomic and proteomic data from the same time-point can be used for AOP improvement. This data will promote the development of biomarkers for the presence of neonicotinoid insecticides or chemicals with a similar mechanism of action in soils.

Sub-chronic administration of fluoxetine does not alter prey-capture or predator avoidance behaviors in adult South African clawed frogs (Xenopus laevis).

Animals will halt foraging efforts and engage defensive behaviors in response to predator cues. Some researchers have proposed that the switch from appetitive to avoidance behavior resembles anxiety, but most work on this has been performed in a limited number of animal models, primarily zebrafish and rodents. We used adult South African clawed frogs (Xenopus laevis) to determine if the canonical anxiolytic fluoxetine alters predator-induced changes in appetitive and avoidance behavior in a laboratory-based trade-off task that mimics foraging/predator avoidance tradeoffs in the wild. We hypothesized that sub-chronic fluoxetine treatment (20 d) would not affect baseline behavior but would reverse predator-induced changes in food intake, appetitive and avoidance behavior, and the abundance of anxiety related gene transcripts in the optic tectum, a brain area central to ecological decision making in frogs. We found that fluoxetine significantly reduced baseline locomotion compared to vehicle-treated animals. Fluoxetine had no effect on appetitive and avoidance behaviors that were sensitive to predator cues in this assay and did not alter any of the anxiety-related transcripts in the tectum. We conclude that while peripheral sub-chronic administration of fluoxetine significantly reduces locomotion, it does not modify predator-induced changes in approach and avoidance behaviors in this assay. Our findings are not consistent with visual predator cues causing state anxiety in adult frogs.

The effect of age and FSH stimulation on the ovarian follicular response, nuclear maturation, and gene expression of cumulus-oocyte complexes in prepubertal gilts

This study investigated the effects of age and FSH treatment on the ovarian response, follicular fluid (FF) biochemical composition, nuclear maturation, and molecular profile of cumulus-oocytes complexes (COCs) recovered from prepubertal gilts. Thirty-five prepubertal gilts were separated according to age [140 (n = 20) or 160 (n = 15) days], and within each age, the gilts were allotted to receive either 100 mg of FSH [treated; G140+FSH (n = 10) and G160+FSH (n = 7)] or saline solution [control; G140+control (n = 10) and G160+control (n = 8)]. Thus, four experimental groups were included in this study. In the FSH-treated gilts, the percentage of medium follicles increased (P < 0.0001) in the same proportion with which the percentage of small follicles decreased (P < 0.0001). In addition, the glucose concentration in the FF obtained from medium follicles increased (P < 0.05), while that of triglycerides decreased (P < 0.05) in the FSH-treated gilts. The FSH stimulation also improved (P < 0.05) the number of grade I COCs obtained from medium follicles and the meiotic maturation and BCB + rates. FSH treatment only upregulated (P < 0.05) HMGCR expression in immature COCs from prepubertal gilts. The metaphase II and BCB + rates, FF glucose and plasma IGF-1 levels were greater (P < 0.05) in prepubertal gilts at 160 than at 140 days of age. Age had no effect (P > 0.05) on the transcript abundance of the target genes in immature COCs. Hence, oocytes obtained from 140-day-old prepubertal gilts appeared less meiotically competent than those of 160-day-old prepubertal gilts. Our study suggests a possible strategy of using FSH treatment to improve oocyte quantity, quality, and nuclear maturation in 140 and 160-day-old prepubertal gilts.

Postharvest chilling diminishes melon flavor via effects on volatile acetate ester biosynthesis.

In postharvest handling systems, refrigeration can extend fruit shelf life and delay decay via slowing ripening progress; however, it selectively alters the biosynthesis of flavor-associated volatile organic compounds (VOCs), which results in reduced flavor quality. Volatile esters are major contributors to melon fruit flavor. The more esters, the more consumers enjoy the melon fruit. However, the effects of chilling on melon flavor and volatiles associated with consumer liking are yet to be fully understood. In the present study, consumer sensory evaluation showed that chilling changed the perception of melon fruit. Total ester content was lower after chilling, particularly volatile acetate esters (VAEs). Transcriptomic analysis revealed that transcript abundance of multiple flavor-associated genes in fatty acid and amino acid pathways was reduced after chilling. Additionally, expression levels of the transcription factors (TFs), such as NOR, MYB, and AP2/ERF, also were substantially downregulated, which likely altered the transcript levels of ester-associated pathway genes during cold storage. VAE content and expression of some key genes recover after transfer to room temperature. Therefore, chilling-induced changes of VAE profiles were consistent with expression patterns of some pathway genes that encode specific fatty acid- and amino acid-mobilizing enzymes as well as TFs involved in fruit ripening, metabolic regulation, and hormone signaling.

Concentrations of vitamin B12 and folate in maternal serum and fetal fluids, metabolite interrelationships, and hepatic transcript abundance of key folate and methionine cycle genes: the impacts of maternal nutrition during the first 50 d of gestation.

Adequate maternal nutrition is key for proper fetal development and epigenetic programming. One-carbon metabolites (OCM), including vitamin B12, folate, choline, and methionine, play a role in epigenetic mechanisms associated with developmental programming. This study investigated the presence of B12 and folate in maternal serum, allantoic fluid (ALF), and amniotic fluid (AMF), as well as how those concentrations in all three fluids correlate to the concentrations of methionine-folate cycle intermediates in heifers receiving either a control (CON) or restricted (RES) diet for the first 50 d of gestation and fetal hepatic gene expression for methionine-folate cycle enzymes. Angus cross heifers (n = 43) were estrus synchronized, bred via artificial insemination with semen from a single sire, and randomly assigned to one of two nutrition treatments (CON = 20, RES = 23). Heifers were ovariohysterectomized on either day 16 (n = 14), 34 (n = 15), or 50 of gestation (n = 14), where samples of maternal serum (n = 42), ALF (n = 29), and AMF (n = 11) were collected and analyzed for concentrations of folate and B12. Concentrations of B12 and folate in ALF were greater (P < 0.05) in RES compared to CON. For ALF, folate concentrations were also greater (P < 0.01) on day 34 compared to day 50. There was a significant (P = 0.04) nutrition × fluid interaction for B12 concentrations where concentrations were greatest in restricted ALF, intermediate in control ALF, and lowest in CON and RES serum and AMF. Folate concentrations were greatest (P < 0.01) in ALF, intermediate in serum, and lowest in AMF. Additionally, positive correlations (P < 0.05) were found between ALF and AMF folate concentrations and AMF concentrations of methionine, serine, and glycine. Negative correlations (P < 0.05) between AMF folate and serum homocysteine were also observed. Both positive and negative correlations (P < 0.05) depending on the fluid evaluated were found between B12 and methionine, serine, and glycine concentrations. There was a downregulation (P = 0.05) of dihydrofolate reductase and upregulation (P = 0.03) of arginine methyltransferase 7 gene expression in RES fetal liver samples compared with CON fetal liver on day 50. Combined, these data show restricted maternal nutrition results in increased B12 and folate concentrations present in fetal fluids, and increased expression of genes for enzymes within one-carbon metabolism.

Improvement of Seed Germination under Salt Stress via Overexpressing Caffeic Acid O-methyltransferase 1 (SlCOMT1) in Solanum lycopersicum L.

Melatonin (MT) is a phytohormone-like substance and is profoundly involved in modulating nearly all aspects of plant development and acclimation to environmental stressors. However, there remain no studies about the effects of MT on tomato seed germination under salt stress. Here we reported that the overexpression of caffeic acid O-methyltransferase 1 (SlCOMT1) significantly increased both MT content and salt tolerance in the germinated seeds of a transgenic tomato relative to wild type (WT) samples. Physiological investigation showed higher amylase activity in the stressed overexpression seeds than WT, leading to the promoted starch decomposition and enhanced soluble sugar content. The stimulated production of osmolytes and enhanced activities of SOD, POD, and CAT, together with the significant reduction in H2O2 and O2·- accumulation, were revealed in the stressed overexpression seeds relative to WT, largely accounting for their lower membrane lipid peroxidation. qPCR assay showed that, upon salt stress, the transcript abundance of hub genes related to germination (SlCYP707A1, SlABA1, SlGA3ox2 and SlGA2ox4) and stress tolerance (SlCDPK1, SlWRKY33 and SlMAPK1) were distinctly altered in the overexpression samples when compared to WT, providing a molecular basis for MT-mediated improvement of seed salt tolerance. Altogether, our observations shed new insights into biological functions of SlCOMT1 and could expand its utilization in genetic improvement of tomato salt tolerance in future.

1-Methylcyclopropene retards pak choi (Brassica rapasubsp. chinensis) yellowing via BcNAC055-, BcMYB44-, and BcOBF1-mediated regulation of the key chlorophyll degrading geneBcNYC1during storage at 20 °C

AbstractThe objective of this study was to investigate the molecular regulatory mechanism of 1-methylcyclopropene (1-MCP) treatment on leaf yellowing of pak choi during storage at 20 °C. In the present study, compared with the control and 10 μL/L ethylene-treated pak choi, 5.0 μL/L 1-MCP fumigation alleviated the yellowing process of pak choi as proved by the maintenance of higher levels of colour, chlorophyll content, and appearance. 1-MCP treatment decreased chlorophyll breakdown by downregulating the activity of chlorophyll-degrading enzymes (chlorophyll b reductase (CBR) and pheophytinase (PPH)) and the expression of their related genes (BcNYC1 and BcPPH). The application of 1-MCP also inhibited the transcript abundance of ethylene biosynthesis and signal transduction genes (BcACO, BcEIN3, and BcERF) and reduced ethylene production, thus helping to maintain the green colour of pak choi; however, ethylene treatment exerted the opposite function. Meanwhile, three transcription factors (BcNAC055, BcMYB44, and BcOBF1) were successfully cloned. Experiments performed using yeast one-hybrid assay, dual-luciferase reporter system and in vivo bioluminescence imaging assay, confirmed that BcNAC055, BcMYB44, and BcOBF1 directly bound to the BcNYC1 promoter. 1-MCP inhibited the expression levels of BcNAC055, BcMYB44, BcOBF1, and BcNYC1 during storage, while the application of ethylene activated their expression. In conclusion, 1-MCP downregulated the expression of the key chlorophyll degradation gene BcNYC1 by inhibiting the transcript level of its three positive regulators (BcNAC055, BcMYB44, and BcOBF1) and lowered the activity of chlorophyll-degrading enzyme CBR, thereby helping mitigate chlorophyll breakdown to retard leaf yellowing in pak choi. The results of this work extend our understanding of 1-MCP-delayed yellowing in postharvest pak choi.

Roles of plastid-located phosphate transporters in carotenoid accumulation.

Enhanced carotenoid accumulation in plants is crucial for the nutritional and health demands of the human body since these beneficial substances are acquired through dietary intake. Plastids are the major organelles to accumulate carotenoids in plants and it is reported that manipulation of a single plastid phosphate transporter gene enhances carotenoid accumulation. Amongst all phosphate transport proteins including phosphate transporters (PHTs), plastidial phosphate translocators (pPTs), PHOSPHATE1 (PHO1), vacuolar phosphate efflux transporter (VPE), and Sulfate transporter [SULTR]-like phosphorus distribution transporter (SPDT) in plants, plastidic PHTs (PHT2 & PHT4) are found as the only clade that is plastid located, and manipulation of which affects carotenoid accumulation. Manipulation of a single chromoplast PHT (PHT4;2) enhances carotenoid accumulation, whereas manipulation of a single chloroplast PHT has no impact on carotenoid accumulation. The underlying mechanism is mainly attributed to their different effects on plastid orthophosphate (Pi) concentration. PHT4;2 is the only chromoplast Pi efflux transporter, and manipulating this single chromoplast PHT significantly regulates chromoplast Pi concentration. This variation subsequently modulates the carotenoid accumulation by affecting the supply of glyceraldehyde 3-phosphate, a substrate for carotenoid biosynthesis, by modulating the transcript abundances of carotenoid biosynthesis limited enzyme genes, and by regulating chromoplast biogenesis (facilitating carotenoid storage). However, at least five orthophosphate influx PHTs are identified in the chloroplast, and manipulating one of the five does not substantially modulate the chloroplast Pi concentration in a long term due to their functional redundancy. This stable chloroplast Pi concentration upon one chloroplast PHT absence, therefore, is unable to modulate Pi-involved carotenoid accumulation processes and finally does affect carotenoid accumulation in photosynthetic tissues. Despite these advances, several cases including the precise location of plastid PHTs, the phosphate transport direction mediated by these plastid PHTs, the plastid PHTs participating in carotenoid accumulation signal pathway, the potential roles of these plastid PHTs in leaf carotenoid accumulation, and the roles of these plastid PHTs in other secondary metabolites are waiting for further research. The clarification of the above-mentioned cases is beneficial for breeding high-carotenoid accumulation plants (either in photosynthetic or non-photosynthetic edible parts of plants) through the gene engineering of these transporters.

Comparison of N2O-reducing abilities and genome features of two nosZ-containing denitrifying bacteria, Pseudomonas veronii DM15 and Pseudomonas frederiksbergensis DM22.

The study systematically compared the N2O-reducing functional performances and the genomic features of two N2O-reducing isolates, aimed to screen out effective N2O-reducing bacteria with strong environmental adaption, and explore the possible regulation. Two N2O reducers, namely, Pseudomonas veronii DM15 (DM15) and Pseudomonas frederiksbergensis DM22 (DM22), isolated from paddy soil were selected. Their N2O-reducing abilities, and nosZ gene transcript abundance were determined under different temperatures (20°C, 30°C, 40°C) and oxygen concentrations (0%, 10%, 21%), and the whole genomes were sequenced by Illumina sequencing. The results showed that both DM15 and DM22 exhibited the strongest N2O reducing activity at 30°C and under anaerobic conditions. In comparison, DM15 generally exhibited significantly higher N2O reducing abilities and nosZ gene expression than DM22 under all tested conditions. In addition, DM15 possessed obviously higher expression potentials (codon adaptation index (CAI) value) of nos genes than DM22, and the nos cluster of the former contained a transcriptional regulator gene of dnr, while the latter did not. The results indicate that DM15 showed obviously stronger N2O-reducing abilities than DM22 under various conditions, which might be closely associated with its dnr transcriptional regulator, and thus promoting the higher transcriptional activities of nos genes. Although anaerobic conditions were the optimal conditions for N2O reduction in both strains, DM15 still reduced a certain amount of N2O even under aerobic conditions.

A case study: temporal trends of environmental stressors and reproductive health of smallmouth bass (Micropterus dolomieu) from a site in the Potomac River Watershed, Maryland, USA

Decades of poor reproductive success and young-of-the-year survival, combined with adult mortality events, have led to a decline in the smallmouth bass (SMB; Micropterus dolomieu) population in sections of the Potomac River. Previous studies have identified numerous biologic and environmental stressors associated with negative effects on SMB health. To better understand the impact of these stressors, this study was conducted at the confluence of Antietam Creek and the Potomac River from 2013 to 2019 to identify temporal changes associated with SMB reproductive health. Surface water samples were collected and analyzed for over 300 organic contaminants, including pesticides, phytoestrogens, pharmaceuticals, hormones and total estrogenicity (E2Eq). Adult SMB were collected and sampled for multiple endpoints, including gene transcripts associated with reproduction (molecular), histopathology (cellular), and organosomatic indices (tissue). In males, biomarkers of estrogenic endocrine disruption, including testicular oocytes (TO) and plasma vitellogenin (Vtg) were assessed. Numerous agriculture-related contaminants or land use patterns were associated with gene transcript abundance in both male and female SMB. Positive associations between pesticides in the immediate catchment with TO severity and E2Eq with plasma Vtg in males were identified. In males, the prevalence of TO and detectable levels of plasma Vtg, liver vitellogenin transcripts (vtg) and testis vtg were high throughout the study. Peaks of complex mixtures of numerous contaminants occurred during the spring/early summer when spawning and early development occurs and to a lesser extent in fall/winter during recrudescence. Management practices to reduce exposure during these critical and sensitive periods may enhance reproductive health of these economically important sportfishes.