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Abstract Background Thraustochytrids have gained attention as a potential source for the production of docosahexaenoic acid (DHA), where DHA is predominantly stored in the form of triacylglycerol (TAG). The TAG biosynthesis pathways, including the acyl-CoA-dependent Kennedy pathway and the acyl-CoA-independent pathway, have been predicted in thraustochytrids, while the specific details regarding their roles are currently uncertain. Results Phospholipid:diacylglycerol acyltransferase (PDAT) plays a key role in the acyl-CoA-independent pathway by transferring acyl-group from phospholipids (PL) to diacylglycerol (DAG) to from TAG. In thraustochytrid Aurantiochytrium sp. SD116, an active AuPDAT was confirmed by heterologous expression in a TAG-deficient yeast strain H1246. Analysis of AuPDAT function in vivo revealed that deletion of AuPDAT led to slow growth and a significant decrease in cell number, but improved PL content in the single cell during the cell growth and lipid accumulation phases. Interestingly, deletion of AuPDAT did not affect total lipid and TAG content, but both were significantly increased within a single cell. Moreover, overexpression of AuPDAT also resulted in a decrease in cell number, while the total lipid and cell diameter of a single cell were markedly increased. Altogether, both up-regulation and down-regulation of AuPDAT expression affected the cell number, which further associated with the total lipid and TAG content in a single cell. Conclusions Our study demonstrates that AuPDAT-mediated pathway play a minor role in TAG synthesis, and that the function of AuPDAT may be involved in regulating PL homeostasis by converting PL to TAG in a controlled manner. These findings expand our understanding of lipid biosynthesis in Aurantiochytrium sp. and open new avenues for developing “customized cell factory” for lipid production.

Mucor circinelloides has the ability to use a wide range of carbon sources to accumulate lipids. The different carbon sources have significantly influenced the total lipids through different metabolic pathways in oleaginous organisms. In the present study, the effects of exogenous oil (soybean oil) at different concentrations as an alternative carbon source on its fungal growth and lipid accumulation were investigated without changing fermentation culture conditions and the original C/N ratio, while using 100% glucose as a control. Total lipids and their fatty acid composition were determined by gas chromatography (GC). Moreover, the expression levels of desaturases genes involved in fatty acid biosynthesis were investigated in the studied strains using real-time fluorescence quantitative PCR. The results showed that a concentration of 35 g/L of glucose mixed with 24.3 g/L of soybean oil were the optimal carbon sources with constant carbon to nitrogen ratio. The biomass, lipid content and lipid yield of M. circinelloides grown in this medium increased rapidly in the early stage of fermentation, and the growth trend gradually slowed down in the late stage of fermentation, decreasing slightly after 72 h of fermentation. It was found that the highest biomass, total lipids, and lipid yields were significantly increased at 19.60 g/L, 42.62%, and 8.35 g/L (33.15%, 23.8%, and 65.10%), respectively. In contrast, the control group (those cultured on glucose as a single carbon source) was only 14.72 g/L, 34.41%, and 5.06 g/L, respectively. The critical step was in which α-linolenic acid (ALA) was stored from the exogenous oil in fungal cells supplying enough soybean oil with mixed substrates for fatty acids synthesis. The overexpression of the △12-desaturase gene in WJ11 significantly (P

Soil microorganisms play an important role in maintaining soil quality and function, although the response of soil microbial biodiversity to heavy metals has been extensively investigated, the microbe-microbe associations under the influence of both native plant species and extremely high heavy metal contamination are not well documented. We examined the diversity and composition of microbial communities and the physicochemical properties in the rhizosphere of three native plant species, Carex breviculmis, Buddleja davidii, and Artemisia annua growing on and around a Pb-Zn waste heap with a nearly 100-year history of natural recovery. Both plant species and heavy metals influence soil microbial diversity and composition. C. breviculmis and A. annua showed a prominent advantage in increasing rhizosphere microbial diversity and richness as well as network complexity compared with plant Buddleja davidii at severely contaminated soil, which was mainly related to the accumulation of soil nutrients such as soil organic carbon (SOC), total nitrogen ammonium nitrogen and nitrate nitrogen rather than a reduction in heavy metal concentrations. Moreover, the heavy metal concentration and soil nutrient levels significantly affected the microbial groups affiliated with Proteobacteria, Chloroflexi, Ascomycota, and Basidiomycota, in which those affiliated with Chloroflexi and Ascomycota were positively associated with heavy metals. Soil microbial network on the Pb-Zn waste heap exhibited higher average degree and a higher proportion of positive links than those around the waste heap, and thus soil microbial structure became more complexity and unstable with increasing heavy metal pollution.

The predominant use of junk food in our societies is continuously held responsible for the obese body physiques and overweight among the kids and adolescents. The current supportive environments where organic foods are limited, and new processed foods have been brought to the market with more variant tastes and acceptability for the kids and adolescents that have diverged their eating patterns. It has significantly contributed to the health issues and growth discrepancies of the users. However, the awareness of the food contents is an important milestone for understanding the risks associated with the usage of junk foods. A quantitative approach has been used in this study to measure the effect of perceived severity, vulnerability and fear on the junk food eating behaviors and ultimately on the obesity. The moderating role of product knowledge hiding has also been measured on the relationship of junk food eating and obesity. Structural equation modeling is used using the software Smart-PLS for measuring the hypothesis with a sample size of 228 selected through purposive sampling. The sample consisted of kids and adolescents who were reached on purpose for data collection. The current study has explored the role of perceived severity, vulnerability and the fear of using junk foods which have been found as a negative effect on junk food eating behavior which is positively associated with obesity among the kids and adolescents. The result of study shows that perceived threat has a negative effect on the junk food eating behavior of the adolescents. However, the positive relationship of junk food eating behavior with obesity can be decreased if the information about the products is not hidden. This study will be useful for making the consumers aware of the product knowledge hiding of the junk food usage. Moreover, it will help the users in creating understanding of risks allied with the use of junk food which may be addressed in order to avoid obesity issues in the kids and adolescents globally.

The citrate transporter protein (CTP) plays an important role in citrate efflux from the mitochondrial matrix to cytosol that has great importance in oleaginous fungi. The cytoplasmic citrate produced after citrate efflux serves as the primary carbon source for the triacylglycerol and cholesterol biosynthetic pathways. Because of the CTP's importance, our laboratory has extensively studied its structure/function relationships in Mucor circinelloides to comprehend its molecular mechanism. In the present study, the tricarboxylate citrate transporter (Tct) of M. circinelloides WJ11 has been cloned, overexpressed, purified, kinetically, and structurally characterized. The Tct protein of WJ11 was expressed in Escherichia coli, isolated, and functionally reconstituted in a liposomal system for kinetic studies. Our results showed that Tct has a high affinity for citrate with Km 0.018 mM. Furthermore, the tct overexpression and knockout plasmids were created and transformed into M. circinelloides WJ11. The mitochondria of the tct-overexpressing transformant of M. circinelloides WJ11 showed a 49% increase in citrate efflux, whereas the mitochondria of the tct-knockout transformant showed a 39% decrease in citrate efflux compared to the mitochondria of wild-type WJ11. To elucidate the structure-function relationship of this biologically important transporter a 3D model of the mitochondrial Tct protein was constructed using homology modeling. The overall structure of the protein is V-shaped and its 3D structure is dimeric. The transport stability of the structure was also assessed by molecular dynamics simulation studies. The activity domain was identified to form hydrogen bond and stacking interaction with citrate and malate upon docking. Tricarboxylate citrate transporter has shown high binding energy of −4.87 kcal/mol to citric acid, while −3.80 kcal/mol to malic acid. This is the first report of unraveling the structural characteristics of WJ11 mitochondrial Tct protein and understanding the approach of the transporting toward its substrate. In conclusion, the present findings support our efforts to combine functional and structural data to better understand the Tct of M. circinelloides at the molecular level and its role in lipid accumulation.

Canthaxanthin is a reddish-orange xanthophyll with strong antioxidant activity and higher bioavailability than carotenes, primarily used in food, cosmetics, aquaculture, and pharmaceutical industries. The spiking market for natural canthaxanthin promoted researchers toward genetic engineering of heterologous hosts for canthaxanthin production. Mucor circinelloides is a dimorphic fungus that produces β-carotene as the major carotenoid and is considered as a model organism for carotenogenic studies. In this study, canthaxanthin-producing M. circinelloides strain was developed by integrating the codon-optimized β-carotene ketolase gene (bkt) of the Haematococcus pluvialis into the genome of the fungus under the control of strong promoter zrt1. First, a basic plasmid was constructed to disrupt crgA gene, a negative regulator of carotene biosynthesis resulted in substantial β-carotene production, which served as the building block for canthaxanthin by further enzymatic reaction of the ketolase enzyme. The genetically engineered strain produced a significant amount (576 ± 28 μg/g) of canthaxanthin, which is the highest amount reported in Mucor to date. Moreover, the cell dry weight of the recombinant strain was also determined, producing up to more than 9.0 g/L, after 96 h. The mRNA expression level of bkt in the overexpressing strain was analyzed by RT-qPCR, which increased by 5.3-, 4.1-, and 3-folds at 24, 48, and 72 h, respectively, compared with the control strain. The canthaxanthin-producing M. circinelloides strain obtained in this study provided a basis for further improving the biotechnological production of canthaxanthin and suggested a useful approach for the construction of more valuable carotenoids, such as astaxanthin.

Interactions between plant and soil communities have important implication for plant competition, development and succession. In order to explore the internal mechanism behind natural succession of abandoned farmland, we test the effect of plant–soil interaction on plant growth and competitive ability through performing a pot experiment, which included three grasses in different successional stages on the Loess Plateau of China (Setaria viridis, Stipa bungeana, and Bothriochloa ischaemum) in monoculture and all possible two- and three-way combinations, along with a plant-free control pot. The plants were harvested after about 4 months, and the rhizospheric soil was collected. The bacterial communities of the soils were analyzed by high-throughput sequencing of the 16S rRNA gene. Plant competition affected richness of bacterial communities. Proteobacteria and Bacteroidetes were generally higher and Actinobacteria and Acidobacteria were lower in relative abundance in the mixed treatments associated with B. ischaemum. Photosynthetic bacterium, Genus Rhodobacter family Rhodospirillaceae, affected the growth condition and increased the competitive ability of B. ischaemum. Differences in the amounts of soil organic carbon, water-soluble organic carbon and nitrate nitrogen and available phosphorus drove the differences in bacterial communities. Our study has an important significance for understanding the trend of natural succession on the abandoned farmland on the Loess Plateau of China.

Background Long-term fertilisation has a large influence on soil physical and chemical properties in agro-ecosystems. The effects on the distribution of aggregates, however, are not fully understood. We determined the dynamic change of the distribution of aggregates and soil organic carbon (SOC) content over time in a long-term field experiment established in 1998 on the Loess Plateau of China and illustrated the relationship between them. Methods We determined SOC content and the distribution of aggregates in nine fertiliser treatments: manure (M); nitrogen (N); phosphorus (P); M and N; M, N, and P; M and P; N and P; bare land; and an unfertilised control. These parameters were then used for a path analysis and to analyse the fractal dimension (Dv). Results The organic fertiliser increased SOC content. The proportions of 0.1–0.25 mm microaggregates and 0.25–0.5 mm macroaggregates were higher and the proportion of the 0.01–0.05 mm size class of the silt + clay fraction was lower in the treatments receiving organic fertiliser (M, MN, MNP, and MP) than that in the control, indicating that the addition of organic fertiliser promoted aggregation. The distribution of aggregates characterised by their fractal dimension (Dv), however, did not differ among the treatments. Discussion Dv was strongly correlated with the proportion of the

Analyzing the dynamic patterns of species diversity and spatial heterogeneity of vegetation in grasslands during secondary succession could help with the maintenance and management of these ecosystems. Here, we evaluated the influence of secondary succession on grassland plant diversity and spatial heterogeneity of abandoned croplands on the Loess Plateau (China) during four phases of recovery: 1–5, 5–10, 10–20, and 20–30 years. The species composition and dominance of the grassland vegetation changed markedly during secondary succession and formed a clear successional series, with the species assemblage dominated by Artemisia capillaris→ Heteropappus altaicus→ A. sacrorum. The diversity pattern was one of low–high–low, with diversity peaking in the 10–20 year phase, thus corresponding to a hump-backed model in which maximum diversity occurring at the intermediate stages. A spatially aggregated pattern prevailed throughout the entire period of grassland recovery; this was likely linked to the dispersal properties of herbaceous plants and to high habitat heterogeneity. We conclude that natural succession was conducive to the successful recovery of native vegetation. From a management perspective, native pioneer tree species should be introduced about 20 years after abandoning croplands to accelerate the natural succession of grassland vegetation.

Background. Long-term fertilisation has a large influence on soil physical and chemical properties in agro-ecosystems. The effects on the distribution of aggregates, however, are not fully understood. We determined the dynamic change of the distribution of aggregates and soil organic carbon (SOC) content over time in a long-term field experiment established in 1998 on the Loess Plateau of China and illustrated the relationship between them. Methods. We determined SOC content and the distribution of aggregates in nine fertiliser treatments: manure (M); nitrogen (N); phosphorus (P); M and N; M, N, and P; M and P; N and P; bare land; and an unfertilised control. These parameters were then used for a path analysis and to analyse the fractal dimension (Dv). Results. The organic fertiliser increased SOC content. The proportions of 0.1–0.25 mm microaggregates and 0.25–0.5 mm macroaggregates were higher and the proportion of the 0.01–0.05 mm size class of the silt + clay fraction was lower in the treatments receiving organic fertiliser (M, MN, MNP, and MP) than that in the control, indicating that the addition of organic fertiliser promoted aggregation. The distribution of aggregates characterised by their fractal dimension (Dv), however, did not differ among the treatments. Discussion. Dv was strongly correlated with the proportion of the <0.002 mm size class of the silt + clay fraction that did not differ significantly among the treatments. The change in the distribution of aggregates was strongly correlated with SOC content, which could produce organic polymer binding agents to increase the proportion of larger particles. Long-term application of organic fertiliser is thus necessary for the improvement and maintenance of soil quality in semi-arid agricultural land when residues are removed. [ABSTRACT FROM AUTHOR]