Nutrient-sufficient Conditions Research Articles

Overexpression of SLIM1 transcription factor accelerates vegetative development in Arabidopsis thaliana.

The transcription factor Sulfur Limitation 1 (SLIM1) belongs to the plant-specific Ethylene Insenstive3-Like transcription factor family and is known to coordinate gene expression in response to sulfur deficiency. However, the roles of SLIM1 in nutrient-sufficient conditions have not been characterized. Employing constitutive SLIM1 overexpression (35S::SLIM1) and CRISPR/Cas9 mutant plants (slim1-cr), we identified several distinct phenotypes in nutrient-sufficient conditions in Arabidopsis thaliana. Overexpression of SLIM1 results in plants with approximately twofold greater rosette area throughout vegetative development. 35S::SLIM1 plants also bolt earlier and exhibit earlier downregulation of photosynthesis-associated genes and earlier upregulation of senescence-associated genes than Col-0 and slim1-cr plants. This suggests that overexpression of SLIM1 accelerates development in A. thaliana. Genome-wide differential gene expression analysis relative to Col-0 at three time points with slim1-cr and two 35S::SLIM1 lines allowed us to identify 1,731 genes regulated directly or indirectly by SLIM1 in vivo.

Constitutive basis of root system architecture: uncovering a promising trait for breeding nutrient- and drought-resilient crops

Root system architecture (RSA) plays a pivotal role in efficient uptake of essential nutrients, such as phosphorous (P), nitrogen (N), and water. In soils with heterogeneous nutrient distribution, root plasticity can optimize acquisition and plant growth. Here, we present evidence that a constitutive RSA can confer benefits for sorghum grown under both sufficient and limiting growth conditions. Our studies, using P efficient SC103 and inefficient BTx635 sorghum cultivars, identified significant differences in root traits, with SC103 developing a larger root system with more and longer lateral roots, and enhanced shoot biomass, under both nutrient sufficient and deficient conditions. In addition to this constitutive attribute, under P deficiency, both cultivars exhibited an initial increase in lateral root development; however, SC103 still maintained the larger root biomass. Although N deficiency and drought stress inhibited both root and shoot growth, for both sorghum cultivars, SC103 again maintained the better performance. These findings reveal that SC103, a P efficient sorghum cultivar, also exhibited enhanced growth performance under N deficiency and drought. Our results provide evidence that this constitutive nature of RSA can provide an avenue for breeding nutrient- and drought-resilient crops.

HCMV-encoded viral protein US12 promotes autophagy by inducing autophagy flux

The human cytomegalovirus (HCMV)-encoded US12 gene family is a group of ten predicted seven-transmembrane domain proteins that are structurally similar to G-protein-coupled receptors or transmembrane Bax inhibitor-1 motif-containing proteins; however, the roles of US12 family proteins in virus–host interactions remain to be discovered. Here, we suggest a new function of the US12 protein in regulating cellular autophagy. US12 is predominantly located to the lysosome and interacts with the lysosomal membrane protein 2 (LAMP2). A liquid chromatography–mass spectrometry (MS)/MS-based targeted proteomics analysis shows that US12 is tightly correlated with autophagy. US12 induces autophagy via upregulating ULK1 phosphorylation and subsequent LC3-II conversion, thereby accelerating autophagic flux. Moreover, HeLa cells overexpressing US12 displays intense LC3-specific staining and autolysosome formation even under nutrient-sufficient conditions. Furthermore, the physical interaction of p62/SQSTM1 with US12 is involved in the resistance to the degradation of p62/SQSTM1 by autophagy, despite the induction of both autolysosome formation and autophagic flux. Although the effect of US12 expression in HCMV infection on autophagy remains undetermined, these findings provide new insights into the viral drivers of host autophagy during HCMV evolution and pathogenesis.

Effect of nutrients deficiency on biofilm formation and single cell protein production with a purple non-sulphur bacteria enriched culture

Purple non-sulphur bacteria (PNSB) are of interest for biorefinery applications to create biomolecules, but their production cost is expensive due to substrate and biomass separation costs. This research has utilized fuel synthesis wastewater (FSW) as a low-cost carbon-rich substrate to produce single-cell protein (SCP) and examines PNSB biofilm formation using this substrate to achieve a more efficient biomass-liquid separation. In this study, PNSB were grown in Ca, Mg, S, P, and N-deficient media using green shade as biofilm support material. Among these nutrient conditions, only N-deficient and control (nutrient-sufficient) conditions showed biofilm formation. Although total biomass growth of the control was 1.5 times that of the N-deficient condition and highest overall, the total biofilm-biomass in the N-deficient condition was 2.5 times greater than the control, comprising 49% of total biomass produced. Total protein content was similar between these four biomass samples, ranging from 35.0 ± 0.2% to 37.2 ± 0.0%. The highest protein content of 44.7 ± 1.3% occurred in the Mg-deficient condition (suspended biomass only) but suffered from a low growth rate. Overall, nutrient sufficient conditions are optimal for overall protein productivity and dominated by suspended growth, but where fixed growth systems are desired for cost-effective harvesting, N-deficient conditions provide an effective means to maximize biofilm production without sacrificing protein content.

Lipid production under a nutrient-sufficient condition outperforms starvation conditions due to a natural polarization of lipid content in algal biofilm

Attached algae cultivation for biofuel production has received proliferated attention given its advantage to reduce harvesting/dewatering costs and contamination risk. In this study, the effects of various nutrient starvation strategies on lipid production through the attached cultivation of Ettlia sp. YC001 was investigated. A detailed study on compositional variation over the depth of biofilms was followed to understand the phenomena within the biofilms under various nutrient conditions. As a result, the nutrient starvation strategy enhanced the lipid content of biofilms by 3–7 %, however, it limited biomass growth significantly. Ultimately, lipid production with sufficient nutrient supply outperformed those with nutrient starvations, reaching up to the maximum lipid productivity of 6.5 ± 0.1 g/m2/day. Compositional analyses over the depth of biofilms identified that high lipid accumulation was induced in the top layer of the biofilm regardless of medium condition as the top layers were naturally under stress from nutrient deficiency and high light exposure.

Differential metabolic rearrangements in the roots and leaves of Cicer arietinum caused by single or double nitrate and/or phosphate deficiencies.

Nitrate (NO3 - ) and phosphate (Pi) deficiencies are the major constraints for chickpea productivity, significantly impacting global food security. However, excessive fertilization is expensive and can also lead to environmental pollution. Therefore, there is an urgent need to develop chickpea cultivars that are able to grow on soils deficient in both NO3 - and Pi. This study focused on the identification of key NO3 - and/or Pi starvation-responsive metabolic pathways in the leaves and roots of chickpea grown under single and double nutrient deficiencies of NO3 - and Pi, in comparison with nutrient-sufficient conditions. A global metabolite analysis revealed organ-specific differences in the metabolic adaptation to nutrient deficiencies. Moreover, we found stronger adaptive responses in the roots and leaves to any single than combined nutrient-deficient stresses. For example, chickpea enhanced the allocation of carbon among nitrogen-rich amino acids (AAs) and increased the production of organic acids in roots under NO3 - deficiency, whereas this adaptive response was not found under double nutrient deficiency. Nitrogen remobilization through the transport of AAs from leaves to roots was greater under NO3 - deficiency than double nutrient deficiency conditions. Glucose-6-phosphate and fructose-6-phosphate accumulated in the roots under single nutrient deficiencies, but not under double nutrient deficiency, and higher glycolytic pathway activities were observed in both roots and leaves under single nutrient deficiency than double nutrient deficiency. Hence, the simultaneous deficiency generated a unique profile of metabolic changes that could not be simply described as the result of the combined deficiencies of the two nutrients.

A Model to Incorporate the bHLH Transcription Factor OsIRO3 within the Rice Iron Homeostasis Regulatory Network.

Iron (Fe) homeostasis in plants is governed by a complex network of regulatory elements and transcription factors (TFs), as both Fe toxicity and deficiency negatively impact plant growth and physiology. The Fe homeostasis network is well characterized in Arabidopsis thaliana and remains poorly understood in monocotyledon species such as rice (Oryza sativa L.). Recent investigation of the rice Fe homeostasis network revealed OsIRO3, a basic Helix–Loop–Helix (bHLH) TF as a putative negative regulator of genes involved in Fe uptake, transport, and storage. We employed CRISPR-Cas9 gene editing to target the OsIRO3 coding sequence and generate two independent T-DNA-free, loss-of-function iro3 mutants in rice cv. Nipponbare. The iro3 mutant plants had similar phenotype under nutrient-sufficient conditions and had stunted growth under Fe-deficient conditions, relative to a T-DNA free, wild-type control (WT). Under Fe deficiency, iro3 mutant shoots had reduced expression of Fe chelator biosynthetic genes (OsNAS1, OsNAS2, and OsNAAT1) and upregulated expression of an Fe transporter gene (OsYSL15), relative to WT shoots. We place our results in the context of the existing literature and generate a model describing the role of OsIRO3 in rice Fe homeostasis and reinforce the essential function of OsIRO3 in the rice Fe deficiency response.

Effect of csn2 gene deficiency on the starvation tolerance and the synthesis of extracellular polysaccharides in oligotrophic environment of Streptococcus mutans

Objective: To explore the effect of csn2 gene deficiency on starvation tolerance and extracellular polysaccharides (EPS) synthesis in an oligotrophic environment of Streptococcus mutans (Sm). Methods: The csn2 gene deletion strains and complementary strains of Sm were cultivated and then an oligotrophic growth environment for Sm growth by setting different concentration gradient media were created. Cell growth in oligotrophic environment was detected by growth curve. Biofilm volume was measured by crystalline violet staining. Scanning electron microscopy (SEM) and laser confocal microscope were performed to observe the biofilm structure of Sm. The synthesis of EPS was measured by the anthrone-sulfuric acid method. The expression of genes related to EPS synthesis was evaluated by quantitative real-time PCR (qRT-PCR). Results: The growth curve results showed that the deletion of csn2 gene inhibited the growth of Sm under starvation stress. Furthermore, the results of laser confocal microscope showed that the biofilm EPS/bacteria ratios produced by the wild-type strain, csn2 gene-deficient strain and complement strains under nutrient sufficient culture conditions were 0.44±0.07, 1.05±0.13 and 0.57±0.08 respectively, while the ratios of EPS/bacteria in an oligotrophic environment were 0.93±0.24, 3.05±0.21 and 1.32±0.46 respectively, indicating that the deletion of csn2 gene enhanced the ability of extracellular polysaccharide synthesis of Sm in the oligotrophic environment. The expression levels of EPS synthesis-related genes gtfB and gtfC were up-regulated by 2.5 fold and 1.8 fold respectively and the expression level of gtfD was down-regulated by two-thirds. Conclusions: The csn2 gene deficiency showed multiple effects on the physiological functions and virulence characteristics of Sm, including starvation tolerance and EPS synthesis. These changes might be related to the shift of the complex regulative network caused by csn2 gene deletion.

Acaulospora as the Dominant Arbuscular Mycorrhizal Fungi in Organic Lowland Rice Paddies Improves Phosphorus Availability in Soils

Flooding in rainfed lowlands greatly impairs the mutualistic relationship between indigenous arbuscular mycorrhizal fungi (AMF) and rice. In flooded soils, root colonization by AMF is arrested, but some AMF genera, defined as the core AMF, remain present. However, the core AMF in rainfed lowlands and their symbiotic roles remain unknown. Here, we showed that Acaulospora fungi were the core AMF in rice seedling roots of the Sangyod Muang Phatthalung (SMP) landrace rice variety grown in non-flooded and flooded paddy soils. Subsequently, indigenous Acaulospora spores were propagated by trap cultures using maize as the host plants. Therefore, to clarify the roles of cultured Acaulospora spores in a symbiotic partnership, the model japonica rice variety Nipponbare was grown in sterile soil inoculated with Acaulospora spores, and recolonized with a native microbial filtrate from the organic rice paddy soil. Our data demonstrated that the inoculation of Acaulospora spores in well-drained soil under a nutrient-sufficient condition for six weeks enabled 70 percent of the rice roots to be colonized by the fungi, leading to higher phosphate (Pi) accumulation in the mycorrhizal roots. Unexpectedly, the growth of rice seedlings was significantly suppressed by inoculation while photosynthetic parameters such as fractions of incoming light energy and relative chlorophyll content were unaltered. In the soil, the Acaulospora fungi increased soil phosphorus (P) availability by enhancing the secretion of acid phosphatase in the mycorrhizal roots. The findings of this work elucidate the symbiotic roles of the dominant Acaulospora fungi from lowland rice paddies.

Poly-β-Hydroxybutyrate Production by Rhodopseudomonas sp. Grown in Semi-Continuous Mode in a 4 L Photobioreactor

The synthesis of polyhydroxybutyrate (PHB) by photosynthetic non-sulfur bacteria is a potential approach for producing biodegradable plastics. In this work, acetate was used as a single carbon source to study the effect on PHB formation in Rhodopseudomonas sp. cultured in a cylindrical four-liter photobioreactor under semi-continuous mode. The cultivation process is divided into a symmetrical growth phase and a PHB accumulation phase separated temporally. The symmetrical growth phase (nutrient sufficient conditions) was followed by a sulfur-limited phase to promote PHB accumulation. The main novelty is the progressive lowering of the sulfur concentration into Rhodopseudomonas culture, which was obtained by two concomitant conditions: (1) sulfur consumption during the bacterial growth and (2) semi-continuous growth strategy. This caused a progressive lowering of the sulfur concentration into Rhodopseudomonas culturedue to the sulfur-free medium used to replace 2 L of culture (50% of the total) that was withdrawn from the photobioreactor at each dilution. The PHB content ranged from 9.26% to 15.24% of cell dry weight. At the steady state phase, the average cumulative PHB was >210 mg/L. Sulfur deficiency proved to be one of the most suitable conditions to obtain high cumulative PHB in Rhodopseudomonas culture.

Quercetin enhances fatty acid β-oxidation by inducing lipophagy in AML12 hepatocytes

Recent evidence demonstrated that chronic intake of quercetin attenuated hepatic fat accumulation in various animal models of obesity and diabetes. However, whether quercetin has the ability to enhance energy metabolism in hepatocytes and its exact mechanisms have yet to be identified. In the present study, we investigated whether quercetin directly enhanced the energy metabolism of cultured hepatocytes by focusing on lipophagy, involving selective autophagic degradation of lipid droplets. As an indicator of mitochondrial respiration, oxygen consumption was measured following 12-h treatment with quercetin or its related flavonoids, isorhamnetin and rutin (10 μM) using an extracellular flux analyzer. Treatment of alpha mouse liver 12 (AML12) hepatocytes with quercetin enhanced mitochondrial respiration, but isorhamnetin and rutin did not. Results of a palmitate-bovine serum albumin fatty acid oxidation assay showed that quercetin significantly increased the oxygen consumption of AML12 hepatocytes, suggesting enhanced fatty acid β-oxidation. However, as expression levels of mitochondrial oxidative phosphorylation proteins were unaltered by quercetin, we explored whether lipophagy contributed to enhanced fatty acid β-oxidation. Increased colocalization of lipid droplets and lysosomes confirmed that quercetin promoted lipophagy in AML12 hepatocytes. Furthermore, pharmacological inhibition of the autophagy–lysosomal pathway abolished the enhancement of fatty acid β-oxidation induced by quercetin in AML12 hepatocytes, suggesting that the enhancement of lipophagy by quercetin contributed to increased fatty acid β-oxidation. Finally, we showed that quercetin could activate AMPK signaling, which regulates autophagy even under nutrient-sufficient conditions. Our findings indicate that quercetin enhanced energy metabolism by a potentially novel mechanism involving promotion of lipophagy to produce the substrate for fatty acid β-oxidation in mitochondria through activation of AMPK signaling. Our results suggest the possibility that nutrient-induced lipophagy might contributes to the reduction of fat in hepatocytes.

A redundant hydraulic function of root hairs in barley plants grown in hydroponics.

The root hair-less brb of Hordeum vulgare L. (bald root barley) mutant was used to assess the significance that root hairs have for the hydraulic properties of roots and response to a limited supply of mineral nutrients in plants grown on hydroponics. The barley brb mutant and its parent wild-type (H. vulgare cv. Pallas) were grown under nutrient sufficient control conditions, and under conditions of low supply of P and N. Plants were analysed when they were 14-18 days old. Root hydraulic conductivity (Lp) was determined for excised root systems and intact transpiring plants, and cell Lp was determined through cell pressure probe measurements. The formation of Casparian bands and suberin lamellae was followed through staining of cross-sections. The presence or absence of root hairs had no effect on the overall hydraulic response of plants to nutritional treatments. Root and cell Lp did not differ between the two genotypes. The most apparent difference between brb and wild-type plants was the consistently reduced formation of apoplastic barriers in brb plants. Any hydraulic function of root hairs can be redundant in barley, at least under the hydroponic conditions tested.

Bioplastic production by feeding the marine Rhodovulum sulfidophilum DSM-1374 with four different carbon sources under batch, fed-batch and semi-continuous growth regimes

In the present study, the ability of the marine bacterium Rhodovulum sulfidophilum DSM-1374 to convert, via photo-fermentative process, certain organic acids such as single carbon source (acetate, lactate, malate and succinate) into polyhydroxyalkanoate accumulations within bacterial cells is evaluated. The main goal of the investigation was poly-3-hydroxybutyrate (P3HB) synthesis by a photo-fermentative process. Of the four carbon sources, only succinate simultaneously produced P3HB and H2 (268 mg/L and 1085 mL/L respectively). Malate was the least productive source for P3HB; the other carbon sources (acetate and lactate) produced a significant amount of polymer (596 mg P3HB/L for acetate and 716 mg P3HB/L for lactate) when R. sulfidophilum was cultured in batch growth conditions. Cumulative P3HB increased significantly when the bacterium was grown under two steps: nutrient sufficient conditions (step 1) followed by macronutrient deficient conditions (step 2). The highest cumulative P3HB was observed at the end of step 2 (1000 mg/L) when R. sulfidophilum was fed with lactate under phosphorus starvation. When grown over 1200 h, under a semi-continuous regimen, the harvested dry-biomass reached a constant content of P3HB (39.1 ± 1.6 % of cell dry-weight), in the semi-steady state condition. Since lactate is an abundant byproduct of world industries, it can be used to mitigate the environmental impact in a modern circular bio-economy.

Human pancreatic cancer cells under nutrient deprivation are vulnerable to redox system inhibition

Large regions in tumor tissues, particularly pancreatic cancer, are hypoxic and nutrient-deprived because of unregulated cell growth and insufficient vascular supply. Certain cancer cells, such as those inside a tumor, can tolerate these severe conditions and survive for prolonged periods. We hypothesized that small molecular agents, which can preferentially reduce cancer cell survival under nutrient-deprived conditions, could function as anticancer drugs. In this study, we constructed a high-throughput screening system to identify such small molecules and screened chemical libraries and microbial culture extracts. We were able to determine that some small molecular compounds, such as penicillic acid, papyracillic acid, and auranofin, exhibit preferential cytotoxicity to human pancreatic cancer cells under nutrient-deprived compared with nutrient-sufficient conditions. Further analysis revealed that these compounds target to redox systems such as GSH and thioredoxin and induce accumulation of reactive oxygen species in nutrient-deprived cancer cells, potentially contributing to apoptosis under nutrient-deprived conditions. Nutrient-deficient cancer cells are often deficient in GSH; thus, they are susceptible to redox system inhibitors. Targeting redox systems might be an attractive therapeutic strategy under nutrient-deprived conditions of the tumor microenvironment.

Deletion of a small, secreted and cysteine-rich protein Cpl1 leads to increased invasive growth of Cryptococcus neoformans into nutrient agar

Invasive growth of yeast cells into nutrient agar is induced by different stresses and contributes to the survival of yeast cells under several adverse conditions. The mechanism of invasive growth of Saccharomyces cerevisiae has been extensively investigated. However, there is very little information about the mechanism of invasive growth of another human pathogen yeast Cryptococcus neoformans. Here, we report that deletion of a small and secreted cysteine-rich protein Cpl1 in C. neoformans JEC21 leads to increased adhesive and invasive growth into nutrient agar. The increased adhesive and invasive growth does not depend on the only known adhesion protein Cfl1 and its main controller Znf2. Cpl1Δ accumulates significantly higher level of intracellular labile zinc ion, leading to increased glucose uptake, higher level of mitochondrial membrane potential, ATP and Reactive Oxygen Species(ROS) production. Higher level of ROS activates Snf1, leading to invasive growth of Cpl1Δ. Three cysteine residues at the N-terminals of the cysteine-rich domain controls the increased invasive growth under nutrient sufficient conditions. This is the first report that a small and secreted cysteine-rich protein negatively regulates invasive growth of C. neoformans through regulating the intracellular labile zinc ion level. The function of this cysteine-rich domain was systematically investigated by site-directed mutagenensis in C. neoformans. The work contributes to understanding the function of this protein family and the invasive growth mechanism in C. neoformans.

Effects of Temperature and Nutrient Supply on Resource Allocation, Photosynthetic Strategy, and Metabolic Rates of Synechococcus sp.

Temperature and nutrient supply are key factors that control phytoplankton ecophysiology, but theirrole is commonly investigated in isolation. Their combined effect on resource allocation, photosynthetic strategy, and metabolism remains poorly understood. To characterize the photosynthetic strategy and resource allocation under different conditions, we analyzed the responses of a marine cyanobacterium (Synechococcus PCC 7002) to multiple combinations of temperature and nutrient supply. We measured the abundance of proteins involved in the dark (RuBisCO, rbcL) and light (Photosystem II, psbA) photosynthetic reactions, the content of chlorophyll a, carbon and nitrogen, and the rates of photosynthesis, respiration, and growth. We found that rbcL and psbA abundance increased with nutrient supply, whereas a temperature-induced increase in psbA occurred only in nutrient-replete treatments. Low temperature and abundant nutrients caused increased RuBisCO abundance, a pattern we observed also in natural phytoplankton assemblages across a wide latitudinal range. Photosynthesis and respiration increased with temperature only under nutrient-sufficient conditions. These results suggest that nutrient supply exerts a stronger effect than temperature upon both photosynthetic protein abundance and metabolic rates in Synechococcus sp. and that the temperature effect on photosynthetic physiology and metabolism is nutrient dependent. The preferential resource allocation into the light instead of the dark reactions of photosynthesis as temperature rises is likely related to the different temperature dependence of dark-reaction enzymatic rates versus photochemistry. These findings contribute to our understanding of the strategies for photosynthetic energy allocation in phytoplankton inhabiting contrasting environments.

Competitive interactions between two allelopathic algal species: Heterosigma akashiwo and Phaeodactylum tricornutum

ABSTRACTBoth Heterosigma akashiwo and Phaeodactylum tricornutum have been reported to produce allelochemicals capable of inhibiting the growth of co-occurring microalgae. Here, potential allelopathy between H. akashiwo and P. tricornutum was evaluated using bi-algal culture, cell-free culture filtrate and a no-contact co-culturing system in nutrient-replete media. Experiments were also conducted in the no-contact co-culturing system under nutrient-limited conditions. In nutrient-replete bi-algal culture, the growth of P. tricornutum and H. akashiwo each tended to be strongly suppressed when the other species was inoculated at high cell densities. A mathematical model was used to simulate the growth interactions of the two species in bi-algal culture and showed that P. tricornutum outcompeted H. akashiwo over time with different initial cell densities under nutrient-sufficient conditions, indicating that P. tricornutum was more potent in allelopathy. Heterosigma akashiwo growth was inhibited both in the P. tricornutum culture filtrate and no-contact co-culturing system. This confirmed that the extracellular allelopathic compounds released by P. tricornutum were one of the sources of the H. akashiwo growth inhibition. Nutrient deficiency did not increase the extent of allelopathic activity of allelochemicals.

Elucidating the influence of environmental factors on biogas-based polyhydroxybutyrate production by Methylocystis hirsuta CSC1

The valorization of biogas as a feedstock for the generation of added-value bioproducts will play a key role on the sustainability of anaerobic digestion. The present work assessed the influence of key environmental parameters (O2:CH4 ratio, temperature and nitrogen source) on the growth and polyhydroxybutyrate (PHB) synthesis under nitrogen limiting conditions of the type II methanotroph Methylocystis hirsuta CSC1 using biogas as a feedstock. The O2:CH4 ratios tested (1:1, 1.5:1 and 2:1) did not affect significantly M. hirsuta CSC1 growth yields (~5 g TSS mol−1 CH4), although lower CH4 removal rates were reached under O2-limiting conditions (ratio 1:1). The highest PHB content (45 wt%) was achieved at a ratio 2:1 and was threefold higher than those obtained at lower ratios (~15 wt%). The increase in temperature from 15 to 25 °C resulted in increases in the growth yield (from 5 to 6 g TSS mol−1 CH4) and PHB content (from 32 to 40 wt%). Conversely, the lowest PHB content (30 wt%) was reached at 37 °C, together with a negligible growth under nutrient sufficient conditions. The nitrogen source also played a key role on both M. hirsuta CSC1 growth and PHB synthesis. Thus, ammonium resulted in the highest growth yield (7 g TSS mol−1 CH4), although the maximum PHB content was achieved when biomass was previously grown in nitrate as the nitrogen source (41 wt%). Nitrite exerted an inhibitory effect on M. hirsuta CSC1 growth.

Poly-3-hydroxybutyrate and H2 production by Rhodopseudomonas sp. S16-VOGS3 grown in a new generation photobioreactor under single or combined nutrient deficiency

The main goal of this investigation was setting up a growth strategy to separate H2 evolution from P3HB synthesis in order to increase cumulative P3HB in Rhodopseudomonas cells. The accumulation of poly-3-hydroxybutyrate (P3HB) was investigated culturing Rhodopseudomonas sp. S16-VOGS3 with three carbon substrates either as acetate, butyrate or lactate and with two nitrogen sources either as ammonium or glutamate. The investigation was carried out under several stress conditions caused by single or double nutrient deficiency. The content of P3HB in cell dry weight (CDW) was 21.8% with lactate; 24.6% with acetate and 27.6% with butyrate under sulfur deficient conditions. The P3HB content increased significantly culturing Rhodopseudomonas sp. S16-VOGS3 with butyrate following three phases of growth: phase-1, nutrient sufficient conditions; phase-2, nitrogen-deficiency and phase-3, sulfur-deficient conditions. Under this last phase, the highest P3HB content was achieved (34.4% of CDW). A combined production of P3HB and molecular H2 was obtained when Rhodopseudomonas sp. S16-VOGS3 was cultured with either acetate or butyrate under nitrogen sufficiency (glutamate) or nitrogen deficiency.

A homolog of Arabidopsis SDP1 lipase in Nannochloropsis is involved in degradation of de novo-synthesized triacylglycerols in the endoplasmic reticulum

Organisms of the microalgal genus Nannochloropsis produce high levels of triacylglycerols (TAGs), an efficient raw material for biofuels. A complete understanding of the TAG-breakdown pathway is critical for improving the productivity of TAGs to meet future needs. Among a number of lipases annotated as TAG lipase in the genomes of every organism, Arabidopsis SUGAR-DEPENDENT 1 (AtSDP1) lipases are characterized as a type of crucial TAG lipase in plants, similar to ScTgl3–5 in Saccharomyces cerevisiae. Homologs of the AtSDP1 TAG lipases are universally found in the genomes of plants, fungi, and algae. Here we identified two homologs of AtSDP1 TAG lipases in the oleaginous microalga species Nannochloropsis oceanica, NoTGL1 and NoTGL2. We generated single- and double-knockout strains for these lipases by homologous recombination. Whereas overall TAG content in the NoTGL2 single-knockout mutant was identical to that of wild type, the NoTGL1 knockout showed a two-fold increase in TAG content per cell in early log phase under nutrient-sufficient conditions without affecting growth. Homologs of AtSDP1 in S. cerevisiae are localized to the surface of lipid droplets, and AtSDP1 is transported from peroxisomes to the surface of lipid droplets. In contrast, NoTGL1 localized to the endoplasmic reticulum in both Nannochloropsis and yeast. We suggest that homologs of AtSDP1 lipases in Nannochloropsis modulate de novo TAG biosynthesis in the endoplasmic reticulum, unlike the roles of these lipases in other organisms. These results provide important insights into the mechanisms of TAG metabolism catalyzed by homologs of AtSDP1 lipase, which are highly conserved across species.