Effects Of Nitrogen Limitation Research Articles

Combined effects of nitrogen limitation and overexpression of malic enzyme gene on lipid accumulation in Dunaliella parva

Overexpression of Dunaliella parva (D. parva) malic enzyme (ME) gene (DpME) significantly increased DpME expression and ME enzyme activity in transgenic D. parva. Nitrogen limitation had an inhibitory effect on protein content, and DpME overexpression could improve protein content. Nitrogen limitation increased carbohydrate content, and Dunaliella parva overexpressing malic enzyme gene under nitrogen limitation (DpME-N−) group showed the lowest starch content among all groups. Dunaliella parva overexpressing malic enzyme gene under nitrogen sufficient condition (DpME) and DpME-N− groups showed considerably high mRNA levels of DpME. ME activity was significantly enhanced by DpME overexpression, and nitrogen limitation caused a smaller increase. DpME overexpression and nitrogen limitation obviously enhanced lipid accumulation, and DpME overexpression had more obvious effect. Compared with control (wild type), lipid content (68.97%) obviously increased in DpME-N− group. This study indicated that the combination of DpME overexpression and nitrogen limitation was favorable to the production of microalgae biodiesel.

Nitrogen limitation affects carbon and nitrogen metabolism in mung bean (Vigna radiata L.)

Studying the effects of nitrogen limitation on carbon, nitrogen metabolism, and nutrient uptake of mung bean is a scientific issue. In this study, urea (CO(NH2)2, 125 kg hm−2) was applied at the V2, V6, R1, R2, and R4 stages, respectively, to ensure sufficient N resources during the growth process of mung beans. This study found that nitrogen limitation inhibited mung bean photosynthesis and reduced photosynthetic efficiency, which was manifested by reducing Pn (net photosynthetic rate), Gs (stomatal conductance), Tr (transpiration rate), and Ci (intercellular carbon dioxide concentration). Second, nitrogen limitation reduced N metabolism-related enzyme activity, such as NR (nitrate reductase), GOGAT (glutamate synthase), and GDH (glutamate dehydrogenase), indicating that nitrogen limitation inhibited the process of nitrogen metabolism, reducing nitrogen assimilation. Meanwhile, topdressing N fertilizer can promote the P and K uptake, and improve the partial factor productivity of P and K, which suggests that nitrogen limitation reduced P and K use efficiency. In addition, this study found that Lvfeng5 responded more significantly to nitrogen fertilizers, and had higher nitrogen use efficiency or better adaptability compared with Lvfeng2. This study provided valuable insights into the physiological and metabolic responses of mung beans to nutrient deficiency.

The effect of nitrogen limitation on carbohydrates and β-glucan accumulation in Nannochloropsis oculata

Nitrogen (N) is the second most abundant element in microalgae biomass, following carbon. Microalgae have a high carbon and nitrogen metabolism system, making nitrogen limitation (N-limitation) a crucial factor that modifies their biochemical composition. In Nannochloropsis genus cultures, N-limitation can cause lipid or carbohydrate accumulation. This research aims to evaluate the effect of N-limitation on carbohydrate and β-glucan accumulation in Nannochloropsis oculata during maximum biomass production (i.e., finished logarithmic phase and early start stationary phase in batch culture) in the two-stage process context. To achieve this goal, we tested the effect of five levels of N availability (treatments 100, 75, 50, 25, and 0 % of N, concerning F medium) in batch culture using sodium nitrate (NaNO3) as the N source. The initial availability of N in each treatment was 1.7, 1.3, 0.8, 0.4, and 0 mmol∙l−1. The results showed that the highest concentration of N. oculata biomass, carbohydrate, and β-glucans content was produced during the logarithmic phase at 0.8 and 1.32 mmol∙l−1 N initial batch culture concentration (50 and 75 % N treatments), without a significant difference (P > 0.05) from the F medium used. Total carbohydrates obtained were 15.2 %, 19.7 %, 17.2 %, 12.7 %, and 14.2 % (dry weight) for the 100 %, 75 %, 50 %, 25 %, and 0 % N treatments, respectively. Similarly, the β-glucans in microalgal biomass were 5.0 %, 7.4 %, 6.7 %, 4.4 %, and 1.7 % (dry weight) for treatments 100 %, 75 %, 50 %, 25 %, and 0 % N, respectively. The 25 % and 0 % N treatments had a negative effect (P < 0.05) on cell population growth, carbohydrates, and β-glucans accumulation. Based on these results, we recommended conducting studies on the effect of N-limitation in N. oculata culture primarily at 50 % and 75 % N treatments, during the stationary phase and/or second stage of the two-stage process context, such as a chronic or and long-term effect, along with a partial supply of N or testing other inductors might also be explored. This knowledge can help to optimize the industrial production of microalgae β-glucans.

Influence of Substrate on the Fermentation Characteristics and Culture-Dependent Microbial Composition of Water Kefir

Water kefir is a sparkling fermented beverage produced by fermenting water kefir grains in a sucrose solution containing dried fruits or fruit extracts. The objective of this study was to investigate the influence of substrate composition on the fermentation kinetics and culture-dependent microbial composition of water kefir. First, the impact of different fruit substrates and nitrogen limitation was examined. Fermentation of different fruit-based media with a single water kefir culture demonstrated that the substrate mainly influenced the type and ratio of the organic acids produced. These organic acid profiles could be linked to the culture-dependent microbial composition. In addition, the microbial composition and the associated dominant microorganisms observed were influenced by the water kefir fermentation conditions. Investigation of the effect of nitrogen limitation on the fermentation kinetics of several water kefir cultures showed that under such conditions, the fermentative capacity of the cultures declined. However, this decline was not immediate, and specific water kefir microorganisms may have enabled some cultures to maintain a higher fermentative capacity for longer. Thus, the water kefir fermentation kinetics and characteristics could be linked to the substrate composition, microorganisms present, and the process conditions under which the fermentations were performed.

Effects of nitrogen limitation on Leymus Chinensis clonal growth form and bud bank

Nitrogen is an important element which affects the growth and clonal growth of plant species in grassland. Leymus chinensis (Trin.) Tzvel is a dominant species in grasslands with a high nutritional quality; however, data on the effects of low nitrogen concentration on L. chinensis clonal growth strategy are limited. The aim of this study is to evaluate the response of clonal growth of L. chinensis to low nitrogen concentrations. The growth form and biomass allocation ratios of L. chinensis under different low nitrogen concentrations were evaluated, and the quantitative characteristics of three bud types were analyzed. The proportion of spreading ramets was favored by low nitrogen supply, indicating that L. chinensis under low nitrogen conditions primarily exhibits a guerrilla growth form, meaning a long mean spacer length and production of spreading ramets. Under limited nitrogen conditions, biomass distribution was mainly concentrated in underground organs (including the roots and rhizomes), while the high proportion of axillary rhizome buds suggested that L. chinensis grows primarily by occupying effective living spaces under low nitrogen conditions. The balance between two clonal growth form and the change in the proportion of different types of buds is an important strategy for L. chinensis to cope with nutrient limitations.

Minimum nitrogen cell quota for maximal growth rate in cycloturbidostat cultures of Picochlorum oklahomense

Replicate cycloturbidostats cultures of Picochlorum oklahomense were manipulated to quantify the minimum nitrogen cell quota supporting maximum growth rate and to study the effects of nitrogen limitation on cycloturbidostat cultures growing at near-μmax under steady-state, balanced growth conditions. Results show that cyclical diurnal neutral lipid accumulation and ultradian growth are not mutually exclusive, and that cycloturbidostat cultures may be maintained near or at the threshold of N-limitation to control cell composition and modulate biomass and lipid productivity. Using this novel approach, we have identified the threshold of N limitation in P. oklahomense and investigated the effects of N limitation on replicate cycloturbidostat cultures. Mildly N-limited cultures grew rapidly (87 % μmax) and maintained near optimal physiology, while also accumulating neutral lipids during the photoperiod. Flow cytometry revealed differences in temporal patterns between treatments, demonstrating the importance of the cell-based approach in manipulating microalgal cultures. This study highlights the utility of turbidostats in experimental phycology and defines a scantly explored concept in the study of N limitation, the threshold of N cell quota necessary to support μmax below which cells are forced to re-allocate resources and slow growth, denoted QN, T.

Biochemical and Morphological Changes Triggered by Nitrogen Stress in the Oleaginous Microalga Chlorella vulgaris.

Oleaginous microalgae have been considered promising sources of biodiesel due to their high lipid content. Nitrogen limitation/starvation is one of the most prominent strategies to induce lipid accumulation in microalgae. Nonetheless, despite numerous studies, the mechanism underlying this approach is not well understood. The aim of this study was to investigate the effect of nitrogen limitation and starvation on biochemical and morphological changes in the microalga Chlorella vulgaris FACHB-1068, thereby obtaining the optimal nitrogen stress strategy for maximizing the lipid productivity of microalgal biomass. The results showed that nitrogen limitation (nitrate concentration < 21.66 mg/L) and starvation enhanced the lipid content but generally decreased the biomass productivity, pigment concentration, and protein content in algal cells. Comparatively, 3-day nitrogen starvation was found to be a more suitable strategy to produce lipid-rich biomass. It resulted in an increased biomass production and satisfactory lipid content of 266 mg/L and 31.33%, respectively. Besides, nitrogen starvation caused significant changes in cell morphology, with an increase in numbers and total size of lipid droplets and starch granules. Under nitrogen starvation, saturated fatty acids (C-16:0, C-20:0, and C-18:0) accounted for the majority of the total fatty acids (~80%), making C. vulgaris FACHB-1068 a potential feedstock for biodiesel production. Our work may contribute to a better understanding of the biochemical and morphological changes in microalgae under nitrogen stress. Besides, our work may provide valuable information on increasing the lipid productivity of oleaginous microalgae by regulating nitrogen supply.

Effect of nitrogen limitation and soil biophysics on Holocene greening of the Sahara

Abstract. The so-called Green Sahara (GS), which was a wet and vegetative Sahara region in the early to mid-Holocene, provides useful information on our climate simulation because it is a consequence of complex interaction between biophysical and climatic processes. It is still a challenge to simulate the GS in terms of vegetative extent and precipitation using current climate models. This study attempts to simulate the Green Sahara 8000 years ago by using the state-of-the-art Earth system model CESM that incorporates the nitrogen cycle and the soil–precipitation feedbacks. Our study puts more emphasis on the impact of soil biophysical properties (e.g., bare-soil albedo, porosity, heat capacity, and hydraulic conductivity) and soil nitrogen influenced by soil organic matter on the simulation of the GS. In this coupled simulation, vegetation interacts with changes in soil properties and soil organic matter by phenology, decomposition, and allocation of carbon and nitrogen. With changes in the Earth's orbit and dust in the early to mid-Holocene, the model simulates increased precipitation in North Africa but does not capture the extent of the GS. Our analysis shows that the Holocene greening is simulated better if the amount of soil nitrogen and soil texture is properly modified for the humid and vegetative GS period. Soil biochemical and physical properties increase precipitation and vegetation cover in North Africa through their influence on photosynthesis and surface albedo as well as their consequent enhanced albedo–precipitation and evapotranspiration–precipitation feedbacks. Our findings suggest that future climate simulation needs to consider consequent changes in soil nitrogen and texture with changes in vegetation cover and density for proper climate simulations.

Effect of nitrogen limitation on growth, biochemical composition, and cell ultrastructure of the microalga Picocystis salinarum

In recent years biodiesel production has attracted worldwide attention due to the awareness of fossil fuel depletion and microalgae biomass is considered a promising raw material for its formulation. The present study evaluated the effects of different levels of nitrogen limitation (37.5, 18.75, 9.375 mg L−1 NaNO3) on the growth, cell ultrastructure, and biochemical composition of a halophilic native strain of the green alga Picocystis salinarum as a potential raw material source for biodiesel. During a culture period of 20 days, growth measurements and photosynthetic pigments were estimated. Cell density, dry weight, and chlorophylls a, b content decreased with time as nitrogen limitation increase; however, carotenoid content increased. In addition, nitrogen limitation caused an progressive increase in the lipid and carbohydrate yield and a decrease in protein. The high N limitation (9.375 mg L−1) had a significant effect on the accumulation of total lipid content (33.87% dry weight). Carbohydrate content (30.98% dry weight) and protein content (1.89% dry weight) decrease. The lipid content showed a differential FAME profile with high saturated fatty acid values (996.08 μg g−1 dry weight) mainly palmitic acid, compare with the unsaturated ones that showed low values under high N limitation. The gradual increase of lipid content was also corroborated by transmission electron microscopy images with a single large lipid droplet cell formation. Therefore, evaluation of the algal culture conditions such as N limitation, as a strategy to maximize lipid content and improve the fatty acid profile in unexplored strain of P. salinarum, showed a potential biomass yield as a suitable candidate for biodiesel production.

Long-term effects of elevated CO2 on the population dynamics of the seagrass Cymodocea nodosa: Evidence from volcanic seeps

Population reconstruction techniques was used to assess for the first time the population dynamics of a seagrass, Cymodocea nodosa, exposed to long-term elevated CO2 near three volcanic seeps and compared them with reference sites away from the seeps. Under high CO2, the density of shoots and of individuals (apical shoots), and the vertical and horizontal elongation and production rates, were higher than at the reference sites. Nitrogen limitation effects on rhizome elongation and production rates and on biomass were more evident than CO2 as these were highest at the location where the limitation of nitrogen was highest. At the seep where the availability of CO2 was highest and nitrogen lowest, density of shoots and individuals were highest, probably due to CO2 effects on shoot differentiation and induced reproductive output, respectively. At the three seeps, there was higher short- and long-term shoot recruitment than at the reference sites, and growth rates was around zero, indicating that elevated CO2 increases the turnover of C. nodosa shoots.

Effects of Nitrogen Limitation on Phytoplankton Physiology in the Western Arctic Ocean in Summer

AbstractPhytoplankton in the Arctic Ocean are subject to nitrogen limitation in the summer, however, how severely the nitrogen stress affects phytoplankton physiology remains largely unknown. In the summers of 2015–2018, we examined the distribution of phytoplankton photophysiological properties across two contrasting regions of the Arctic Ocean with distinctly different levels of nitrogen availability in the upper water column. We quantified the extent of nitrogen stress using a highly sensitive fluorescence induction and relaxation system to obtain continuous underway measurements and via discrete sample analyses of phytoplankton physiology, as well as nutrient enrichment incubations. The results revealed vast regions in the Chukchi Sea where phytoplankton photosynthesis was severely nitrogen‐stressed. Thereby, the maximum quantum yield of photochemistry in photosystem II showed only a small decrease (12 ± 9%) relative to its nutrient‐replete values, while the maximum photosynthetic electron transport rates under saturating irradiance were impaired to a greater extent (40 ± 17%). This phytoplankton photosynthesis response is indicative of a severe nitrogen limitation, which results in dramatic reduction in growth and net primary production rates. Nutrient enrichment incubations also revealed a marked increase in large‐size phytoplankton growth (&gt;10 μm) after the nitrogen stress was alleviated, suggesting that the larger cells were more susceptible to nitrogen stress. These results are important for understanding how regional nitrogen fluxes control variability in the primary production and phytoplankton community structure and how these processes might change with rapid climate changes in the Arctic Ocean.

Using chlorophyll fluorescence kinetics to determine photosynthesis in aquatic ecosystems

AbstractVariable fluorescence techniques are increasingly used to assess phytoplankton photosynthesis. All fluorescence techniques and models for photosynthetic electron transport rates (ETRs) are amplitude‐based and are subject to errors, especially when phytoplankton growth is nutrient‐limited. Here we develop a new, kinetic‐based approach to measure, directly and in absolute units, ETRs and to estimate growth rates in phytoplankton. We applied this approach to investigate the effects of nitrogen limitation on phytoplankton photophysiology and growth rates. Nutrient stress leads to a decrease in the quantum yield of photochemistry in Photosystem II (Fv/Fm); however, the relationship between Fv/Fm and growth rates is highly nonlinear, which makes it impossible to quantify the reduction in phytoplankton growth rates from Fv/Fm alone. In contrast, the decline in growth rates under nitrogen stress was proportional to the decrease in kinetic‐based photosynthetic rates. Our analysis suggests the kinetic fluorescence measurements markedly improve the accuracy of ETR measurements, as compared to classical amplitude‐based measurements. Fluorescence‐based methods for primary production rely on measurements of ETRs and then conversion to carbon fixation rates by using the electron yields of carbon fixation. The electron yields exhibit 10‐fold variability in natural phytoplankton communities and are strongly affected by nutrient limitation. Our results reveal that a decrease in the growth rates and the electron yields of carbon fixation is driven by, and can be quantified from, a decrease in photosynthetic turnover rates. We propose an algorithm to deduce the electron yields of carbon fixation, which greatly improve fluorescence‐based measurements of primary production and growth rates.

Cogongrass (Imperata cylindrica) Leaf Extract Counteracts the Effects of Nitrogen Limitation and Induces Changes in Biomass, Lipid, and Fatty Acid Profile in Chlorella vulgaris UMT-M1 and Scenedesmus sp. KS-2

This study assessed the feasibility of unhydrolyzed cogongrass as a medium fortificant for cultivation of Chlorella vulgaris UMT-M1 and Scenedesmus sp. KS-2. Nitrogen-limited media supplemented with 0.1–0.5% (w/v) cogongrass leaf extract induced changes in lipid content and produced effects that appear to counteract the effects of nitrogen depletion, restoring lipid and fatty acid content to levels similar to nitrogen-replete media while biomass was decreased. While nitrogen-limited media led to an increase in total lipid, oleic acid (C18:1) and monounsaturated fatty acid (MUFA) content and a decrease in palmitic acid (C16:0), polyunsaturated fatty acid (PUFA) and saturated fatty acid (SFA) content, the presence of cogongrass extract evidently cancelled this effect. As a sole ingredient, cogongrass reduced biomass, C16:0, PUFA and SFA, but increased total oil, C18:1 and MUFA to levels similar to N-limited media. Unexpectedly, a 45.8% increase in lipid productivity in Scenedesmus sp. under N-limitation was observed, despite a reduction in biomass. This study demonstrates that the extract from cogongrass and likely from other plant sources could potentially be used as a medium fortificant for microalga cultivation for lipid production. It could also be used as part of a cultivation strategy towards the production of desired fatty acid composition for biodiesel applications. In addition, this study could spark new research for alternative medium fortificants for microalga cultivation from various unutilized or waste plant biomass resources.

Production of carbohydrates, lipids and polyunsaturated fatty acids (PUFA) by the polar marine microalga Chlamydomonas malina RCC2488

Polar microalgae that are highly productive in cold climates can produce large amounts of biomass and polyunsaturated fatty acids (PUFA). The polar Chlamydomonas malina RCC2488, grows at low temperatures and produces high amounts of lipids, which are mainly composed of PUFA. However, not much is known about its phylogenetic relationship with other strains within the order Chlamydomonadales and the optimum growth conditions for maximum biomass productivity have not yet been identified. In this study, a phylogenetic analysis was performed to determine the closest relatives of C. malina within the Chlamydomonadales order. To select the best growth conditions for maximum biomass productivities in cultivations performed at 8 °C, different salinities (0–80) and light intensities (70–500 μmol photons m−2 s−1) were tested, using bubble column and flat-panel photobioreactors. The effect of nitrogen limitation was tested to determine if C. malina can accumulate energy reserve metabolites (carbohydrates and lipids). Phylogenetic analysis confirmed that C. malina, which belongs to the Chlamydomonales order, is closely related to the psychrophilics Chlamydomonas sp. UWO 241 and Chlamydomonas sp. SAG 75.94, as well as to the mesophilic C. parkeae MBIC 10599. The highest biomass (527 mg L−1 day−1), lipid (161.3 mg L−1 day−1) and polyunsaturated fatty acids (PUFA; 85.4 mg L−1 day−1) productivities were obtained at a salinity of 17.5, light intensity of 250 μmol photons m−2 s−1 and nitrogen replete conditions. Strikingly, the marine C. malina can grow even in fresh water, but the biomass productivity was reduced. While the intracellular lipid content remained unchanged under nitrogen deprivation, the carbohydrate content increased (up to 49.5% w/w), and the protein content decreased. The algal lipids were mainly comprised of neutral lipids, which were primarily composed of PUFA. Chlamydomonas malina RCC2488 is a polar marine microalga suitable for high biomass, carbohydrate, lipid and PUFA productivities at low temperatures.

Effect of nitrogen limitation on antioxidant qualities is highly associated with genotypes of lettuce (Lactuca sativa L.)

Antioxidants are highly beneficial to human health, and their accumulation in lettuce, one of the most popular leafy vegetables, depends on both genetic and environmental factors. Nitrogen (N) availability plays an essential role in regulating antioxidant accumulation, but the influence of genotype × N interactions on the antioxidant qualities of lettuce is poorly understood. Therefore, the present study investigated the variation of growth and antioxidant qualities of 20 lettuce (Lactuca sativa L.) genotypes (10 green lettuce genotypes and 10 red lettuce genotypes) under limited N (low N, LN) conditions and standard N (high N, HN) conditions. For all 20 genotypes, LN conditions reduced shoot (i.e., leaf) growth, but increased plant concentrations of vitamin C, glutathione, and phenolic compounds, with the exception of carotenoids, compared with HN conditions. Because of reduced biomass under LN conditions, not all lettuce genotypes exhibited increased antioxidant yields or total antioxidant capacity yield. The variation in antioxidant quality was primarily genetically determined. Generally, the green lettuce genotypes exhibited more pronounced increases in antioxidant yields and total antioxidant capacity yield than the red lettuce genotypes under LN conditions. These results suggest that even though LN conditions generally tend to improve the antioxidant qualities of lettuce, the extent of this effect is highly dependent on genotype. Therefore, genotype should be given priority in future studies that aim to improve antioxidant qualities in lettuce through N management.

Boosting algal lipids: Diurnal shifts in temperature exceed the effects of nitrogen limitation

Algal lipids have been observed to increase during autumn conditions (low light, low mean temperature, and diurnal shift in temperature), in large‐scale outdoor photobioreactors. In this paper, we tested the effect of diurnal shifts in temperature (DS) and nitrogen (N) limitation on algal BODIPY lipid fluorescence cell−1 (BPF). We show that DS increased BPF in algal biomass up to 28% more compared with N limitation, the standard stressor to boost neutral lipids (NL) in commercial production. Biomass yield was constant, regardless the DS range (6°C‐12°C). A combination of both stressors had an additive effect on algal BPF. A polyculture from an outdoor photobioreactor was cultivated under controlled conditions at different regimes of light, temperature, and N limitation. DSs were mimicking autumn conditions with a difference of 6°C, 10°C, and 12°C between day and night. Biomass and BPF were monitored over one to two weeks, and NLs were stained with a fluorescent marker (BODIPY) and detected with flow cytometry. Results indicate that, during autumn conditions, daily heating and cooling processes in contrast to N limitation do not challenge the trade‐off between biomass production and BPF. During seasons when day temperature is still relatively high, DSs are rapid BPF boosting stressors, while N limitation could be applied to boost BPF further during other seasons.

Factors Controlling Seasonal Phytoplankton Dynamics in the Delaware River Estuary: an Idealized Model Study

Phytoplankton biomass in estuaries is controlled by complex biological and chemical processes that control growth and mortality, and physical processes that control transport and dilution. The effects of these processes on phytoplankton blooms were systematically analyzed, focusing on identifying the dominant controlling factors out of river-induced flushing, tidal dispersion, nutrient limitation, and light limitation. To capture the physical processes related to flow and sediment dynamics, we used the idealized width-averaged iFlow model. The model was extended with a nutrient-phytoplankton module to capture the essential biological-chemical processes. The model was applied to the Delaware River Estuary for the productive months of March to November. Model results were compared with field observations. It was found that phytoplankton blooms cannot form in the lower bay due to tidal dispersion, as water from the estuary and coastal ocean mix in early spring, and due to local effects of nitrogen limitation in summer. In the middle to upper bay, sediment-induced deterioration of the light climate limits the growth but allows for blooms in the mid bay, while no blooms can form in the turbidity maximum zone in the upper estuary. Further upstream in the tidal river, the effects of river-induced flushing dominate in early spring and prevent bloom formation. In the summer and fall, lower river discharges and higher growth rates at higher temperatures allow blooms to form and persist. Analysis of the connectivity between mid bay and tidal river blooms showed that coastal ocean phytoplankton may contribute to mid bay blooms, but do not penetrate beyond the turbidity maximum zone.

Effect of nitrogen limitation on polyhydroxyalkanoates production efficiency, properties and microbial dynamics using a soil-derived mixed continuous culture

ABSTRACTIn the present study, a soil-derived mixed microbial consortium was developed for polyhydroxyalkanoates (PHAs) production via nutrient limitation. The enhanced consortium was then cultured continuously in sequential batch and the effect of different nitrogen to carbon ratios (N:C) on the yields and properties of produced PHAs and on the changes of the microbial population were investigated. In all cases, the produced polymers were identified as blends or co-polymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) units, with 3HB being the dominant monomer. The degradation profile and the transition temperatures of the produced PHAs were further assessed and compared, as well as the molecular weights which ranged from 77.104 Da to 180.104 Da. In order to further investigate the effect of culturing conditions on the development and alteration of the microbial consortium, the Nile blue live staining was applied in the mixed cultures at the end of each operational period. Various fluorescent single colonies were selected based on differences in their morphology and were identified. Finally, the alterations in microbial diversity and community composition assessed via the RISA profiling method (rRNA Intergenic Spacer Analyses), resulted in the identification of a number of dominant organisms.

Genotoxic, Metabolic, and Oxidative Stresses Regulate the RNA Repair Operon of Salmonella enterica Serovar Typhimurium.

The σ54 regulon in Salmonella enterica serovar Typhimurium includes a predicted RNA repair operon encoding homologs of the metazoan Ro60 protein (Rsr), Y RNAs (YrlBA), RNA ligase (RtcB), and RNA 3'-phosphate cyclase (RtcA). Transcription from σ54-dependent promoters requires that a cognate bacterial enhancer binding protein (bEBP) be activated by a specific environmental or cellular signal; the cognate bEBP for the σ54-dependent promoter of the rsr-yrlBA-rtcBA operon is RtcR. To identify conditions that generate the signal for RtcR activation in S Typhimurium, transcription of the RNA repair operon was assayed under multiple stress conditions that result in nucleic acid damage. RtcR-dependent transcription was highly induced by the nucleic acid cross-linking agents mitomycin C (MMC) and cisplatin, and this activation was dependent on RecA. Deletion of rtcR or rtcB resulted in decreased cell viability relative to that of the wild type following treatment with MMC. Oxidative stress from peroxide exposure also induced RtcR-dependent transcription of the operon. Nitrogen limitation resulted in RtcR-independent increased expression of the operon; the effect of nitrogen limitation required NtrC. The adjacent toxin-antitoxin module, dinJ-yafQ, was cotranscribed with the RNA repair operon but was not required for RtcR activation, although YafQ endoribonuclease activated RtcR-dependent transcription. Stress conditions shown to induce expression the RNA repair operon of Escherichia coli (rtcBA) did not stimulate expression of the S Typhimurium RNA repair operon. Similarly, MMC did not induce expression of the E. colirtcBA operon, although when expressed in S Typhimurium, E. coli RtcR responds effectively to the unknown signal(s) generated there by MMC exposure.IMPORTANCE Homologs of the metazoan RNA repair enzymes RtcB and RtcA occur widely in eubacteria, suggesting a selective advantage. Although the enzymatic activities of the eubacterial RtcB and RtcA have been well characterized, the physiological roles remain largely unresolved. Here we report stress responses that activate expression of the σ54-dependent RNA repair operon (rsr-yrlBA-rtcBA) of S Typhimurium and demonstrate that expression of the operon impacts cell survival under MMC-induced stress. Characterization of the requirements for activation of this tightly regulated operon provides clues to the possible functions of operon components in vivo, enhancing our understanding of how this human pathogen copes with environmental stressors.

Optimization of Culture Condition for Ganoderic Acid Production in Ganoderma lucidum Liquid Static Culture and Design of a Suitable Bioreactor.

Ganoderma lucidum, a famous medicinal mushroom used worldwide, is a rich source of triterpenoids which, together with polysaccharides, are believed to be the main effective constituents of G. lucidum. With the increase of market demand, the wild resource is facing serious limitations, and the quality of cultivated fruiting bodies can be seriously affected by the availability of wood resources and by cultivation management practices. In the present study, we aimed to develop an alternative way to produce useful triterpenoids from G. lucidum. We cultured the strain using a two-stage liquid culture strategy and investigated the effects of nitrogen limitation, carbon supply, static culture volume and air supply in the static culture stage on the accumulation of five triterpenoids (GA-P, GA-Q, GA-T, GA-S, GA-R). Our results showed that, under optimized condition, the total yield of the five triterpenoids reached 963 mg/L (as determined by HPLC). Among the five triterpenoids, GA-T accounted for about 75% of the total yield. Besides, a bioreactor suitable for fungal liquid static culture with a 10 L extensible plastic bag shaped culture unit was designed and in which the maximum total yield of the five GAs reached 856.8 mg/L, and the GAs content reached 5.99%. Our results demonstrate the potential of industrial application of G. lucidum culture for the production of triterpenoids, especially GA-T. Air supply significantly improved the accumulation of triterpenoids, and this will provide important clues to understand why more triterpenoids are produced in the mycelia mat under static liquid culture conditions.