Nitrogen-deficient Conditions Research Articles

Genome-Wide Identification and Expression Analysis of PkNRT Gene Family in Korean Pine (Pinus koraiensis)

The utilization of nitrogen (N) is crucial for the optimal growth and development of plants. As the dominant form of nitrogen in temperate soil, nitrate (NO3−) is absorbed from the soil and redistributed to other organs through NO3− transporters (NRTs). Therefore, exploration of the role of NRTs in response to various NO3− conditions is crucial for improving N utilization efficiency (NUE). Here, we present a comprehensive genome-wide analysis and characterization of the NRT gene family in Korean pine, an invaluable tree species cultivated extensively in northeastern China. A total of 76 PkNRTs were identified in Korean pine and further divided into three subfamilies (NRT1/NPF, NRT2, and NRT3) based on phylogenetic analysis. All PkNRTs were distributed on 11 chromosomes, with multiple tandem duplications observed. The tissue-specific expression analysis indicated that most PkNRTs showed differential expression in six vegetative tissues. Furthermore, a significantly greater number of lateral roots was observed in seedlings under nitrogen-deficient conditions, accompanied by an increase in both total root biomass and root length. The temporal expression profiles of 16 PkNRTs in seedling roots revealed that four PkNRTs, PkNPF5.6, PkNPF5.13, PkNPF6.1, and PkNPF6.2, exhibited significantly upregulated expression under the NO3− deficiency condition, whereas robust induction was observed for PkNPF1.1, PkNRT2.6, and PkNRT3.3 upon the NO3− sufficiency condition. The expression patterns of the PkNRTs suggest their potential diverse roles as key participants in root NO3− uptake under varying NO3− conditions during root development. These findings would provide a theoretical foundation for further investigations into the functions of PkNRTs in Korean pine.

Comparative metabolomic analysis of Haematococcus pluvialis during hyperaccumulation of astaxanthin under the high salinity and nitrogen deficiency conditions.

Revealing the differences of metabolite profiles of H. pluvialis during hyperaccumulation of astaxanthin under the high salinity and nitrogen deficiency conditions was the key issues of the present study. To investigate the optimum NaCl and NaNO3 concentration and the corresponding metabolic characteristic related to the astaxanthin accumulation in H. pluvialis, a batch culture experiment was conducted. The results indicated that 7.5g·L- 1 and 0g·L- 1 (nitrogen deficiency) were the optimum NaCl and NaNO3 levels for the astaxanthin accumulation respectively, under which the highest astaxanthin contents reached up to 7.51mg·L- 1 and 5.60mg·L- 1. A total of 132 metabolites were identified using LC-MS/MS technique, among which 30 differential metabolites with statistical significance were highlighted. Subsequently, 18 and 10 differential metabolic pathways in the high salinity (HS) and nitrogen-deficient (ND) treatments were extracted and annotated respectively. The values of Fv/Fm, Yield and NPQ were all at the highest level in the ND group during the experiment. The levels of the metabolites in the ND group were almost lower than those both in the control (CK) and HS group, while which in the HS group were substantially at the higher or close levels compared to the CK group. Finally, 7 metabolic markers related to the astaxanthin accumulation were highlighted in the HS and ND group respectively. L-Proline, L-Aspartate, Uridine 5'-monophosphate (UMP), Succinate, L-2-Hydroxygluterate, L-Valine and Inosine 5'-monophosphate (IMP) were identified as the metabolic markers in the HS group, whose fold change were 0.85, 4.14, 0.31, 0.66, 3.10, 1.32 and 0.30. Otherwise, the metabolic markers were Glyceric acid, Thymine, sn-Glycerol 3-phosphate, Glycine, Allantoic acid, L-Valine and IMP in the ND group, with the fold change 0.23, 2.11, 0.38, 0.41, 0.50 and 2.96 respectively. The results provided the comparative metabolomic view of astaxanthin accumulation in H. pluvialis under the different cultivation conditions, moreover showed a novel insights into the astaxanthin production.

Integrated transcriptomics and metabolomics analyses provide new insights into cassava in response to nitrogen deficiency

Nitrogen deficiency is a key constraint on crop yield. Cassava, the world’s sixth-largest food crop and a crucial source of feed and industrial materials, can thrive in marginal soils, yet its yield is still significantly affected by limited nitrogen availability. Investigating cassava’s response mechanisms to nitrogen scarcity is therefore essential for advancing molecular breeding and identifying nitrogen-efficient varieties. This research undertook a comprehensive analysis of cassava seedlings’ physiological, gene expression, and metabolite responses under low nitrogen stress. Findings revealed that nitrogen deficiency drastically suppressed seedling growth, significantly reduced nitrate and ammonium transport to aerial parts, and led to a marked increase in carbohydrate, reactive oxygen species, and ammonium ion levels in the leaves. Transcriptomic and metabolomic analyses further demonstrated notable alterations in genes and metabolites linked to carbon and nitrogen metabolism, flavonoid biosynthesis, and the purine metabolic pathway. Additionally, several transcription factors associated with cassava flavonoid biosynthesis under nitrogen-deficient conditions were identified. Overall, this study offers fresh insights and valuable genetic resources for unraveling cassava’s adaptive mechanisms to nitrogen deprivation.

Nitrogen stress-induced modulation of antimicrobial and antioxidant activities in Aphanothece sp.: Insights from phytochemical profiling and molecular docking

The modulation of nutrient availability represents a critical factor influencing the metabolic pathways and production of bioactive compounds in algae. This study aimed to examine the impact of nitrogen stress on the antimicrobial and antioxidant properties, phytochemical composition, and molecular docking assessments of the cyanobacterium Aphanothece sp. The results demonstrated that Aphanothece sp. when cultivated under conditions of nitrogen deficiency, exhibited markedly elevated antimicrobial and antioxidant activity compared to the control. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of the extracts obtained from algae on E. coli ATCC 25922 were found to be 25 mg mL−1 when the sample was grown in nitrogen medium and 6.25 mg mL−1 when grown in a nitrogen-free medium. The IC50 value obtained from DPPH free radical scavenging assay changed from 137.30 ± 16.22 mg mL−1 to 20.07 ± 1.55 mg mL−1 in nitrogen-free medium. According to the results of GC–MS analysis, the major components in the extract obtained from the algae grown in the nitrogen-free environment were 13,16-Octadecadiynoic acid, methyl ester (35.10 %) and Hexadecanoic acid, methyl ester (22.65 %), while the major components in the extract obtained from the algae grown in nitrogen-containing medium were Protopine (65.68 %) and 3,6,9,12,15-Pentaoxabicyclo(15.3.1)henicosa-1(21),17,19-triene-2,16-dione (12.15 %). The findings suggest a range of binding energies and interaction profiles for the compounds in the extracts with their molecular targets, which may have implications for their potential significance in therapeutic applications. The results of this study indicate that nitrogen stress can be employed as a strategic tool to enhance the production of valuable bioactive compounds in Aphanothece sp., thereby underscoring its potential applications in biotechnology and drug development.

Deciphering nitrogen dynamics in aeroponics: physio-biochemical and enzymatic responses influencing nitrogen use efficiency in contrasting potato genotypes

Excessive use of nitrogen (N) in crops, such as potatoes, can lead to economic and environmental repercussions. We hypothesized that potato genotypes with resilient root systems and high genetic capabilities for nitrogen-use efficiency (NUE) could effectively mitigate these challenges. Consequently, we investigated intraspecific variations and characteristics within six distinct potato genotypes exhibiting diverse NUEs in response to varying nitrogen levels in an aeroponic system. The morpho-physiological and biochemical properties showed significant genotypic variations, especially related to the N-assimilating enzyme levels and root characteristics. Notably, the root systems of all genotypes demonstrated greater responsiveness to low nitrogen levels, with genotype C17 showcasing the most substantial root system irrespective of nitrogen concentration. Root morphological traits displayed robust positive correlations with NUtE, primarily influenced by genotype rather than nitrogen concentration. Conversely, nitrogen levels, displaying positive correlations with NUpE, influenced growth and activities of N-assimilating enzymes. Based on their distinct root systems, metabolic activities, and NUE profiles, genotypes C17 and C11 were determined to be N-efficient and N-inefficient, respectively. This study provides novel insights into the physiological and biochemical mechanisms underlying nitrogen use efficiency in potato genotypes under aeroponic conditions, offering potential targets for breeding programs, optimizing fertilizer management and cultivation strategies to improve crop performance under nitrogen-deficient conditions. Future investigations, employing multi-omics approaches, will elucidate key genes and pathways in nitrogen metabolism, potentially offering avenues to enhance root architecture and improve NUE.

High light and nitrogen deficiency synergistically lead to biofuel and biomass accumulation in diatoms

As a clean energy source, the lipids from microalgae serve as alternatives to fossil oils. There are significant differences in the biofuel and biomass accumulation of microalgae under different environmental conditions. This study investigated the response of biofuel accumulation, photosynthetic capacity, and growth of diatom Thalassiosira pseudonana under stress conditions of high light and nitrogen deficiency, aiming to explore the features of biofuel and biomass accumulation in microalgae under special environmental conditions. The results indicated that the synthesis of chlorophyll was inhibited, leading to reduced light absorption and less damage to the photosynthetic system, with an improvement in electron transport efficiency. Lipid metabolism was more active during the dark cycle, with a 2.89-fold accumulation of fatty acids and a significant increase in the content of neutral lipids (2.39 times). The cell cycle progressed faster, resulting in a marked increase in biomass (1.41 times). These processes consumed a considerable amount of energy, affecting the synthesis of ribosomes. These findings suggest that increasing light exposure and reducing nitrogen supply can achieve the co-accumulation of biofuel and biomass in T. pseudonana.

Nitrogen Nutrition-Induced Changes in Macronutrient Content and Their Indirect Effect on N-Metabolism Via an Impact on Key N-Assimilating Enzymes in Bread Wheat (Triticum aestivum L.)

Judicious application of nitrogen (N) fertilizers in crop production is critical for reducing the nitrate pollution of groundwater and greenhouse gas emissions. It is, thus, important to improve the nitrogen use efficiency under the reduced application of nitrogen. A genotypic variation in N-uptake and N-use efficiency particularly under low N-input conditions exists across crops that can be deciphered and exploited for environmentally sustainable farming without any significant penalty of yield and quality. The present research conducted under the nutrient solution culture aimed to explore the inherent variability in the growth response of ten genetically diverse wheat varieties to low fertilizer N-application (N-, 10 μM N) in comparison to N sufficient control (N+, 8.5 mM N) viz., a viz., the activity of various key N-assimilating enzymes and to delineate the indirect effect of low N on uptake and partitioning of other major macronutrients viz., P, K, S, which may indirectly regulate the N-use efficiency. A notable increase in sulfur, potassium, and phosphorus content was observed under nitrogen-deficient conditions. Varieties such as Carnamah and HD 2824 exhibit a significant increase in shoot phosphorus content, emphasizing their potential to optimize phosphorus acquisition and utilization efficiency under nutrient-limited conditions. The findings highlight the complex interplay between nutrient availability and plant responses, showcasing varietal-specific adaptations to nitrogen limitations.

Metabolic profiling reveal changes in shoots and roots of nitrogen-deficient tea plants (Camellia sinensis cv. Jinxuan)

Tea plants require high levels of nitrogen for optimal growth. However, delayed or insufficient nitrogen supply often induces nitrogen deficiency stress. This stress can adversely affect the plant's growth, development, and metabolic processes. Therefore, studying how tea plants respond to nitrogen deficiency conditions is crucial for enhancing our understanding and management of their growth environment. In this study, we utilized High-Performance Liquid Chromatography (HPLC), Gas Chromatography-Mass Spectrometry (GC–MS), and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS) techniques to analyze the non-volatile and volatile components of both shoots and roots of the 'Jing Xuan (JX)' tea variety under nitrogen deprivation for 30 days (Nd). The results revealed that nitrogen deprivation led to a deceleration in shoot growth of 'JX', accompanied by reduced chlorophyll and water-soluble extract contents, with nitrogen accumulating predominantly in the roots. Significant decreases were observed in caffeine content in shoots and in the total amounts of theanine and catechins in both shoots and roots, while epigallocatechin gallate (EGCG) accumulation increased in shoots. Moreover, we identified 65 volatile components in shoots, including a notable decrease in floral and fruity compounds such as trans-β-Ionone, Geranylacetone, and α-Ionone. The biosynthesis of amino acids and phenylpropane metabolism are more active pathways under nitrogen stress. Nitrogen deficiency led to reduced levels of amino acids, carboxylic acids, flavonoids, steroids, and their derivatives in roots, with the most notable decline observed in carboxylic acids, typified by Leucine, l-Aspartate, and l-histidine. In contrast, shoots accumulate carbohydrates and phenolic compounds, mainly sucrose. Additionally, metabolites such as Myristic acid and 3-Dehydroshikimate could be a critical indicator of stress levels. These findings enhance our understanding of tea plant growth and metabolism under nitrogen deficiency, providing valuable insights for breeding nitrogen-efficient tea varieties.

Modification of Growth Media for Scenedesmus and Nitchzer Species for Fatty Acid Methyl Esters Production in Raceway Pond

The aim of this study was to modify the growth media for Scenedesmus and Nitchzia species for fatty acid methyl esters production in raceway pond. Native Scenedesmus and Nitchzia species were cultivated in raceway pond in three different growth media (Nitrogen-sufficient, Nitrogen-deficient and Blue Green 11) media at different periods to assess their quantitative parameters (biomass concentration (g/L), biomass productivity (g/L/d), lipid content (%), and lipid productivity (mg/L/d) as well as their fatty acid methyl esters composition. The quantitative parameters were determined following standard methods and the fatty acid methyl esters composition was analysed by Gas Chromatography Mass Spectrometry (GC-MS). The results of the quantitative parameters obtained from Scenedesmus species were highest in the Blue Green 11 (control) media as compared to the experimental media. Statistically, there is significant difference (p<0.05) in the lipid content obtained from Scenedesmus species between the Blue Green 11 media and the experimental media. The biomass concentration and lipid content obtained from Nitchzia species were also found to be highest in Blue Green 11 media as compared to the other two media. However, the lipid content obtained from Nitchzia species in the three different media showed insignificant difference (p>0.05). Scenedesmus and Nitchzia species did not respond to nitrogen deficiency condition. The Gas Chromatography Mass Spectrometry analysis of the fatty acid methyl esters composition revealed the presence of similar compounds in different proportion. The dominant compound of the fatty acid methyl esters of the two species in the three different media are Hexadecanoic acid methyl ester ((palmitic acid)), 9-octadecenoic acid (Z) methyl ester ((oleic acid) and 9, 12-octadecadienoic acid methyl ester (linolenic acid) among others. The Carbon16-Carbon18 (C16-C18) chain in Scenedesmus and Nitchzia species ranges from 74.14-61.84% and 63.81-61.98% respectively. These could suggest that both species have a great potential for the production of fatty acid methyl esters in large scale cultivation.

Heterologous glucose-6-phosphate dehydrogenase empowers the biofuel potential of Tetradesmus obliquus via generating lipogenic NADPH

Large-scale production of biofuels from oleaginous microalgal feedstocks in a commercially feasible manner is hampered by their relatively slow growth rates and overall lipid yield than the rapidly proliferating algal species. Therefore, it is pivotal to obviate this bottleneck by augmenting lipid overproduction in fast-growing microalgal species by heterologous overexpression lipogenic genes from model oleaginous species. In this regard, the pentose phosphate pathway of Tetradesmus obliquus was perturbed by heterologous overexpression of glucose-6-phosphate dehydrogenase (PtG6PD) from the model pennate diatom Phaeodactylum tricornutum, known for its oleaginicity. The role of PtG6PD in increasing lipid production was reported in the heterologous T. obliquus. Total lipid content was increased by 2.09- and 2.31-fold in transgenic lines than WT by elevating the lipogenic NADPH, without any negative impact on their growth rates. Interestingly, lipid yields in the transgenic strains reached up to 93 mg/L and 103 mg/L which showed a further 2-fold enhancement under nitrogen-deficient conditions. Furthermore, it was demonstrated that PtG6PD overexpression also altered fatty acid composition, with a specific increment in total monounsaturated fatty acids. Our results uncovered the pivotal role of the pentose phosphate pathway in supplying lipogenic NADPH in T. obliquus and identified key genetic targets for lipid biosynthesis enhancement in alternative hosts. Altogether, these findings open avenues for metabolic engineering in non-model and fast-growing non-oleaginous microalgal species, thereby presenting a promising strategy for empowering commercially viable photosynthetic cell factories for biofuel production.

Identification of novel marker-trait associations and candidate genes for combined low phosphorus and nitrogen-deficient conditions in rice at seedling stage

Identification of novel marker-trait associations and candidate genes for combined low phosphorus and nitrogen-deficient conditions in rice at seedling stage

Transcriptional, post-transcriptional, and post-translational regulation of polyunsaturated fatty acid synthase genes in Aurantiochytrium limacinum strain BL10: Responses to nitrogen starvation

Under nitrogen deficient conditions, the Aurantiochytrium limacinum strain BL10 greatly increases the production of docosahexaenoic acid (DHA) and n-6 docosapentaenoic acid. Researchers have yet to elucidate the mechanism by which BL10 promotes the activity of polyunsaturated fatty acid synthase (Pfa), which plays a key role in the synthesis of polyunsaturated fatty acid (PUFA). Analysis in the current study revealed that in nitrogen-depleted environments, BL10 boosts the transcription and synthesis of proteins by facilitating the expression of pfa genes via transcriptional regulation. It was also determined that BL10 adjusts the lengths of the 5′- and 3′-untranslated regions (suggesting post-transcriptional regulation) and modifies the ratio of two Pfa1 isoforms to favor PUFA production via post-translational regulation (ubiquitination). These findings clarify the exceptional DHA production of BL10 and provide additional insights into the regulatory mechanisms of PUFA biosynthesis in Aurantiochytrium.

Nitrogen starvation causes lipid remodeling in Rhodotorula toruloides

BackgroundThe oleaginous yeast Rhodotorula toruloides is a promising chassis organism for the biomanufacturing of value-added bioproducts. It can accumulate lipids at a high fraction of biomass. However, metabolic engineering efforts in this organism have progressed at a slower pace than those in more extensively studied yeasts. Few studies have investigated the lipid accumulation phenotype exhibited by R. toruloides under nitrogen limitation conditions. Consequently, there have been only a few studies exploiting the lipid metabolism for higher product titers.ResultsWe performed a multi-omic investigation of the lipid accumulation phenotype under nitrogen limitation. Specifically, we performed comparative transcriptomic and lipidomic analysis of the oleaginous yeast under nitrogen-sufficient and nitrogen deficient conditions. Clustering analysis of transcriptomic data was used to identify the growth phase where nitrogen-deficient cultures diverged from the baseline conditions. Independently, lipidomic data was used to identify that lipid fractions shifted from mostly phospholipids to mostly storage lipids under the nitrogen-deficient phenotype. Through an integrative lens of transcriptomic and lipidomic analysis, we discovered that R. toruloides undergoes lipid remodeling during nitrogen limitation, wherein the pool of phospholipids gets remodeled to mostly storage lipids. We identify specific mRNAs and pathways that are strongly correlated with an increase in lipid levels, thus identifying putative targets for engineering greater lipid accumulation in R. toruloides. One surprising pathway identified was related to inositol phosphate metabolism, suggesting further inquiry into its role in lipid accumulation.ConclusionsIntegrative analysis identified the specific biosynthetic pathways that are differentially regulated during lipid remodeling. This insight into the mechanisms of lipid accumulation can lead to the success of future metabolic engineering strategies for overproduction of oleochemicals.

Genotype-specific nonphotochemical quenching responses to nitrogen deficit are linked to chlorophyll a to b ratios

Non-photochemical quenching (NPQ) protects plants from photodamage caused by excess light energy. Substantial variation in NPQ has been reported among different genotypes of the same species. However, comparatively little is known about how environmental perturbations, including nutrient deficits, impact natural variation in NPQ kinetics. Here, we analyzed a natural variation in NPQ kinetics of a diversity panel of 225 maize (Zea mays L.) genotypes under nitrogen replete and nitrogen deficient field conditions. Individual maize genotypes from a diversity panel exhibited a range of changes in NPQ in response to low nitrogen. Replicated genotypes exhibited consistent responses across two field experiments conducted in different years. At the seedling and pre-flowering stages, a similar portion of the genotypes (∼33%) showed decrease, no-change or increase in NPQ under low nitrogen relative to control. Genotypes with increased NPQ under low nitrogen also showed greater reductions in dry biomass and photosynthesis than genotypes with stable NPQ when exposed to low nitrogen conditions. Maize genotypes where an increase in NPQ was observed under low nitrogen also exhibited a reduction in the ratio of chlorophyll a to chlorophyll b. Our results underline that since thermal dissipation of excess excitation energy measured via NPQ helps to balance the energy absorbed with energy utilized, the NPQ changes are the reflection of broader molecular and biochemical changes which occur under the stresses such as low soil fertility. Here, we have demonstrated that variation in NPQ kinetics resulted from genetic and environmental factors, are not independent of each other. Natural genetic variation controlling plastic responses of NPQ kinetics to environmental perturbation increases the likelihood it will be possible to optimize NPQ kinetics in crop plants for different environments.

Overexpression of rice OsNRT1.1A/OsNPF6.3 enhanced the nitrogen use efficiency of wheat under low nitrogen conditions.

Overexpression of OsNRT1.1A promotes early heading and increases the tolerance in wheat under nitrogen deficiency conditions. The application of inorganic nitrogen (N) fertilizers is a major driving force for crop yield improvement. However, the overuse of fertilizers significantly raises production costs and leads to environmental problems, making it critical to enhance crop nitrogen use efficiency (NUE) for the sake of sustainable agriculture. In this study, we created a series of transgenic wheat lines carrying the rice OsNRT1.1A gene, which encodes a nitrate transporter, to investigate its possible application in improving NUE in wheat. The transgenic wheat exhibited traits such as early maturation that were highly consistent with the overexpression of OsNRT1.1A in Arabidopsis and rice. However, we also observed that overexpression of the OsNRT1.1A gene in wheat can facilitate the growth of roots under low N conditions but has no effect on other aspects of growth and development under normal N conditions. Thus, it may lead to the improvement of wheat low N tolerance,which is different from the effects reported in other plants. A field trial analysis showed that transgenic wheat exhibited increased grain yield per plant under low N conditions. Moreover, transcriptome analysis indicated that OsNRT1.1A increased the expression levels of N uptake and utilization genes in wheat, thereby promoting plant growth under low N conditions. Taken together, our results indicated that OsNRT1.1A plays an important role in improving NUE in wheat with low N availability.

Physiological and Transcriptional Characteristics of Banana Seedlings in Response to Nitrogen Deficiency Stress

Nitrogen is a crucial element for the growth and development of plants, directly affecting crop growth and yield. To investigate the physiological and molecular mechanism of nitrogen-deficiency stress, we conducted an investigation into the effects of different nitrogen levels on the growth, photosynthetic characteristics, and gene transcription levels of banana seedlings. Compared with the control group with normal nitrogen levels (NN), the height of plants receiving Reduced-N (NR), Low-N (LN), and N-Free (NF) treatments was decreased by 0.45 cm, 2.5 cm, and 3.25 cm, respectively. Their dry weight was reduced by 1.63 g, 2.99 g, and 2.88 g, respectively. Conversely, the dry weight of the underground plant part in the LN and NF treatment groups exhibited an increase of 0.13 g and 0.16 g, respectively. Regarding photosynthetic characteristics, the Specialty Products Agricultural Division (SPAD) values of the NR, LN, and NF treatments showed reductions of 15.5%, 30.4%, and 35.9%, respectively, compared with those of the control treatments. The values of maximum photosynthetic efficiency (Fv/Fm), actual photosynthetic efficiency (Y(Ⅱ)), and relative electron transfer (ETR) of the banana seedlings decreased to different degrees after NR, LN, and NF treatment, and their values were positively correlated with N levels. Gene transcription analysis showed that N transport-related proteins, including NRT1.7, NRT2.3a, NRT2.3b, and NRT2.5, were significantly up-regulated to increase the nitrogen absorption capacity of plant roots. On the other hand, various transcription factors including GRAS, MYB, and WRKY were notably up-regulated, facilitating root growth and the expanding root absorption area, thereby enhancing nitrogen uptake. Furthermore, genes associated with endogenous hormone metabolic pathways such as gibberellin (GA), strigolactone (SL), and brassinosteroids (BR) were activated in banana plants subjected to low nitrogen stress, enhancing the plant’s ability to adapt to nitrogen-deficient conditions. These findings offer valuable insights into understanding the transcriptional regulatory mechanisms governing banana responses to low nitrogen stress and breeding new varieties with improved nutrient utilization.

A new isolate cold-adapted Ankistrodesmus sp. OR119838: influence of light, temperature, and nitrogen concentration on growth characteristics and biochemical composition using the two-stage cultivation strategy.

Natural-based chemicals from microalgae such as lipids and pigments are the interests in industries and the bioeconomy. Cold-adapted Ankistrodesmus sp. OR119838, an isolated strain from Cheshmeh-Sabz Lake in northeastern Iran, was cultivated using a two-stage culture strategy under different environmental conditions. With doubling the nitrate concentration at the vegetative stage (170mg/L) and increasing the light intensity (180µmol photons/m2/s) the highest specific growth rate (0.61 ± 0.02 per day) and biomass productivity (121.1 ± 7.2 mg/L/day) were observed at 25°C. In the optimal growth condition Chl a and Chl b contents of Ankistrodesmus sp. OR119838 reached the highest amount (11.07 ± 0.14 and 11.23 ± 0.29 µg/mL, respectively) at 25°C. While carotenoid content correlated negatively with optimum biomass productivity (-0.708) and had the best value (12.23 ± 0.29µg/mL) in nitrogen deficiency (42 mg/L) and intense light conditions (180µmol photons/m2/s) at 15°C. Lipid content was increased with declined nitrate concentration (42mg/L), high light intensity, and 180µmol photons/m2/s at 25°C. The highest percentage of polyunsaturated fatty acids (71.94%) and α-linolenic acid (57.73 ± 6.63%) was observed in conditions with 170mg/L nitrate concentration and low light intensity (40µmol photons/m2/ s) at the low temperature (15°C). While saturated fatty acids content (43.27%) and palmitic acid reached the highest amount under 40µmol photons/m2/s, 42mg/L nitrate at 25°C (35.02 ± 5.33%). Biomass productivity of Ankistrodesmus sp. OR119838, as a cold-adapted strain, decreased by only 8.2% with a 10-degree decline in temperature. Therefore, this strain has good potential to grow in open ponds by tolerating the daily temperature fluctuations.

Effects of two types of Coccomyxa sp. KJ on in vitro ruminal fermentation, methane production, and the rumen microbiota.

Coccomyxa sp. KJ is a unicellular green microalga that accumulates abundant lipids when cultured under nitrogen-deficient conditions (KJ1) and high nitrogen levels when cultured under nitrogen-sufficient conditions (KJ2). Considering the different characteristics between KJ1 and KJ2, they are expected to have different effects on rumen fermentation. This study aimed to determine the effects of KJ1 and KJ2 on in vitro ruminal fermentation, digestibility, CH4 production, and the ruminal microbiome as corn silage substrate condition. Five treatments were evaluated: substrate only (CON) and CON + 0.5% dry matter (DM) KJ1 (KJ1_L), 1.0% DM KJ1 (KJ1_H), 0.5% DM KJ2 (KJ2_L), and 1.0% DM KJ2 (KJ2_H). DM degradability-adjusted CH4 production was inhibited by 48.4 and 40.8% in KJ2_L and KJ2_H, respectively, compared with CON. The proportion of propionate was higher in the KJ1 treatments than the CON treatment and showed further increases in the KJ2 treatments. The abundances of Megasphaera, Succiniclasticum, Selenomonas, and Ruminobacter, which are related to propionate production, were higher in KJ2_H than in CON. The results suggested that the rumen microbiome was modified by the addition of 0.5-1.0% DM KJ1 and KJ2, resulting in increased propionate and reduced CH4 production. In particular, the KJ2 treatments inhibited ruminal CH4 production more than the KJ1 treatments. These findings provide important information for inhibiting ruminal CH4 emissions, which is essential for increasing animal productivity and sustaining livestock production under future population growth.

Bacterial endosymbionts of a nitrogen-fixing yeast Rhodotorula mucilaginosa JGTA-S1 - insights into a yet unknown micro-ecosystem.

Rhodotorula mucilaginosa JGTA-S1 is a yeast strain capable of fixing nitrogen and improving nitrogen nutrition in rice plants because of its nitrogen-fixing endobacteria, namely Stutzerimonas (Pseudomonas) stutzeri and Bradyrhizobium sp. To gain a deeper understanding of yeast endosymbionts, we conducted a whole-genome shotgun metagenomic analysis of JGTA-S1 cells grown under conditions of nitrogen sufficiency and deficiency. Our results showed that the endosymbiont population varied depending on the nitrogen regime. Upon mechanical disruption of yeast cells, we obtained endosymbionts in culturable form viz. Bacillus velezensis and Staphylococcus sp. under nitrogen-replete conditions and Lysinibacillus telephonicus., Brevibacillus sp., and Niallia circulans under nitrogen-depleted conditions. S. stutzeri and Bradyrhizobium sp. the previously reported endosymbionts remained unculturable. The culturable endosymbionts Staphylococcus sp. and Bacillus velezensis appear to possess genes for dissimilatory nitrate reduction (DNRA), an alternative pathway for ammonia synthesis. However, our findings suggest that these endosymbionts are facultative as they survive outside the host. The fitness of the yeast was not affected by curing of these microbes. Curing the yeast diazotrophic endosymbionts took a toll on its fitness. Our results also showed that the populations of S. stutzeri and B. velezensis increased significantly under nitrogen-depleted conditions compared to nitrogen-sufficient conditions. The importance of DNRA and nitrogen fixation is also reflected in the metagenomic reads of JGTA-S1.

Photofermentative Biohydrogen Production Enhancement with Nanoparticle

This study will explore the use of a biological pathway known as photo fermentation for the production of biohydrogen under anoxic and nitrogen deficient conditions due to its high hydrogen production, and to emphasize the role of a dominating enzyme, the nitrogenase enzyme, in catalyzing the process of hydrogen evolution from a medium composed of Purple Non-Sulfur Bacteria (Rhodopseudomonas Palustris) with glycerol as the substrate. For Purple Non-Sulphur Bacteria, the optimal H2 production temperature ranges between 30 °C to 36 °C. Simultaneously, illumination is applied by LED lights and is varied from no string LED illumination, to single string illumination (240 W/m2), and double string illumination (480 W/m2) to simulate and examine performance in outdoor conditions and achieved a maximum of 428% increase in hydrogen productivity compared to previous studies. Consequently, this suggests that it is economically appealing to use the non-growing R. palustris bacterium as the chosen biocatalyst for continuous hydrogen production. These preliminary results constitute the development of continuous H2 production which evolves into the green fuel of the future production.