Biotechnological strategies overcoming limitations to H. pluvialis-derived astaxanthin production and Morocco's potential.

Haematococcus pluvialis is the richest source of natural astaxanthin, but the production of H. pluvialis-derived astaxanthin is usually limited by its slow cell proliferation and astaxanthin accumulation. Efforts to enhance biomass productivity, astaxanthin accumulation, and extraction are ongoing. This review highlights different approaches that have previously been studied in microalgal species for enhanced biomass productivity, as well as optimized methods for astaxanthin accumulation and extraction, and how these methods could be combined to bypass the challenges limiting natural astaxanthin production, particularly in H. pluvialis, at all stages (biomass production, and astaxanthin accumulation and extraction). Biotechnological approaches, such as overexpressing low CO2 inducible genes, utilizing complementary carbon sources, CRISPR-Cas9 bioengineering, and the use of active compounds, for biomass productivity are outlined. Direct astaxanthin extraction from H. pluvialis zoospores and Morocco's potential for microalgal-based astaxanthin production are equally discussed. This review emphasizes the need to engineer an optimized H. pluvialis-derived astaxanthin production system combining two or more of these strategies for increased growth, and astaxanthin productivity, to compete in the larger, lower-priced market in aquaculture and nutraceutical sectors.

Necessity of stirring for outdoor microalgal-bacterial granular sludge process.

As a green process, microalgal-bacterial granular sludge (MBGS) process shows talents in achieving pollutant removal, resource recovery and carbon neutralization. However, when it comes to application, the adequate mixing of MBGS and substrate should be adopted theoretically. Therefore, this study devoted to address the necessity of stirring for MBGS in municipal wastewater treatment. Outdoor performances showed that stirring significantly enhanced both of the photosynthetic efficiency and biomass productivity of MBGS with almost 2-fold increase as compared to non-stirred MBGS, while the average pore size and microalgae-to-bacteria ratio also increased. Consequently, stirring acted as a pivotal role in accelerating pollutants removal, with removals of organics (89.89% COD) and nutrients (99.22% NH4+-N, 92.15% PO43--P) reaching peak levels at 2 h and 6 h, respectively, while removals of organics (87.50% COD) and nutrients (86.11% NH4+-N, 86.76% PO43--P) removal peaked at 8 h for non-stirred MBGS. The improved granule characteristics and microbial compositions due to the stirring were found to be favorable for MBGS to adapting to the changeable weather. Based on the above results, the possible underlying mechanisms of stirring for improving MBGS were illustrated. Overall, stirring positively impacted the photosynthetic efficiency, biomass productivity, pollutant removal and microbial structure for MBGS. This study gains knowledge on stirred MBGS process under outdoor conditions for its future practical application.

Innovative and sustainable cultivation strategy for the production of Spirulina platensis using anaerobic digestates diluted with residual geothermal water.

The following study investigates the possibility of growing the Spirulina platensis (S. platensis) cyanobacteria on two agro-industrial anaerobic digestion (AD) digestates diluted with geothermal water. The two digestates (FAWD: Food and Agricultural Wastes Digestate and CDD: Cheese Diary Digestate) were selected based on their different chemical characteristics, attributed to the type of feedstock and the operating conditions used during the AD process. In the first part of the study, a screening experiment was performed in 200 mL glass tubes to evaluate the appropriate dilution factor to generate the maximum S. platensis growth using both AD digestates individually and geothermal water as sustainable alternative dilution agent. Based on the different growth parameters measured, dilution rates of 5x and 40x were chosen for CDD and FAWD respectively, as a trade-off between growth performances and quantity of water to use. Volumetric productivities of 33 ± 1 mg/L/d and 56 ± 8 mg/L/d combined with maximal concentrations of 0.52 ± 0.02 g/L and 0.69 ± 0.02 g/L were achieved when cultivating S. platensis on CDD and FAWD, respectively. In the second part, the selected experimental results were scaled-up to 6 L flat panels bioreactors and S. platensis biomass productivities of 71 and 101 mg/L/d were obtained for CDD and FAWD, respectively using sodium bicarbonate as inorganic carbon source. When regulating the pH to 8.5 with carbon dioxide (CO2) injection, cultures were able to produce up to 1.13 g/L and 0.79 g/L of S. platensis corresponding to biomass productivities of 81 and 136 mg/L/d for CDD and FAWD, respectively. In addition, S. platensis properly assimilated the ammonium present in the digestate-based culture media, with removal efficiency up to 98% in the case of the CDD substrate. The characterization of the final S. platensis biomass revealed the presence of high concentration of carbohydrates (48.6-70.3 % of dry weight) in the culture supplemented with both AD digestates. The experimental findings show the potential of reusing liquid digestate, CO2 as well as geothermal water for the sustainable production of carbohydrate-rich S. platensis biomass.

Long term outdoor microalgal phycoremediation of anaerobically digested abattoir effluent.

Sufficient and reliable long-term field data on the growth, productivity and nutrient removal rates of microalgal based wastewater treatment system is essential to validate its overall techno-economic feasibility. Here, we investigated the semi-continuous microalgal cultivation of Scenedesmus sp. in anaerobically digested abattoir effluent (ADAE) for 13 months in outdoor raceway ponds operated at 20 cm depth. This study was initiated with three different cultures consisting of 1) monocultures of Chlorella sp., 2) Scenedesmus sp., and 3) an equal mixed concentration of both microalgae species. However, after 15 weeks, Scenedesmus sp. was found to be the most dominant microalgae species in all the different cultures, even completely taking over the Chlorella sp. monoculture. Over the course of summer and early autumn, the average weekly biomass productivity of Scenedesmus sp. cultures was 12.5 ± 0.6 g m-2 d-1 which was 16% and 30% higher than productivities recorded in spring and winter, respectively. All available ammoniacal nitrogen (NH3-N) was found to be exhausted during each growth period with an average 33.6% nitrogen assimilation rate. The average rate of phosphate and COD (chemical oxygen demand) removals were 85.2% and 37.5% throughout the cultivation period. No significant differences were found in carbohydrate, lipid and protein content of Scenedesmus sp. during different seasons of the year. Over 53% increase in biomass productivity can be achieved if CO2 is added to control culture pH at pH 6.5. Here, we successfully demonstrated reliability of continuous long-term cultivation of microalgae in ADAE for simultaneous wastewater treatment and algal biomass production.

The impact of livestock grazing on the evapotranspiration-vegetation biomass relationship in a Southern Hemisphere salt marsh, Buenos Aires (Argentina).

Among the ecosystem services provided by salt marshes is the use of their natural vegetation as pastures for livestock production. As a result, the prediction of biomass productivity can be of great interest for the sustainable management of these environments. Evapotranspiration is one of the variables most used to estimate the yield of green biomass in pastures and crops, which to date has not been examined for natural environments such as salt marshes. We studied the aboveground biomass and species cover variability for two categories (erect and sward plants) in three plots affected by low, moderate, and high cattle grazing. Erect biomass was associated only with Spartina densiflora while for sward plants it gathered a diverse set of prostrate and stoloniferous species with high seasonal turnover. The evapotranspiration was estimated with a coupled surface resistance-Penman-Monteith model developed for these environments. The biomass of the plant categories shows different growth response according to livestock impact. S. densiflora has a slow-growing response after cattle consumption, even with high evapotranspiration. On the other hand, sward plants respond with biomass overproduction to livestock consumption, and a significantly positive relationship to evapotranspiration rate.

Macrophyte biomass productivity for heavy metal adsorption.

The search for low cost adsorbents that have metal-binding capacities has intensified in the last decades. Some natural aquatic macrophytes have been studied as adsorbents to remove heavy metals. Macrophytes ease to propagate converts them into plague for many ecosystems while they are also considered by some activities as a residue; therefore its resignification implies positive environmental effects. Whereas these macrophytes can be obtained from water bodies where they develop naturally, controlled production in greenhouses may be more appropriate for its use as filling in fixed-bed reactors that must operate continuously throughout the year. This work focused on obtaining the macrophytes growth parameters in order to calculate the most proper greenhouse containers dimensions and their required cultivation periods to be employed in a determined fixed-bed reactor with a certain effluent flow to filter. These parameters include: the biomass yield, the area required to obtain certain dry weight of a given biosorbent, and the relative growth and propagation rates. Selected macrophytes species were Azolla pinnata, Salvinia molesta, Limnobium leavigatum, Lemna minor and Pistia stratiotes. The characterization of each biomass and the study of their performance as biosorbents were conducted for the removal of Cu (II), Pb (II) and Cr (VI) ions from aqueous solutions, always procuring the WHO guidelines for drinking water. Azolla pinnata resulted in the species with the highest percentage of dry weight (6.56%), and the lowest values of relative growth and propagation rates. This species was as well the most efficient in removal of Cu and Pb (96.7% and 99.4%, respectively), while Pistia stratiotes was better adsorbent for Cr with a removal of 58.8%. The possibility of metal recovery and macrophyte biomasses reuse was also proven. Given their natural abundance, elemental growth conditions and propagation rates, macrophytes represent a low cost alternative to the most efficient commercial adsorbents.

A comprehensive study on the effect of light quality imparted by light-emitting diodes (LEDs) on the physiological and biochemical properties of the microalgal consortia of Chlorella variabilis and Scenedesmus obliquus cultivated in dairy wastewater.

The effect of light wavelengths on the physiological, biochemical and lutein content of the microalgal consortia Chlorella variabilis and Scenedesmus obliquus was evaluated using different light sources. Among different light treatments, cool-white fluorescent light produced the highest biomass of 673 mg L-1 with a specific growth rate of 0.75 day-1 followed by blue (500 mg L-1; 0.73 day-1). The chlorophyll content was enhanced under blue light (10.7 mg L-1) followed by cool fluorescent light (9.3 mg L-1), whereas the lutein productivity was enhanced under cool fluorescent light (7.22 mg g-1). Protein content of the microalgal consortia was enhanced under all light treatments with the highest protein accumulation under cool-white fluorescent light (~56% of dry mass) closely followed by amber light (52% of dry mass), whereas the carbohydrate content was higher under amber light (~35% of dry mass). The results revealed that the consortia could grow well on diluted dairy wastewater thereby reducing the cost of algal production when compared with the use of inorganic media and a two-phase culture process utilizing cool fluorescent and amber light could be employed for maximizing algal biomass and nutrient composition with enhanced lutein production. The study also emphasizes on the economic efficiency of LED lights in terms of biomass produced based on the modest electricity consumed and the importance of using amber light for cultivating microalgae for its nutrient content which has seldom been studied.

Strategic implementation of phosphorus repletion strategy in continuous two-stage cultivation of Chlorella sp. HS2: Evaluation for biofuel applications.

Lipid production in microalgae under nitrogen (N) starved condition can be enhanced by excess phosphorus (P) supply in the second stage of two-stage cultivation strategy. However, implementing two-stage cultivation is difficult in large-scale cultivation system as it requires high energy of transferring large algal biomass from first stage to second stage. To address this problem, we have optimized a continuous two-stage (CTS) cultivation strategy using Chlorella sp. HS2, where nitrogen in the growth environment is depleted naturally via consumption. To enhance both biomass and lipid productivity this strategy explored supplementation of additional P from 50% to 2500% of the initial concentration at the start of N-limited second stage of growth. The results of the optimization study in photobioreactor (PBR) showed that supplementing 500% of initial P and 100% of initial other nutrients (O) (N0-P500-O100) on 5th day showed the maximum biomass productivity of 774.4 mg L-1 d-1. It was observed that Chlorella sp. HS2 grown in PBR yielded higher biomass (3.8 times), lipid (6.1 times) and carbohydrate (5.5 times) productivity in comparison to the open raceway ponds (ORP) study, under optimum nutrient and carbon supply condition. The maximum lipid (289.6 mg L-1 d-1) and carbohydrate (219.2 mg L-1 d-1) productivities were obtained in TPBR-3, which were 1.9 and 1.3 times higher than that of TPBR-2 (+ve control) and 9.6 and 3.7 times higher than that of TPBR-1 (-ve control), respectively. Fatty acid mainly composed of C16/C18 (84.5%-85.7%), which makes the microalgal oil suitable for biofuel production. This study concluded that feeding excess amount of P is an effective and scalable strategy to improve the biomass and lipid productivity of CTS cultivation.

Biological nutrient recovery from human urine by enriching mixed microalgal consortium for biodiesel production.

Utilization of waste resources is necessary to harness the long-term sustainability of algal technology. The study focused on the use of human urine as the basic nutrient source for culturing native microalgal consortium and further optimized the process parameters using response surface methodology. A full factorial, central composite rotatable design (CCRD) with three variables: urine concentration (1-10% vol of urine/vol of distil water [%v/v]), pH (6.5-9) and light intensity (50-350 µmolphotonsm-2sec-1) was used to evaluate the microalgal biomass and lipid content. Results indicated that at 95% confidence limits, the selected factors influence the biomass and lipid productivity. The maximum biomass productivity of 211.63 ± 1.40 mg l-1 d-1 was obtained under optimized conditions with 6.50% v/v of urine, pH of 7.69 and at light intensity of 205.40 µmolphotonsm-2sec-1. The lipid content was found to increase from 18.96 ± 1.30% in control media to 26.27 ± 1.94% under optimal conditions. The interactive effect of variables over the microalgal biomass and lipid content has also been elucidated. The data obtained were comparable to the BG11 media (control). Optimized diluted urine media in the presence of ammonium ions and under limited nitrate showed better lipid yields. Significant lipid biomolecules were detected in the algal oil extracts obtained from the diluted urine media characterized by Fourier transform infrared spectroscopy (FTIR) and Nuclear magnetic resonance (NMR). Gas chromatography-mass spectrometry (GCMS) revealed the presence of several monounsaturated and polyunsaturated fatty acids in the transesterified algal oil. Such studies would aid in technically realizing the field scale cultivation of microalgae for biofuels.

Comparative study on attached-growth photobioreactors under blue and red lights for treatment of septic tank effluent.

Attached-growth photobioreactors (AG-PBRs) employing low-cost attached-growth media were applied to treat septic tank effluent which contained abundant organic and nutrient matters as well as pathogenic microorganisms. This study investigated effects of blue and red LED lights on organic, nutrient and pathogenic removals, biomass productivity and compositions of microbial community in the AG-PBR system. The experimental results showed the blue AG-PBR to be more effective in removing chemical oxygen demand (COD), total nitrogen (TN) and ammonia nitrogen (NH3-N) and generating biomass productivity than those of the red AG-PBR (P < 0.05). Mass balance analysis indicated that the TN and total phosphorus (TP) were removed mainly by assimilation into the biomass. The TN removal rates via nitrification and denitrification processes in the blue AG-PBR were found to be higher than that of the red AG-PBR, corresponding to the observed results of bacterial biomass and abundances of nitrifying and denitrifying bacterial species in the treatment systems. The maximal areal algal biomass productivity of 47 gDW/(m2. d) in the blue AG-PBRs was found to be higher than those of other algal attached-growth systems. Although, the red and blue AG-PBR systems could effectively treat the septic tank effluent to meet the national and international discharge standards, based on treatment efficiencies and biomass productivity, the blue AG-PBR is recommended for treatment of septic tank effluent.

Drought sensitivity of aboveground productivity in Leymus chinensis meadow steppe depends on drought timing.

There is limited understanding of the combined effects of discrete climate extremes and chronic environmental changes on ecosystem processes and functioning. We assessed the interactions of extreme drought timing (45 days, in spring or summer) and nitrogen (N) addition in a full factorial field experiment in a Leymus chinensis-dominated meadow steppe in northeast China. We evaluated the resistance and recovery of the grassland (calculated in terms of aboveground biomass) to these two drought events. The spring drought reduced aboveground biomass by 28% in the unfertilized plots and by 33% in the fertilized plots, and the effects persisted during the subsequent post-drought period within the same growing season; however, the summer drought had no significant influence on aboveground biomass. Although there were no significant interactive effects between drought timing and N addition, we observed a potential trend of N addition increasing the proportion of aboveground biomass suppressed by spring drought but not summer drought. Moreover, the drought resistance of the aboveground biomass was positively correlated with the response of the belowground biomass to drought. One year after the extreme drought events, the spring drought effects on aboveground and belowground biomass were negligible. Our results indicate that the drought sensitivity of productivity likely depends on the phenological and morphological traits of the single highly dominant species (Leymus chinensis) in this meadow steppe.

Influence of nutrient supplementation and starvation conditions on the biomass and lipid productivities of Micractinium reisseri grown in wastewater for biodiesel production.

Generation of biodiesel from microalgae grown in wastewater can offer a cost-effective approach, whilst wastewaters usually do not contain the optimum concentrations of the essential nutrients and carbon sources that result in lowering the productivities of biomass and lipid. This study aimed to overcome this limitation by manipulating wastewater with various concentrations of nutrients (NO3-, PO43-, Cl- and SO42-) and three carbon sources either individually or in combination to cultivate Micractinium reisseri for biodiesel production. Initially, various dilutions of wastewater were tested and a concentration up to 75% of wastewater showed the highest biomass productivity (0.076 g L-1 d-1) and lipid productivity (0.014 g L-1 d-1). The optimum manipulating conditions for maximum lipid production and the highest productivity required 50% decrease in phosphorous from the concentration of the control medium and supplementation with 1.0 g L-1 of glucose. Under this condition, biomass and lipid productivities increased by 1.7 and 4-folds, respectively, compared to those observed in the control. Furthermore, phosphorous starvation condition in the presence of glucose significantly improved fatty acid profile in the biomass and biodiesel quality related parameters.

Functional traits determine tree growth and ecosystem productivity of a tropical montane forest: Insights from a long-term nutrient manipulation experiment.

Trait-response effects are critical to forecast community structure and biomass production in highly diverse tropical forests. Ecological theory and few observation studies indicate that trees with acquisitive functional traits would respond more strongly to higher resource availability than those with conservative traits. We assessed how long-term tree growth in experimental nutrient addition plots (N, P, and N + P) varied as a function of morphological traits, tree size, and species identity. We also evaluated how trait-based responses affected stand scale biomass production considering the community structure. We found that tree growth depended on interactions between functional traits and the type or combination of nutrients added. Common species with acquisitive functional traits responded more strongly to nutrient addition, mainly to N + P. Phosphorous enhanced the growth rates of species with acquisitive and conservative traits, had mostly positive effects on common species and neutral or negative effects in rare species. Moreover, trees receiving N + P grew faster irrespective of their initial size relative to trees in control or to trees in other treatment plots. Finally, species responses were highly idiosyncratic suggesting that community processes including competition and niche dimensionality may be altered under increased resource availability. We found no statistically significant effects of nutrient additions on aboveground biomass productivity because acquisitive species had a limited potential to increase their biomass, possibly due to their generally lower wood density. In contrast, P addition increased the growth rates of species characterized by more conservative resource strategies (with higher wood density) that were poorly represented in the plant community. We provide the first long-term experimental evidence that trait-based responses, community structure, and community processes modulate the effects of increased nutrient availability on biomass productivity in a tropical forest.

Effect of different CO2 concentrations on biomass, pigment content, and lipid production of the marine diatom Thalassiosira pseudonana.

The marine diatom Thalassiosira pseudonana grown under air (0.04% CO2) and 1 and 5% CO2 concentrations was evaluated to determine its potential for CO2 mitigation coupled with biodiesel production. Results indicated that the diatom cultures grown at 1 and 5% CO2 showed higher growth rates (1.14 and 1.29 div day-1, respectively) and biomass productivities (44 and 48 mgAFDWL-1 day-1) than air grown cultures (with 1.13 div day-1 and 26 mgAFDWL-1 day-1). The increase of CO2 resulted in higher cell volume and pigment content per cell of T. pseudonana. Interestingly, lipid content doubled when air was enriched with 1-5% CO2. Moreover, the analysis of the fatty acid composition of T. pseudonana revealed the predominance of monounsaturated acids (palmitoleic-16:1 and oleic-18:1) and a decrease of the saturated myristic acid-14:0 and polyunsaturated fatty acids under high CO2 levels. These results suggested that T. pseudonana seems to be an ideal candidate for biodiesel production using flue gases.

A mathematical model to guide genetic engineering of photosynthetic metabolism.

The optimization of algae biomass productivity in industrial cultivation systems requires genetic improvement of wild type strains isolated from nature. One of the main factors affecting algae productivity is their efficiency in converting light into chemical energy and this has been a major target of recent genetic efforts. However, photosynthetic productivity in algae cultures depends on many environmental parameters, making the identification of advantageous genotypes complex and the achievement of concrete improvements slow. In this work, we developed a mathematical model to describe the key factors influencing algae photosynthetic productivity in a photobioreactor, using experimental measurements for the WT strain of Nannochloropsis gaditana. The model was then exploited to predict the effect of potential genetic modifications on algae performances in an industrial context, showing the ability to predict the productivity of mutants with specific photosynthetic phenotypes. These results show that a quantitative model can be exploited to identify the genetic modifications with the highest impact on productivity taking into full account the complex influence of environmental conditions, efficiently guiding engineering efforts.

Isolation and characterization of Leptolyngbya sp. KIOST-1, a basophilic and euryhaline filamentous cyanobacterium from an open paddle-wheel raceway Arthrospira culture pond in Korea.

AIMS: Cyanobacteria have been used as sustainable bioresource producers for foods, feeds and other valuable natural products. However, selection of a new species (other than Arthrospira), with advantageous properties for alimentary purposes, continues to be a challenge due to potential toxicity and low biomass productivity. In this study, we report a valuable filamentous cyanobacterium isolated from Korea. METHODS AND RESULTS: Morphological and phylogenetic analyses demonstrated that the isolate belongs to the genus Leptolyngbya, and consequently designated Leptolyngbya sp. KIOST-1. Interestingly, Leptolyngbya sp. KIOST-1 possessed numerous advantageous characteristics for biomass production, similar to Arthrospira. The isolate readily propagated in SOT medium with efficient biomass productivity, and its optimum growth was observed at 30°C under alkaline and saline conditions. Moreover, more than half of the cellular components in Leptolyngbya sp. KIOST-1 were composed of protein, with approx. 40% of essential amino acids. Most importantly, no significant cytotoxicity was detected in the isolate. CONCLUSIONS: Leptolyngbya sp. KIOST-1 has a number of advantageous characteristics for alimentary purposes due to its efficient productivity, high protein content and lack of potential cytotoxicity. SIGNIFICANCE AND IMPACT OF THE STUDY: Leptolyngbya sp. KIOST-1 may be considered a potential candidate for industrial biomass production, similar to Arthrospira.

Seasonal isolation of microalgae from municipal wastewater for remediation and biofuel applications.

AIMS: The objective of the study was to isolate the microalgae strains from treated municipal wastewater in both summer and winter seasons in order to identify strains better suited for nutrient remediation and biofuel production under either cooler or warmer temperatures. METHODS AND RESULTS: Fifty-six strains in total were isolated and identified by DNA sequencing from effluent samples collected from a local wastewater treatment plant during the summer and winter of 2011. Screening of 41 isolates based on the fatty acid productivity at either 22 or 10°C resulted in the selection of 12 strains organized into two groups of 6-the M (mild) and C (cool) groups, respectively. Four of the C-group strains were isolated from the winter sample, while four of the M-group isolates were isolated from the summer sample. Fatty acid pools in M-group strains were heavily regulated in response to growth temperature while C-group strains were more insensitive. In three of the six C-group strains, the rates of biomass and fatty acid productivity at 10°C exceeded the corresponding rates at 22°C. Conversely, M group were always more productive at 22 compared to 10°C. Mixotrophic strategies to enhance productivity were generally unsuccessful in M-group strains at 22°C but proved to be more effective in C-group cultures at 10°C. CONCLUSIONS: In general, C-group strains appeared better suited for growth in municipal wastewater at 10°C, while M-group strains were better suited at 22°C. On balance, C-group isolates were more likely to come from winter wastewater samples while M-group strains were more likely to come from the summer sample. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results demonstrate that the effects of temperature on microalgal growth for wastewater remediation can be mitigated somewhat by isolation and careful selection of strains adapted to seasonal wastewater conditions.

Transgenic sugarcane overexpressing Glyoxalase III improved germination and biomass production at formative stage under salinity and water-deficit stress conditions.

The glyoxalase system, involving Glyoxalase I (GlyI) and Glyoxalase II (Gly II), plays a vital role in abiotic stress tolerance in plants. A novel enzyme Glyoxalase III (Gly III) was found recently from bacteria, yeast, and plant species. This enzyme provides a new way to detoxify Methylglyoxal (MG), a cytotoxic α-oxoaldehyde, which, in excess, can cause complete cell destruction by forming Reactive Oxygen Species (ROS) and Advanced Glycation End products (AGEs) or DNA/RNA mutation. In this background, the current study examined sugarcane transgenic events that exhibit an increase in expression of EaGly III, to assess their performance in terms of germination and biomass production during formative stage under stress conditions. Southern blot analysis outcomes confirmed the integration of transgene in the transgenic plants. The results from quantitative RT-PCR analyses confirmed high expression levels of EaGly III in transgenic events compared to wild type (WT) under salinity (100 and 200 mM NaCl) and drought (withholding watering) conditions. Transgenic events exhibited enhanced biomass productivity ranged between 0.141 Kg/pot and 0.395 Kg/pot under 200 mM salinity and 0.262 Kg/pot and 0.666 Kg/pot under drought stress. Further, transgenic events observed significantly higher germination rates under salinity and drought conditions compared to that of WT. Subcellular localization prediction by EaGlyIII-GFP fusion expression in sugarcane callus showed that it is distributed across the cytoplasm, thus indicating its widespread activity within the cell. These results strongly suggest that enhancing EaGly III activity is a useful strategy to improve the salinity and drought-tolerance in sugarcane as well as other crops.

Response of varying combined nutrients on biomass and biochemical composition of marine diatoms Chaetoceros gracilis and Thalassiosira weissflogii.

Marine diatoms have high adaptability and are known to accumulate lipids under nutrient stress conditions. The present study involves determining the effect of varying macro and micronutrients on growth kinetics and metabolite production of oleaginous marine diatoms, Thalassiosira weissflogii and Chaetoceros gracilis. The results highlighted that C. gracilis and T. weissflogii showed maximum biomass yield of 0.86 ± 0.06 g/L and 0.76 ± 0.01 g/L in the 2f and f supplemented medium respectively. A 2.5-fold increase in cellular lipid content was recorded in the 2f culture setup of both strains ranging from 20 % to 26.7 % (w/w). The study also reveals that high eutrophic nutrient media (f, 2f and 4f) triggered biomass productivity as well as total protein and carbohydrate content in both strains. Thus, providing a reproducible insight of trophic flexibility of diatoms, concomitant with the increment in multiple commercially valuable products.

Improvement of N and P ratio for enhanced biomass productivity and sustainable cultivation of Chlorella vulgaris microalgae.

Microalgae cultivation could contribute to the achievement of several sustainable development goals (SDGs). However, cultivating Chlorella vulgaris, like any other microalgae, is challenging due to various biotic, abiotic and process related factors that can affect its growth and biomass productivity. Nutrient availability, particularly N and P, and their ratio play a crucial role in building cellular structures and maintaining metabolic processes, determining basically the maximum achievable biomass productivity under given circumstances. The present article aims to improve the N and P ratio to enhance the biomass productivity of Chlorella vulgaris microalgae as well as to characterize the biomass growth kinetics that can be used for prediction purposes. The results showed that the nutrient solutions prepared with increased nitrate concentration (T1 - N:P = 55:1 and T3 - N:P = 28:1) promoted chlorophyll formation and significantly outperformed the control sample (BG-11 - N:P = 35:1) with 192% and 183%, leading to higher biomass productivity with 1160 µg L-1 and 1103 µg L-1, respectively. Moreover, a strong positive correlation was revealed (0.81) between phosphate concentration and microalgae activity rate, indicating the role of phosphorous in energy transfer, resulted in stimulated microalgae activity rates with 71.2% and 70.66% in the phosphate-increased nutrient solutions (T2 - N:P = 14:1 and T3 - N:P = 28:1). In addition, an exponential equation was introduced to characterize the biomass growth kinetics, of which the theoretically achievable maximum chlorophyll concentration (CTAM) and the theoretical cultivation time (tcultivation) were determined for the tested nutrient solutions with variable N:P ratio. It was concluded, that the higher the N:P ratio, the higher the CTAM is, nevertheless the absolute concentration of these nutrients need to be considered as well. The introduced two key parameters could provide valuable information for decision makers regarding the optimization of growth conditions, nutrient supplementation, and harvesting, additionally decreasing the production costs and making the cultivation cycles more effective and sustainable.