Final Biomass Yield Research Articles

Differences in dry matter production and grain yield of maize (Zea mays L.) cultivars with contrasting nitrogen efficiency

ABSTRACT We aimed to investigate the differences in dry matter accumulation (DMA), distribution, remobilisation, and yield of maize cultivars with contrasting nitrogen efficiency responses to nitrogen application rates. Nitrogen fertiliser increased DMA, crop growth rate (CGR), redistribution of pre-silking dry matter, and accumulation of post-silking dry matter while reducing the root/shoot biomass ratio to substantially increase biomass yield and final maize grain yield. Higher final theoretical biomass yield, maximum increasing rate (MIR), and average increasing rate, later days to MIR, longer duration of rapid growth period, higher ratio of vegetative organs, and rational dry matter distribution aboveground and belowground contribute to higher DMA and CGR in the later growth stage, and higher assimilated dry matter into grain and biomass yield to finally obtain higher grain yield for N-efficient maize cultivar. The N-efficient cultivar had obvious advantages on grain yield compared with N-inefficient cultivar caused by higher dry matter average increasing rate and biomass and kernels per ear, while the advantages increased first, then decreased with higher N rates, and the highest value was observed at 247.5 kg N ha−1. Therefore, selecting N-efficient maize improves maize yield per unit area while reducing N fertiliser supply, especially at low N application rates.

An integrated novel plasma-microalgae approach for landfill leachate treatment using a high-ammonia tolerant strain of Chlorella vulgaris

Landfilling is the most common method worldwide to deal with the produced municipal solid wastes (MSW). Nonetheless, one major drawback of landfilling is the production of landfill leachate (LL). Various methods for LL treatment exist which can mainly be divided into biological and physical-chemical methods, however integrating both approaches result in more effective treatment. In this study, an indigenous Chlorella vulgaris was isolated from a landfill leachate treatment site, purified, and genetically identified for leachate treatment purposes. One replicate, C.V.M* (CCAP 211/141), of the tested Chlorella vulgaris showed an outstanding growth in 20 % LL (v/v) with a significant ammonia removal (p < 0.05). To confirm these findings, both of the Chlorella vulgaris replicates (C.V.M* and C.V.N) were further tested in 20 % LL (v/v). A dramatic increase in the growth of C.V.M* by 19-fold over its peer C.V.N with a 75 % ammonia removal (starting from concentration 290.73 mg/L) was observed. This percentage is further improved when injecting plasma activated microbubbles as a pre-treatment step. The plasma pre-treatment resulted in LL decolourisation after 3 h and reduced ammonia-N concentration by 1.9-fold. C.V.M* showed the highest growth in 20 % LL pre-treated by plasma compared to the untreated LL with a final biomass yield of 0.38 g/L, increase in the total ammonia-N removal (79 %) and a significant decrease in the pH value (8.6–6.67). Whole genome sequencing for both replicates revealed differences in genotypes between them which might indicate the possibility of a developed mutation or sexual reproduction that might have increased the ability of replicate/strain C.V.M* to tolerate harsher conditions and higher ammonia-N concentrations in 20 % diluted LL. These results might pave the way for a possible powerful candidate in LL treatment using a high-ammonia tolerant algal strain which when coupled with plasma pre-treatment might provide an effective, eco-friendly, and sustainable LL treatment approach.

Cannabidiol industrial hemp growth, biomass, and temporal cannabinoids accumulation under different planting dates in southern Florida

AbstractDue to limited information, identifying suitable cannabidiol (CBD) dominant industrial hemp (Cannabis sativa L.) cultivars and optimal planting date are critical for hemp commercialization for CBD production in Florida. Two field trials were conducted with seven cultivars planted on three different dates from late April to late June. The cultivars were received from two different suppliers (Kentucky and Colorado) representing their adaptation. Plant growth parameters such as plant height and green canopy cover were recorded at 2−3 weeks intervals during crop growth. Temporal CBD and Δ‐9‐tetrahydrocannabinol (THC) concentrations in developing flowers were analyzed weekly beginning two weeks after flowering until maturity. The final floral biomass yield was measured at harvest. CBD and THC concentration curve over crop growing season varied among the tested cultivars. High positive correlations between CBD and THC concentrations (R2 = 0.88−0.98) were observed in all cultivars. This resulted in similar CBD/THC ratios across all planting conditions. Earlier planting resulted in higher CBD, THC, and floral biomass yield in a few cultivars, with no significant effect in most cultivars. Cultivars adapted in Kentucky performed better compared to Colorado in floral biomass and CBD yield. However, THC concentrations in the cultivars adapted in Kentucky mostly exceeded the acceptable legal threshold (0.3% THC) at harvest. Results indicate cultivar performance was affected more by genetics compared to planting date. Screening more cultivars adapted to regions of similar latitudes and environmental conditions is necessary to identify suitable hemp cultivars for Florida or similar climatic conditions.

Cultivation of the PHB-producing cyanobacterium Synechococcus leopoliensis in a pilot-scale open system using nitrogen from waste streams

PHB-producing cyanobacteria may provide the raw material for bio-based and biodegradable plastics. To commercialize photoautotrophic PHB production, their cultivation needs to be scaled up in open systems and to reduce costs and increase sustainability, nutrients must be obtained from waste streams. Here, the feasibility of these steps was verified. Different PHB-producing cyanobacteria were compared in laboratory-scale cultivations using either water from recirculating aquaculture systems or pre-processed liquid digestate as nutrient sources. Then, Synechococcus leopoliensis was cultivated in an open thin-layer photobioreactor (18 m2, 200 L), were growth in mineral Z-medium was again compared to said waste streams. All cultivations were successful. Cultivation in mineral medium resulted in both the highest final biomass yield (6 g L−1) and productivity (0.7 g L−1 d−1). Both waste stream-based media showed lower biomass yields and productivities (2 g L−1 and 0.25–0.3 g L−1 d−1). However, due to differences in the cultivation conditions (e.g., temperature, nutrient supply), final biomass yield and productivity do not represent the performance of the cultivations adequately. Relative parameters such as nitrogen and energy conversion ratios indicate that cultivation with aquaculture water suffered from insufficient nitrogen supply to the culture, whereas the use pre-processed liquid digestate resulted in a substantial loss of nitrogen due to volatilization. All cultivations in the open system were continued in the laboratory, where cultures were starved for ten days under nutrient-depleted conditions (without nitrogen, phosphorus, or both). While PHB accumulation occurred, concentrations were comparatively low (< 1%dw). The comparison of the results suggests that PHB yields were influenced more by the initial cultivation condition than by the specific type of nutrient depletion. Thus, while the cultivation with waste streams in an open system is feasible, environmental parameters seem to influence PHB yields considerably and must be considered for the optimization of the complete process.

Removal of primary nutrient degraders reduces growth of soil microbial communities with genomic redundancy.

Understanding how microorganisms within a soil community interact to support collective respiration and growth remains challenging. Here, we used a model substrate, chitin, and a synthetic Model Soil Consortium (MSC-2) to investigate how individual members of a microbial community contribute to decomposition and community growth. While MSC-2 can grow using chitin as the sole carbon source, we do not yet know how the growth kinetics or final biomass yields of MSC-2 vary when certain chitin degraders, or other important members, are absent. To characterize specific roles within this synthetic community, we carried out experiments leaving out members of MSC-2 and measuring biomass yields and CO2 production. We chose two members to iteratively leave out (referred to by genus name): Streptomyces, as it is predicted via gene expression analysis to be a major chitin degrader in the community, and Rhodococcus as it is predicted via species co-abundance analysis to interact with several other members. Our results showed that when MSC-2 lacked Streptomyces, growth and respiration of the community was severely reduced. Removal of either Streptomyces or Rhodococcus led to major changes in abundance for several other species, pointing to a comprehensive shifting of the microbial community when important members are removed, as well as alterations in the metabolic profile, especially when Streptomyces was lacking. These results show that when keystone, chitin degrading members are removed, other members, even those with the potential to degrade chitin, do not fill the same metabolic niche to promote community growth. In addition, highly connected members may be removed with similar or even increased levels of growth and respiration. Our findings are critical to a better understanding of soil microbiology, specifically in how communities maintain activity when biotic or abiotic factors lead to changes in biodiversity in soil systems.

Efficient Green Light Acclimation of the Green Algae Picochlorum sp. Triggering Geranylgeranylated Chlorophylls.

In analogy to higher plants, eukaryotic microalgae are thought to be incapable of utilizing green light for growth, due to the “green gap” in the absorbance profiles of their photosynthetic pigments. This study demonstrates, that the marine chlorophyte Picochlorum sp. is able to grow efficiently under green light emitting diode (LED) illumination. Picochlorum sp. growth and pigment profiles under blue, red, green and white LED illumination (light intensity: 50–200 μmol m−2 s−1) in bottom-lightened shake flask cultures were evaluated. Green light-treated cultures showed a prolonged initial growth lag phase of one to 2 days, which was subsequently compensated to obtain comparable biomass yields to red and white light controls (approx. 0.8 gDW L−1). Interestingly, growth and final biomass yields of the green light-treated sample were higher than under blue light with equivalent illumination energies. Further, pigment analysis indicated, that during green light illumination, Picochlorum sp. formed unknown pigments (X1-X4). Pigment concentrations increased with illumination intensity and were most abundant during the exponential growth phase. Mass spectrometry and nuclear magnetic resonance data indicated, that pigments X1-X2 and X3-X4 are derivatives of chlorophyll b and a, which harbor C=C bonds in the phytol side chain similar to geranylgeranylated chlorophylls. Thus, for the first time, the natural accumulation of large pools (approx. 12 mg gDW −1) of chlorophyll intermediates with incomplete hydrogenation of their phytyl chains is demonstrated for algae under monochromatic green light (Peak λ 510 nm, full width at half maximum 91 nm). The ability to utilize green light offers competitive advantages for enhancing biomass production, particularly under conditions of dense cultures, long light pathways and high light intensity. Green light acclimation for an eukaryotic microalgae in conjunction with the formation of new aberrant geranylgeranylated chlorophylls and high efficiency of growth rates are novel for eukaryotic microalgae. Illumination with green light could enhance productivity in industrial processes and trigger the formation of new metabolites–thus, underlying mechanisms require further investigation.

New insights into phenotypic heterogeneity for the distinct lipid accumulation of Schizochytrium sp. H016

BackgroundSchizochytrium sp. is a marine heterotrophic protist and an important sustainable resource for high value-added docosahexaenoic acid in the future. The production of different phenotypes during the continuous subculture of Schizochytrium sp. results in a serious reduction in lipid yield and complicates the used of this strain in scientific research and industrial production. Hence, obtaining an improved understanding of the phenotypic differences and molecular mechanisms underlying the cell-to-cell heterogeneity of Schizochytrium sp. is necessary.ResultsAfter continuous culture passage, Schizochytrium sp. H016 differentiated into two subpopulations with different morphologies and showed decreased capacity for lipid production. The presence of cell subpopulations with degraded lipid droplets led to a substantial decrease in overall lipid yield. Here, a rapid screening strategy based on fluorescence-activated cell sorting was proposed to classify and isolate subpopulations quickly in accordance with their lipid-producing capability. The final biomass and lipid yield of the subpopulation with high cell lipid content (i.e., H016-H) were 38.83 and 17.22 g/L, respectively, which were 2.07- and 5.38-fold higher than those of the subpopulation with low lipid content (i.e., H016-L), respectively. Subsequently, time‑resolved transcriptome analysis was performed to elucidate the mechanism of phenotypic heterogeneity in different subpopulations. Results showed that the expression of genes related to the cell cycle and lipid degradation was significantly upregulated in H016-L, whereas the metabolic pathways related to fatty acid synthesis and glyceride accumulation were remarkably upregulated in H016-H.ConclusionThis study innovatively used flow cytometry combined with transcriptome technology to provide new insights into the phenotypic heterogeneity of different cell subpopulations of Schizochytrium sp. Furthermore, these results lay a strong foundation for guiding the breeding of oleaginous microorganisms with high lipid contents.

Simulating Maize Productivity under Selected Climate Smart Agriculture Practices Using AquaCrop Model in a Sub-humid Environment

Crop models are crucial in assessing the reliability and sustainability of soil water conservation practices. The AquaCrop model was tested and validated for maize productivity under the selected climate smart agriculture (CSA) practices in the rainfed production systems. The model was validated using final biomass (B) and grain yield (GY) data from field experiments involving seven CSA practices (halfmoon pits, 2 cm thick mulch, 4 cm thick mulch, 6 cm thick mulch, 20 cm deep permanent planting basins (PPB), and 30 cm deep) and the control (conventional practice) where no CSA was applied. Statistics for coefficient of determination (R2), Percent bias (Pbias), and Nash–Sutcliffe (E) for B and GY indicate that the AquaCrop model was robust to predict crop yield and biomass as illustrated by the value of R2 &gt; 0.80, Pbias −1.52–1.25% and E &gt; 0.68 for all the CSA practices studied. The relative changes between the actual and simulated water use efficiency (WUE) of grain yield was observed in most of the CSA practices. However, measured WUE was seemingly better in the 2 cm thick mulch, indicating a potential for water saving and yield improvement. Therefore, the AquaCrop model is recommended as a reliable tool for assessing the effectiveness of the selected CSA practices for sustainable and improved maize production; although, the limitations in severely low soil moisture conditions and water stressed environments should be further investigated considering variations in agroecological zones.

Improving subtropical cotton production by using late winter sowing to reduce climatic risk

• Late-winter sowing to avoid coinciding boll development and harvest with unfavourable climatic factors was explored. • Crop establishment was successful despite temperatures being well below accepted cold stress thresholds. • Earlier sowing coincided peak boll demand with improved conditions maximising growth and reproductive partitioning. • Lint yield increased over 40 % with late-winter sowing compared to traditional spring sowing. Climate variability is a challenge to the reliable production of high yielding cotton, Gossypium hirsutum L. in the subtropics. Shifting the growing season earlier by sowing during late winter (August) to reduce the risk of exposure to unfavourable radiation, temperature and rainfall during mid-summer offered the potential to improve stagnating and variable yields. This study aimed to determine the effect of late winter sowing on establishment, growth, development and lint yield of cotton. Accepted knowledge from temperate climates suggested cold temperature stress would impede crop establishment, so the application of clear oxodegradable film to the soil surface at sowing to mitigate cold temperature stress was also explored. Crop establishment, biomass accumulation and partitioning, light interception and lint yield were measured for cotton sown with and without film at intervals from early August to mid-September in four consecutive experiments (2013–2016). The application of film provided no benefits for establishment and only marginal improvement for crop growth and yield despite minimum temperatures being well below accepted chilling injury thresholds in temperate environments. Greater daily solar radiation levels both solarised the soil and shortened the duration of daily minimum temperatures, enabling successful crop establishment. Early sowing did not hinder crop production, and in 3 of the 4 experiments, lint yield was significantly higher ( P < 0.001) for August (2314−3305 kg ha −1 ) than September-sown crops (2053−2290 kg ha −1 ). The impact of sowing time was further explored through the measurement of biomass and yield of 29 high input commercial crops sown between August and December in 2016–2019. Final biomass and lint yield declined ( P < 0.001) by 2.89 g m -2 and 7.23 kg ha −1 for each day that sowing occurred after August 1. Combined analyses of the experiments and commercial crop data confirmed that late-winter sown crops had the highest daily growth rate between flowering and maturity, declining by 37 % for summer-sown crops. RUE g (glucose equivalent) of 2.27 g MJ −1 PAR was not changed by sowing time. Late winter sowing best matched flowering and boll development with the summer solstice and a lower incidence of cloud or heat stress. An earlier harvest also reduced the risk of pre-harvest weathering, collectively enabling over 40 % higher yields than traditional spring sowing without additional crop inputs. Spring and later sowings coincided unfavourable climate with the more susceptible growth phases of late boll filling or early flowering respectively and declining solar radiation and temperatures prevented yield recovery for the latest summer-sown crops. As the occurrence of excessive temperature-related weather events is expected to increase with climate change, particularly for the subtropics, this study demonstrates how a knowledge of crop response can be used to evolve sowing time strategies to better mitigate climatic risk.

Mild water and salt stress improve water use efficiency by decreasing stomatal conductance via osmotic adjustment in field maize

Water and salt stress often occur simultaneously in heavily irrigated arid agricultural areas, yet they are usually studied in isolation. To understand the physiological bases of water use efficiency (WUE) of field-grown maize (Zea mays) at multi-scales under combined water and salt stress, we investigated the joint effects of water and salt stress on physiology, growth, yield, and WUE of two genotypes (XY335 and ZD958). We measured leaf stomatal conductance (gs), net photosynthesis rate (A) and hydraulic traits, whole-plant growth and water use (ET), and final biomass and grain yield. Leaf osmotic adjustment was a key trait of the physiological differences between XY335 and ZD958 under water and salt stress. Although the responses of the two genotypes were different, mild water and salt stress improved intrinsic water use efficiency (iWUE = A/gs) by (i) decreasing gsvia increasing osmotic adjustment and hydraulic resistance, and (ii) declining A via increasing stomatal limitations rather than reducing photosynthetic capacity. Joint water and salt stress had a synergistic effect on reproductive growth and grain formation of maize. Mild water and salt stress reduced ET, stabilized grain yield, and improved grain WUE via declining gs, maintaining photosynthetic capacity, and improving harvest index. Collectively, our study provides a novel insight into the physiological mechanisms of WUE and demonstrates an approach for the efficient management of water and salt by using a growth stage-based deficit irrigation strategy or/and selecting genotypes with strong osmotic adjustment capacity and high harvest index.

Engineering adipic acid metabolism in Pseudomonas putida

Bio-upcycling of plastics is an upcoming alternative approach for the valorization of diverse polymer waste streams that are too contaminated for traditional recycling technologies. Adipic acid and other medium-chain-length dicarboxylates are key components of many plastics including polyamides, polyesters, and polyurethanes. This study endows Pseudomonas putida KT2440 with efficient metabolism of these dicarboxylates. The dcaAKIJP genes from Acinetobacter baylyi, encoding initial uptake and activation steps for dicarboxylates, were heterologously expressed. Genomic integration of these dca genes proved to be a key factor in efficient and reliable expression. In spite of this, adaptive laboratory evolution was needed to connect these initial steps to the native metabolism of P. putida, thereby enabling growth on adipate as sole carbon source. Genome sequencing of evolved strains revealed a central role of a paa gene cluster, which encodes parts of the phenylacetate metabolic degradation pathway with parallels to adipate metabolism. Fast growth required the additional disruption of the regulator-encoding psrA, which upregulates redundant β-oxidation genes. This knowledge enabled the rational reverse engineering of a strain that can not only use adipate, but also other medium-chain-length dicarboxylates like suberate and sebacate. The reverse engineered strain grows on adipate with a rate of 0.35 ± 0.01 h−1, reaching a final biomass yield of 0.27 ± 0.00 gCDW gadipate−1. In a nitrogen-limited medium this strain produced polyhydroxyalkanoates from adipate up to 25% of its CDW. This proves its applicability for the upcycling of mixtures of polymers made from fossile resources into biodegradable counterparts.

Assessment of Chlorella sorokiniana Growth in Anaerobic Digester Effluent.

Microalgae are considered a potential source of valuable compounds for multiple purposes and are potential agents for bioremediation of aquatic environments contaminated with different pollutants. This work evaluates the use of agricultural waste, unsterilized and anaerobically digested, to produce biomass from a strain of Chlorella sorokiniana. Furthermore, the presence of bacteria in these wastes was investigated based on the bacterial 16S rRNA gene sequencing. The results showed a specific growth rate ranging between 0.82 and 1.45 day−1, while the final biomass yield in different digestate-containing treatments (bacterial-contaminated cultures) ranged between 0.33 and 0.50 g L−1 day−1. Besides, substantial amounts of ammonium, phosphate, and sulfate were consumed by C. sorokiniana during the experimental period. The predominant bacteria that grew in the presence of C. sorokiniana in the effluent-containing treatments belonged to the genera Chryseobacterium, Flavobacterium, Sphingomonas, Brevundimonas, Hydrogenophaga, Sphingobacterium, and Pseudomonas. Therefore, this microalga can tolerate and grow in the presence of other microorganisms. Finally, these results show that anaerobically digested agricultural waste materials are a good substitute for growth media for green microalgae; however, phosphate and sulfate levels must also be controlled in the media to maintain adequate growth of microalgae.

Evaluation of pyrolysis chars derived from marine macroalgae silage as soil amendments.

Pyrolysis char residues from ensiled macroalgae were examined to determine their potential as growth promoters on germinating and transplanted seedlings. Macroalgae was harvested in May, July and August from beach collections, containing predominantly Laminaria digitata and Laminaria hyperborea; naturally seeded mussel lines dominated by Saccharina latissima; and lines seeded with cultivated L. digitata. Material was ensiled, pressed to pellets and underwent pyrolysis using a thermo‐catalytic reforming (TCR) process, with and without additional steam. The chars generated were then assessed through proximate and ultimate analysis. Seasonal changes had the prevalent impact on char composition, though using mixed beach‐harvested material gave a greater variability in elements than when using the offshore collections. Applying the char at 5% (v/v)/2% (w/w) into germination or seedling soils was universally negative for the plants, inhibiting or delaying all parameters assessed with no clear advantage in harvesting date, species or TCR processing methodology. In germinating lettuce seeds, soil containing the pyrolysis chars caused a longer germination time, poorer germination, fewer true leaves to be produced, a lower average plant health score and a lower final biomass yield. For transplanted ryegrass seedlings, there were lower plant survival rates, with surviving plants producing fewer leaves and tillers, lower biomass yields when cut and less regrowth after cutting. As water from the char‐contained plant pots inhibited the lettuce char control, one further observation was that run‐off water from the pyrolysis char released compounds which detrimentally affected cultivated plant growth. This study clearly shows that pyrolysed macroalgae char does not fit the standard assumption that chars can be used as soil amendments at 2% (w/w) addition levels. As the bioeconomy expands in the future, the end use of residues and wastes from bioprocessing will become a genuine global issue, requiring consideration and demonstration rather than hypothesized use.

Comparison of the Photoautotrophic Growth Regimens of Chlorella sorokiniana in a Photobioreactor for Enhanced Biomass Productivity

Microalgae have a wide industrial potential because of their high metabolic diversity and plasticity. Selection of optimal cultivation methods is important to optimize multi-purpose microalgal biotechnologies. In this research, Chlorella sorokiniana AM-02 that was isolated from a freshwater lake was cultured under various high photosynthetic photon flux density (PPFD) conditions and CO2 gas levels in standard Bold’s basal medium (BBM). Furthermore, a wide range of nitrate levels (180–1440 mg L−1) was tested on the growth of C. sorokiniana. Microalgae growth, pigment concentration, medium pH, exit gas composition, as well as nitrate, phosphate, and sulfate levels were measured during an experimental period. The preferred high PPFD and optimal CO2 levels were found to be 1000–1400 μmol photons m−2 s−1 and 0.5–2.0% (v/v), respectively. The addition of nitrate ions (up to 1440 mg L−1) to the standard growth medium increased final optical density (OD750), cell count, pigment concentration, and total biomass yield but decreased the initial growth rate at high nitrate levels. Our findings can serve as the basis for a robust photoautotrophic cultivation system to maximize the productivity of large-scale microalgal cultures.

The effect of irrigation strategies and nitrogen fertilizer rates on maize growth and grain yield

In North Florida, increasing nitrogen loads and water quality declines have become a major concern, in part as result of anthropogenic non-point source activities such as agriculture. The main objective of this study was to investigate the effect of irrigation strategies and nitrogen (N) fertility rates on maize biomass, yield and water productivity in sandy soils. The field experiment was conducted 2015–2017 in Live Oak, Florida using a randomized complete block with a split plot design and four replicates. Treatments evaluated five irrigation strategies: (i) GROW, mimicking grower irrigation practices in the region, (ii) SWB, using a soil water balance to schedule irrigation; (iii) SMS, using soil moisture sensors to schedule irrigation; (iv) RED, applying 60% of the GROW treatment; and (v) NON, non-irrigated, and three N fertility rates: (i) low (157 kg N/ha), (ii) medium (247 kg N/ha), and (iii) high (336 kg N/ha). In comparison to GROW, the SWB, SMS and RED irrigation treatments showed no differences in final biomass, N uptake nor grain yield; however, these treatments achieved on average 41, 47, and 36% irrigation reduction, respectively, without impacts on yield during the three maize seasons evaluated. For most of the variables, statistical differences were found between the low and the high N rates, but no differences compared to the medium N rate. A 26% reduction of N fertilizer was achieved using the medium N rate without negative impact on N uptake, biomass nor yield in comparison to the high N fertilization rate. During this experiment, maize N uptake reached a plateau; thus, potential N losses resulted from applications exceeding recommended rates. Furthermore, the implementation of these more efficient irrigation and N fertilizer management strategies reduced irrigation and N fertilizer applications without negative impacts in yield. Thus, these practices may prevent potential N leaching to waterbodies while improving profits.

Inoculation concentration modulating the secretion and accumulation pattern of exopolysaccharides in desert cyanobacterium Microcoleus vaginatus.

Cyanobacterial exopolysaccharides (EPS) accumulated during microalgal cultivation have significant application potential in antioxidation, pharmaceutical products, and so on. Inoculation concentration strongly affects the cultivation cost, biomass, and EPS accumulation. In this study, a high-EPS-excreted desert cyanobacterium Microcoleus vaginatus was isolated, and the effects of inoculation concentration on biomass, photosynthetic activity, and EPS accumulation were explored. The results showed that the original fluorescence (Fo ) provided a good indication to cyanobacterial biomass, when Chl-a concentration was lower than 10mgL-1 . Inoculation concentration significantly affected cyanobacterial biomass and EPS concentration (P<0.001), whereas did not affect photosynthetic activity (Fv /Fm ; P>0.05). The two fractions of EPS, capsular exopolysaccharides (CPS) and released exopolysaccharides (RPS) were strongly affect by inoculation concentration. Other than forming thick sheath (CPS) surrounded the filaments, M. vaginatus excreted higher proportions of RPS to culture medium, and the ratio of RPS to CPS ranged from 1.08 to 1.58 depending on the inoculation concentration. Additionally, although the biomass and EPS accumulation increased with inoculation concentration, the increasing inoculation concentration did not bring to the proportionate increase of the final biomass and EPS yield. Altogether, comprehensively considering the EPS yield and productivity, inoculation concentration of 0.04mgChl-aL-1 is recommended for M. vaginatus to produce EPS, with an EPS yield of 94.32mgL-1 and EPS productivity of 184.86mg (mgChl-a)-1 L-1 d-1 at the end of experiment.

Increasing growth and yield by altering carbon metabolism in a transgenic leaf oil crop.

SummaryEngineering high biomass plants that produce oil (triacylglycerol or TAG) in vegetative rather than seed‐related tissues could help meet our growing demand for plant oil. Several studies have already demonstrated the potential of this approach by creating transgenic crop and model plants that accumulate TAG in their leaves and stems. However, TAG synthesis may compete with other important carbon and energy reserves, including carbohydrate production, and thereby limit plant growth. The aims of this study were thus: first, to investigate the effect of TAG accumulation on growth and development of previously generated high leaf oil tobacco plants; and second, to increase plant growth and/or oil yields by further altering carbon fixation and partitioning. This study showed that TAG accumulation varied with leaf and plant developmental stage, affected leaf carbon and nitrogen partitioning and reduced the relative growth rate and final biomass of high leaf oil plants. To overcome these growth limitations, four genes related to carbon fixation (encoding CBB cycle enzymes SBPase and chloroplast‐targeted FBPase) or carbon partitioning (encoding sucrose biosynthetic enzyme cytosolic FBPase and lipid‐related transcription factor DOF4) were overexpressed in high leaf oil plants. In glasshouse conditions, all four constructs increased early growth without affecting TAG accumulation while chloroplast‐targeted FBPase and DOF4 also increased final biomass and oil yields. These results highlight the reliance of plant growth on carbon partitioning, in addition to carbon supply, and will guide future attempts to improve biomass and TAG accumulation in transgenic leaf oil crops.

Engineering the Osmotic State of Pseudomonas putida KT2440 for Efficient Cell Disruption and Downstream Processing of Poly(3-Hydroxyalkanoates).

In the last decade, the development of novel programmable cell lytic systems based on different inducible genetic constructs like the holin–endolysin and lysozyme appears as a promising alternative to circumvent the use of costly enzymes and mechanical disrupters for downstream processing of intracellular microbial products. Despite the advances, upon activation of these systems the cellular disruption of the biocatalyst occurs in an extended period, thus delaying the recovery of poly(3-hydroxyalkanoate) (PHA). Herein the osmotic state of Pseudomonas putida KT2440 was engineered by inactivating the inner-membrane residing rescue valve MscL, which is responsible mainly for circumventing low-osmolarity challenges. Then the major outer membrane porin OprF and the specific porin OprE were overproduced during PHA producing conditions on decanoate-grown cells. The engineered P. putida strains carrying each porin showed no impairment on growth rate and final biomass and PHA yield after 48 h cultivation. Expression of both porins in tandem in the mutant strain KTΔmscL-oprFE led to a slight reduction of the biomass synthesis (∼10%) but higher PHA accumulation (%wt) relative to the cell dry mass. Each strain was then challenged to an osmotic upshift for 1 h and subsequently to a rapid passage to a hypotonic condition where the membrane stability of the KTΔmscL-oprFE suffered damage, resulting in a rapid reduction of cell viability. Cell disruption accounted for >95% of the cell population within 3 h as reported by colony forming units (CFU), FACS analyses, and transmission electron microscopy. PHA recovery yielded 94.2% of the biosynthesized biopolymer displaying no significant alterations on the final monomer composition. This study can serve as an efficient genetic platform for the recovery of any microbial intracellular compound allowing less unit operation steps for cellular disruption.

Evaluation of dietary protein requirement for the growth performance of minor carp,Cirrhinus reba(Hamilton, 1822) fingerlings

An experiment was designed to evaluate the requirement of dietary protein for Reba carp, Cirrhinus reba fingerlings using semi-purified diet. Five isocaloric diets were prepared containing 20%, 25%, 30%, 35% and 40% level of protein using casein, gelatin as a major protein source. The experiment was conducted for 105 days in triplicates to evaluate the growth, survival, feed utilization, carcass composition of C. reba. Mean weight gain was highest (p < .05) in T3. Specific growth rate was highest in T3 and lowest in T1. Similarly, significantly (p < .05) highest net fish yield were recorded in T3. No significant difference was recorded in survivability. Feed conversion ratio, protein efficiency ratio, feed conversion efficiency and protein conversion efficiency were significantly (p < .05) higher for T3 compared with other treatments. Highest carcass protein and lipid values were recorded in T3. So, conclusion can be drawn that 30% crude protein is optimum for overall performance. Furthermore, broken-line analysis for specific growth rate, final biomass and net fish yield indicated that dietary protein level of 28.148 to 28.253% is optimum for the best growth performance of fingerling C. reba.

Optimization of Recombinant Membrane Protein Production in the Engineered Escherichia coli Strains SuptoxD and SuptoxR.

Membrane proteins (MPs) execute a wide variety of critical biological functions in all living organisms and constitute approximately half of current targets for drug discovery. As in the case of soluble proteins, the bacterium Escherichia coli has served as a very popular overexpression host for biochemical/structural studies of membrane proteins as well. Bacterial recombinant membrane protein production, however, is typically hampered by poor cellular accumulation and severe toxicity for the host, which leads to low levels of final biomass and minute volumetric yields. In previous work, we generated the engineered E.coli strains SuptoxD and SuptoxR, which upon coexpression of the effector genes djlA or rraA, respectively, can suppress the cytotoxicity caused by MP overexpression and produce enhanced MP yields. Here, we systematically looked for gene overexpression and culturing conditions that maximize the accumulation of membrane-integrated and well-folded recombinant MPs in these strains. We have found that, under optimal conditions, SuptoxD and SuptoxR achieve greatly enhanced recombinant production for a variety of MP, irrespective of their archaeal, eubacterial, or eukaryotic origin. Furthermore, we demonstrate that the use of these engineered strains enables the production of well-folded recombinant MPs of high quality and at high yields, which are suitable for functional and structural studies. We anticipate that SuptoxD and SuptoxR will become broadly utilized expression hosts for recombinant MP production in bacteria.