The ppk-expressing transgenic rice floating bed improves P removal in slightly polluted water.

With significant economic advantages, the plant floating bed has been widely utilized in the ecological remediation of eutrophic water because of the excessive phosphorus (P) and nitrogen discharge in China. Previous research has demonstrated that polyphosphate kinase (ppk)-expressing transgenic rice (Oryza sativa L. ssp. japonica) (ETR) can increase the P absorption capacity to support rice growth and boost rice yield. In this study, the floating beds of ETR with single copy line (ETRS) and double copy line (ETRD) are built to investigate their capacity to remove aqueous P in slightly polluted water. Compared with the wild type Nipponbare (WT) floating bed, the ETR floating beds greatly reduce the total P concentration in slightly polluted water though the ETR floating beds have the same removal rates of chlorophyll-a, NO3--N, and total nitrogen in slightly polluted water. The P uptake rate of ETRD on the floating bed is 72.37% in slightly polluted water, which is higher than that of ETRS and WT on the floating beds. Polyphosphate (polyP) synthesis is a critical factor for the excessive phosphate uptake of ETR on the floating beds. The synthesis of polyP decreases the level of free intracellular phosphate (Pi) in ETR on the floating beds, simulating the phosphate starvation signaling. The OsPHR2 expression in the shoot and root of ETR on the floating bed increased, and the corresponding P metabolism gene expression in ETR was changed, which promoted Pi uptake by ETR in slightly polluted water. The Pi accumulation further promoted the growth of ETR on the floating beds. These findings highlight that the ETR floating beds, especially ETRD floating bed, have significant potential for P removal and can be exploited as a novel method for phytoremediation in slightly polluted water.

SWATH-MS-Based Proteomics Reveals the Regulatory Metabolism of Amaryllidaceae Alkaloids in Three Lycoris Species.

Alkaloids are a class of nitrogen-containing alkaline organic compounds found in nature, with significant biological activity, and are also important active ingredients in Chinese herbal medicine. Amaryllidaceae plants are rich in alkaloids, among which galanthamine, lycorine, and lycoramine are representative. Since the difficulty and high cost of synthesizing alkaloids have been the major obstacles in industrial production, particularly the molecular mechanism underlying alkaloid biosynthesis is largely unknown. Here, we determined the alkaloid content in Lycoris longituba, Lycoris incarnata, and Lycoris sprengeri, and performed a SWATH-MS (sequential window acquisition of all theoretical mass spectra)-based quantitative approach to detect proteome changes in the three Lycoris. A total of 2193 proteins were quantified, of which 720 proteins showed a difference in abundance between Ll and Ls, and 463 proteins showed a difference in abundance between Li and Ls. KEGG enrichment analysis revealed that differentially expressed proteins are distributed in specific biological processes including amino acid metabolism, starch, and sucrose metabolism, implicating a supportive role for Amaryllidaceae alkaloids metabolism in Lycoris. Furthermore, several key genes collectively known as OMT and NMT were identified, which are probably responsible for galanthamine biosynthesis. Interestingly, RNA processing-related proteins were also abundantly detected in alkaloid-rich Ll, suggesting that posttranscriptional regulation such as alternative splicing may contribute to the biosynthesis of Amaryllidaceae alkaloids. Taken together, our SWATH-MS-based proteomic investigation may reveal the differences in alkaloid contents at the protein levels, providing a comprehensive proteome reference for the regulatory metabolism of Amaryllidaceae alkaloids.

Amplified cyanobacterial bloom is derived by polyphosphate accumulation triggered by ultraviolet light.

Cyanobacterial blooms appear more strongly, constantly and globally, yet the positive effect of surface solar ultraviolet radiation (UV) on cyanobacterial bloom in natural freshwater habitats is largely ignored. Here in-situ and laboratory studies were jointly designed to probe the mechanism of cyanobacterial bloom promoted by solar UV light. The results showed that solar UV light is a key trigger factor for the accumulation of total phosphorus, dissolved inorganic phosphorus and polyphosphate (polyP) in blooming cyanobacterial cells. The increase of UV dose induces polyP accumulation to result in the excessive phosphorus uptake of blooming cyanobacteria, which provides sufficient phosphorus for cyanobacterial growth in suitable environment. Solar UV light also can promote the contents of phycocyanin, allophycocyanin, and phycoerythrin, producing sufficient ATP by photosynthesis for polyP synthesis in cyanobacterial cells in lake enviroment. The frequent variations of UV irradiance exposure prompts cyanobacteria to absorb excessive phosphorus from suspended solid or sediment. Cyanobacterial intracellular phosphorus is accumulated for their growth. UV light promotes polyP accumulation in blooming cyanobacterial cells to avoid damage. The adsorption amount of phosphorus increases for exuberant growth and then more surface blooming cyanobacteria are exposed to UV light to absorb ample phosphorus. Thus, the positive feedback occurs in lake water bodies with abundant phosphorus. This amplified cycle of cyanobacterial density and phosphorus due to solar UV light in eutrophic water bodies is analogous to a triode to amplify visible photosynthesis by UV light as a base electric current in the energy flow process in lake environment, therefore, "Cyanobacterial Phosphorus Assimilation Ultraviolet Effect" is used to describe this phenomenon. A new explanation is provided for the continuing proliferating mechanism of cyanobacterial bloom. Besides, a new perspective appears on the outbreak of cyanobacterial blooms in natural eutrophic lake water bodies worldwide.

Deactivation of cyanobacteria blooms and simultaneous recovery phosphorus through electrolysis method.

A novel method for remediating eutrophic lakes through electrolysis was made possible by one titanium (Ti) mesh, which serves as a cathode and two anodes of Ti mesh coated with ruthenium (IV) oxide and iridium (IV) oxide (RuO2-IrO2/Ti). Once the three-electrode components RuO2-IrO2/Ti and Ti are stabilized, they can carry out electrolytic reaction to control cyanobacteria blooms and assist with the remediation of eutrophic water. The order of influence on the theoretical energy consumption involved in removing algae is as follows: The electrode spacing was more effective than electrode voltage, which proved more effective than electrolysis time through the orthogonal test method. Thus, an electrode spacing of 60 mm, an electrode voltage of 30 V, and an electrolysis time of 12 h are the optimal electrolysis methods used to remove cyanobacterial blooms. The strong acidic environment produced by the anode increased the concentration of hydroxyl radical (•OH) and other strong oxidizing substances, which were the main roles that made cyanobacteria bloom inactivation. The electrolysis reaction was conducive to the transformation of organophosphorus in cyanobacterial blooms to dissolved inorganic phosphorus (DIP) in water. Some DIP was most deposited on the cathode after electro-depositing enhanced the removal of P in water with the 12-h prolonged electrolysis time. Meanwhile, it was beneficial to reduce the total nitrogen (TN) and ammonia nitrogen (NH3-N) in the water. Thus, electrolysis proved to be an effective way to the inactivation of cyanobacteria blooms and simultaneously recover P as the concentration became higher.

Novel simulation of aqueous total nitrogen and phosphorus concentrations in Taihu Lake with machine learning.

This study demonstrates the utility of internal nutrient loads as an additional parameter to improve the performance of machine learning models in predicting the temporal variations of aqueous TN and TP concentrations in Taihu Lake, a large shallow lake. Internal loads, as a potential input parameter for machine learning models, were estimated using a mass balance calculation. The results showed that between 2011 and 2018 the maximum monthly internal loads of nitrogen and phosphorus in Taihu Lake were 4200 t and 178 t, respectively. Monthly changes in the aqueous TN and TP concentrations of Taihu Lake did not correlate significantly with inflow loads whereas the correlations with estimated internal loads were positive and significant. Long short-term memory (LSTM), random forest (RF), and gradient boosting regression tree (GBRT) models were built, and for all of them the inclusion of internal loads in the input parameters improved their performance. LSTM model III, whose input parameters included both inflow loads and internal loads, had the best performance, based on a testing root mean square error of 0.11 mg TN/L and 0.017 mg TP/L. A 28 % decrease in the annual aqueous TP concentration in Taihu Lake in 2018 simulated by LSTM model III was achieved by lowering the average water level from 3.29 m to 2.99 m, suggesting a possible strategy to control the TP concentration in the lake. In summary, our study showed that aqueous TN and TP concentrations in shallow lakes can be simulated using machine learning, with LSTM models outperforming RF and GBRT models; in these models, internal loads should be included as an input parameter. Additionally, our study identified the water level as an important factor affecting the aqueous TP concentration in Taihu Lake.

Effects of biochar on the growth of Vallisneria natans in surface flow constructed wetland.

To improve the nitrogen and phosphorus removal efficiency of surface flow constructed wetlands (SFCWs), biochar was added to an SFCW matrix. The effects of adding different amounts of biochar on water purification, the growth of Vallisneria natans (V. natans), and microbial mechanisms were explored through SFCW simulation experiments. The results showed that through the joint action of biochar and V. natans, the concentrations of total nitrogen, total phosphorus, and ammonia nitrogen in the effluent significantly decreased. The total biomass, relative growth rate, and chlorophyll content of V. natans were significantly reduced by adding biochar (≥20%, v/v), as the root activity and the root to leaf biomass ratio slightly increased at first and then decreased. The carbon and nitrogen contents of V. natans slightly increased with the addition of biochar (≥10%, v/v), but the phosphorus content slightly decreased. Moreover, the nitrogen content of the matrices decreased significantly over time (P<0.05), and the phosphorus content in the matrix showed an increasing trend in the same period. In addition, the microbial 16S rDNA sequencing results indicated that the diversity and abundance of the microbial community in the matrix of the biochar-added SFCW tended to decrease. Nevertheless, the abundance of functional bacteria related to nitrogen and phosphorus removal (i.e., Pseudomonas and Dechloromonas) slightly increased, which would benefit denitrification and dephosphorization in the SFCW. Hence, the addition of biochar to the SFCW matrix facilitated the improvement of effluent water quality, while excessive biochar addition (≥10%, v/v) restrained the growth of V. natans but did not cause death. This conclusion provides valid data support regarding the ability of biochar-added SFCW to purify lightly contaminated water.

Spatiotemporal differences in phosphorus release potential of bloom-forming cyanobacteria in Lake Taihu.

The abnormal elevation of cyanobacterial density and total phosphorus concentration after the reduction of exogenous pollutants in Lake Taihu is still an open question. An in-situ light-dark bottle method was used to investigate the spatiotemporal differences of phosphorus release potential of bloom-forming cyanobacteria (BFC) in Lake Taihu. Generalized additive model analysis (GAM) of field data revealed that the phosphorus release potential of BFC increased with the upregulation of Chlorophyll a (Chl-a) content per cell, which was further validated by the laboratory experiment results. We deduced that the accumulation of Chl-a content per cell might be an essential index of high phosphorus release potential of BFC. The phosphorus release potential of BFC was much higher in summer and autumn than that in spring and winter, while the phosphorus absorption potential increased with the rising of temperature. The distinct physiological status of BFC at different seasons brought about their variation in phosphorus release potential. Additionally, high phosphorus release potential of BFC region mainly concentrated in the eastern and the central, northwest, western, and the south of Lake Taihu in spring, summer, autumn, and winter, respectively. Further studies showed that the spatial differences in phosphorus release potential of BFC were most probably due to the horizontal drift of BFC driven by the prevailing wind. Collectively, the synergism of BFC's physiological status and horizontal drift determined the spatiotemporal differences of phosphorus release potential of BFC in Lake Taihu. Moreover, apparent spatiotemporal differences in phosphorus release potential of BFC were essential factors that induced the distinct distribution of total phosphorus in Lake Taihu. This study provides insight for exploring the reason for the constant increase of total dissolved phosphorus concentration and cyanobacterial density in Lake Taihu for the past 5 years.

Electrolysis-enhanced ecological floating bed and its factors influencing nitrogen and phosphorus removal in simulated hyper-eutrophic water.

To enhance ammonia nitrogen (NH3-N) and phosphate (PO43--P) removal in hyper-eutrophic water, electrolysis-enhanced ecological floating bed (EEEFB) was designed with a Mg-Al alloy anode, a Ir-Ta-Ti metal oxide-coated titanium anode, and an Fe anode with the same graphite cathode. The results showed that the Mg-Al alloy anode with graphite cathode had a better ability to enhance NH3-N and PO43--P removal. When the current density was 0.37 mA·cm-2, the electrolysis time was 24 h/d, and the net removal rates of NH3-N and PO43--P were 62% and 99.4%, respectively. In winter, the purification efficiencies of NH3-N and PO43--P were as high as 7388.4 mg·m-2 and 4297.5 mg·m-2, respectively, by EEEFBs which were significantly higher than the traditional ecological floating bed (p < 0.05). Scanning electron microscopy (SEM) and X-ray spectrometry confirmed that the PO43--P was deposited in the sediment of EEEFBs with Mg-Al alloy anode and Fe anode.

Dark accelerates dissolved inorganic phosphorus release of high-density cyanobacteria.

Bloom-forming cyanobacteria dramatically influence nutrient cycling in eutrophic freshwater lakes. The phosphorus (P) assimilation and release of bloom-forming cyanobacteria significantly may also affect the phosphorus source and amounts in water. To understand the phosphorus release process of bloom-forming cyanobacteria below the accumulated surface and sedimentary bloom-forming cyanobacteria, the degradation of bloom-forming cyanobacteria dominated by Microcystis spp. at different cell density in the dark was investigated over a 25-day microcosm experiment. The dissolved inorganic phosphorus (DIP) and dissolved total phosphorus (DTP) contents increased with the increment of cyanobacterial density, and the dark status markedly increased the proportion of DIP in water during the decline period of bloom-forming cyanobacteria. Meanwhile, the process of cyanobacterial apoptosis accompanied by the changes of malondialdehyde (MDA) and phosphatase (AKP) contents, and the increases of superoxide dismutase (SOD) and catalase (CAT) activities of cyanobacteria in the dark, especially in low-density groups (5.23×108 cells L-1), which further affect the physicochemical water parameters. Moreover, the DIP release from high-density cyanobacteria (7.86×107 cells L-1~5.23×108 cells L-1) resulted from the relative abundance of organophosphorus degrading bacteria in the dark. Therefore, the fast decay of cyanobacteria in the dark could accelerate DIP release, the high DIP release amount from accumulated bloom-cyanobacteria provide adequate P quickly for the sustained growth of cyanobacteria.

Transcriptome Analysis of Different Tissues Reveals Key Genes Associated With Galanthamine Biosynthesis in Lycoris longituba.

L ycoris longituba is a traditional medicinal plant containing the bioactive compound galanthamine (Gal), a type of Amaryllidaceae alkaloid and can be used to treat Alzheimer's disease. However, research on its genome or transcriptome and associated genes in the biosynthetic pathway is incomplete. In this study, we estimated the nuclear genome size of this species to be 29.33 Gb by flow cytometry. Then, RNA sequencing of the leaves, roots, and bulbs of L. longituba was carried out. After de novo assembly, 474,589 all-transcripts and 333,440 all-unigenes were finally generated. In addition, the differentially expressed genes (DEGs) were identified, and genes involved in the galanthamine metabolic pathway encoding tyrosine decarboxylase (TYDC), phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), p-coumarate 3-hydroxylase (C3H), norbelladine synthase (NBS), norbelladine 4'-O-methyltransferase (OMT), and noroxomaritidine synthase (CYP96T1) were detected and validated by real-time quantitative PCR analysis. One candidate gene, Lycoris longituba O-Methyltransferase (LlOMT), was identified in the proposed galanthamine biosynthetic pathway. Sequence analysis showed that LlOMT is a class I OMT. LlOMT is localized in the cytoplasm, and biochemical analysis indicated that the recombinant LlOMT catalyzes norbelladine to generate 4'-O-methylnorbelladine. The protoplast transformation result showed that the overexpression of LlOMT could increase the Gal content. Our results indicate that LlOMT may play a role in galanthamine biosynthesis in L. longituba. This work provides a useful resource for the metabolic engineering of Amaryllidaceae alkaloids.

Highly efficient removal of nitrogen and phosphorus in an electrolysis-integrated horizontal subsurface-flow constructed wetland amended with biochar.

Electrolysis combined with biochar (BC) was used in a constructed wetland to intensify nitrogen (N) and phosphorus (P) removal from wastewater simultaneously. A pilot study was conducted using an electrolysis-integrated, BC-amended, horizontal, subsurface-flow, constructed wetland (E-BHFCW). The research results showed that both electrolysis and BC substrate played important roles in the intensified, constructed wetland. The electrolysis combined BC substrate greatly enhanced the removal rates of nitrate (49.54%) and P (74.25%) when the E-BHFCW operated under the lower current density of 0.02 mA/cm2 and an electrolysis time of 24 h. Improved N removal was accomplished with the electrochemical denitrification of iron cathodes; the autotrophic denitrification bacteria appeared to remove nitrate which was adsorbed on the BC substrate because hydrogen gas was produced by cathodes in the E-BHFCW. Less nitrate was taken directly by wetland plants and microbes. The in-situ formation of ferric ions from a sacrificial iron anode, causing P chemical sedimentation and physical adsorption, improved P removal. BC, modified by iron ions from an iron anode to adsorb the nitrate and P, was a good material to improve effluent water quality. It can also serve as a favorable microbial carrier to bio-transform nitrate to N gas. This is because there were abundant and diverse bacterial communities in the biofilm on the BC substrate in the E-BHFCW. Thus, electrolysis integrated with BC in a constructed wetland is a novel, feasible and effective technique for enhancing wastewater N and P removal.

Highly Effective Polyphosphate Synthesis, Phosphate Removal, and Concentration Using Engineered Environmental Bacteria Based on a Simple Solo Medium-Copy Plasmid Strategy.

Microbial polyphosphate (polyP) production is vital to the removal of phosphate from wastewater. However, to date, engineered polyP synthesis using genetically accessible environmental bacteria remains a challenge. This study develops a simple solo medium-copy plasmid-based polyphosphate kinase (PPK1) overexpression strategy for achieving maximum intracellular polyphosphate accumulation by environmental bacteria. The polyP content of the subsequently engineered Citrobacter freundii (CPP) could reach as high as 12.7% of its dry weight. The biomass yield of CPP was also guaranteed because of negligible metabolic burden effects resulting from the medium plasmid copy number. Consequently, substantial removal of phosphate (Pi) from the ambient environment was achieved simultaneously. Because of the need for exogenous Pi for in vivo ATP regeneration, CPP could thoroughly remove Pi from synthetic municipal wastewater when it was applied for the "one-step" removal of Pi with a bench-scale sequence batch membrane reactor. Almost all the phosphorus except for that assimilated by CPP for cellular growth could be recovered in the form of more concentrated Pi. Overall, engineering environmental bacteria to overexpress PPK1 via a solo medium-copy plasmid strategy may represent a valuable general option for not only biotechnological research based on sufficient intracellular polyP production but also removal of Pi from wastewater and Pi enrichment.

Cadmium toxicity to Microcystis aeruginosa PCC 7806 and its microcystin-lacking mutant.

The adverse effects of microcystin (MC) produced by cyanobacteria have drawn considerable attention from the public. Yet it remains unclear whether MC confers any benefits to the cyanobacteria themselves. One suggested function of MC is complexation, which may influence the bioaccumulation and toxicity of trace metals. To test this hypothesis, we examined Cd toxicity to wild-type Microcystis aeruginosa PCC 7806 (WT) and its MC-lacking mutant (MT) under nutrient-enriched (+NP), phosphorus-limited (-P), and nitrogen-limited (-N) conditions. The accumulation of Cd and the biochemical parameters associated with its detoxification [total phosphorus (TP), inorganic polyphosphate (Poly-P), and glutathione (GSH) in the cells as well as intra- and extra-cellular carbohydrates] were quantified. Although the -P cyanobacteria accumulated less Cd than their +NP and -N counterparts, the different nutrient-conditioned cyanobacteria were similarly inhibited by similar free ion concentration of Cd in the medium ([Cd2+]F). Such good toxicity predictability of [Cd2+]F was ascribed to the synchronous decrease in the intracellular concentrations of Cd and TP. Nevertheless, Cd toxicity was still determined by the intracellular Cd to phosphorus ratio (Cd/P), in accordance with what has been reported in the literature. On the other hand, the concentrations of TP, Poly-P, and carbohydrates went up, but GSH concentration dropped down with the enhancement of [Cd2+]F, indicating their association with Cd detoxification. Although the inactivation of MC peptide synthetase gene had some nutrient and Cd concentration dependent effects on the parameters above, both cyanobacterial strains showed the same Cd accumulation ability and displayed similar Cd sensitivity. These results suggest that MC cannot affect metal toxicity either by regulating metal accumulation or by altering the detoxification ability of the cyanobacteria. Other possible functions of MC need to be further investigated.

Effects of nitrogen and phosphorus on arsenite accumulation, oxidation, and toxicity in Chlamydomonas reinhardtii.

We studied arsenite (iAs(III)) accumulation, oxidation, and toxicity in the freshwater green alga Chlamydomonas reinhardtii under nutrient-enriched (+NP), phosphorus-limited (-P), and nitrogen-limited (-N) conditions. The -P alga (55.1 µM) had a Michaelis constant (Kd) for uptake approximately one tenth of the +NP (419 µM) and -N (501 µM) cells, indicating iAs(III) uptake inhibition by extracellular phosphate. This conclusion was supported by the hyperbolic reduction in iAs(III) uptake rate (V) from 9.2 to 0.8 µmol/g-dw/h when the extracellular phosphate concentration went up from 0 to 250 µM. The maximal iAs(III) uptake rate (Vmax) of the -N alga (24.3 µmol/g-dw/h) was twice as much as that of the +NP (12 µmol/g-dw/h) and -P (8.1 µmol/g-dw/h) cells. It implies that more arsenic transporters were synthesized under the -N condition. Once accumulated, iAs(III) was oxidized and a higher proportion of arsenate (iAs(V)) was observed at lower [As]dis or under nutrient-limited conditions. Nevertheless, iAs(III) oxidation mainly occurred outside the cells with the extent of oxidation reciprocal to [As]dis. Based on the logistic modeling of the concentration-response curves in the +NP, -P, and -N toxicity tests, iAs(III) had an [As]dis-based EC50 of 1763, 13.1, and 1208 µM and an intracellular arsenic concentration based EC50 of 35.6, 28.8, and 195 µmol/g-dw, respectively. Higher iAs(III) toxicity to the -P cells occured because of their increased iAs(III) accumulation, whereas the underlying mechanisms why the -N alga was more tolerant need to be further revealed. Overall, both N and P had remarkable effects on the behavior and effects of iAs(III), which cannot be disregarded in the biogeochemical cycling research of arsenic.

[Characteristics and influencing factors of phosphorus adsorption on sediment in Lake Taihu and Lake Hulun].

Lake Taihu and Lake Hulun in southern and northern China were selected for the investigation of differences in the characteristics and influencing factors of phosphorus (P) adsorption on the sediments of these two lakes by laboratory simulation experiment. The results showed that: (1) The P adsorption capacity of sediment in Lake Taihu was much higher than that in Lake Hulun, and the maximum adsorption capability for the sediments in these two lakes was 1428.57 mg x kg(-1) and 56.81 mg x kg(-1), respectively; (2) Compared with the sediments in Lake Hulun, the particle diameters from sediments in Lake Taihu were much smaller, and their specific surface areas were much larger, so their P adsorption capacity were much higher; (3) The equilibrium adsorbed amounts in these two lakes were correlated with the total amounts of Al, Fe, Mn (P < 0.05) and significantly correlated to the contents of active Al, Fe, Si, Mn (P < 0.01). However, there was significant negative correlation between the equilibrium adsorbed amounts and total Si contents for the sediments in these two lakes (P < 0.01); (4) The P adsorption capacity decreased as pH values in the overlying water increased, and the increase of pH values affected the adsorption behavior of phosphorus on the sediments in Lake Taihu much more obviously. Therefore, the characteristics of sediments and the variations of pH values in the overlying water affected the adsorption behavior of P on sediments in lake water body.

Seasonal variation of phytoplankton nutrient limitation in Lake Taihu, China: a monthly study from year 2011 to 2012.

Lake Taihu is the third largest freshwater lake in China with severe eutrophication issues. However, it remains ambiguous how its phytoplankton growth is limited by various nutrients in different seasons. A series of bottle-enrichment assays in Meiliang Bay was thus performed once a month from July, 2011 to June, 2012 in the present study. The initial chlorophyll a concentration and phytoplankton cell density ranged from 4.70 to 34.6 µg/l and from 1.25×10(6) to 6.72×10(8) cells/l with three peaks in July, November, and March. Although Cyanophyta was dominant (30.9-99.2 percent) in most cases, other phyla like Chlorophyta, Bacillariophyta, and Cryptophyta could account for as much as 69.1 percent of total phytoplankton in cold seasons. The microcystin-LR content in the particulate phase followed a similar seasonal pattern as Cyanophyta. It further went up exponentially with the proportion of cyanobacteria in phytoplankton suggesting more toxigenic species and (or) upregulated microcystin synthesis when the contribution of Cyanophyta was enhanced. On the other hand, the dissolved concentrations of various nitrogen and phosphorus species reached their maxima in late spring and autumn, respectively. According to its growth response to nutrient addition, phytoplankton in Meiliang Bay was restricted by nitrogen in August, October, and November. No nutrient limitation occurred in July, September, and April, whereas phosphorus deficiency prevailed in the other months. Overall, nutrient limitation in Lake Taihu and possibly other aquatic ecosystems worldwide may be more dynamic than what we thought before, which should be considered to eliminate eutrophication.

[Analysis on the forms and release potential of nutrients in sediments from lakes in the West Jiangsu Province].

To probe the effect of nitrogen and phosphorus (P) in sediment on aqueous nutrient concentrations, forms of phosphorus, sediment water interface diffusion fluxes and phosphorus release and adsorption of 10 lakes in the west Jiangsu province were studied. Sediments were sequentially extracted by the modified Ruttenberg's method, and the result showed that inorganic phosphorus in sediments of the lakes was the major fraction of total phosphorus, but the proportion of bioavailable phosphorus was relatively low. Compared with the lakes in northern Jiangsu, the Fe-P was relatively high in sediments of lakes in southern Jiangsu. The diffusion fluxes of lakes were mainly based on NH4(+) -N diffusion in the west Jiangsu, and ions of most of lakes diffused from overlying water to sediment, NO3(-) -N diffused from sediment to overlying water in lakes except for Lake Hongze, Lake Shijiu and Lake Xuanwu. The condition leaning to acidic (pH < 4) or alkaline (pH > 10) was beneficial to phosphorus release, and except for Lake Xuanwu, the least amount of phosphorus released form sediment was higher in acidic condition than that in alkaline condition. The amount of phosphorus adsorption was increasing with temperature increase. Considering the phosphorus adsorption on native phosphorus polluted (NAP-polluted) sediments. All data of P adsorption on sediment samples collected from 7 lakes were fitted to the modified Langmuir model. And the values of the maximum adsorption capacity (Q(m)), phosphorus binding constant (k), native adsorbed exchangeable phosphorus (Q(NAP)), were subsequently obtained. The results showed that the adsorption amount of P in Lake Hongze and Lake Xuanwu was relatively low, which were liable to release P from sediments.

[Regional heterogeneity of lake eutrophication effects in China].

Although biomass of algae (Chl-a) were in variant levels between different lake regions in China under the same nutrients conditions, it demonstrated that efficiencies of TN/TP used by algae had regional differences. In order to clarify the differences, curve estimation in SPSS was used to analyze the linear relationship between Ig Chl-a and Ig TN/lg TP. The slopes of these linear equations were identified as the efficiencies of TN/TP used by algae. The slopes of linear equations from Mengxin Plateau, Yungui Plateau, Northeast Mountain-Plain, lower reach of Yangtze River Plain and North Plain were 1.002, 0.817, 0.761, 0.545, 0.250, orderly. The efficiencies of TN used by algae ranged from the highest to the lowest were lower reach of Yangtze River Plain, Yungui Plateau, North Plain, Northeast Mountain-Plain, Mengxin Plateau, and the slopes of linear equations were 1.401, 1.058, 0.447, 0.239, 0. 099, respectively. Consequently, in Northeast Mountain-Plain, Mengxin Plateau the efficiencies of TP used by algae were higher than those of TN, and in Yangtze River Plain, Yungui Plateau, North Plain, the efficiencies of TN used by algae were higher than those of TP. On the other hand, in order to describe the effects of algae on transparency in different lakes, the relationships between Chl-a and SD were analyzed. The results showed that in Yungui Plateau the effect of algae on transparency was the most obvious as the variation of SD explained by Chl-a was the highest, and Northeast Mountain-Plain, Mengxin Plateau and North Plain followed. However, in lower reach of Yangtze River Plain, the relationship between Chl-a and SD was not significant.

Accumulation kinetics of arsenic in Daphnia magna under different phosphorus and food density regimes.

In the present study, the dissolved uptake, dietary assimilation, and elimination of arsenic (initially added as arsenate) in the freshwater cladoceran Daphnia magna were examined. A biphasic correlation between the arsenic uptake rate and its ambient concentration, as well as a two-saturation-site arsenic uptake competition with phosphate was observed. The calculated uptake rate constant, as influenced by the ambient phosphorus concentration, ranged from 0.035 to 0.35 L/g/d. Food concentration substantially decreased (by 23.2-64.4%) the arsenic assimilation efficiency with the incipient limiting algal food concentration of 3.86 mg/L dry weight. Arsenic assimilation by the daphnids was independent of their own phosphorus status, but was lower when their algal diet was phosphorus-limited and thus contained a higher proportion of arsenite due to the enhanced biotransformation. Arsenic efflux rate constant ranged from 0.34 to 0.44 d(-1) with increased food concentration slightly facilitating its loss. Excretion, accounting for 51.3 to 60.6% of total loss, was the dominant pathway for arsenic elimination with a remarkable contribution from offspring production (24.7-29.8%), whereas molting (3.64-4.05%) and egestion (7.9-11.9%) had minor roles only. According to the well-established biokinetic model, dietary assimilation was predicted to be the main pathway for arsenic bioaccumulation in the daphnids, and arsenic has a great potential to be biodiminished along the food chain.

Simultaneous removal of phosphorus and nitrogen in a sequencing batch biofilm reactor with transgenic bacteria expressing polyphosphate kinase.

To improve phosphorus removal from wastewater, we constructed a high-phosphate-accumulating microorganism, KTPPK, using Pseudomonas putida KT2440 as a host. The expression plasmid was constructed by inserting and expressing polyphosphate kinase gene (ppk) from Microcystis aeruginosa NIES-843 into broad-host-range plasmid, pBBR1MCS-2. KTPPK was then added to a sequencing batch biofilm reactor (SBBFR) using lava as a biological carrier. The results showed that SBBFR with KTPPK not only efficiently removed COD, NH(3)-N, and NO(3)(-)-N but also had a high removal capacity for PO(4)(3-)-P, resulting in a low phosphorus concentration remaining in the outflow of the SBBFR. The biofilm increased by 30-53% on the lava in the SBBFR that contained KTPPK after 11 days when compared with the reactor that contained P. putida KT2440. Real-time quantitative polymerase chain reaction confirmed that the copy of ppk was maintained at about 3.5 × 10(10) copies per µL general DNA in the biofilm after 20 days. Thus, the transgenic bacteria KTPPK could maintain a high density and promote phosphorus removal in the SBBFR. In summary, this study indicates that the use of SBBFR with transgenic bacteria has the potential to remove phosphorus and nitrogen from wastewater.