Down-regulation of iron/zinc ion transport and toxin synthesis in Microcystis aeruginosa exposed to 5,4'-dihydroxyflavone.

Flavonoids, common natural polyphenolic compounds from plants, have been proposed as highly effective and safe algicides. However, the molecular mechanism of flavonoids inhibiting Microcystis aeruginosa remains unclear. This study aims in exploring the global transcriptional changes and molecular docking in cyanobacterial cells in response to flavonoids. Transcriptomic analysis revealed that 5,4'-dihydroxyflavone (DHF) primarily affected the genes transcription of iron and zinc ion transport, resulting in the blockage of transport for iron (II), iron (III) and zinc (II), which eventually led to a decrease in intracellular iron and zinc content. 5,4'-DHF can also interfere with iron and zinc transport by binding to metal ion transport-related proteins, leading to eliminated biological activities in M. aeruginosa. Meanwhile, 5,4'-DHF inhibit microcystin synthesis and reduce the content of intercellular toxin by inhibiting the transcription of mcyC and binding with McyC protein, implying that 5,4'-DHF have potential to reduce the risk of microcystins in the environment. Moreover, iron starvation and down-regulation of photosynthesis-related genes transcription led to the inhibition of electron transport in photosynthetic system. These results provide more information for the inhibitory mechanism of flavonoids, and the inhibition of flavonoids on metal ion transmembrane transport provides a new perspective for the development of allelochemical algicides.

Natural Algaecide Sphingosines Identified in Hybrid Straw Decomposition Driven by White-Rot Fungi.

Harmful algal blooms (HABs), which are promoted by eutrophication and intensified by global warming, occur worldwide. Allelochemicals, which are natural chemicals derived from plants or microbes, are emerging weapons to eliminate these blooms. However, the cost and technical challenges have limited the discovery of novel antialgal allelochemicals. Herein, the decomposition of agricultural straws is manipulated by white-rot fungi and achieved elevated antialgal efficiency. The transcriptomic analysis reveals that nutrient limitation activated fungal decomposition. By using a comparative nontarget metabolomics approach, a new type of allelochemical sphingosines (including sphinganine, phytosphingosine, sphingosine, and N-acetylsphingosine) is identified. These novel natural algaecides exhibit superior antialgal capability, with as high as an order of magnitude lower effective concentration on blooming species than other prevalent allelochemicals. The co-expression relationship between transcriptomic and metabolomic results indicate that sphinganine is strongly correlated with the differentially expressed lignocellulose degradation unigenes. The algal growth suppression is triggered by the activation of programmed cell death, malfunction of algal photosystem and antioxidant system, the disruption on CO2 assimilation and light absorption. The sphingosines reported here are a new category of allelochemicals in addition to the well-known antialgal natural chemicals, which are potential species-specific agents for HABs control identified by multi-omics methodology.

Programmed cell death process in freshwater Microcystis aeruginosa and marine Phaeocystis globosa induced by a plant derived allelochemical.

Harmful algal blooms (HAB) are a serious problem worldwide. Allelochemicals from natural plants were recently thought to be promising anti-algaecide in controlling harmful algae. However, the programmed cell death (PCD) process of algae under allelopathic pressure induced by 5,4'-dihydroxyflavone (5,4'-DHF) was poorly understood. In this study, two common and worldwide distributed microalgae, Microcystis aeruginosa and Phaeocystis globosa were selected as target algae, and the PCD processes induced by 5,4'-DHF were cross-compared between the two species. Both algae species were inhibited significantly by 5,4'-DHF with the relative sensitivity of 0.11. To uncover the PCD progress systematically, signals for PCD triggering, antioxidant enzyme activity, photosynthetic ability variation, caspase-like activities and typical indicators were investigated. In both species, typical indicators of PCD - phosphatidylserine externalization and chromatin condensation - were detected. The intracellular reactive oxygen species (ROS), nitric oxide (NO) and H2O2 were the potential signal molecules to stimulate PCD, and caspase-like activities were activated with an elevation of cytochrome c indicating the initiation of PCD in both species. However, P. globosa responded to 5,4'-DHF immediately after 3 h with the elevation of ROS and not in M. aeruginosa. Antioxidant enzyme activities of superoxide dismutase (SOD) and catalase (CAT) in M. aeruginosa and P. globosa also showed different patterns on day 3. Specifically, SOD activity in M. aeruginosa increased significantly while it decreased significantly in P. globosa, CAT activity in M. aeruginosa decreased significantly while it increased significantly in P. globosa (p < 0.05). Malondialdehyde (MDA) content in P. globosa increased significantly (p < 0.001) while it showed no variation in M. aeruginosa. Overall, this study is one of the earliest studies to explore the inhibition and action mechanism of plant derived flavonoids on harmful algae from the perspective of PCD, and provide new insights into the antialgal mechanism of allelochemicals.

3-Indoleacrylic acid from canola straw as a promising antialgal agent - Inhibition effect and mechanism on bloom-forming Prorocentrum donghaiense.

Harmful algal blooms (HABs) have induced severe damage worldwide. A novel high-efficient antialgal natural chemical, 3-indoleacrylic acid (3-IDC) with a 5-day half-maximal inhibitory concentration (IC50, 5d), was discovered from canola straw, and its algal inhibition mechanism was investigated. Adverse effects were observed on the growth of P. donghaiense with 3-IDC addition, following an increase in reactive oxygen species (ROS) production. 3-IDC also hindered the photosynthetic mechanism of P. donghaiense cells. Transcriptional results showed 3-IDC inhibiting the functions of all the nutrient assimilating genes, down-regulated ribulose-1,5-bisphosphate carboxylase/oxygenase II, and cytochrome f genes. The expression of heat shock protein (HSP) 70 and 90 and rhodopsin genes were also suppressed. The binding affinity of investigated receptors was observed. The conformational changes induced by the spatial microstructural alteration through 3-IDC may further contribute to the perturbation of those enzyme catalytic activities. The present results provide new insights on controlling HABs using 3-IDC.

New Insights into the Microplastic Enrichment in the Blue Carbon Ecosystem: Evidence from Seagrass Meadows and Mangrove Forests in Coastal South China Sea.

Microplastics were recently found to aggregate in the blue carbon ecosystems (BCEs), which are known for their ability to store carbon by slowing down the water flow. However, evidence is largely lacking on how the accumulation of microplastics is related to carbon sequestration in BCEs and if this trap effect is driven by its biological characteristics. In this study, the trap effect of microplastics by BCEs was evaluated for various seagrasses (Zostera japonica, Halophila ovalis, and Halophila beccarii) and mangroves (Aegiceras corniculatum and Avicennia marina). Significant accumulation was found in the seagrass meadow dominated by H. beccarii and the mangrove forest dominated by A. marina, with microplastics enriched by 1.3 to 17.6 times compared to their corresponding unvegetated sites. The abundance of microplastics varied greatly from 17.68 ± 8.10 to 611.75 ± 81.52 particles per kg of dry sediment, with the highest abundance in A. marina mangrove sediments. A strong positive correlation was found between the abundance of microplastics and the particulate organic carbon content at all study sites (Pearson, R = 0.86, p < 0.01). Higher diversity of microplastic colors and size was found in the H. beccarii meadow, and higher diversity of shapes was found in the A. marina forest. Our results added new insights to the understanding of the mechanism of microplastic trapping by BCEs and coupled the behavior of microplastics with the organic carbon in the sediment.

Effects of polyacrylamide molecular weight and mass concentration on water transport characteristics of iron tailings.

Iron tailings have few macropores which severely inhibit infiltration and transport of soil water. Polyacrylamide (PAM) can regulate soil water, but it is rarely used when remediating tailings matrix. In this research, PAM of four molecular weights of 300w, 600w, 800w, and 1000w were selected as amendments, and were each applied at five mass concentrations of 0% (CK), 0.01%, 0.04%, 0.08%, and 0.16% to observe their effects on water transport in iron tailings using column simulations in the laboratory. After adding PAM, the water retention and saturated water content of iron tailings increased significantly (P < 0.05). With increases in PAM molecular weight and mass concentration, the saturated hydraulic conductivity showed a downward trend, but the saturated hydraulic conductivity increased after a dry-wet cycle. With the increase of PAM mass concentration, adding PAM of 1000w molecular weight to iron tailing decreased infiltration capacity, but treatments of other molecular weights all initially increased then decreased infiltration capacity. The greatest improvement on infiltration capacity of iron tailings was observed with the addition of PAM of 300w molecular weight and 0.01% mass concentration. Adding PAM increased the vertical depth of the saturation zone of iron tailings (P < 0.05) with a maximum depth of 20.83 cm. The Kostiakov model more accurately simulated water infiltration of iron tailings compared with the Horton and Philip models. On the whole, when PAM of low molecular weight and concentration was added to iron tailings, PAM increased stable infiltration, saturated water content, and water retention. It also inhibited saturated hydraulic conductivity of iron tailings. Therefore, in practice, it is necessary to select the appropriate molecular weight and mass concentration of PAM according to the dominant limiting factors and remediation needs of the matrix.

Seaweed farms provide refugia from ocean acidification.

Seaweed farming has been proposed as a strategy for adaptation to ocean acidification, but evidence is largely lacking. Changes of pH and carbon system parameters in surface waters of three seaweed farms along a latitudinal range in China were compared, on the weeks preceding harvesting, with those of the surrounding seawaters. Results confirmed that seaweed farming is efficient in buffering acidification, with Saccharina japonica showing the highest capacity of 0.10 pH increase within the aquaculture area, followed by Gracilariopsis lemaneiformis (ΔpH = 0.04) and Porphyra haitanensis (ΔpH = 0.03). The ranges of pH variability within seaweed farms spanned 0.14-0.30 unit during the monitoring, showing intense fluctuations which may also help marine organisms adapt to enhanced pH temporal variations in the future ocean. Deficit in pCO2 in waters in seaweed farms relative to control waters averaged 58.7 ± 15.9 µatm, ranging from 27.3 to 113.9 µatm across farms. However, ΔpH did not significantly differ between day and night. Dissolved oxygen and Ωarag were also elevated in surface waters at all seaweed farms, which are benefit for the survival of calcifying organisms. Seaweed farming, which unlike natural seaweed forests, is scalable and is not dependent on suitable substrate or light availability, could serve as a low-cost adaptation strategy to ocean acidification and deoxygenation and provide important refugia from ocean acidification.

LncRNA NEAT1 Knockdown Inhibits Retinoblastoma Progression by miR-3619-5p/LASP1 Axis.

Retinoblastoma (RB) is the most common intraocular tumor in childhood. Long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NTAT1) has been reported to be related to RB progression. This study aims to study the molecular mechanism of NEAT1 in regulating cell cycle, proliferation, apoptosis, migration, and invasion in RB. The expression levels of NEAT1 and miR-3619-5p were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The protein expression of LIM and SH3 domain protein 1 (LASP1) was measured by western blot. The proliferation of RB cells was analyzed by cell counting kit-8 (CCK-8) and cell colony formation assays. Cell migration and invasion were evaluated by transwell assay. Cell cycle and apoptosis were assessed by flow cytometry analysis. The association between miR-3619-5p and NEAT1 or LASP1 was predicted by starBase 3.0 database and identified by dual-luciferase reporter assay. The effects of NEAT1 knockdown on the tumor growth in vivo were detected by in vivo tumor formation assay. NEAT1 expression was dramatically up-regulated, and miR-3619-5p expression was obviously downregulated in RB tissues and cells compared with control groups. The protein level of LASP1 was obviously increased in RB tissues or cells relative to paracancerous normal tissues or cells, respectively. Functionally, NEAT1 silencing inhibited RB cell migration, invasion, and proliferation, whereas induced cell apoptosis and cell cycle arrest in RB; this phenomenon was partially abolished by miR-3619-5p inhibitor. Mechanistically, NEAT1 acted as a sponge of miR-3619-5p, and miR-3619-5p was associated with LASP1. In addition, NEAT1 knockdown decreased the volume and weight of RB tumor in vivo. Together, NEAT1 silencing repressed cell migration, invasion, and proliferation, whereas induced cell apoptosis and cycle arrest by sponging miR-3619-5p to inhibit LASP1 expression in RB cells. This study may provide a theoretical basis for RB therapy.

New insights into the harmful algae inhibition by Spartina alterniflora: Cellular physiology and metabolism of extracellular secretion.

This study investigated the response of three toxic harmful algal blooms (HABs) species, Heterosigma akashiwo, Chattonella marina, and Alexandrium tamarense to coastal invasive plant, Spartina alterniflora. In this study, the growth of three HABs species were suppressed significantly by S. alterniflora extracts, showing a dose-response relationship. The raphidophyte H. akashiwo and C. marina exhibited higher inhibitory response with EC50,7d decreased by 14% and 75% as compared to the dinoflagellate A. tamarense. C. marina was the most sensitive among the three species. S. alterniflora extracts disrupted algal cellular integrity and photosynthesis. Furthermore, the extracellular organic matters were detected by fluorescence excitation-emission matrix. Algal metabolites, protein-like substances (tyrosine-like peak and tryptophan-like peak) decreased as time prolonged and the humic-like substances (UVA marine humic-like peak) increased when algal cells were exposed to S. alterniflora extracts. These results provide new insights to the inhibition mechanism of S. alterniflora extracts on HABs species.

Seagrass beds acting as a trap of microplastics - Emerging hotspot in the coastal region?

Microplastics is an emerging environmental problem in the world. However, presence and fate of microplastics in seagrass meadows are barely known. In this study, the abundance and diversity of microplastic from Enhalus acodoides vegetated sites and bare sites were quantified and characterized in Xincun bay and Li'an bay, Hainan, China. Microplastics ranged from 80.0 to 884.5 particles per kg of dry sediment, and fibers were the dominant shape. The most frequent colors of microplastics were blue, transparent and black. The dominant size of microplastics was in the range of 125-250 µm. And the seagrass sediments were enriched in microplastics 2.1 and 2.9 times for Xincun bay and Li'an bay, respectively. The trap effect of seagrass was non-selective regarding the shape, color and size of microplastics. High anthropogenic pollution and poor beach management may contribute to higher concentrations of microplastics in Li'an bay.

Inhibition of harmful algae Phaeocystis globosa and Prorocentrum donghaiense by extracts of coastal invasive plant Spartina alterniflora.

Harmful algal blooms (HABs) and the invasion of smooth cordgrass (Spartina alterniflora) have become important environmental problems in intertidal zones of China, which caused serious damage on the coastal ecological systems. By using S. alterniflora as algaecide, this helps to utilize this invasive plant, in addition, is expected to help to control the expansion of S. alterniflora. The potential of S. alterniflora in HABs mitigation was investigated on controlling Phaeocystis globosa (haptophyceae) and Prorocentrum donghaiense (dinophyceae). The growth of both HABs species was significantly inhibited at high concentrations, and P. globosa was more sensitive than P. donghaiense. Furthermore, the extracts of S. alterniflora reduced the effective quantum yield, photosynthetic efficiency, and relative maximal electron transport rate of both algal species at high concentrations, which implies a disruption on their photosynthetic system. Flavonoids, which were previously known as antialgal chemicals, were found to be abundant in the extracts of S. alterniflora by UPLC-MS detection. Our results revealed that the potential of S. alterniflora as a novel antialgal agent for controlling HABs, simultaneously, resource utilization possibility for the invasive plant S. alterniflora.

Understanding the inhibitory mechanism of antialgal allelochemical flavonoids from genetic variations: Photosynthesis, toxin synthesis and nutrient utility.

Flavonoids are natural polyphenolic compounds from plants. As a new biotechnological algaecide, the molecular mechanism of plant flavonoids on the inhibition of Microcystis aeruginosa is still unknown. Therefore, in this study, we analyzed the variation of expressions of photosynthesis-related genes, microcystin synthesis-related genes and the genes involved in N and P acquisition in M. aeruginosa under the flavonoids stress. The results showed that the expression of psbD1, psaB and rbcL related to photosynthesis were influenced by three flavonoids but with different changing tendencies. The transcription of mcyA, mcyD and mcyH related to microcystin synthesis were decreased after 5-d of exposure, which could block microcystin synthesis. Meanwhile, flavonoids treatments resulted in the inhibition of N and P acquisition related genes transcription to affect the absorption of N and P in algal cells, and further influenced the physiological metabolic process of M. aeruginosa.

Teleconnection between phytoplankton dynamics in north temperate lakes and global climatic oscillation by time-frequency analysis.

We are still facing the knowledge gap of how the water-quality extremes (i.e. phytoplankton blooms), their causes, severity or occurrence could be directly related to the climatic oscillation. Considering that the climatic and phytoplankton concentration time series are highly non-stationary, we applied the advanced time-frequency analysis - Ensemble Empirical Mode Decomposition (EEMD), Hilbert-Huang Spectrum (HHS) and Wavelet Analysis (WA) - to examine the variability of long term phytoplankton dynamics from 1986 to 2014 in five North Temperate Lakes (NTLs). These analysis techniques isolated five separate time series for the surface Chlorophyll a concentrations (CHL) of the five NTLs and a time series for the global climate oscillation (denoted by multivariate ENSO index, MEI), and showed that these time series generally operated at similar time scales. The long-term residual trends of decreasing were found in three lakes (i.e., BM, SP and TR lakes), which are the same to global climate dynamics (MEI). The wavelet analysis reveals strong coherency between MEI and CHL data sets for all lakes, with a periodicity of 64-months. Intuitive associations between the CHL and MEI data set showed that two types of ENSO (El Nino and La Nina) differ in their influences to CHL. Potential mechanisms relating the phytoplankton dynamics in NTLs to climatic oscillation (ENSO) were also discussed.