The present study investigated the physiological responses, photosynthetic activity, and microbial community structure of leaf-associated biofilms on the microphyte Vallisneria natans during a harmful algal bloom. Results of the physiological and photosynthetic indices (Fᵥ/Fₘ ratios [maximum quantum yield of photosystem II (PSII)]; malondialdehyde content; total chlorophyll; and activities of superoxide dismutase, catalase and peroxidase) indicated that algal blooms could cause inhibition of photosynthesis, oxidative stress and an antioxidant system stress response in Vallisneria natans leaf-associated biofilms. Multifractal analysis suggested that allelochemicals or algal organic matter released by cyanobacteria could reduce the surface roughness of the leaf. Microbial diversity analysis of the biofilms showed that algal blooms slightly altered the microbial community structure while the richness and evenness of the microbial composition remained stable. This study provided useful information to better understand the adverse effects of algal blooms on submerged macrophytes.

Harmful microalgal blooms are a threat to aquatic organisms, ecosystems and human health. Toxic dinoflagellates of the genus Alexandrium are known to produce paralytic shellfish toxins and to release bioactive extracellular compounds (BECs) with potent cytotoxic, hemolytic, ichtyotoxic and allelopathic activity. Negative allelochemical interactions refer to the chemicals that are released by the genus Alexandrium and that induce adverse effects on the physiology of co-occurring protists and predators. Releasing BECs gives the donor a competitive advantage that may help to form dense toxic blooms of phytoplankton. However BECs released by Alexandrium minutum are uncharacterized and it is impossible to quantify them using classical chemical methods. Allelochemical interactions are usually quantified through population growth inhibition or lytic-activity based bioassays using a secondary target organism. However these bioassays require time (for growth or microalgal counts) and/or are based on lethal effects. The use of pulse amplitude modulation (PAM) fluorometry has been widely used to assess the impact of environmental stressors on phytoplankton but rarely for allelochemical interactions. Here we evaluated the use of PAM and propose a rapid chlorophyll fluorescence based bioassay to quantify allelochemical BECs released from Alexandrium minutum. We used the ubiquitous diatom Chaetoceros muelleri as a target species. The bioassay, based on sub-lethal effects, quantifies allelochemical activity from different samples (filtrates, extracts in seawater) within a short period of time (2 h). This rapid bioassay will help investigate the role of allelochemical interactions in Alexandrium bloom establishment. It will also further our understanding of the potential relationship between allelochemical activities and other cytotoxic activities from BECs. While this bioassay was developed for the species A. minutum, it may be applicable to other species producing allelochemicals and may provide further insights into the role and impact of allelochemical interactions in forming dense algal blooms and structuring marine ecosystems.

To elucidate the origin of the wide-spread contaminations of plant derived commodities with various alkaloids, we employed co-cultures of pyrrolizidine alkaloid (PA) containing Senecio jacobaea plants with various alkaloid free acceptor plants. Our analyses revealed that all plants grown in the vicinity of the Senecio donor plants indeed contain significant amounts of the PAs, which previously had been synthesized in the Senecio plants. These findings illustrate that typical secondary metabolites, such as pyrrolizidine alkaloids, are commonly transferred and exchanged between living plants. In contrast to the broad spectrum of alkaloids in Senecio, in the acceptor plants nearly exclusively jacobine is accumulated. This indicates that this alkaloid is exuded specifically by the Senecio roots. Although the path of alkaloid transfer from living donor plants is not yet fully elucidated, these novel insights will extend and change our understanding of plant-plant interactions and reveal a high relevance with respect to the widespread alkaloidal contaminations of plant-derived commodities. Moreover, they could be the basis for the understanding of various so far not fully understood phenomena in cultivation of various crops, e.g. the beneficial effects of crop rotations or the co-cultivation of certain vegetables.

In this study, we aim to explore the allelochemicals of marine brown algae Sargassum fusiforme and the mechanism of allelopathy effect on Heterosigma akashiwo. Six potential allelopathic substances, palmitic acid, arachidonic acid, α-linolenic acid (ALA), fucosterol, 24-hydroperoxy-24-vinylcholesterol, and saringosterowere, were isolated and identified from S. fusiforme by column chromatography, NMR, TOF-MS and GC‐MS analyses. The ALA and 24-hydroperoxy-24-vinylcholesterol showed remarkable inhibitory activities on H. akashiwo growth. Besides, the allelopathic mechanism between the ALA and H. akashiwo was preliminarily investigated. The results indicated that the activities of SOD and POD of H. akashiwo gradually decreased under high ALA concentration. The reduction of photosynthetic pigments and chlorophyll fluorescence parameters, as well as apparent electron transport rate in H. akashiwo cells, aggravated gradually with increasing the concentration of the ALA. In summary, this study revealed the responses of morphology and physiology of H. akashiwo when exposed by ALA, while revealing the potential of macroalgae in red tide control.

The blooms of harmful microalgae represent a prominent threat to fisheries, public health, and economies throughout the world. Recent studies have shown that certain macroalgae release allelochemicals that can inhibit the growth of bloom-forming microalgae. In this study, we found that the macroalga Pyropia haitanensis significantly inhibited growth of the harmful bloom-forming microalgae Pseudo-nitzschia pungens and Pseudo-nitzschia multiseries. The inhibitory-effect of the live thali of P. haitanensis was highest, followed by that of dry powder, water-soluble extract, and culture medium filtrate. The Pseudo-nitzschia species died 96 h after exposure to 5–10 g fresh-weight L⁻¹ of P. haitanensis live thalli. Furthermore, an aqueous extract of P. haitanensis suppressed the growth of P. pungens and P. multiseries, thereby indicating that P. haitanensis contains stable allelopathic substances that cause the observed inhibitory-effects. On the basis of these findings, we conclude that the macroalga P. haitanensis would have potential utility in controlling the blooms of Pseudo-nitzschia species.

The dinoflagellate Karlodinium veneficum has worldwide distribution and is associated with harmful algal blooms through the production of karlotoxins. We investigated the allelopathy and toxicity to explore the potential ecological implications. Prorocentrum donghaiense was inhibited significantly when grown either in co-cultures or in culture filtrate of K. veneficum. In addition, the effect of the co-occurring microalga species (P. donghaiense) on the hemolytic activity of K. veneficum was also evaluated. P. donghaiense did not inhibit the growth of K. veneficum but increased the hemolytic activity. The culture of K. veneficum was loaded onto an RP-C18 column and eluted with different percentages of aqueous methanol solution. 80% methanol fraction not only inhibited the growth of P. donghaiense by allelopathy but also exhibited strong hemolytic activity, indicating that the allelochemicals and toxins of K. veneficum might be the same components. Furthermore, KmTx 3 (C₆₈H₁₂₄O₂₄) was identified using HPLC-HRMS from this fraction.

The microorganisms and allelochemicals in Pontederia cordata planting water may have a synergistic inhibitory effect on algae. To study this synergy, an algae-inhibiting organism was isolated and identified, and its growth and feeding characteristics were studied. The organism was identified as Poterioochromonas malhamensis yzs924 based on both its morphology and molecular barcoding employing 18S rDNA gene sequences.The growth and feeding of P. malhamensis were affected by environmental factors and the state of its prey. (1) P. malhamensis is a mixotrophic flagellate. Its heterotrophic growth was the fastest in a wheat grain medium, and its growth rate in this study reached 2.5 day⁻¹. (2) Within a short period of time (2 days), P. malhamensis growth was slower under continuous dark conditions than under alternating light and dark conditions, but it fed on Microcystis aeruginosa more rapidly under dark conditions. (3) High pH was disadvantageous to the growth and grazing of P. malhamensis. When the pH was kept stable at 9, P. malhamensis could not grow continuously. (4) When the initial density of M. aeruginosa was 5 × 10⁷ cells/mL or is in a period of decline, P. malhamensis could not remove all M. aeruginosa. The combined use of P. malhamensis and allelochemicals may represent a method of M. aeruginosa control, but this approach requires further research.

Egeria densa release allelochemicals harmful to other species, including cyanobacteria. Under natural conditions, E. densa and Microcystis aeruginosa can be found in various aquatic systems with different light levels. The effect that the coexistence of E. densa has on M. aeruginosa was tested under different photosynthetically active radiation (PAR) intensities. The growth, chlorophyll-a, oxidative stress, and antioxidant activities of M. aeruginosa were quantified after 7 days of coexistence with E. densa. A control experiment without E. densa was conducted for M. aeruginosa under the same conditions to distinguish the effect of the PAR intensity from the allelopathic effect. The allelopathy of E. densa on M. aeruginosa is significantly influenced by the PAR intensity. PAR intensities lower or higher than the favorable range further suppressed M. aeruginosa under coexistence. Research has revealed that the allelopathic effect of E. densa can be utilized to biologically suppress M. aeruginosa under various light conditions. Strong second-order polynomial distributions between the optical density (OD) and cellular H₂O₂ content of M. aeruginosa were found for both the control and coexistence conditions. In the practical application of E. densa allelopathy, OD can be adopted as a convenient method to obtain an approximate stress status on M. aeruginosa.

A continuous flow filtration system was designed to identify and quantify the removal mechanisms of Cyanobacteria (Microcystis aeruginosa) by hydroponic biofilters of Phalaris arundinacea compared to synthetic filters. The filtration units were continuously fed under plug-flow conditions with Microcystis grown in photobioreactors. Microcystis cells decreased at the two flow rates studied (1.2 ± 0.2 and 54 ± 3 cm³ minˉ¹) and results suggested physical and chemical/biological removal mechanisms were involved. Physical interception and deposition was the main removal mechanism with packing density of the media driving the extent of cell removal at high flow, whilst physical and chemical/biological mechanisms were involved at low flow. At low flow, the biofilters decreased Microcystis cell numbers by 70% compared to the controls. The decrease in cell numbers in the biofilters was accompanied by a chlorotic process (loss of green colour), suggesting oxidative processes by the release of allelochemicals from the biofilters.

Green tides have been increasing in frequency and severity in coastal areas in recent years, and thus, the adverse effects of green tides on the environment have attracted much research attention. Allelopathy is one of the most significant effects of green tide algae on the surrounding organisms. In this study, a series of experiments were developed to fully investigate the allelopathic effects of the green tide alga Ulva linza on two common coastal phytoplankton species and to isolate and identify the chemical compositions of the allelochemicals. Our results indicated that the fresh tissue, dry powder, and aqueous extracts all inhibited the population growth of the two experimental phytoplankton species, indicating the occurrence of allelopathy; further analysis of the allelochemicals identified multiple kinds of fatty acids as allelopathic compounds, in which the most abundant ones were hexadecanoic acid; 9Z, 12Z, 15Z-octadecatetrienoic acid; and 9E-octadecenoic acid. Our findings enrich the database for research on allelopathy between marine green tide algae and phytoplankton. Both algae and phytoplankton could increase their own competitive abilities during bloom formation, thereby changing the phytoplankton community structure.