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Recent studies have shown that seagrasses could possess potential applications in the treatment of inflammatory disorders. Five seagrass species (Zostera muelleri, Halodule uninervis, Cymodocea rotundata, Syringodium isoetifolium, and Thalassia hemprichii) from the Great Barrier Reef (QLD, Australia) were thus collected, and their preliminary antioxidant and anti-inflammatory activities were evaluated. From the acetone extracts of five seagrass species subjected to 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging antioxidant assay, the extract of Z. muelleri had the highest activity (half minimal concentration of inhibition (IC50) = 138 µg/mL), with the aerial parts (IC50 = 119 µg/mL) possessing significantly higher antioxidant activity than the roots (IC50 ≥ 500 µg/mL). A human peripheral blood mononuclear cells (PBMCs) assay with bacterial lipopolysaccharide (LPS) activation and LEGENDplex cytokine analysis showed that the aerial extract of Z. muelleri significantly reduced the levels of inflammatory cytokines tumour necrosis factor alpha (TNF-α), interleukin (IL)-1β, and IL-6 by 29%, 74%, and 90%, respectively, relative to the LPS treatment group. The aerial extract was thus fractionated with methanol (MeOH) and hexane fraction, and purification of the MeOH fraction by HPLC led to the isolation of 4-hydroxybenzoic acid (1), luteolin (2), and apigenin (3) as its major constituents. These compounds have been previously shown to reduce levels of TNF-α, IL-1β, and IL-6 and represent some of the major bioactive components of Z. muelleri aerial parts. This investigation represents the first study of the antioxidant and anti-inflammatory properties of Z. muelleri and the first isolation of small molecules from this species. These results highlight the potential for using seagrasses in treating inflammation and the need for further investigation.

Genetic diversity can modulate a population\'s response to a changing environment and plays a critical role in its ecological function. While multiple processes act to maintain genetic diversity, sexual reproduction remains the primary driving force. Eelgrass (Zostera marina) is an important habitat-forming species found in temperate coastal ecosystems across the globe. Recent increases in sea surface temperatures have resulted in shifts to a mixed-annual life-history strategy (i.e., displaying characteristics of both annual and perennial meadows) at its southern edge-of-range. Given that mating systems are intimately linked to standing levels of genetic variation, understanding the scope of sexual reproduction can illuminate the processes that shape genetic diversity. To characterize edge-of-range eelgrass mating systems, developing seeds on flowering Z. marina shoots were genotyped from three meadows in Topsail, North Carolina. In all meadows, levels of multiple mating were high, with shoots pollinated by an average of eight sires (range: 3-16). The number of fertilized seeds (i.e., reproductive success) varied significantly across sires (range: 1-25) and was positively correlated with both individual heterozygosity and self-fertilization. Outcrossing rates were high (approx. 70%) and varied across spathes. No clones were detected, and kinship among sampled flowering shoots was low, supporting observed patterns of reproductive output. Given the role that genetic diversity plays in enhancing resistance to and resilience from ecological disturbance, disentangling the links between life history, sexual reproduction, and genetic variation will aid in informing the management and conservation of this key foundation species.

BACKGROUND: Breast cancer is the most common type of cancer diagnosed in women. Finding novel therapeutic agents with significant cytotoxic action and minimal adverse impact on normal cells becomes crucial. Today, natural anticancer agents present an unconventional method of treating cancer, either as a curative or preventative agent, with considerable concern for marine organisms.
METHODS: The anticancer effect of the alcoholic extract of different Red Sea Seagrasses on MCF-7 human breast cancer cell line has been investigated. Seagrasses were collected from Wadi El Gamal, Red Sea and extracted. Qualitative HPLC analysis was performed on the extracts for the identification of their active biomarkers. This study was aimed to explore the cytotoxic impact of Thalassia hemprichii (Ehren.) and Enhalus acoroides (L.f.) Royle on MCF-7 and their mode of action. Their anti-proliferative effects on cancer cells were performed using Neutral red assay. On the other hand, their apoptotic effect and their capacity to induce cell cycle arrest were investigated by flow cytometry assay. The effect of Seagrasses on the mitochondrial membrane potential (ΔψM) was studied by using JC-1 mitochondrial membrane potential assay kit in Seagrasses treated cancer cells to Δψ Caspases 3/7activity was examined using the colorimetric method. Gene expression analysis and quantitative real time RT-PCR for the sea grasses on MCF-7 was performed. Immune-blotting technique for Bcl-2 and p53 was investigated.
RESULTS: HPLC analysis demonstrated that the extracts contained mainly flavonoids and polyphenols such as Caffeic acid, Chlorogenic acids, catechin and kaempferol that might be responsible for these anticancer effects. Seagrasses alcoholic crude extract markedly suppressed the growth and expansion of MCF-7 cells concentration-dependently with no toxicity against normal human skin fibroblast HSF. Thalassia hemprichii and Enhalus acoroides trigger mode of cell death primarily via apoptosis as confirmed by the flow cytometry. Additionally, they have ability to induce G0/S cell cycle arrest in MCF-7. The data showed the depletion in mitochondrial membrane potential (ΔψM) in the treated cells dose-dependently Caspases 3/7activities markedly increased following 24 h treatment. Finally, Gene expression analysis showed a marked reduction in Bcl-2, Survivin and CDC2 gene expression levels and a significant increase in the expression of p53 and CC2D1A as compared to control cells.
CONCLUSIONS: In summary, the Methanolic extract of seagrass, Thalassia hemperchii and Enhalus ocoroides are able to induce concentration-dependent cytotoxic effects in human MCF-7 cells through intrinsic pathway of apoptosis in MCF-7 cells. This study reveals the beneficial importance of sea grasses as a source of anticancer agents. Further in vivo study is recommended for the active isolated biomolecules.

Turtlegrass virus X, which infects the seagrass Thalassia testudinum, is the only potexvirus known to infect marine flowering plants. We investigated potexvirus distribution in seagrasses using a degenerate reverse transcription polymerase chain reaction (RT-PCR) assay originally designed to capture potexvirus diversity in terrestrial plants. The assay, which implements Potex-5 and Potex-2RC primers, successfully amplified a 584 nt RNA-dependent RNA polymerase (RdRp) fragment from TVX-infected seagrasses. Following validation, we screened 74 opportunistically collected, apparently healthy seagrass samples for potexviruses using this RT-PCR assay. The survey examined the host species T. testudinum, Halodule wrightii, Halophila stipulacea, Syringodium filiforme, Ruppia maritima, and Zostera marina. Potexvirus PCR products were successfully generated only from T. testudinum samples and phylogenetic analysis of sequenced PCR products revealed five distinct TVX sequence variants. Although the RT-PCR assay revealed limited potexvirus diversity in seagrasses, the expanded geographic distribution of TVX shown here emphasizes the importance of future studies to investigate T. testudinum populations across its native range and understand how the observed fine-scale genetic diversity affects host-virus interactions.

This study investigated the physiological responses of two tropical seagrass species, Halophila ovalis and Thalassia hemprichii, to heat stress under varying light conditions in a controlled 5-day experiment. The experimental design included four treatments: control, saturating light, heat stress under sub-saturating light, and heat stress under saturating light (combined stress). We assessed various parameters, including chlorophyll fluorescence, levels of reactive oxygen species (ROS), antioxidant enzyme activities, and growth rates. In H. ovalis, heat stress resulted in a significant reduction in the maximum quantum yield of photosystem II (Fv/Fm) regardless of the light condition. However, the effects of heat stress on the effective quantum yield of photosystem II (ɸPSII) were more pronounced under saturating light conditions. In T. hemprichii, saturating irradiance exacerbated the heat stress effects on Fv/Fm and ɸPSII, although the overall photoinhibition was less severe than in H. ovalis. Heat stress led to ROS accumulation in H. ovalis and reduced the activity of superoxide dismutase (SOD) and ascorbate peroxidase in the sub-saturating light condition. Conversely, T. hemprichii exhibited elevated SOD activity under saturating light. Heat stress suppressed the growth of both seagrass species, regardless of the light environment. The Biomarker Response Index indicated that H. ovalis displayed severe effects in the heat stress treatment under both light conditions, while T. hemprichii exhibited moderate effects in sub-saturating light and major effects in saturating light conditions. However, the Effect Addition Index revealed an antagonistic interaction between heat stress and high light in both seagrass species. This study underscores the intricate responses of seagrasses, emphasizing the importance of considering both local and global stressors when assessing their vulnerability.

High concentrations of copper can pollute coastal waters, primarily from agricultural runoff and mining activities, which can harm marine organisms, including seagrasses. The molecular mechanism of copper toxicity to seagrass currently remains unclear. To determine the response to copper, physiological and multi-omic analyses were conducted to explore the molecular mechanism by which copper affects the global threatened seagrass Halophila beccarii Asch. Excessive copper stress causes oxidative damage and stimulates the activity of the antioxidant enzyme system to remove excess reactive oxygen species (ROS), thereby reducing the damage caused by copper stress. Cu increases the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7), ascorbate peroxidase (EC 1.11.1.11), glutathione peroxidase (EC 1.11.1.9), ascorbate oxidase (EC 1.10.3.3), glutathione reductase (EC 1.6.4.2), and dehydroascorbate reductase (EC 1.8.5.1) and the content of malondialdehyde and reduces the activity of monodehydroascorbate reductase (EC 1.6.5.4). Under copper stress, H. beccarii upregulates the metabolic pathways of steroid biosynthesis and cutin, suberin, and wax biosynthesis, downregulates the metabolic pathways of arginine and proline metabolism and fructose and mannose metabolism; the levels of expression of the ribosome-related genes; upregulates the levels of expression of circadian rhythm-related proteins and downregulates the levels of glutathione metabolism and the proteins related to carbon fixation. This study provides new insights into the response of seagrass to copper stress and reports potential candidate metabolites, genes, and proteins that can be considered as biomarkers to improve the protection and management of seagrass meadows.

The increase of marine heat waves (MHWs) occurrence is exacerbated in Mediterranean Sea and temperature resilience-enhancing strategies on key species, such as the seagrass Posidonia oceanica, need to be investigated. \"Priming\" describes a stimulus that prepares an organism for an improved response to upcoming environmental changes by triggering a memory that remains during a lag-phase. The aim of this study, conducted in Sardinia (Italy), was to investigate whether the development of thermo-primed P. oceanica seedlings is affected by a field simulated MHW depending on the duration of the lag-phase. After the thermo-priming stimulus, seedlings had a 0, 7 or 14 days lag-phase and after that, for each lag-phase group, half of the seedlings experienced a simulated MHW (the other half served as controls). Some other seedlings did not experience either the priming stimulus or the lag-phase. Results did not show any evidence of a memory triggered by the priming stimulus, but they highlighted the importance of an acclimation phase before the highest temperature: seedlings that experienced a gradual increase of temperature had a higher number of leaves and shorter leaf necrosis length compared to seedlings that had a lag-phase between two heat events. Regardless the priming stimulus, MHWs slowed down the development of the leaf and root length. Considering the increase of temperature fluctuations, testing different intensities of priming and different length of lag-phase is necessary to provide information about the adaptive success of the species.

Zostera marina, a critical keystone marine angiosperm species in coastal seagrass meadows, possesses a photosensitive oxygen evolving complex (OEC). In harsh environments, the photoinactivation of the Z. marina OEC may lead to population declines. However, the factors underlying this photosensitivity remain unclear. Therefore, this study was undertaken to elucidate the elements contributing to Z. marina OEC photosensitivity. Our results demonstrated a gradual decrease in photosystem II performance towards shorter wavelengths, especially blue light and ultraviolet radiation. This phenomenon was characterized by a reduction in Fv/Fm and the rate of O2 evolution, as well as increased fluorescence at 0.3 ms on the OJIP curve. Furthermore, exposure to shorter light wavelengths and longer exposure durations significantly reduced the relative abundance of the OEC peripheral proteins, indicating OEC inactivation. Analyses of light-screening substances revealed that carotenoids, which increased most notably under 420 nm light, might primarily serve as thermal dissipators instead of efficient light filters. In contrast, anthocyanins reacted least to short-wavelength light, in terms of changes to both their content and the expression of genes related to their biosynthesis. Additionally, the levels of aromatically acylated anthocyanins remained consistent across blue-, white-, and red-light treatments. These findings suggest that OEC photoinactivation in Z. marina may be linked to inadequate protection against short-wavelength light, a consequence of insufficient synthesis and aromatic acylation modification of anthocyanins.

Seagrass meadows serve as critical marine habitats, offering numerous benefits to both humans and wildlife. Taiping Island, situated in the South China Sea, has been historically known for its abundant seagrass meadows. However, satellite imagery suggested there was a low density of seagrasses around Taiping Island. On the other hand, many green turtles (Chelonia mydas) were observed from the shore. To investigate this phenomenon, we conducted drone surveys of the shallow reefs and discovered a density of 902 ± 601 (mean ± SD) individual turtles per square kilometer during high tides. In addition, we conducted a cage experiment to test the hypothesis that large herbivores are impacting seagrass abundance negatively. The results indicated that the blade lengths of seagrasses in cages were significantly longer than those outside. It is likely that large herbivores such as green sea turtles are the key consumers of seagrass on the shallow reef flats of Taiping Island. Accordingly, further research and management should take into account that the increasing number of sea turtles may deplete the seagrasses and have an impact on the seagrass ecosystems.