Phenolic chemistry of the seagrass Zostera marina Linnaeus: First assessment of geographic variability among populations on a broad spatial scale.

The variability of the phenolic content of thirteen populations of Zostera marina L. (six narrow-leaved and seven wide-leaved ecotypes) from different geographical zones, i.e., Baltic Sea, Mediterranean, East and West Atlantic, and East Pacific coasts was evaluated. Depending on the location, three to five phenolic acids and nine to fourteen flavonoids were identified of which an undescribed flavonoid sulfate. The phenolic concentrations of the thirteen populations differ among countries and among sites within countries. However, the same individuals were found almost everywhere. Substantial phenolic concentrations were found at all study sites with the exception of Puck Bay (Baltic Sea). Some geographical differences in the flavonoid content were observed. The highest phenolic diversity was found with specimens from the French Atlantic coast and the lowest with the Northeastern American sample (Cape Cod, MA). Regardless of their leaf width, the content of phenolic compounds was found to be similar and mainly characterized by rosmarinic acid and luteolin 7,3'-disulfate. The results demonstrate that geographic origin influences the phenolic composition of Z. marina primarily in terms of concentration, but not in terms of individual compound identity, despite the large geographic scale and the contrasting climatic and environmental conditions associated with it. This work is the first study to consider the spatial variability of phenolic compounds for a seagrass species on a spatial scale covering four bioregions. This is also the first to compare the phenolic chemistry of the two ecotypes of Z. marina.

Phenolic fingerprints of the Pacific seagrass Phyllospadixtorreyi - Structural characterization and quantification of undescribed flavonoid sulfates.

Four undescribed flavonoid sulfates were isolated from Phyllospadix torreyi S. Watson foliar tissue. In addition, nine known flavonoid sulfates and three phenolic acids were isolated from the same extract, of which seven had never been reported for the genus Phyllospadix. Structural elucidation of individual phenolics was assigned using complementary informations from their spectral evidence (HPLC-DAD, LC-MS, NMR, and UV) and chemical behavior. The inter-annual variation in phenolic concentrations was determined by quantitative HPLC-DAD over a three-year period. The results showed a relative constancy of phenolic content over time and the high prevalence of flavonoid disulfates (70-90% of the total flavonoids detected). All samples were found dominated by the unreported nepetin 7, 3'-disulfate and 5-methoxyluteolin 7, 3'-disulfate, followed by luteolin 7, 3'-disulfate. Considering the economic potential of flavonoid sulfates in the pharmaceutical and nutraceutical segments, a sample of detrital leaves was also analyzed. The same phenolic pattern was found and the concentration of the individuals, although lower than in fresh material, makes this abundant biomass of interest for dietary and pharmaceutical applications.

Seasonal variation of phenolic compounds in Zostera marina (Zosteraceae) from the Baltic Sea.

Seasonal variations of phenolic compounds, in leaves of Zostera marina L. from the Baltic Sea near Kiel/Germany were investigated. Dominant compounds were mono- and disulfated flavonoids and phenylpropanoic acids, in particular luteolin 7,3'-O-disulfate and diosmetin 7-O-sulfate as well as rosmarinic acid, a dimeric phenylpropanoid. All detected sulfated flavones showed similar seasonal trends: there were two significant concentration peaks in June and November. Moreover, two geographically distinct flavonoid chemotypes were identified based on their respective main flavonoid; one chemotype was characterized by the prevalence of luteolin 7,3'-O-disulfate (German Baltic Sea), and the other by the prevalence of diosmetin 7-O-sulfate (Norwegian North Sea). Furthermore, an undescribed tetrameric phenylpropanoid, 7'',8''-didehydrosalvianolic acid B, was isolated and its structure was established by extensive NMR, MS, and CD experiments. This compound inhibited activity of Na+/K+-ATPase in the micro-molar range without any cytotoxic effects against human cancer and normal cells.

Erratum: Comparative Chloroplast Genomes of Zosteraceae Species Provide Adaptive Evolution Insights Into Seagrass.

[This corrects the article DOI: 10.3389/fpls.2021.741152.].

Comparative Chloroplast Genomes of Zosteraceae Species Provide Adaptive Evolution Insights Into Seagrass.

Seagrasses are marine flowering plants found in tropical and sub-tropical areas that live in coastal regions between the sea and land. All seagrass species evolved from terrestrial monocotyledons, providing the opportunity to study plant adaptation to sea environments. Here, we sequenced the chloroplast genomes (cpGenomes) of three Zostera species, then analyzed and compared their cpGenome structures and sequence variations. We also performed a phylogenetic analysis using published seagrass chloroplasts and calculated the selection pressure of 17 species within seagrasses and nine terrestrial monocotyledons, as well as estimated the number of shared genes of eight seagrasses. The cpGenomes of Zosteraceae species ranged in size from 143,877 bp (Zostera marina) to 152,726 bp (Phyllospadix iwatensis), which were conserved and displayed similar structures and gene orders. Additionally, we found 17 variable hotspot regions as candidate DNA barcodes for Zosteraceae species, which will be helpful for studying the phylogenetic relationships and interspecies differences between seagrass species. Interestingly, nine genes had positive selection sites, including two ATP subunit genes (atpA and atpF), two ribosome subunit genes (rps4 and rpl20), two DNA-dependent RNA polymerase genes (rpoC1 and rpoC2), as well as accD, clpP, and ycf2. These gene regions may have played key roles in the seagrass adaptation to diverse environments. The Branch model analysis showed that seagrasses had a higher rate of evolution than terrestrial monocotyledons, suggesting that seagrasses experienced greater environmental pressure. Moreover, a branch-site model identified positively selected sites (PSSs) in ccsA, suggesting their involvement in the adaptation to sea environments. These findings are valuable for further investigations on Zosteraceae cpGenomes and will serve as an excellent resource for future studies on seagrass adaptation to sea environments.

Phenolic chemistry of the seagrass Zostera noltei Hornem. Part 1: First evidence of three infraspecific flavonoid chemotypes in three distinctive geographical regions.

The flavonoid content of Zostera noltei leaves was investigated over a broad spatial scale using chromatographic and spectroscopic techniques (HPLC-DAD, LC/MS and NMR). Samples were collected at fifteen localities covering Mediterranean Sea and NE Atlantic coast, and representative of three types of coastal ecosystems: mesotidal bays, coastal lagoons, and open-sea. Three geographically distinct flavonoid chemotypes were identified on the basis of their respective major compound. One is characterized by apigenin 7-sulfate (Eastern part of Gulf of Cadiz), one by diosmetin 7-sulfate (French Atlantic coast and Mediterranean Sea), and the third contained similar quantities of the above two compounds (Mauritania and South Portugal). Our results show that metabolomic profiling using a combination of analytical techniques is a tool of choice to characterize chemical phenotype accurately. This work emphasizes for the first time the spatial variability in the flavonoid chemistry of Z. noltei throughout Atlantic and Mediterranean range, and constitutes the first report of chemical races in the Zosteraceae family. This infraspecific chemical differentiation should be considered when dealing with the role of Z. noltei in coastal ecosystems or in the selection of the best population donor for Z. noltei beds restoration. Combined with molecular identification, phenolic fingerprinting might be helpful to elucidate the evolutionary history of Z. noltei.

Secondary metabolites of seagrasses (Alismatales and Potamogetonales; Alismatidae): Chemical diversity, bioactivity, and ecological function.

Seagrasses are the only higher plants living in fully marine environments; they play a significant role in coastal ecosystems. Seagrasses inhabit the coastal shelves of all continents except Antarctica and can grow in depths of up to 90 m. Because of their eminent ecological importance, innumerous studies have been dedicated to seagrasses and their ecology. However, the phytochemistry has not been equally well investigated yet and many of the existing studies in chemical ecology are only investigating the chemistry at the level of compound classes, e.g. phenolics, and not at the level of chemically defined metabolites. In the present review, the existing literature on secondary metabolites of seagrasses, their known source seagrasses, their bioactivity, and ecological function are compiled and critically assessed. Moreover, research gaps are highlighted and avenues for future research are discussed. Currently, a total of 154 chemically defined natural products have been reported from the about 70 seagrass species known worldwide. Compounds reported include simple phenols derivatives (four compounds), phenylmethane derivatives (14 compounds), phenylethane derivatives (four compounds), phenylpropane derivatives including their esters and dimers (20 compounds), chalkones (four compounds), flavonoids including catechins (57 compounds), phenylheptanoids (four compounds), one monoterpene derivative, one sesquiterpene, diterpenoids (13 compounds), steroids (31 compounds), and one alkaloid. Most of the existing bioactivity studies of seagrass metabolites and extracts have been directed to potential cytotoxic, antimicrobial, or antimacrofouling activity. Antimicrobial studies have been performed towards panels of both human pathogens and ecologically relevant pathogens. In the antimacrofouling studies, investigations of the potential of zosteric acid from the genus Zostera are the most numerous and have yielded so far the most interesting results. Studies on the chemical ecology of seagrasses often have been focused on variation in phenolic compounds and include but are not limited to studies on variation due to abiotic factors, seasonal variation, variation in response to grazing by fish or sea urchins, or following microbial attack.

Development of microsatellite loci for the endangered seagrass Zostera japonica (Zosteraceae).

PREMISE OF THE STUDY: New microsatellite markers were developed for the Asian endangered seagrass Zostera japonica (Zosteraceae) to assess genetic diversity and population structure of this species. In China, Z. japonica populations have drastically decreased since the 1970s. METHODS AND RESULTS: A total of 12 polymorphic tetranucleotide microsatellite loci were isolated and characterized in Z. japonica. The number of alleles per locus ranged from one to 11. The expected and observed heterozygosity ranged from 0 to 0.772 and from 0 to 1.000, respectively. CONCLUSIONS: The new microsatellites will be useful in evaluating clonality and population structure of Z. japonica and aiding in conservation and management of the endangered seagrass in Asia.