The Use of Invasive Algae Species as a Source of Secondary Metabolites and Biological Activities: Spain as Case-Study.

In the recent decades, algae have proven to be a source of different bioactive compounds with biological activities, which has increased the potential application of these organisms in food, cosmetic, pharmaceutical, animal feed, and other industrial sectors. On the other hand, there is a growing interest in developing effective strategies for control and/or eradication of invasive algae since they have a negative impact on marine ecosystems and in the economy of the affected zones. However, the application of control measures is usually time and resource-consuming and not profitable. Considering this context, the valorization of invasive algae species as a source of bioactive compounds for industrial applications could be a suitable strategy to reduce their population, obtaining both environmental and economic benefits. To carry out this practice, it is necessary to evaluate the chemical and the nutritional composition of the algae as well as the most efficient methods of extracting the compounds of interest. In the case of northwest Spain, five algae species are considered invasive: Asparagopsis armata, Codium fragile, Gracilaria vermiculophylla, Sargassum muticum, and Grateulopia turuturu. This review presents a brief description of their main bioactive compounds, biological activities, and extraction systems employed for their recovery. In addition, evidence of their beneficial properties and the possibility of use them as supplement in diets of aquaculture animals was collected to illustrate one of their possible applications.

Independent and synergistic activity of the flavonoids of Gracilaria corticata as promising antidiabetic agents

The therapeutic approaches for Type 2 Diabetes Mellitus rely most on the usage of oral hypoglycaemic drugs. These drugs have adverse side effects and hence alternative medicines are continuously explored. The present study intends to investigate the antidiabetic potential of the flavonoids present in Gracilaria corticata. The flavonoids were isolated (FEGC) and their inhibitory activity on the carbohydrate hydrolysing enzymes such as α-amylase and α-glucosidase was analysed. The flavonoids were found to inhibit α-amylase and α-glucosidase with an IC50 value of 302 µg and 75 µg respectively. The synergistic effect of FEGC and luteolin was also investigated and the results show that both FEGC and luteolin inhibited synergistically at half their IC50 values. The observations of this study reveal that the flavonoids of G. corticata have potential antidiabetic activity and can act independently or synergistically in the management of Type 2 Diabetes Mellitus

Unraveling the nuclear and chloroplast genomes of an agar producing red macroalga, Gracilaria changii (Rhodophyta, Gracilariales).

Agar and agarose have wide applications in food and pharmaceutical industries. Knowledge on the genome of red seaweeds that produce them is still lacking. To fill the gap in genome analyses of these red algae, we have sequenced the nuclear and organellar genomes of an agarophyte, Gracilaria changii. The partial nuclear genome sequence of G. changii has a total length of 35.8Mb with 10,912 predicted protein coding sequences. Only 39.4% predicted proteins were found to have significant matches to protein sequences in SwissProt. The chloroplast genome of G. changii is 183,855bp with a total of 201 open reading frames (ORFs), 29 tRNAs and 3 rRNAs predicted. Five genes: ssrA, leuC and leuD CP76_p173 (orf139) and pbsA were absent in the chloroplast genome of G. changii. The genome information is valuable in accelerating functional studies of individual genes and resolving evolutionary relationship of red seaweeds.

Genetic and morphological analyses of Gracilaria firma and G. changii (Gracilariaceae, Rhodophyta), the commercially important agarophytes in western Pacific.

Many studies classifying Gracilaria species for the exploitation of agarophytes and the development of the agar industry were conducted before the prevalence of molecular tools, resulting in the description of many species based solely on their morphology. Gracilaria firma and G. changii are among the commercially important agarophytes from the western Pacific; both feature branches with basal constrictions that taper toward acute apices. In this study, we contrasted the morpho-anatomical circumscriptions of the two traditionally described species with molecular data from samples that included representatives of G. changii collected from its type locality. Concerted molecular analyses using the rbcL and cox1 gene sequences, coupled with morphological observations of the collections from the western Pacific, revealed no inherent differences to support the treatment of the two entities as distinct taxa. We propose merging G. changii (a later synonym) into G. firma and recognize G. firma based on thallus branches with abrupt basal constrictions that gradually taper toward acute (or sometimes broken) apices, cystocarps consisting of small gonimoblast cells and inconspicuous multinucleate tubular nutritive cells issuing from gonimoblasts extending into the inner pericarp at the cystocarp floor, as well as deep spermatangial conceptacles of the verrucosa-type. The validation of specimens under different names as a single genetic species is useful to allow communication and knowledge transfer among groups from different fields. This study also revealed considerably low number of haplotypes and nucleotide diversity with apparent phylogeographic patterns for G. firma in the region. Populations from the Philippines and Taiwan were divergent from each other as well as from the populations from Malaysia, Thailand, Singapore and Vietnam. Establishment of baseline data on the genetic diversity of this commercially important agarophyte is relevant in the context of cultivation, as limited genetic diversity may jeopardize the potential for its genetic improvement over time.

Complete chloroplast genome of Gracilaria firma (Gracilariaceae, Rhodophyta), with discussion on the use of chloroplast phylogenomics in the subclass Rhodymeniophycidae.

BACKGROUND: The chloroplast genome of Gracilaria firma was sequenced in view of its role as an economically important marine crop with wide industrial applications. To date, there are only 15 chloroplast genomes published for the Florideophyceae. Apart from presenting the complete chloroplast genome of G. firma, this study also assessed the utility of genome-scale data to address the phylogenetic relationships within the subclass Rhodymeniophycidae. The synteny and genome structure of the chloroplast genomes across the taxa of Eurhodophytina was also examined. RESULTS: The chloroplast genome of Gracilaria firma maps as a circular molecule of 187,001 bp and contains 252 genes, which are distributed on both strands and consist of 35 RNA genes (3 rRNAs, 30 tRNAs, tmRNA and a ribonuclease P RNA component) and 217 protein-coding genes, including the unidentified open reading frames. The chloroplast genome of G. firma is by far the largest reported for Gracilariaceae, featuring a unique intergenic region of about 7000 bp with discontinuous vestiges of red algal plasmid DNA sequences interspersed between the nblA and cpeB genes. This chloroplast genome shows similar gene content and order to other Florideophycean taxa. Phylogenomic analyses based on the concatenated amino acid sequences of 146 protein-coding genes confirmed the monophyly of the classes Bangiophyceae and Florideophyceae with full nodal support. Relationships within the subclass Rhodymeniophycidae in Florideophyceae received moderate to strong nodal support, and the monotypic family of Gracilariales were resolved with maximum support. CONCLUSIONS: Chloroplast genomes hold substantial information that can be tapped for resolving the phylogenetic relationships of difficult regions in the Rhodymeniophycidae, which are perceived to have experienced rapid radiation and thus received low nodal support, as exemplified in this study. The present study shows that chloroplast genome of G. firma could serve as a key link to the full resolution of Gracilaria sensu lato complex and recognition of Hydropuntia as a genus distinct from Gracilaria sensu stricto.

Delimitating cryptic species in the Gracilaria domingensis complex (Gracilariaceae, Rhodophyta) using molecular and morphological data.

Species in the genus Gracilaria that display conspicuously flattened vegetative morphologies are a taxonomically challenging group of marine benthic red algae. This is a result of their species richness, morphological similarity, and broad phenotypic plasticity. Within this group, the Gracilaria domingensis complex is one of the most common, conspicuous, and morphologically variable species along the tropical western Atlantic Ocean. Previous research has identified that members of this complex belong to two distantly related clades. However, despite this increased phylogentic resolution, species delimitations within each of these clades remain unclear. Our study assessed the species diversity within this difficult complex using morphological and molecular data from three genetic markers (cox1, UPA, and rbcL). We additionally applied six single-marker species delimitation methods (SDM: ABGD, GMYCs, GMYCm, SPN, bPTP, and PTP) to rbcL, which were largely in agreement regarding species delimitation. These results, combined with our analysis of morphology, indicate that the G. domingensis complex includes seven distinct species, each of which are not all most closely related: G. cervicornis; a ressurected G. ferox; G. apiculata subsp. apiculata; a new species, Gracilaria baiana sp. nov.; G. intermedia subsp. intermedia; G. venezuelensis; and G. domingensis sensu stricto, which includes the later heterotypic synonym, G. yoneshigueana. Our study demonstrates the value of multipronged strategies, including the use of both molecular and morphological approaches, to decipher cryptic species of red algae.

Effects of galactose adaptation in yeast for ethanol fermentation from red seaweed, Gracilaria verrucosa.

A total monosaccharide concentration of 39.6 g/L, representing 74.0 % conversion of 53.5 g/L total carbohydrate from 80 g dw/L (8 % w/v) Gracilaria verrucosa slurry, was obtained by thermal acid hydrolysis and enzymatic saccharification. G. verrucosa hydrolysate was used as a substrate for ethanol production by 'separate hydrolysis and fermentation' (SHF). The ethanol production and yield (Y EtOH) from Saccharomyces cerevisiae KCCM 1129 with and without adaptation to high galactose concentrations were 18.3 g/L with Y EtOH of 0.46 and 13.4 g/L with Y EtOH of 0.34, respectively. Relationship between galactose adaptation effects and mRNA transcriptional levels were evaluated with GAL gene family, regulator genes of the GAL genetic switch and repressor genes in non-adapted and adapted S. cerevisiae. The development of galactose adaptation for ethanol fermentation of G. verrucosa hydrolysates allowed us to enhance the overall ethanol yields and obtain a comprehensive understanding of the gene expression levels and metabolic pathways involved.

Molluscs associated with the macroalgae of the genus Gracilaria (Rhodophyta): importance of algal fronds as microhabitat in a hypersaline mangrove in Northeastern Brazil / Moluscos associados a macroalgas do gênero Gracilaria (Rhodophyta): importância das frondes algais como microhabitat em um manguezal hipersalino no Nordeste do Brasil

The fronds of marine macroalgae play an important role in coastal ecosystems because the algae banks are utilized as a microhabitat by different taxa, including molluscs, one of the most abundant and diverse animals of marine ecosystems. In this study, we characterized the malacofauna associated with the macroalgae Gracilaria domingensis (Kützing) Sonder ex Dickie 1874 and Gracilaria cuneata Areschoug 1854 of a hypersaline mangrove on the northern coast of the state of Rio Grande do Norte, Northeastern Brazil. The first alga dominates in the rainy season and it is substituted by second one in the dry period. A total of 1,490 molluscs were surveyed, representing 56 species in 29 families: 1,081 were associated with G. domingensis and 409 with G. cuneata, the latter showing the greater diversity (H′=1.25). Columbellidae, Neritidae, Pyramidellidae and Cerithiidae were among the most representative families in the number of species and individuals. The micromolluscs were dominant in the algal microhabitat, constituting 74.63% of the malacofauna recorded. The columbellid Parvanachis obesa (C. B. Adams, 1845) was the dominant species followed by the neritid Neritina virginea (Linnaeus, 1758) in both algae. In spite of the annual alternated succession of the algae species, at least 15 mollusc species are common for these algae. Furthermore, juveniles of P. obesa were recorded in both seasons, indicating a continuous reproduction. Possible reasons for difference in abundance, diversity and dominance of molluscs living on these algae are discussed. Both species of substrate-algae represent an important microhabitat for refuge, feeding and the reproduction of small-sized mollusc species during rainy and dry seasons.

As frondes de macroalgas marinhas exercem um importante papel nos ecossistemas costeiros, tendo em vista que os bancos de algas são utilizados como microhabitat por diferentes táxons, incluindo os moluscos, que estão entre os mais abundantes e diversos animais dos ecossistemas marinhos. Neste estudo, caracterizou-se a malacofauna associada às macroalgas Gracilaria domingensis (Kützing) Sonder ex Dickie 1874 e Gracilaria cuneata Areschoug 1854 de um manguezal hipersalino da costa norte do estado do Rio Grande do Norte, Nordeste do Brasil. A primeira alga domina no período chuvoso e é substituída pela segunda no período seco. Foram inventariados 1.490 moluscos, representando 56 espécies em 29 famílias: 1.081 estavam associados a G. domingensis e 409 a G. cuneata, esta última apresentando maior diversidade (H′=1,25). Columbellidae, Neritidae, Pyramidellidae e Cerithiidae estiveram entre as famílias mais representativas em número de espécies e indivíduos. Os micromoluscos foram dominantes no microhabitat algal, constituindo 74,63% da malacofauna registrada. O columbelídeo Parvanachis obesa (C. B. Adams, 1845) foi a espécie dominante, seguida pelo neritídeo Neritina virginea (Linnaeus, 1758) em ambas as algas. Apesar da sucessão alternada anual das espécies de algas, pelo menos 15 espécies de moluscos são comuns a estas algas. Além disso, os jovens de P. obesa foram registrados em ambas as estações, indicando uma reprodução contínua. São discutidas possíveis razões para a diferença na abundância, diversidade e dominância dos moluscos habitantes destas algas. Ambas as espécies de alga-substrato representam um importante microhabitat para refúgio, alimentação e reprodução para pequenas espécies de moluscos durante as estações chuvosa e seca.

Development of chloroplast simple sequence repeats (cpSSRs) for the intraspecific study of Gracilaria tenuistipitata (Gracilariales, Rhodophyta) from different populations.

BACKGROUND: Gracilaria tenuistipitata is an agarophyte with substantial economic potential because of its high growth rate and tolerance to a wide range of environment factors. This red seaweed is intensively cultured in China for the production of agar and fodder for abalone. Microsatellite markers were developed from the chloroplast genome of G. tenuistipitata var. liui to differentiate G. tenuistipitata obtained from six different localities: four from Peninsular Malaysia, one from Thailand and one from Vietnam. Eighty G. tenuistipitata specimens were analyzed using eight simple sequence repeat (SSR) primer-pairs that we developed for polymerase chain reaction (PCR) amplification. FINDINGS: Five mononucleotide primer-pairs and one trinucleotide primer-pair exhibited monomorphic alleles, whereas the other two primer-pairs separated the G. tenuistipitata specimens into two main clades. G. tenuistipitata from Thailand and Vietnam were grouped into one clade, and the populations from Batu Laut, Middle Banks and Kuah (Malaysia) were grouped into another clade. The combined dataset of these two primer-pairs separated G. tenuistipitata obtained from Kelantan, Malaysia from that obtained from other localities. CONCLUSIONS: Based on the variations in repeated nucleotides of microsatellite markers, our results suggested that the populations of G. tenuistipitata were distributed into two main geographical regions: (i) populations in the west coast of Peninsular Malaysia and (ii) populations facing the South China Sea. The correct identification of G. tenuistipitata strains with traits of high economic potential will be advantageous for the mass cultivation of seaweeds.

Molluscs associated with the macroalgae of the genus Gracilaria (Rhodophyta): importance of algal fronds as microhabitat in a hypersaline mangrove in Northeastern Brazil.

The fronds of marine macroalgae play an important role in coastal ecosystems because the algae banks are utilized as a microhabitat by different taxa, including molluscs, one of the most abundant and diverse animals of marine ecosystems. In this study, we characterized the malacofauna associated with the macroalgae Gracilaria domingensis (Kützing) Sonder ex Dickie 1874 and Gracilaria cuneata Areschoug 1854 of a hypersaline mangrove on the northern coast of the state of Rio Grande do Norte, Northeastern Brazil. The first alga dominates in the rainy season and it is substituted by second one in the dry period. A total of 1,490 molluscs were surveyed, representing 56 species in 29 families: 1,081 were associated with G. domingensis and 409 with G. cuneata, the latter showing the greater diversity (H'=1.25). Columbellidae, Neritidae, Pyramidellidae and Cerithiidae were among the most representative families in the number of species and individuals. The micromolluscs were dominant in the algal microhabitat, constituting 74.63% of the malacofauna recorded. The columbellid Parvanachis obesa (C. B. Adams, 1845) was the dominant species followed by the neritid Neritina virginea (Linnaeus, 1758) in both algae. In spite of the annual alternated succession of the algae species, at least 15 mollusc species are common for these algae. Furthermore, juveniles of P. obesa were recorded in both seasons, indicating a continuous reproduction. Possible reasons for difference in abundance, diversity and dominance of molluscs living on these algae are discussed. Both species of substrate-algae represent an important microhabitat for refuge, feeding and the reproduction of small-sized mollusc species during rainy and dry seasons.