Vaibhav A. Mantri

Fatty acid profiling of tropical marine macroalgae: An analysis from chemotaxonomic and nutritional perspectives

The lipid and fatty acid (FA) compositions for 100 marine macroalgae were determined and discussed from the context of chemotaxonomic and nutritional perspectives. In general, the lipid contents in macroalgae were low (2.3-20 mg/g fr. wt.) but with substantially high amounts of nutritionally important polyunsaturated fatty acids (PUFAs) such as LA, ALA, STA, AA, EPA and DHA, that ranged from 10% to 70% of TFAs. More than 90% of the species showed nutritionally beneficial n6/n3 ratio (0.1:1-3.6:1) (p≤0.001). A closer look at the FA data revealed characteristic chemotaxonomic features with C18 PUFAs (LA, ALA and STA) being higher in Chlorophyta, C20 PUFAs (AA and EPA) in Rhodophyta while Phaeophyta depicted evenly distribution of C18 and C20 PUFAs. The ability of macroalgae to produce long-chain PUFAs could be attributed to the coupling of chloroplastic FA desaturase enzyme system from a photosynthetic endosymbiont to the FA desaturase/elongase enzyme system of a non-photosynthetic eukaryotic protist host. Further, the principal component analysis segregated the three macroalgal groups with a marked distinction of different genera, families and orders, Hierarchical cluster analyses substantiated the phylogenetic relationships of all orders investigated except for those red algal taxa belonging to Gigartinales, Ceramiales, Halymeniales and Rhodymeniales for which increased sampling effort is required to infer a conclusion. Also, the groups deduced from FA compositions were congruent with the clades inferred from nuclear and plastid genome sequences. This study further indicates that FA signatures could be employed as a valid chemotaxonomic tool to differentiate macroalgae at higher taxonomic levels such as family and orders.

Seaweed protoplasts: status, biotechnological perspectives and needs

Protoplasts are living plant cells without cell walls which offer a unique uniform single cell system that facilitates several aspects of modern biotechnology, including genetic transformation and metabolic engineering. Extraction of cell wall lytic enzymes from different phycophages and microbial sources has greatly improved protoplast isolation and their yield from a number of anatomically more complex species of brown and red seaweeds which earlier remained recalcitrant. Recently, recombinant cell wall lytic enzymes were also produced and evaluated with native ones for their potential abilities in producing viable protoplasts from Laminaria. Reliable procedures are now available to isolate and culture protoplasts from diverse groups of seaweeds. To date, there are 89 species belonging to 36 genera of green, red and brown seaweeds from which successful protoplast isolation and regeneration has been reported. Of the total species studied for protoplasts, most belonged to Rhodophyta with 41 species (13 genera) followed by Chlorophyta and Phaeophyta with 24 species each belonging to 5 and 18 genera, respectively. Regeneration of protoplast-to-plant system is available for a large number of species, with extensive literature relating to their culture methods and morphogenesis. In the context of plant genetic manipulation, somatic hybridization by protoplast fusion has been accomplished in a number of economically important species with various levels of success. Protoplasts have also been used for studying foreign gene expression in Porphyra and Ulva. Isolated protoplasts are also exploited in numerous miscellaneous studies involving membrane function, cell structure, bio-chemical synthesis of cell walls etc. This article briefly reviews the status of various developments in seaweed protoplasts research and their potentials in genetic improvement of seaweeds, along with needs that must to be fulfilled for effective realization of the objectives envisaged for protoplast research.

Ultrasound assisted methods for enhanced extraction of phycobiliproteins from marine macro-algae, Gelidium pusillum (Rhodophyta)

Extraction of phycobiliproteins (R-phycoerythrin, R-PE and R-phycocyanin, R-PC) from macro-algae is difficult due to the presence of large polysaccharides (agar, cellulose etc.) present in the cell wall which offer major hindrance for cell disruption. The present study is aimed at developing most suitable methodology for the primary extraction of R-PE and R-PC from marine macro-algae, Gelidium pusillum(Stackhouse) Le Jolis. Such extraction of phycobiliproteins by using ultrasonication and other conventional methods such as maceration, maceration in presence of liquid nitrogen, homogenization, and freezing and thawing (alone and in combinations) is reported for the first time. Standardization of ultrasonication for different parameters such as ultrasonication amplitude (60, 90 and 120µm) and ultrasonication time (1, 2, 4, 6, 8 and 10mins) at different temperatures (30, 35 and 40°C) was carried out. Kinetic parameters were estimated for extraction of phycobiliproteins by ultrasonication based on second order mass transfer kinetics. Based on calorimetric measurements, power, ultrasound intensity and acoustic power density were estimated to be 41.97W, 14.81W/cm2 and 0.419W/cm3, respectively. Synergistic effect of ultrasonication was observed when employed in combination with other conventional primary extraction methods. Homogenization in combination with ultrasonication resulted in an enhancement in efficiency by 9.3% over homogenization alone. Similarly, maceration in combination with ultrasonication resulted in an enhancement in efficiency by 31% over maceration alone. Among all the methods employed, maceration in combination with ultrasonication resulted in the highest extraction efficiency of 77 and 93% for R-PE and R-PC, respectively followed by homogenization in combination with ultrasonication (69.6% for R-PE and 74.1% for R-PC). HPLC analysis was carried out in order to ensure that R-PE was present in the extract and remained intact even after processing. Microscopic studies indicated a clear relation between the extraction efficiency of phycobiliproteins and degree of cell disruption in a given primary extraction method. These combination methods were found to be effective for extraction of phycobiliproteins from rigid biomass of Gelidium pusillum macro-algae and can be employed for downstream processing of biomolecules also from other macro-algae.

Seaweed floristic studies along tsunami affected Indian coasts: A litmus test scenario after 26th December 2004

On 26th December 2004, the world witnessed the devastating power of tsunami, affecting many countries, bordering the Indian Ocean region. This has caused significant changes in the shallow and intertidal regions of the Indian coast, especially the Andaman and Nicobar Islands, Tamil Nadu, Kerala and Pondicherry. The baseline data on biomass availability and distribution of benthic intertidal seaweed species were collected immediately after this catastrophic event by spot surveying 11 selected localities of the above-mentioned regions. In all, 45 species belonging to 31 genera were recorded during the present survey, the maximum number of seaweed species were recorded at Thirumullavarum, Kerala with the minimum at Car Nicobar, Andaman and Nicobar Islands. A very different trend was observed in the case of biomass availability at some locations which was due to the influence of habitat suitability over the tsunami damage. The details of this study have been provided in the present communication

Integration of protein extraction with a stream of byproducts from marine macroalgae: A model forms the basis for marine bioeconomy.

The present study describes an advanced biorefinery model for marine macroalgae that assumes significant importance in the context of marine bio-economy. The method investigated in this study integrates the extraction of crude proteins with recovery of minerals rich sap, lipids, ulvan and cellulose from fresh biomass of Ulva lactuca. The protein content extracted was 11±2.12% on dry weight basis with recovery efficiency of 68.75±4.01%. The amino acid composition of crude protein fraction showed iso-leucine as the most abundant amino acid with 16.51±0.03% followed by histidine, arginine, tyrosine, serine, aspartic acid, threonine, phenyl alanine, leucine, alanine, lysine, glycine and glutamic acid (0.22±0.24%). The digestibility of protein was as high as 85.86±5.92% indicating its suitability for use in food supplements. The protein production with co-recovery of other products would not only result in effective utilisation marine macroalgal resources but also forms the basis for marine bio-economy.

Reproductive Processes in Red Algal Genus Gracilaria and Impact of Climate Change

The genus Gracilaria forms a basis for the multimillion-dollar phycocolloid industry. This genus has a tropical origin and is represented by about 110 species throughout the world. Despite increasing interest in agar as a commercial product, relatively little is known about the overall reproductive processes in genus the Gracilaria. The typical life history of Gracilaria follows a basic pattern known as “Polysiphonia type,” which is triphasic in nature; however, various alternating strategies are also known. The ecophysiological differences have been reported in different life-cycle stages. Biomass production as well as induction of reproduction have been significantly correlated with seawater temperature. The studies using microsatellite DNA markers have shown that the large-scale and continuous farming practices in Gracilaria chilensis have significantly modified the important life-history traits when compared with wild populations, which could have important consequences on the evolution of the haploid–diploid life cycle. The phase- and sex-related genetic markers have been identified in G. lemaneiformis by RAPD method. The population structure in this genus is determined by the vital rates of spore mortality, recruitments, and reproduction associated with the different stages of the life cycle. The spore production and survival is environmentally controlled in a majority of the Gracilaria species. However, the spore dispersal has not been affected by the size of the spore, and haploid spores have better dispersal abilities than diploid ones owing to their higher longevity and buoyancy in the water column under in vitro conditions. The performance of nonmotile spermatia has been evaluated under field conditions in G. gracils through microsatellite markers; the results have shown that the fertilization success depends on the distance traveled by spermatia, male–male competition, and female choice. Intrinsic factors such as life cycle and mating system are shown to be responsible for genetic structure within the population in G. gracils. The germination of spore follows the Dumontia-type of cell division in many Gracilaria species. The germinating spores have shown coalescing nature, which is ecologically as well as economically important. The overall understanding of the climatically driven environmental changes is much more complex and includes the species-tolerance limit toward the particular change, alternative dispersal or reproductive strategy, physiological adaptation, biotic interactions, etc. The effect of such a change on reproductive processes of the genus Gracilaria is mostly unpredictable, since results of only a few laboratory-based studies are insufficient to draw a conclusion.