Seaweed micropropagation techniques and their potentials: an overview

The seaweed industry worldwide uses 7.5–8.0 million tonnes of wet seaweeds annually with a majority of it derived from cultivated farms, as the demand for seaweed based-products exceeds the supply of seaweed raw material from natural stocks. The main advantage of cultivation is that it not only obviates overexploitation of natural populations but also facilitates the selection of germplasm with desired traits. To enhance the economic prospects of seaweed cultivation, varied practices, such as simple and cost effective cultivation methods, use of select germplasm as seed stock coupled with good farm management practices, etc., are adopted. Nevertheless, in vitro cell culture techniques have also been employed as they facilitate development and propagation of genotypes of commercial importance. There are more than 85 species of seaweeds for which tissue culture aspects have been reported. Although the initial aim of these techniques focuses mostly on genetic improvement and clonal propagation of seaweeds for mariculture, recently the scope of these techniques has been extended for use in bioprocess technology for production of high value chemicals of immense importance in the pharmaceutical and nutraceutical sectors. Recently, there has been a phenomenal interest in intensifying seaweed tissue and cell culture research to maximize the add-on value of seaweed resources. This paper deals with the status of seaweed micropropagation techniques and their applications in the context of the marine biotech industry. Further, it also provides an analysis of the problems to be resolved for removing the barriers that are impeding the true realization of potentials offered by these techniques for sustainable development and utilization of seaweed resources.     

Seaweed production: overview of the global state of exploitation,farming and emerging research activity

ABSTRACT

The use of seaweeds has a long history, as does the cultivation of a select and relatively small group of species. This review presents several aspects of seaweed production, such as an update on the volumes of seaweeds produced globally by both extraction from natural beds and cultivation. We discuss uses, production trends and economic analysis. We also focus on what is viewed as the huge potential for growing industrial-scale volumes of seaweeds to provide sufficient, sustainable biomass to be processed into a multitude of products to benefit humankind. The biorefinery approach is proposed as a sustainable strategy to achieve this goal. There are many different technologies available to produce seaweed, but optimization and more efficient developments are still required. We conclude that there are some fundamental and very significant hurdles yet to overcome in order to achieve the potential contributions that seaweed cultivation may provide the world. There are critical aspects, such as improving the value of seaweed biomass, along with a proper consideration of the ecosystem services that seaweed farming can provide, e.g. a reduction in coastal nutrient loads. Additional considerations are environmental risks associated with climate change, pathogens, epibionts and grazers, as well as the preservation of the genetic diversity of cultivated seaweeds. Importantly, we provide an outline for future needs in the anticipation that phycologists around the world will rise to the challenge, such that the potential to be derived from seaweed biomass becomes a reality.     

Prospects and challenges for industrial production of seaweed bioactives

Large‐scale seaweed cultivation has been instrumental in globalizing the seaweed industry since the 1950s. The domestication of seaweed cultivars (begun in the 1940s) ended the reliance on natural cycles of raw material availability for some species, with efforts driven by consumer demands that far exceeded the available supplies. Currently, seaweed cultivation is unrivaled in mariculture with 94% of annual seaweed biomass utilized globally being derived from cultivated sources. In the last decade, research has confirmed seaweeds as rich sources of potentially valuable, health‐promoting compounds. Most existing seaweed cultivars and current cultivation techniques have been developed for producing commoditized biomass, and may not necessarily be optimized for the production of valuable bioactive compounds. The future of the seaweed industry will include the development of high value markets for functional foods, cosmeceuticals, nutraceuticals, and pharmaceuticals. Entry into these markets will require a level of standardization, efficacy, and traceability that has not previously been demanded of seaweed products. Both internal concentrations and composition of bioactive compounds can fluctuate seasonally, geographically, bathymetrically, and according to genetic variability even within individual species, especially where life history stages can be important. History shows that successful expansion of seaweed products into new markets requires the cultivation of domesticated seaweed cultivars. Demands of an evolving new industry based upon efficacy and standardization will require the selection of improved cultivars, the domestication of new species, and a refinement of existing cultivation techniques to improve quality control and traceability of products.     

Mass cultivation of seaweeds: current aspects and approaches

A word-wide overview is presented of the current state of mass cultivation of seaweeds. In comparison with a total annual commercial production of fish, crustaceans and molluscs of about 120 × 10⁶t, of which one-third is produced by aquaculture, the production of seaweeds is about 10 × 10⁶t wet weight; the majoirty of this comes from culture-based systems. The Top Ten Species List is headed by the kelp Laminaria japonica with 4.2 × 10⁶t fresh weight cultivated mainly in China. The productivity of a well-developed, multi-layered, perennial seaweed vegetation is as high as dense terrestrial vegetation, and even higher annual values for productivity have been reported for tank cultures of macroalgae. Epiphytes provide a major problem for the seaweed cultivator, but can be controlled by growing plants at high densities in rope cultures in the sea, or, more easily, in seaweed tank cultures on land. The main environmental problem of animal (fed) aquaculture is the discharge of nutrient loads into coastal waters, e.g., 35 kg N and 7 kg P t^–1 aquacultured fish. Integration of fish and seaweed farming may help to solve this problem, since seaweeds can remove up to 90% of the nutrient discharge from an intensive fish farm. Mass culture of commercially valuable seaweed species is likely to play an increasingly important role as a nutrient-removal system to alleviate eutrophication problems due to fed aquaculture.     

A review of the bladed Bangiales (Rhodophyta) in China: history,culture and taxonomy

In this paper, we review cultural history, mariculture and taxonomic work to date for Porphyra sensu lato (bladed Bangiales) in China. The bladed Bangiales are a red seaweed group with high species biodiversity and economic value. In China, species occur along the length of the coast and are highly integrated into the country’s culture. Chinese people have used the bladed Bangiales as food and pharmaceuticals for about 1700 years with many references to these seaweeds in ancient books. The mariculture of bladed Bangiales in China also has a long history and an industry has been established based on some species, notably Pyropia yezoensis. The scientific study of the taxonomy of the bladed Bangiales in China began in the late 1920s and to date, based on morphological identification, 25 species and five varieties have been recorded for China, of which 12 species are considered to be endemic to the country. The majority of species have distribution data showing evidence of possible changes due to increasing water temperatures along the coast. The global biodiversity of the bladed Bangiales has been revealed using molecular approaches. This points to the need for molecular taxonomy of Chinese material to document species diversity and distribution, particularly as it includes the wild stocks for seaweed cultivation and because coastal habitats are increasingly impacted by the increasing human population and an expanding mariculture industry. There is a considerable body of literature on the bladed Bangiales in China, but much of it is Chinese and in obscure publications, so we review it here for the benefit of readers worldwide.     

Cultivation of red seaweeds: a Latin American perspective

The Latin American seaweed industry plays an important role at a global scale as 17 % of all seaweeds and 37 % of red seaweeds for the phycocolloid industry comes from this region. Increased market demand for algal raw materials has stimulated research and development into new cultivation technologies, particularly in those countries with economically important seaweed industries such as Argentina, Brazil, Chile, México, and Peru. The marine area of Latin America includes almost 59,591 km² of coastline ranging in latitude from 30ºN to 55ºS and encompasses four different oceanic domains: Temperate Northern Pacific, Tropical Eastern Pacific, Temperate South America, and Tropical Atlantic. Commercial cultivation of red seaweed in Latin America has been basically centered in the production of Gracilaria chilensis in Chile. Attempts have been made to establish seaweed commercial cultivation in other countries, going from experimental research-oriented studies to pilot community/enterprise based cultivation trials. Some genera such as Kappaphycus and Eucheuma have been studied in Brazil and Mexico, Gracilaria species in Argentina and Brazil, Gracilariopsis in Peru and Venezuela, and Chondracanthus chamissoi in Peru and Chile. In this short review, we address the Latin America perspective on the status and future progress for the cultivation of red seaweeds and their sustainable commercial development, and discuss on the main common problems. Particular emphasis is given to the needs for comprehensive knowledge necessary for the management and cultivation of some of the most valuable red seaweed resources in Latin America.     

Quantitative Sustainability Assessment of Seaweed Biomass as Bioethanol Feedstock

Since terrestrial biomass-based ethanol has environmental and economic vulnerability, seaweed-based bioethanol is emerging as a new biofuel. To investigate the sustainability of seaweeds as bioethanol feedstock, this study quantitatively assesses the energy, freshwater, and fertilizer requirements; land-related carbon balance; and bioethanol productivity of seaweed biomass through comparison with terrestrial biomass. Also, the metal resource potential of seaweeds is assessed because valuable metals can be recovered from seaweed fermentation residue. Compared to corn grain and stover, seaweeds exhibit competitive energy requirements and ethanol productivity. Seaweed cultivation does not incur carbon debt derived from land use change and requires less freshwater than corn grain but more than switchgrass in cultivation and fermentation. Seaweed cultivation also does not require fertilizer application despite the high content of nitrogen and phosphorus. Seaweeds exhibit high resource potential for gold and silver. Therefore, seaweed biomass has high potential as a sustainable bioethanol feedstock.     

Seaweed research and utilization in Chile: moving into a new phase

Two phases have been distinguished classically in the history of Latin American phycological research: the explorer phase characterized by the taxonomic work of mainly European and North American scientists, and the diversification phase marked by the establishment of resident scientists in the area and the training of a new generation of phycologists in subjects other than taxonomy. Over the last 15 years, Chile has entered a third phase, characterized by a significant increase in scientific and economic activity centered around seaweeds. Seaweed cultivation has been commercialized; raw materials are now locally processed and economic returns have more than tripled. In addition, some groups of opportunistic seaweed gatherers have become farmers. Loosely correlated with the above developments has been a significant increase in the number of scientific and technological studies related to seaweeds, in the number of professional phycologists and in the specialization of the various groups. This study first describes these new developments and the conceptual advances achieved in farming and resource management. It also emphasizes some socio-economic differences with seaweed farming in other countries and explores the level of interaction between the local scientific and productive sectors in view of future developments.     

On-land cultivation of functional seaweed products for human usage

Worldwide, there has been much interest in the development and commercialization of human functional products from seaweeds. Novel seaweed compounds with potential applications as bioactive ingredients in natural health products are being isolated in a number of active research programs on this topic. The majority of these research programs do not include cultivation as a critically important component in scaling the discoveries up to commercialization (i.e., economies of scale realized). Many of these seaweeds of interest with potential as functional human products are diminutive in size, sparse in density, and seasonal in occurrence and bioactive efficacy, making commercialization by resource management and harvesting economically challenging and the application of traditional ocean-based production methods risky. Human functional products will require sustainable production coupled with quality assurance and standardized, consistent efficacy. Since humans are the consumers of these types of functional seaweed products, traceability and security of supply are of the utmost importance to successful commercialization. On-land cultivation is essential for commercial success in the development of human functional products from seaweeds at industrial scales. On-land cultivation allows the highest levels of control over quality, efficacy, traceability, and security. On-land cultivation represents the most environmentally acceptable method for the production of biomass from natural resources that could not be economically or sustainably developed any other way. However, on-land cultivation has many associated barriers to development, including high costs associated with capital, operations, maintenance, and cultivar development, and these demands limit industrial scale development of seaweed functional products for human consumption.     

Review: Seaweed tissue culture as applied to biotechnology: Problems,achievements and prospects

Advances have been made in cell and tissue culture of seaweeds to define a unique branch of in vitro techniques; however, they are lagging far behind those of land plants and have limited applications. Explants can be cultivated axenically in enriched or artificial seawater culture media, and regeneration and even callus formation are achieved. In this state of the art technique, seaweed tissue culture may be already useful for certain biotechnological applications, such as clonal propagation of seed material for mariculture. Nevertheless, the absolute control of growth and development as it is exerted in higher plant tissue culture is lacking, and it is required for more complex biotechnological applications in seaweeds. Definitively, we need appropriate cells (competent cells) to induce growth with the most effective chemical regulators in culture medium adjusted towards the addition of carbon sources. Still, free cells and protoplast isolation and regeneration in marine seaweeds constitute the most developed topic in seaweed tissue culture. The regulation of growth and development of seaweed free cell and protoplast cultures may sustain a purposeful use of techniques in the era of genomic applications.     

Ecological engineering in aquaculture: use of seaweeds for removing nutrients from intensive mariculture

Rapid scale growth of intensive mariculture systems can often lead to adverse impacts on the environment. Intensive fish and shrimp farming, being defined as throughput-based systems, have a continuous or pulse release of nutrients that adds to coastal eutrophication. As an alternative treatment solution, seaweeds can be used to clean the dissolved part of this effluent. Two examples of successfully using seaweeds as biofilters in intensive mariculture systems are discussed in this paper. The first example shows that Gracilaria co-cultivated with salmon in a tank system reached production rates as high as 48.9 kg m⁻² a⁻¹, and could remove 50% of the dissolved ammonium released by the fish in winter, increasing to 90–95% in spring. In the second example, Gracilaria cultivated on ropes near a 22-t fish cage farm, had up to 40% higher growth rate (specific growth rate of 7% d⁻¹) compared to controls. Extrapolation of the results showed that a 1 ha Gracilaria culture gave an annual harvest of 34 t (d. wt), and assimilated 6.5% of the released dissolved nitrogen. This production and assimilation was more than twice that of a Gracilaria monoculture. By integrating seaweeds with fish farming the nutrient assimilating capacity of an area increases. With increased carrying capacity it will be possible to increase salmon cage densities before risking negative environmental effects like eutrophication and toxic algal blooms sometimes associated with the release of dissolved nutrients. The potential for using mangroves and/or seaweeds as filters for wastes from intensive shrimp pond farming is also discussed. It is concluded that such techniques, based on ecological engineering, seems promising for mitigating environmental impacts from intensive mariculture; however, continued research on this type of solution is required. This revised version was published online in June 2006 with corrections to the Cover Date.     

Cultivation of Gracilaria in outdoor tanks and ponds

This review deals with the major problems of unattached Gracilaria intensive cultivation in outdoor tanks and ponds. These problems are presented through the main variables affecting the Gracilaria annual yield and the updated solutions evolved. The physical variables include tank and pond structure, seawater characteristics such as velocity, agitation practice, exchange rate, and salinity, light characteristics such as quantity and quality, and temperature modelling. The chemical variables include nutrient composition and regime of application, and inorganic carbon supply with the pH changes involved. The biological variables include seaweed density, epiphyte competition, grazer damage, bacterial disintegration, integrated mariculture and strain selection. The experience gained in the Israeli research on Gracilaria cultivation is discussed in view of other Gracilaria and seaweed intensive cultivation research.     

The use of Gracilaria tikvahiae (Gracilariales,Rhodophyta) as a model system to understand the nitrogen nutrition of cultured seaweeds

Seaweeds have physiological mechanisms to acquire, utilize, and store various forms of nitrogen in environments where nitrogen levels vary tremendously in space and time. Knowledge of the nitrogen relationships of seaweeds is required for the development of successful seaweed mariculture. For example, it would seem at first that continuous nitrogen enrichment would be desirable in such systems because maximal seaweed yields are possible only when growth is not nitrogen-limited. Yet such fertilization is wasteful and can result in yield reductions due to the enhancement of epiphyte growth. Because most seaweeds can rapidly taken up high concentrations of nitrogen, far in excess of what is required for current growth demands, enrichments are needed only when internal nitrogen concentrations fall to near the critical level (i.e., the minimal tissue concentration of nitrogen required for maximal growth). Nutrients are best applied at brief pulses of high nitrogen concentrations.Dedicated to the memory of Bud Brinkhuis, friend and colleague     

Seaweed biogeochemistry: Global assessment of C:N and C:P ratios and implications for ocean afforestation

Algal carbon-to-nitrogen (C:N) and carbon-to-phosphorus (C:P) ratios are fundamental for understanding many oceanic biogeochemical processes, such as nutrient flux and climate regulation. We synthesized literature data (444 species, >400 locations) and collected original samples from Tasmania, Australia (51 species, 10 locations) to update the global ratios of seaweed carbon-to-nitrogen (C:N) and carbon-to-phosphorus (C:P). The updated global mean molar ratio for seaweed C:N is 20 (ranging from 6 to 123) and for C:P is 801 (ranging from 76 to 4102). The C:N and C:P ratios were significantly influenced by seawater inorganic nutrient concentrations and seasonality. Additionally, C:N ratios varied by phyla. Brown seaweeds (Ochrophyta, Phaeophyceae) had the highest mean C:N of 27.5 (range: 7.6–122.5), followed by green seaweeds (Chlorophyta) of 17.8 (6.2–54.3) and red seaweeds (Rhodophyta) of 14.8 (5.6–77.6). We used the updated C:N and C:P values to compare seaweed tissue stoichiometry with the most recently reported values for plankton community stoichiometry. Our results show that seaweeds have on average 2.8 and 4.0 times higher C:N and C:P than phytoplankton, indicating seaweeds can assimilate more carbon in their biomass for a given amount of nutrient resource. The stoichiometric comparison presented herein is central to the discourse on ocean afforestation (the deliberate replacement of phytoplankton with seaweeds to enhance the ocean biological carbon sink) by contributing to the understanding of the impact of nutrient reallocation from phytoplankton to seaweeds under large-scale seaweed cultivation.     

Mesoherbivores reduce net growth and induce chemical resistance in natural seaweed populations

Herbivory on marine macroalgae (seaweeds) in temperate areas is often dominated by relatively small gastropods and crustaceans (mesoherbivores). The effects of these herbivores on the performance of adult seaweeds have so far been almost exclusively investigated under artificial laboratory conditions. Furthermore, several recent laboratory studies with mesoherbivores indicate that inducible chemical resistance may be as common in seaweeds as in vascular plants. However, in order to further explore and test the possible ecological significance of induced chemical resistance in temperate seaweeds, data are needed that address this issue in natural populations. We investigated the effect of grazing by littorinid herbivorous snails (Littorina spp.) on the individual net growth of the brown seaweed Ascophyllum nodosum in natural field populations. Furthermore, the capacity for induced resistance in the seaweeds was assessed by removing herbivores and assaying for relaxation of defences. We found that ambient densities of gastropod herbivores significantly reduced net growth by 45% in natural field populations of A. nodosum. Seaweeds previously exposed to grazing in the field were less consumed by gastropod herbivores in feeding bioassays. Furthermore, the concentration of phlorotannins (polyphenolics), which have been shown to deter gastropod herbivores, was higher in the seaweeds that were exposed to gastropod herbivores in the field. This field study corroborates earlier laboratory experiments and demonstrates that it is important to make sure that the lack of experimental field data on marine mesoherbivory does not lead to rash conclusions about the lack of significant effects of these herbivores on seaweed performance. The results strongly suggest that gastropods exert a significant selection pressure on the evolution of defensive traits in the seaweeds, and that brown seaweeds can respond to attacks by natural densities of these herbivores through increased chemical resistance to further grazing.     

Transforming kelp into a marine bioreactor

The past decade has seen the genetic engineering of various types of seaweed. To date, genetic transformation studies have been carried out in several seaweeds, including the red seaweeds Porphyra, Gracilaria, Grateloupia, Kappaphycus and Ceramium and the green seaweed Ulva. A genetic transformation model system has been established in the most commonly cultivated seaweed, the brown seaweed Laminaria japonica (kelp), based on the transfer of technology used in land plant transformation and also by modulating the seaweed life cycle. This model showed the potential for application of transgenic kelp to the production of valuable products and an indoor cultivation system for transgenic kelp was proposed, taking into account necessary factors for bio-safety. In this review, the establishment at use of the kelp transformation model is introduced, highlighting the potential for transforming kelp into a marine bioreactor.     

IMPACT OF ALGAL RESEARCH IN AQUACULTURE

Algal aquaculture worldwide is estimated to be a $5–6 billion U.S. per year industry. The largest portion of this industry is represented by macroalgal production for human food in Asia, with increasing activity in South America and Africa. The technical foundation for a shift in the last half century from wild harvest to farming of seaweeds lies in scientific research elucidating life histories and growth characteristics of seaweeds with economic interest. In several notable cases, scientific breakthroughs enabling seaweed-aquaculture advances were not motivated by aquaculture needs but rather by fundamental biological or ecological questions. After scientific breakthroughs, development of practical cultivation methods has been accomplished by both scientific and commercial-cultivation interests. Microalgal aquaculture is much smaller in economic impact than seaweed cultivation but is the subject of much research. Microalgae are cultured for direct human consumption and for extractable chemicals, but current use and development of cultured microalgae is increasingly related to their use as feeds in marine animal aquaculture. The history of microalgal culture has followed two main paths, one focused on engineering of culture systems to respond to physical and physiological needs for growing microalgae and the other directed toward understanding the nutritional needs of animals—chiefly invertebrates such as mollusks and crustaceans—that feed upon microalgae. The challenge being addressed in current research on microalgae in aquaculture food chains is to combine engineering and nutritional principles so that effective and economical production of microalgal feed cultures can be accomplished to support an expanding marine animal aquaculture industry.     

Discolored Red Seaweed <Emphasis Type="Italic">Pyropia yezoensis</Emphasis> with Low Commercial Value Is a Novel Resource for Production of Agar Polysaccharides

The red seaweed Pyropia yezoensis has been demonstrated to be a novel resource for the production of high-quality agar. P. yezoensis is grown for the food industry in large-scale Japanese mariculture operations. However, discolored P. yezoensis is mostly discarded as an industrial waste, although it has some kind of utility values. Here, we evaluated the utility of discolored P. yezoensis as a resource for agar production. The quality of agar from the discolored seaweed was comparable to that from normal seaweed. In addition, as a distinguishing characteristic, agar yield was higher from discolored seaweeds than from normal types. Moreover, we successfully used agar from discolored P. yezoensis for bacterial plate media and DNA electrophoresis gels without agarose purification. Thus, our results demonstrate that discolored P. yezoensis is suitable for agar production and use in life science research. Diverting discolored P. yezoensis from disposal to agar production provides a solution to the current industrial waste problem in mariculture, as well as a secure source of agar for research purposes.     

Rapid Survey Technique Using Socio-Economic Indicators to Assess the Suitability of Pacific Island Rural Communities for Kappaphycus Seaweed Farming Development

The literature on economic feasibility of farming seaweeds like Kappaphycus alvarezii in tropical locations is mainly based upon Asian case studies, and often does not take into account social factors in seaweed farming success. Pacific island countries are culturally and economically distinct from Asia, and efforts are now being made to establish seaweed industries here. Past experiences have showed that social factors often outweigh technical factors in determining the success of rural development projects. In addition, Pacific island communities are very diverse in their socio-economic make-up. The particular community chosen for location of a development project is therefore critical to success. Project managers need to recognize in advance the best type of community for seaweed farming development. The objective of this study was to identify socio economic factors that can be used as predictors of project success or failure. Using results of social survey techniques carried out in eight communities within the Fiji Group, a rapid survey technique has been developed which can enable decisions about whether a community is suitable for farming seaweed or not. Though developed from Fiji case studies, the technique can be applied in other rural Asia/Pacific situations.