Alan T. Critchley

Seaweed extracts as biostimulants of plant growth and development.

Marine algal seaweed species are often regarded as an underutilized bioresource, many have been used as a source of food, industrial raw materials, and in therapeutic and botanical applications for centuries. Moreover, seaweed and seaweed-derived products have been widely used as amendments in crop production systems due to the presence of a number of plant growth-stimulating compounds. However, the biostimulatory potential of many of these products has not been fully exploited due to the lack of scientific data on growth factors present in seaweeds and their mode of action in affecting plant growth. This article provides a comprehensive review of the effect of various seaweed species and seaweed products on plant growth and development with an emphasis on the use of this renewable bioresource in sustainable agricultural systems.

Rapid bioassays to evaluate the plant growth promoting activity of Ascophyllum nodosum (L.) Le Jol. using a model plant, Arabidopsis thaliana (L.) Heynh

Ascophyllum nodosum extract products are used commercially in the form of liquid concentrate and soluble powder. These formulations are manufactured from seaweeds that are harvested from natural habitats with inherent environmental variability. The seaweeds by themselves are at different stages of their development life-cycle. Owing to these differences, there could be variability in chemical composition that could in turn affect product consistency and performance. Here, we have tested the applicability of using Arabidopsis thaliana as a model to study the activity of two different extracts from A. nodosum. Three different bioassays: Arabidopsis root-tip elongation bioassay, Arabidopsis liquid growth bioassay and greenhouse growth bioassay were evaluated as growth assays. Our results indicate that both extracts promoted root and shoot growth in comparison to controls. Further, using Arabidopsis plants with a DR5:GUS reporter gene construct, we provide evidence that components of the commercial A. nodosum extracts modulates the concentration and localisation of auxins which could account, at least in part, for the enhanced plant growth. The results suggest that A. thaliana could be used effectively as a rapid means to test the bioactivity of seaweed extracts and fractions.

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.

Cultivation of tropical red seaweeds in the BIMP-EAGA region

The Brunei–Indonesia–Malaysia–Philippines East Asia Growth Area (BIMP-EAGA) is located within the Coral Triangle, known to have the world’s richest biodiversity in marine flora and fauna. This region lies within the 10° N and 10° S of the Equator where natural populations of both Kappaphycus and Eucheuma grow luxuriantly and abundantly. It is in this same region where commercial cultivation of Kappaphycus and Eucheuma began in the Philippines around the mid-1960s. Commercial farming of Kappaphycus (which was originally called Eucheuma) was successful in the Philippines from the early 1970s, after which the technology was transferred to Indonesia and Malaysia in the late 1970s. No seaweed cultivation has been reported in Brunei. At present, carrageenophytes are cultivated in sub-tropical to tropical countries circumferentially around the globe within the 10° N and S of the Equator. However, their combined production is still low as compared to Indonesia, the Philippines, and Malaysia. Notably, few improvements in farming techniques have been made since its first introduction. Some of the major improvements were the introduction of deep-water farming using hanging long lines, multiple rafts, and spider webs in the Philippines; the use of short and long ‘loops’, instead of plastic ‘tie-tie’ in Indonesia; and mechanization in harvesting and use of solar “greenhouse” drying in Malaysia. Commercial cultivation of tropical red seaweeds in the BIMP-EAGA region is dominated by Kappaphycus and Eucheuma (carrageenophytes) and Gracilaria (agarophytes) and the area became the major region for the production of carageenophytes and agarophytes globally. In particular, Indonesia is a major center for the production of Gracilaria. There is an increasing demand for other agarophytes/carrageenophytes in the international market such as Gelidium spp., Pterocladia spp., Porphyroglossum sp., and Ptilophora sp. for paper and ethanol production in Indonesia and Malaysia, and Halymenia for phycoerythrin pigments in the Philippines currently pursued in an experimental stage. A summary of the present status, problems, sustainability, and challenges for the cultivation of tropical red seaweeds in the BIMP-EAGA region are discussed in this paper.

Lipophilic components of the brown seaweed, Ascophyllum nodosum, enhance freezing tolerance in Arabidopsis thaliana

Extracts of the brown seaweed Ascophyllum nodosum enhance plant tolerance against environmental stresses such as drought, salinity, and frost. However, the molecular mechanisms underlying this improved stress tolerance and the nature of the bioactive compounds present in the seaweed extracts that elicits stress tolerance remain largely unknown. We investigated the effect of A. nodosum extracts and its organic sub-fractions on freezing tolerance of Arabidopsis thaliana. Ascophyllum nodosum extracts and its lipophilic fraction significantly increased tolerance to freezing temperatures in in vitro and in vivo assays. Untreated plants exhibited severe chlorosis, tissue damage, and failed to recover from freezing treatments while the extract-treated plants recovered from freezing temperature of −7.5°C in in vitro and −5.5°C in in vivo assays. Electrolyte leakage measurements revealed that the LT50 value was lowered by 3°C while cell viability staining demonstrated a 30–40% reduction in area of damaged tissue in extract treated plants as compared to water controls. Moreover, histological observations of leaf sections revealed that extracts have a significant effect on maintaining membrane integrity during freezing stress. Treated plants exhibited 70% less chlorophyll damage during freezing recovery as compared to the controls, and this correlated with reduced expression of the chlorphyllase genes AtCHL1 and AtCHL2. Further, the A. nodosum extract treatment modulated the expression of the cold response genes, COR15A, RD29A, and CBF3, resulting in enhanced tolerance to freezing temperatures. More than 2.6-fold increase in expression of RD29A, 1.8-fold increase of CBF3 and two-fold increase in the transcript level of COR15A was observed in plants treated with lipophilic fraction of A. nodosum at −2°C. Taken together, the results suggest that chemical components in A. nodosum extracts protect membrane integrity and affect the expression of stress response genes leading to freezing stress tolerance in A. thaliana.

Extracts of the Brown Seaweed Ascophyllum nodosum Induce Gibberellic Acid (GA3)-independent Amylase Activity in Barley

Extracts of the brown seaweed Ascophyllum nodosum have been used as a biostimulant to promote growth and productivity in a number of agricultural production systems. Although the extracts have been shown to improve seedling emergence and vigor in a variety of plants, including barley, the mechanism(s) of this growth-promoting effect is(are) largely unknown. In our study, A. nodosum extract induced amylase activity in barley seed-halves; a significant difference in amylase activity was observed in seeds without an embryo. The addition of activated charcoal to the treatment media negated the bioactivity of the extracts suggesting the organic nature of bioactive compounds in A. nodosum extracts. The extracts induced amylase activity in a gibberellic acid (GA)-deficient barley mutant (grd2). LC-MS-MS analysis failed to detect the presence of GA3 in the extracts. ABA supplementation of the medium caused a significant reduction of amylase activity in GA-treated seeds compared with those treated with the A. nodosum extract. Taken together, our results suggest that the organic components of A. nodosum extract induce amylase activity independent of GA3 and might act in concert with GA-dependent amylase production leading to enhanced germination and seedling vigor in barley. Being derived from a renewable resource, the bioactive compounds from A. nodosum could be used to improve crop productivity in sustainable agricultural systems.

Systematics and genetic variation in commercial shape Kappaphycus and shape Eucheuma (Solieriaceae, Rhodophyta)

The systematics and taxonomy of Kappaphycus and Eucheuma (Solieriaceae) is confused and difficult due to morphological plasticity, lack of adequate characters to identify species and commercial names of convenience. These taxa are geographically widely dispersed through cultivation. Commercial, wild and herbarium sources were analysed; molecular markers provided insights into taxonomy and genetic variation, and where sources of genetic variation may be located. The mitochondrial cox2-3 and plastidal RuBisCo spacers were sequenced. There is a clear genetic distinction between K. alvarezii (“cottonii”) and K. striatum (“sacol”) samples. Kappaphycus alvarezii from Hawaii and some samples from Africa are also genetically distinct. Our data also show that all currently cultivated K. alvarezii from all over the world have a similar mitochondrial haplotype. Within Eucheuma denticulatum (“spinosum”) most African samples are again genetically distinct. Our data also suggest that currently cultivated E. denticulatum may have been “domesticated” several times, whereas this is not evident for the cultivated K. alvarezii. The present markers used do not distinguish all the morpho-types known in cultivation (e.g. var. tambalang, “giant” type) but do suggest that these markers may be useful to assess introductions and species identification in samples.

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.

Low genetic variability of Sargassum muticum (Phaeophyceae) revealed by a global analysis of native and introduced populations.

Sargassum muticum (Yendo) Fensholt is one of the most well‐known invasive species in the world. There have, however, been few genetic investigations on both its introduced and native populations. There are also some questions about the taxonomic status of this species. This study is the first to assess the genetic diversity of S. muticum on a global scale, by utilizing one marker each from the extranuclear genomes, namely, plastidial RUBISCO and mitochondrial TrnW_I spacers, as well as the nuclear internal transcribed spacer 2 (ITS2). Based on the markers investigated, both the invasive as well as the native populations of this species appeared very homogenous, when compared with other invasive and brown macroalgae. No variation in ITS2 and RUBISCO spacer was revealed in S. muticum populations, including those from its native ranges in Asia and the introduced ranges in Europe and North America. Two TrnW_I spacer haplotypes with a fixed two‐nucleotide difference were found between the populations of eastern Japan and the other 15 populations examined. This study confirms that there is no cryptic diversity in the introduced range of this species. All the materials collected globally are indeed S. muticum. Results depicting the distribution range of the two TrnW_I spacer haplotypes also support the earlier suggestion that the source of the introduced S. muticum populations is most likely western and central Japan (Seto Inland Sea), where the germlings of S. muticum were likely to have been transported with the Pacific oysters previously introduced for farming in Canada, UK, and France in earlier years.

A Review of Kappaphycus Farming: Prospects and Constraints

Global warming is of increasing concern worldwide. The question of how to mitigate the CO2 released into the atmosphere is the most topical issue, and sustainable solutions are constantly being sought. Aquaculture has been proposed as one method for the sequestration or immobilization of CO2 through filtration or mechanical/chemical processes for long-term storage (Carlsson et al., 2007). However, the development of new sustainable technologies are but in their infancy, as the aquaculture sector moves to becoming more efficient and sustainable.