Alistair J. McCormick

Photosynthetic biofilms in pure culture harness solar energy in a mediatorless bio-photovoltaic cell (BPV) system

Microbial fuel cells are an emerging technology for converting organic substrates into electrical power. Recent research has shown that biofilms of some bacterial species are capable of self-mediated extracellular electron transfer. The prospect of exploiting this trait in photoautotrophic microbes that do not require an organic substrate has important implications for the future development of renewable solar energy technologies. Here we report on light-driven electrical power generated with biofilms grown from photosynthetic fresh water or marine species without the addition of an artificial electron-shuttling mediator. Green alga (Chlorella vulgaris, Dunaliella tertiolecta) or cyanobacteria (Synechocystis sp. PCC 6803, Synechococcus sp. WH 5701) strains were grown directly on a transparent, conductive anode (indium tin oxide-coated polyethylene terephthalate) and power generation under light and dark conditions was evaluated using a single-chamber bio-photovoltaic cell (BPV) system. Increased power outputs were observed for all strains upon illumination, with the largest light effect observed for Synechococcus (maximum 10.3 mW m−2 total power output recorded under 10 W m−2 white light). Further experiments conducted with Synechococcus and C. vulgaris showed that photosynthetic oxygen evolution rates were consistent with BPV power outputs under different light regimes (red, green and blue light), indicating a direct link between power output and photosynthetic activity. Biofilm power generation in these BPV devices was self-sustained for several weeks and was highly sensitive to ambient light levels. When connected in series, four BPVs (each 0.011 m2) generated enough power to run a commercial digital clock.

Supply and demand: sink regulation of sugar accumulation in sugarcane

Sugarcane (Saccharum spp. hybrids) accumulates sucrose to high concentrations and, as a result, has been the focus of extensive research into the biochemistry and physiology of sucrose accumulation. Despite this, the relationship between source leaf photosynthetic activity and sucrose accumulation in the culm sink is not well understood. The observations that photosynthetic activity declines during culm maturation in commercial cultivars and that high-sucrose-accumulating noble ancestral genotypes (Saccharum officinarum L.) photosynthesize at rates two-thirds of those of low-sucrose ancestors (Saccharum spontaneum L.) indicate that source-sink communication may play a pivotal role in determining sucrose yield. Although maturation of the culm results in a decreased demand for sucrose, recent evidence from partial leaf shading, defoliation, and transgenic studies indicates that sugarcane cultivars are capable of further increases in sugar content. Furthermore, sugarcane leaves appear to retain the capacity to increase the supply of assimilate to culm tissues under conditions of increased assimilate demand. The relationship between source and sink tissues in sugarcane should be viewed within a supply-demand paradigm; an often neglected conceptual approach in the study of this crop. Uncoupling of the signalling pathways that mediate negative feedback between source and sink tissues may result in improved leaf assimilation rates and, consequently, lead to increased sugarcane sucrose yields.

Biophotovoltaics: oxygenic photosynthetic organisms in the world of bioelectrochemical systems

The field of bioelectrochemical system (BES) research includes a wide range of emerging technologies that utilise microbes to catalyze anodic and/or cathodic reactions within a fuel cell setup, and has developed greatly in the last 2–3 years. Although the vast majority of BESs utilise organic substrates as a fuel source (e.g. microbial fuel cells), several systems have been developed that are fuelled by light energy. In this review we focus on and contextualise a specific subset of light-harvesting BESs, which we have called biophotovoltaic systems (BPVs). BPVs utilise oxygenic photosynthetic organisms, such as microalgal and cyanobacterial species, to harvest light energy to generate current, critically, in the absence of an organic feedstock. Here we discuss the state-of-the-art for all light-harvesting BESs and present a novel classification system to illustrate how BPVs integrate into the broad fields of BES and photovoltaic research. We compare and contrast the present understanding of electron transfer pathways in systems that use heterotrophic microbes with those in cyanobacteria-based BPVs. Finally we present, for the first time, an estimate of the achievable power outputs of this emerging technology.

Regulation of photosynthesis by sugars in sugarcane leaves

In sugarcane, increased sink demand has previously been shown to result in increased photosynthetic rates that are correlated with a reduction in leaf hexose concentrations. To establish whether sink limitation of photosynthesis is a result of sugar accumulation in the leaf, excision and cold-girdling techniques were used to modify leaf sugar concentrations in pot-grown sugarcane. In excised leaves that were preincubated in darkness for 3h, sucrose accumulation was reduced but accumulated again upon transfer to the light, while hexose concentrations remained lower than in controls (7.7 micromol mg(-1)FW versus 18.6 micromol mg(-1)FW hexose in controls). These results were associated with a 66% and 59% increase in photosynthetic assimilation (A) and electron transport rate (ETR), respectively, compared to controls maintained in the light. Similar increases in photosynthesis were observed when dark-treated leaves were supplied with 5mM sorbitol, but not when supplied with 5mM sucrose. Further analyses of (14)C-labeled sugars indicated rapid turnover between sucrose and hexose. Cold-girdling (5 degrees C) increased sucrose and hexose levels and resulted in a decline of photosynthetic rates over 5d (48% and 35% decline in assimilation rate and ETR, respectively). These sugar-induced changes in photosynthesis were independent of changes in stomatal conductance. This study demonstrates that the down-regulation of photosynthesis in response to culm sugar accumulation reported previously could be due to the knock-on effect of accumulation of sugar in leaf tissue, and supports the contention that hexose, rather than sucrose, is responsible for the modulation of photosynthetic activity.

Surface morphology and surface energy of anode materials influence power outputs in a multi-channel mediatorless bio-photovoltaic (BPV) system

Bio-photovoltaic cells (BPVs) are a new photo-bio-electrochemical technology for harnessing solar energy using the photosynthetic activity of autotrophic organisms. Currently power outputs from BPVs are generally low and suffer from low efficiencies. However, a better understanding of the electrochemical interactions between the microbes and conductive materials will be likely to lead to increased power yields. In the current study, the fresh-water, filamentous cyanobacterium Pseudanabaena limnetica (also known as Oscillatoria limnetica) was investigated for exoelectrogenic activity. Biofilms of P. limnetica showed a significant photo response during light-dark cycling in BPVs under mediatorless conditions. A multi-channel BPV device was developed to compare quantitatively the performance of photosynthetic biofilms of this species using a variety of different anodic conductive materials: indium tin oxide-coated polyethylene terephthalate (ITO), stainless steel (SS), glass coated with a conductive polymer (PANI), and carbon paper (CP). Although biofilm growth rates were generally comparable on all materials tested, the amplitude of the photo response and achievable maximum power outputs were significantly different. ITO and SS demonstrated the largest photo responses, whereas CP showed the lowest power outputs under both light and dark conditions. Furthermore, differences in the ratios of light : dark power outputs indicated that the electrochemical interactions between photosynthetic microbes and the anode may differ under light and dark conditions depending on the anodic material used. Comparisons between BPV performances and material characteristics revealed that surface roughness and surface energy, particularly the ratio of non-polar to polar interactions (the CQ ratio), may be more important than available surface area in determining biocompatibility and maximum power outputs in microbial electrochemical systems. Notably, CP was readily outperformed by all other conductive materials tested, indicating that carbon may not be an optimal substrate for microbial fuel cell operation.

Introducing an algal carbon-concentrating mechanism into higher plants: location and incorporation of key components

Summary Many eukaryotic green algae possess biophysical carbon‐concentrating mechanisms (CCMs) that enhance photosynthetic efficiency and thus permit high growth rates at low CO 2 concentrations. They are thus an attractive option for improving productivity in higher plants. In this study, the intracellular locations of ten CCM components in the unicellular green alga Chlamydomonas reinhardtii were confirmed. When expressed in tobacco, all of these components except chloroplastic carbonic anhydrases CAH3 and CAH6 had the same intracellular locations as in Chlamydomonas. CAH6 could be directed to the chloroplast by fusion to an Arabidopsis chloroplast transit peptide. Similarly, the putative inorganic carbon (Ci) transporter LCI1 was directed to the chloroplast from its native location on the plasma membrane. CCP1 and CCP2 proteins, putative Ci transporters previously reported to be in the chloroplast envelope, localized to mitochondria in both Chlamydomonas and tobacco, suggesting that the algal CCM model requires expansion to include a role for mitochondria. For the Ci transporters LCIA and HLA3, membrane location and Ci transport capacity were confirmed by heterologous expression and H14 CO 3 ‐ uptake assays in Xenopus oocytes. Both were expressed in Arabidopsis resulting in growth comparable with that of wild‐type plants. We conclude that CCM components from Chlamydomonas can be expressed both transiently (in tobacco) and stably (in Arabidopsis) and retargeted to appropriate locations in higher plant cells. As expression of individual Ci transporters did not enhance Arabidopsis growth, stacking of further CCM components will probably be required to achieve a significant increase in photosynthetic efficiency in this species.

Will an algal CO2-concentrating mechanism work in higher plants?

Many algae use a biophysical carbon concentrating mechanism for active accumulation and retention of inorganic carbon within chloroplasts, with CO2 fixation by RuBisCO within a micro-compartment, the pyrenoid. Engineering such mechanisms into higher plant chloroplasts is a possible route to augment RuBisCO operating efficiency and photosynthetic rates. Significant progress has been made recently in characterising key algal transporters and identifying factors responsible for the aggregation of RuBisCO into the pyrenoid. Several transporters have now also been successfully incorporated into higher plant chloroplasts. Consistent with the predictions from modelling, regulation of higher plant plastidic carbonic anhydrases and some form of RuBisCO aggregation will be needed before the mechanism delivers potential benefits. Key research priorities include a better understanding of the regulation of the algal carbon concentrating mechanism, advancing the fundamental characterisation of known components, evaluating whether higher plant chloroplasts can accommodate a pyrenoid, and, ultimately, testing transgenic lines under realistic growth conditions.

Characterising the structure of photosynthetic biofilms using fluid dynamic gauging

A new configuration of the fluid dynamic gauging technique for measuring soft layers on surfaces was used to monitor the growth of a cyanobacterium, Synechococcus sp. WH 5701, on stainless steel (SS), glass and an indium tinoxide (ITO) on a polyethylene terephthalate (PET) substratum. The biofilm thickness increased steadily over 4weeks and exhibited noticeable changes in microstructure and strength. The biofilms all exhibited a two-layer structure, with a compact layer next to the substratum and a loose layer above. Biofilms on ITO or SS exhibited cohesive failure when removed by fluid shear whereas those on glass exhibited adhesive failure. The technique is able to elucidate various aspects of biofilm behaviour, as illustrated by the action of a biocide (NaOCl) on a mature biofilm.

Progress and challenges of engineering a biophysical CO2-concentrating mechanism into higher plants

Growth and productivity in important crop plants is limited by the inefficiencies of the C3 photosynthetic pathway. Introducing CO2-concentrating mechanisms (CCMs) into C3 plants could overcome these limitations and lead to increased yields. Many unicellular microautotrophs, such as cyanobacteria and green algae, possess highly efficient biophysical CCMs that increase CO2 concentrations around the primary carboxylase enzyme, Rubisco, to enhance CO2 assimilation rates. Algal and cyanobacterial CCMs utilize distinct molecular components, but share several functional commonalities. Here we outline the recent progress and current challenges of engineering biophysical CCMs into C3 plants. We review the predicted requirements for a functional biophysical CCM based on current knowledge of cyanobacterial and algal CCMs, the molecular engineering tools and research pipelines required to translate our theoretical knowledge into practice, and the current challenges to achieving these goals.

Lack of fructose 2,6‐bisphosphate compromises photosynthesis and growth in Arabidopsis in fluctuating environments

The balance between carbon assimilation, storage and utilisation during photosynthesis is dependent on partitioning of photoassimilate between starch and sucrose, and varies in response to changes in the environment. However, the extent to which the capacity to modulate carbon partitioning rapidly through short-term allosteric regulation may contribute to plant performance is unknown. Here we examine the physiological role of fructose 2,6-bisphosphate (Fru-2,6-P2 ) during photosynthesis, growth and reproduction in Arabidopsis thaliana (L.). In leaves this signal metabolite contributes to coordination of carbon assimilation and partitioning during photosynthesis by allosterically modulating the activity of cytosolic fructose-1,6-bisphosphatase. Three independent T-DNA insertional mutant lines deficient in 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (F2KP), the bifunctional enzyme responsible for both the synthesis and degradation of Fru-2,6-P2 , lack Fru-2,6-P2 . These plants have normal steady-state rates of photosynthesis, but exhibit increased partitioning of photoassimilate into sucrose and have delayed photosynthetic induction kinetics. The F2KP-deficient plants grow normally in constant environments, but show reduced growth and seed yields relative to wildtype plants in fluctuating light and/or temperature. We conclude that Fru-2,6-P2 is required for optimum regulation of photosynthetic carbon metabolism under variable growth conditions. These analyses suggest that the capacity of Fru-2,6-P2 to modulate partitioning of photoassimilate is an important determinant of growth and fitness in natural environments.