Rerouting carbon flux to enhance photosynthetic productivity

The bioindustrial production of fuels, chemicals, and therapeutics typically relies upon carbohydrate inputs derived from agricultural plants, resulting in the entanglement of food and chemical commodity markets. We demonstrate the efficient production of sucrose from a cyanobacterial species, Synechococcus elongatus, heterologously expressing a symporter of protons and sucrose (cscB). cscB-expressing cyanobacteria export sucrose irreversibly to concentrations of >10 mM without culture toxicity. Moreover, sucrose-exporting cyanobacteria exhibit increased biomass production rates relative to wild-type strains, accompanied by enhanced photosystem II activity, carbon fixation, and chlorophyll content. The genetic modification of sucrose biosynthesis pathways to minimize competing glucose- or sucrose-consuming reactions can further improve sucrose production, allowing the export of sucrose at rates of up to 36.1 mg liter(-1) h illumination(-1). This rate of production exceeds that of previous reports of targeted, photobiological production from microbes. Engineered S. elongatus produces sucrose in sufficient quantities (up to ～80% of total biomass) such that it may be a viable alternative to sugar synthesis from terrestrial plants, including sugarcane.     

More than just a pair of blue genes: how cyanobacteria adapt to changes in their light environment

Cyanobacteria require light to perform photosynthesis, but not all colors of light are equally useable for them. In particular, blue light-grown cyanobacterial strains, including the well-studied model organism Synechocystis sp. PCC 6803 (Synechocystis), have been observed to exhibit slower growth rates than white or red light-grown cells. In this issue of Physiologia Plantarum, Luimstra et al. (2020) have attempted to understand why cyanobacterial cells suffer under blue light. They measured the molecular and genetic responses of Synechocystis cells to being shifted from white light to blue light. They found that blue light-grown cells make changes that lead to a redistribution of energy flow between the two photosystems that power photosynthesis. These findings could help researchers identify avenues for optimizing photosynthesis in cyanobacterial species, a group of organisms which show great promise as potential solar-powered factories for the production of biofuels and other high-value products.     

An Organic Acid Based Counter Selection System for Cyanobacteria

Cyanobacteria are valuable organisms for studying the physiology of photosynthesis and carbon fixation, as well as metabolic engineering for the production of fuels and chemicals. This work describes a novel counter selection method for the cyanobacterium Synechococcus sp. PCC 7002 based on organic acid toxicity. The organic acids acrylate, 3-hydroxypropionate, and propionate were shown to be inhibitory towards Synechococcus sp. PCC 7002 and other cyanobacteria at low concentrations. Inhibition was overcome by a loss of function mutation in the gene acsA, which is annotated as an acetyl-CoA ligase. Loss of AcsA function was used as a basis for an acrylate counter selection method. DNA fragments of interest were inserted into the acsA locus and strains harboring the insertion were isolated on selective medium containing acrylate. This methodology was also used to introduce DNA fragments into a pseudogene, glpK. Application of this method will allow for more advanced genetics and engineering studies in Synechococcus sp. PCC 7002 including the construction of markerless gene deletions and insertions. The acrylate counter-selection could be applied to other cyanobacterial species where AcsA activity confers acrylate sensitivity (e.g. Synechocystis sp. PCC 6803).     

Density and diversity of protozoa in some arid Australian soils

This is the first extensive study of soil protozoa of arid lands. Twenty-six samples from litters, soils, termitaria, and a cyanobacterial crust, collected from central and south Australian arid lands, were analyzed for numbers and species of gymnamoebae, ciliates, and testacea. Amoebae ranged from 1,000-5,000/g of material, and were two orders of magnitude more abundant than ciliates. Both groups increased in abundance and species richness from bare soils through spinifex to mulga to chenopod vegetations. Testacea ranged 900-5,000/g with similar species richness throughout vegetations, but reached 11,900/g with a doubling of species in a refugium in Kings Canyon. The most prevalent species of amoebae, ciliates, and testacea were taxa associated with ephemeral and disturbed habitats (r-selection). The cyanobacterial crust might be considered a micro-refugium because it contained a number of non-encysting protozoa, including Thecamoeba sp. and Nassula picta, feeding on cyanobacterial filaments. The numbers and species richness of protozoa under shrubs were greater than in bare soils, supporting the resource island hypothesis that desert plants create soil heterogeneity by localizing soil fertility under their canopies.     

古尔班通古特沙漠生物结皮微鞘藻分类学研究

实验研究了从古尔班通古特沙漠生物土壤结皮中分离纯化培养出的11株与微鞘藻(Microcoleus)形态接近的丝状蓝藻,通过形态特征、16S rRNA和ITS二级结构相结合的多相分析方法对其进行分类学研究。研究结果表明,实验藻株隶属于微鞘藻科(Microcoleaceae)的微鞘藻属(Microcoleus)和束脉藻属(Symplocastrum),其中包括2个中国新记录种:斯坦微鞘藻(Microcoleus steenstrupii)和细长束脉藻(Symplocastrum flechtnerii),另外还有具鞘微鞘藻(Microcoleus vaginatus)和类似斯坦微鞘藻的存疑物种。藻丝多少与排列方式、细胞大小与末端细胞形状,以及16S rRNA系统发育位置是确定微鞘藻(Microcoleus)与束脉藻(Symplocastrum)属于不同物种的关键依据, ITS二级结构是区分属内不同物种的重要参考。     

Outdoor cultivation of a nitrogen-fixing marine cyanobacterium,Anabaena sp. ATCC 33047

Optimization of conditions for outdoor production of the nitrogen-fixing cyanobacterium Anabaena sp. ATCC 33047 has been pursued. In open ponds operated under semi-continuous regime biomass productivity values achieved ranged from 9 g (dry weight) m(-2) per day, in winter, to over 20 g m(-2) per day, in summer, provided that key operation parameters, including cell density, were optimized. Under these conditions the efficiency of solar energy conversion by the cells was fairly constant throughout the year, with photosynthetic efficiency values higher than 2%. The cyanobacterial biomass was rich in high-value phycobiliproteins, namely allophycocyanin and phycocyanin, for which open cultures of marine Anabaena represent a most interesting production system. The performance of Anabaena cultures operated under continuous regime in a closed tubular reactor has also been assessed outdoors, in winter. Biomass productivity values similar to those obtained in the ponds have been recorded for the closed system. Additionally, under these conditions, the cells excreted to the medium large amounts of a previously characterized exopolysaccharide, at production rates as high as 35 g m(-2) per day (1.4 g l(-1) per day). Properly operated closed cultures of this strain of Anabaena appear most suitable for outdoor mass production of valuable extracellular polysaccharides.     

Cyanobacterial blooms and water quality in Greek waterbodies

The cyanobacterial species composition of nine Greek waterbodies of different type and trophic status was examined during the warm period of the year (May–October). Cyanobacterial water blooms were observed in all waterbodies. Forty-six cyanobacterial taxa were identified, 11 of which are known to be toxic. Eighteen species are reported for the first time in these waterbodies, 8 of which are known to produce toxins. Toxin producing species were found in all of the waterbodies and were primarily dominant in bloom formations (e.g., Microcystis aeruginosa, Anabaena flos-aquae, Aphanizomenon flos-aquae and Cylindrospermopsis raciborskii). Cosmopolitan species (e.g., M. aeruginosa), pantropic (e.g., Anabaenopsis tanganyikae) and holarctic species (e.g., Anabaena flos-aquae) were encountered. Shallow, eutrophic waterbodies had blooms dominated by Microcystis species and were characterized by phytoplankton association M. Anabaena and Aphanizomenon species of association H were dominant in waterbodies with low dissolved inorganic nitrogen and thermal stratification in the summer. Total cyanobacterial biovolumes (CBV) ranged from 7 to 9,507 cm³ m⁻³ and were higher than Alert Level 2 and Guidance Level 2 (10 cm³ m⁻³; World Health Organization; WHO) in seven of the waterbodies. Chlorophyll a concentrations ranged from 6 to 90,000 mg m⁻³ and were higher than Alert Level 2 and Guidance Level 2 (50 mg m⁻³; WHO) in eight of the waterbodies. There is also an elevated risk of acute toxicosis (Guidance Level 3; WHO) in five waterbodies. Water of an undesirable quality, hazardous to humans and animals occurs in several Greek waterbodies.     

Harmful Cyanobacterial Blooms: Causes, Consequences, and Controls

Cyanobacteria are the Earth’s oldest oxygenic photoautotrophs and have had major impacts on shaping its biosphere. Their long evolutionary history (～3.5 by) has enabled them to adapt to geochemical and climatic changes, and more recently anthropogenic modifications of aquatic environments, including nutrient over-enrichment (eutrophication), water diversions, withdrawals, and salinization. Many cyanobacterial genera exhibit optimal growth rates and bloom potentials at relatively high water temperatures; hence global warming plays a key role in their expansion and persistence. Bloom-forming cyanobacterial taxa can be harmful from environmental, organismal, and human health perspectives by outcompeting beneficial phytoplankton, depleting oxygen upon bloom senescence, and producing a variety of toxic secondary metabolites (e.g., cyanotoxins). How environmental factors impact cyanotoxin production is the subject of ongoing research, but nutrient (N, P and trace metals) supply rates, light, temperature, oxidative stressors, interactions with other biota (bacteria, viruses and animal grazers), and most likely, the combined effects of these factors are all involved. Accordingly, strategies aimed at controlling and mitigating harmful blooms have focused on manipulating these dynamic factors. The applicability and feasibility of various controls and management approaches is discussed for natural waters and drinking water supplies. Strategies based on physical, chemical, and biological manipulations of specific factors show promise; however, a key underlying approach that should be considered in almost all instances is nutrient (both N and P) input reductions; which have been shown to effectively reduce cyanobacterial biomass, and therefore limit health risks and frequencies of hypoxic events.     

The Smallest Known Genomes of Multicellular and Toxic Cyanobacteria: Comparison,Minimal Gene Sets for Linked Traits and the Evolutionary Implications

Cyanobacterial morphology is diverse, ranging from unicellular spheres or rods to multicellular structures such as colonies and filaments. Multicellular species represent an evolutionary strategy to differentiate and compartmentalize certain metabolic functions for reproduction and nitrogen (N₂) fixation into specialized cell types (e.g. akinetes, heterocysts and diazocytes). Only a few filamentous, differentiated cyanobacterial species, with genome sizes over 5 Mb, have been sequenced. We sequenced the genomes of two strains of closely related filamentous cyanobacterial species to yield further insights into the molecular basis of the traits of N₂ fixation, filament formation and cell differentiation. Cylindrospermopsis raciborskii CS-505 is a cylindrospermopsin-producing strain from Australia, whereas Raphidiopsis brookii D9 from Brazil synthesizes neurotoxins associated with paralytic shellfish poisoning (PSP). Despite their different morphology, toxin composition and disjunct geographical distribution, these strains form a monophyletic group. With genome sizes of approximately 3.9 (CS-505) and 3.2 (D9) Mb, these are the smallest genomes described for free-living filamentous cyanobacteria. We observed remarkable gene order conservation (synteny) between these genomes despite the difference in repetitive element content, which accounts for most of the genome size difference between them. We show here that the strains share a specific set of 2539 genes with >90% average nucleotide identity. The fact that the CS-505 and D9 genomes are small and streamlined compared to those of other filamentous cyanobacterial species and the lack of the ability for heterocyst formation in strain D9 allowed us to define a core set of genes responsible for each trait in filamentous species. We presume that in strain D9 the ability to form proper heterocysts was secondarily lost together with N₂ fixation capacity. Further comparisons to all available cyanobacterial genomes covering almost the entire evolutionary branch revealed a common minimal gene set for each of these cyanobacterial traits.     

Photosynthesis of the cyanobacterial soil-crust lichen Collema tenax from arid lands in southern Utah, USA: role of water content on light and temperature responses of CO2 exchange

1. The gelatinous cyanobacterial Collema tenax is a dominant lichen of biotic soil crusts in the western United States. In laboratory experiments, we studied CO₂ exchange of this species as dependent on water content (WC), light and temperature. Results are compared with performance of green-algal lichens of the same site investigated earlier.
2. As compared with published data, photosynthetic capacity of C. tenax is higher than that of other cyanobacterial and green-algal soil-crust species studied. At all temperatures and photon flux densities of ecological relevance, net photosynthesis (NP) shows a strong depression at high degrees of hydration; maximal apparent quantum-use efficiency of CO₂ fixation is also reduced. Water requirements (moisture compensation point, WC for maximal NP) are higher than that of the green-algal lichens. Collema tenax exhibits extreme 'sun plant' features and is adapted to high thallus temperatures.
3. Erratic rain showers are the main source of moisture for soil crusts on the Colorado Plateau, quickly saturating the lichens with liquid water. High water-holding capacity of C. tenax ensures extended phases of favourable hydration at conditions of high light and temperature after the rain for substantial photosynthetic production. Under such conditions the cyanobacterial lichen appears superior over its green-algal competitors, which seem better adapted to habitats with high air humidity, dew or fog as prevailing source of moisture.     

Four-carbon dicarboxylic acid production through the reductive branch of the open cyanobacterial tricarboxylic acid cycle in Synechocystis sp. PCC 6803

Succinate, fumarate, and malate are valuable four-carbon (C4) dicarboxylic acids used for producing plastics and food additives. C4 dicarboxylic acid is biologically produced by heterotrophic organisms. However, current biological production requires organic carbon sources that compete with food uses. Herein, we report C4 dicarboxylic acid production from CO₂ using metabolically engineered Synechocystis sp. PCC 6803. Overexpression of citH, encoding malate dehydrogenase (MDH), resulted in the enhanced production of succinate, fumarate, and malate. citH overexpression increased the reductive branch of the open cyanobacterial tricarboxylic acid (TCA) cycle flux. Furthermore, product stripping by medium exchanges increased the C4 dicarboxylic acid levels; product inhibition and acidification of the media were the limiting factors for succinate production. Our results demonstrate that MDH is a key regulator that activates the reductive branch of the open cyanobacterial TCA cycle. The study findings suggest that cyanobacteria can act as a biocatalyst for converting CO₂ to carboxylic acids.     

蓝藻毒素对底栖动物的毒理学研究进展

近年,由于人类活动加剧,大量氮磷等营养物质流入湖泊等缓流水体,导致水体富营养化。而由此引起有害蓝藻水华的频繁爆发,使生态环境和人类健康受到严重威胁。相关研究表明,蓝藻水华的爆发不仅能够使水体水质恶化,其中一些产毒藻类还会产生大量蓝藻毒素,对水生生物产生重要影响。底栖动物作为水体生态系统的重要组成部分,在食物网中有重要作用,同时其中的许多种类又与人类息息相关,因此关于水华蓝藻毒素对淡水底栖动物的毒理学研究具有重要意义。在介绍蓝藻毒素概况的基础上,综述了蓝藻毒素的致毒机理和对底栖动物的影响,展望了研究方向。     

Bioindustrial manufacturing readiness levels (BioMRLs) as a shared framework for measuring and communicating the maturity of bioproduct manufacturing processes

Readiness level (RL) frameworks such as technology readiness levels and manufacturing readiness levels describe the status of a technology/manufacturing process on its journey from initial conception to commercial deployment. More importantly, they provide a roadmap to guide technology development and scale-up from a ‘‘totality of system’’ approach. Commercialization risks associated with too narrowly focused R&D efforts are mitigated. RLs are defined abstractly so that they can apply to diverse industries and technology sectors. However, differences between technology sectors make necessary the definition of sector specific RL frameworks. Here, we describe bioindustrial manufacturing readiness levels (BioMRLs), a classification system specific to bioindustrial manufacturing. BioMRLs will give program managers, investors, scientists, and engineers a shared vocabulary for prioritizing goals and assessing risks in the development and commercialization of a bioindustrial manufacturing process.     

A Neglected Science: Applying Behavior to Aquatic Conservation

Behavioral theories, insights, and techniques are too frequently ignored by conservation biologists. Yet an animal's survival and reproductive success clearly depend on its behavior. Using examples from marine, freshwater, and terrestrial realms, I assert that behavioral information is invaluable in five conservation areas: (1) managing wild species (e.g., designing marine reserves; reducing animal–human conflicts; understanding and managing species' responses to human-induced environmental stress such as fishing, introduced species, and chemical, visual, and acoustic pollution); (2) actively reversing the decline of imperiled wild species (e.g., reducing bycatch by improving selectivity of fishing gear; re-establishing breeding populations and boosting reproduction); (3) assessing biodiversity (e.g., modeling population viability; censusing and monitoring populations and species); (4) captive breeding and reintroduction programs (e.g., minimizing loss of valuable phenotypes; teaching or maintaining valuable survival skills); and (5) changing human behavior in resource exploitation (e.g., using principles from social psychology). Both realized and potential applications to fishes are stressed. Finally, behavioral diversity, a valuable but neglected element of biodiversity, needs to be explicitly conserved to maintain diverse populations. Arguments are presented that the conservation of species diversity and genetic diversity alone does not necessarily protect important behavioral diversity. The maintenance of both individual and population variability may be essential for the preservation of a species.     

Cyanobacterial dominance in Brazil: distribution and environmental preferences

Based on a literature survey, we evaluated the periods of cyanobacterial dominance in Brazil. We hypothesized that variability of environmental forces along the country will promote or facilitate temporal and spatial mosaic in cyanobacterial dominance. The most striking outcomes are related to the dominance of Cylindrospermopsis, Dolichospermum, and Microcystis. Although they share important adaptive strategies (e.g., aerotopes, large size and toxins production), our findings suggest that they have different environmental preferences. Dolichospermum and Microcystis dominated mainly in warm-rainy periods whereas Cylindrospermopsis was more common during dry periods and in mixed systems, or formed perennial dominance. Maximum phosphorus concentrations were observed in reservoirs dominated by Cylindrospermopsis. Although the main genera reached high biomass levels individually, different abilities to form dominance and co-dominance were observed. The number of co-dominance of Chroococales and Nostocales was almost the same as the individual occurrence of the main genera from these groups. This dataset reveals patterns of dominance of these cyanobacteria and also indicates that physiological features will cause differences in the mechanisms of interactions between species. The understanding of these processes and their relationship to environmental conditions will promote better understanding of cyanobacterial dominance and increase our ability to predict and manage these events.     

PRODUCTION OF SECONDARY METABOLITES BY FILAMENTOUS TROPICAL MARINE CYANOBACTERIA: ECOLOGICAL FUNCTIONS OF THE COMPOUNDS

Cyanobacterial blooms are becoming more common in many reef habitats. The broadly acting feeding deterrent compound ypaoamide, produced by a mixed cyanobacterial assemblage, has been linked to bloom formation and mass fish die-offs ( Siganus argenteus and Siganus spinus ) in Guam. Specific metabolites produced by Lyngbya majuscula Gomont act as both feeding attractants to the specialist herbivore Stylocheilus longicauda , and as effective feeding deterrents to generalist fishes. Two-dimensional TLC (2D-TLC) analysis of cyanobacterial crude extracts was used to select chemically distinct populations (chemotypes) of bloom-forming filamentous cyanobacteria for chemical and ecological evaluation. Crude extracts produced by different species, chemotypes, and chemically distinct Micronesian marine cyanobacterial assemblages deter feeding activity of generalist reef herbivores. The ecological function of cyanobacterial secondary metabolites, especially as related to diversity of compound production and the relationship of metabolite production to bloom formation is discussed.     

Nitrogen Forms Influence Microcystin Concentration and Composition via Changes in Cyanobacterial Community Structure

The eutrophication of freshwaters is a global health concern as lakes with excess nutrients are often subject to toxic cyanobacterial blooms. Although phosphorus is considered the main element regulating cyanobacterial biomass, nitrogen (N) concentration and more specifically the availability of different N forms may influence the overall toxicity of blooms. In this study of three eutrophic lakes prone to cyanobacterial blooms, we examined the effects of nitrogen species and concentrations and other environmental factors in influencing cyanobacterial community structure, microcystin (MC) concentrations and MC congener composition. The identification of specific MC congeners was of particular interest as they vary widely in toxicity. Different nitrogen forms appeared to influence cyanobacterial community structure leading to corresponding effects on MC concentrations and composition. Total MC concentrations across the lakes were largely explained by a combination of abiotic factors: dissolved organic nitrogen, water temperature and ammonium, but Microcystis spp. biomass was overall the best predictor of MC concentrations. Environmental factors did not appear to affect MC congener composition directly but there were significant associations between specific MC congeners and particular species. Based on redundancy analyses (RDA), the relative biomass of Microcystis aeruginosa was associated with MC-RR, M. wesenbergii with MC-LA and Aphanizomenon flos-aquae with MC-YR. The latter two species are not generally considered capable of MC production. Total nitrogen, water temperature, ammonium and dissolved organic nitrogen influenced the cyanobacterial community structure, which in turn resulted in differences in the dominant MC congener and the overall toxicity.     

First report of cyanobacterial diversity and microcystins in a Microcystis strain from Sidi Boughaba,a Moroccan coastal lagoon

The cyanobacterial diversity of Sidi Boughaba, a Moroccan coastal lagoon and Ramsar site, was evaluated and its potentially toxic species were isolated and characterised. This study was the first time that cyanobacterial diversity and cyanotoxin production have been characterised in a Moroccan coastal lagoon. Samples collected in June 2004, July 2008 and August 2013 contained 41 species. Three strains of cyanobacteria were isolated, cultured and assessed for their potentially toxic levels of microcystins. Only Microcystis flos-aquae exhibited the presence of microcystins, at a concentration of 823.41 µg g?1 as MC-LR equivalents. MC-RR and MC-WR were also detected. The presence of toxic Microcystis and other potentially toxic strains, especially in periods of bloom proliferations, could pose environmental and health hazards. Cyanotoxin monitoring of this lagoon is highly recommended.     

Contrasting microcystin production and cyanobacterial population dynamics in two Planktothrix-dominated freshwater lakes

Microcystin concentrations in two Dutch lakes with an important Planktothrix component were related to the dynamics of cyanobacterial genotypes and biovolumes. Genotype composition was analysed by using denaturing gradient gel electrophoresis (DGGE) profiling of the intergenic transcribed spacer region of the rrn operon (rRNA-ITS), and biovolumes were measured by using microscopy. In Lake Tjeukemeer, microcystins were present throughout summer (maximum concentration 30 microg l(-1)) while cyanobacterial diversity was low and very constant. The dominant phototroph was Planktothrix agardhii. In contrast, Lake Klinckenberg showed a high microcystin peak (up to 140 microg l(-1)) of short duration. In this lake, cyanobacterial diversity was higher and very dynamic with apparent genotype successions. Several genotypes derived from DGGE field profiles matched with genotypes from cultures isolated from field samples. The microcystin peak measured in Lake Klinckenberg could be confidently linked to a bloom of Planktothrix rubescens, as microscopic and genotypic analysis showed identity of bloom samples and a toxin-producing P. rubescens culture. Toxin-producing genotypes were detected in the microbial community before they reached densities at which they were detected by using microscopy. Cyanobacterial biovolumes provided additional insights in bloom dynamics. In both lakes, the microcystin content per cell was highest at the onset of the blooms. Our results suggest that while genotypic characterization of a lake can be valuable for detection of toxic organisms, for some lakes a monitoring of algal biomass has sufficient predictive value for an assessment of toxin production.     

Integrating local plant resources and habitat management

The use of wild plant resources as a source of basic needs is an important aspect of multiple-use of land in much of Africa, ranging from vegetation with a low species diversity, high biomass production and resilience to harvesting (e.g.Phragmites reedbeds,Cymbopogon grasslands) through to vegetation with a high diversity of species (and plant life-forms), with a multiple of uses and often low resilience to resource harvesting (e.g. medicinal plants, pole cutting, fuelwood from Afro-montane forest). Complexity and costs of managing sustainable use of wild populations increase markedly with an increasing number of uses and resource users. It is suggested that if the primary objective of core conservation areas is accepted to be the maintenance of habitat and species diversity, then the limited money and manpower available for management of core conservation areas in southern Africa limits sustainable harvesting use of plant resources to low diversity, low conservation priority vegetation types or encroaching species. For high diversity, high conservation priority sites such as Afro-montane or Coastal evergreen forest, the emphasis must be on providing alternative sources of supply to resource users outside of core areas. Botanical gardens staff, with their horticultural experience, can play a very important role through bulking up material for supply to small farmers, herbalists and introducing additional species for agro-forestry. Botanical gardens can play a greater role in the establishment of field gene banks andex situ conservation of vulnerable species. They can also provide a valuable educational and research role on mass production techniques to boost local stocks of threatened and commercially valuable species for cultivation by local people, whether farmers or specialist users (e.g. herbalists) in the country of origin as a means of generating employment and restoring local self-sufficiency.