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At least 271 records · Page 15

Improving Quantitative Laboratory Analysis of Phycobiliproteins to Provide High Quality Validation Data for Ocean Color Remote Sensing Algorithm

Identification and characterization of phytoplankton communities and their physiology is a primary aim of NASA's PACE satellite mission. The concentration and composition of phytoplankton pigments modulate the spectral distribution of light emanating from the ocean, which is measured by ocean color satellites, and thus provide critical information on phytoplankton community composition and physiological parameters. One diagnostic class of pigments not routinely well-characterized is the phycobiliproteins (PBPs), and NASA has a requirement to collect and distribute high quality in situ data in support of data product validation activities for ocean color missions. Phycobiliproteins are light-harvesting proteins that are the predominant photosynthetic pigments in some classes of phytoplankton including cyanobacteria, such as Synechococcus, Trichodesmium, and Microcystis. With the advance of hyperspectral ocean color sensors such as on PACE (expected to launch in late 2022), it is essential that we implement routine analysis of PBPs that satisfies several considerations: reproducible, high extraction efficiency for a variety of environments, and Suitable for large scale analysis. Published techniques for PBP analysis vary in recommendations for: collection, extraction, disruption mode, and analysis; evidence suggests the variation in results may depend at least in part on the species and even strain(s) of interest. Experiments that tested variations in these parameters have drawn very different conclusions regarding extraction efficiency and reproducibility. Cyanobacteria are more difficult to extract than other PBP-containing algae such as cryptophytes, but can be important primary producers. We used a cryptophyte (Rhodomonas salina) and cyanobacterium (Synechococcus sp.) to compare extraction efficiencies of water samples concentrated via centrifugation to filtered samples using two different extraction buffers (phosphate and asolectin-CHAPS). Samples were analyzed on a fluorometer configured for PC and PE detection. The results have important implications for collection and storage of samples for routine analysis; some previous studies (although not all) have suggested that filtered samples have a much lower extraction efficiency than whole water samples.

Kenemer, Christopher↗

The Application of Remote Sensing and Machine Learning to Improve Early Warning Systems for Harmful Algal Events in the Highland Lake Chain, TX

Beginning in 2019, harmful algal events have caused canine deaths in both Lady Bird Lake and Lake Travis located near Austin, Texas. These two reservoirs are part of the larger Highland Lakes chain, managed by the City of Austin Department of Watershed Protection (COA DWP) and the Lower Colorado River Authority (LCRA), which fulfill municipal, commercial, and agricultural water demands. Given the recent increase in favorable environmental conditions for algal events in central Texas, NASA DEVELOP partnered with LCRA and COA DWP to improve monitoring and early detection of algal events, utilizing satellite remote sensing and machine learning. Spatially and temporally varied chlorophyll a concentrations, cyanobacteria detections, turbidity, and water surface temperature products are used as environmental proxies. Landsat 8 Operational Land Imager (OLI) and the Sentinel-2 MultiSpectral Instrument (MSI) data were used to provide a combined revisit time of up to ~3 days and < 30 m per pixel products. Chlorophyll a concentrations were estimated using a pre-trained Mixture Density Network, and cyanobacteria detection was accomplished using the Broad Wavelength Algae Index, which can differentiate algal blooms from algal proliferations (mats). In situ data were used to validate remotely sensed measurements and quantify uncertainties. Preliminary results show a good fit between the modeled output and in situ observations, suggesting that remote sensing data can be used to retrieve biogeochemical properties and/or inherent optical properties (IOPs) of water columns in these inland human-made lakes. Uncertainties were introduced from the sensitivity to atmospheric correction, inherent mismatch between satellite and sampling data, and a relatively lower signal-to-noise ratio over water. The resulting products enable near real-time monitoring of environmental proxies relevant to algal event presence in the Highland Lakes chain, and will ultimately support water management, decision making, and risk communication.

Shuyu Chang↗

Satellites for long-term monitoring of inland U.S. lakes: The MERIS time series and application for chlorophyll-a

Lakes and other surface fresh waterbodies provide drinking water, recreational and economic opportunities, food, and other critical support for humans, aquatic life, and ecosystem health. Lakes are also productive ecosystems that provide habitats and influence global cycles. Chlorophyll concentration provides a common metric of water quality, and is frequently used as a proxy for lake trophic state. Here, we document the generation and distribution of the complete MEdium Resolution Imaging Spectrometer (MERIS; Appendix A provides a complete list of abbreviations) radiometric time series for over 2300 satellite resolvable inland bodies of water across the contiguous United States (CONUS) and more than 5,000 in Alaska. This contribution greatly increases the ease of use of satellite remote sensing data for inland water quality monitoring, as well as highlights new horizons in inland water remote sensing algorithm development. We evaluate the performance of satellite remote sensing Cyanobacteria Index (CI)-based chlorophyll algorithms, the retrievals for which provide surrogate estimates of phytoplankton concentrations in cyanobacteria dominated lakes. Our analysis quantifies the algorithms' abilities to assess lake trophic state across the CONUS. As a case study, we apply a bootstrapping approach to derive a new CI-to-chlorophyll relationship, ChlBS, which performs relatively well with a multiplicative bias of 1.11 (11%) and mean absolute error of 1.60 (60%). While the primary contribution of this work is the distribution of the MERIS radiometric timeseries, we provide this case study as a roadmap for future stakeholders' algorithm development activities, as well as a tool to assess the strengths and weaknesses of applying a single algorithm across CONUS.

MERIS timeseries↗

Lake Champlain Water Resources: Using Earth Observations to Identify Spatial and Seasonal Trends of Harmful Algal Events in Lake Champlain

Lake Champlain provides clean drinking water for 35% of the surrounding watershed and offers recreational opportunities to millions of tourists. However, current levels of cyanobacteria and phosphorus created breeding grounds for harmful algal blooms (HABs). The excess of phosphorus runoff into Lake Champlain over the past decade encouraged toxic cyanobacterial formations, thereby increasing the severity of HABs towards local economy and ecology. In partnership with the Natural Resources Conservation Service (NRCS) Northeast Region, this project utilized Earth observations to identify risk factors associated with toxic algal blooms. The team detected historic algal bloom trends with Sentinel-2 Multispectral Instrument (MSI), Sentinel-3 Ocean and Land Color Instrument (OLCI), Landsat 8 Operational Land Imager (OLI) and Landsat 9 OLI-2. The team also used Sentinel-3 OLCI and the German Aerospace Center’s Earth Sensing Imagery Spectrometer (DESIS) to visualize algal bloom patterns and Landsat 8 OLI, Landsat 9 OLI-2, and Shuttle Radar Topography Mission (SRTM) to identify phosphorus sources within the watershed. The team’s analyses indicated an increase in cyanobacteria blooms during summer months from 2016-2022, with Missisquoi and St. Albans Bay exhibiting the greatest concentrations of toxic events. Furthermore, 16% of the watershed was identified as posing an immediate threat to the lake’s hydrology. The area of greatest concern was the Missisquoi Bay sub-watershed, with 229,044 acres of land prone to excessive phosphorus runoff. Providing this information to the NRCS Northeast Region enabled the organization to quantify risk factors associated with algal blooms and modify mitigation efforts to better target future bloom events.

Brianne Kendall↗

From cytoplasm to lumen—mapping the free pools of protein subunits of three photosynthetic complexes using quantitative mass spectrometry

The phycobilisome (PBS) captures light energy and transfers it to photosystem I (PSI) and photosystem II (PSII). Which and how many copies of protein subunits in PBSs, PSI, and PSII remain unbound in thylakoids are unknown. Here, quantitative mass spectrometry (QMS) was used to quantify substantial pools of free extrinsic subunits of PSII and PSI. Interestingly, the membrane intrinsic PsaL is 3-fold higher than PsaA/B. This scenario complements the static structures of these complexes as revealed by X-ray crystallography and cryo-EM. Furthermore, the ratios of ApcG and photoprotective OCP over PBS indicate a pool of extra ApcG. The 2.5 ratio of CpcG-PBS over CpcL-PBS improves our understanding of these light-harvesting complexes involved in energy capture and photoprotection in cyanobacteria.

cyanobacteria↗

Quantification of microcystin production and biodegradation rates in the western basin of Lake Erie

Abstract Cyanobacterial biomass forecasts currently cannot predict the concentrations of microcystin, one of the most ubiquitous cyanotoxins that threaten human and wildlife health globally. Mechanistic insights into how microcystin production and biodegradation by heterotrophic bacteria change spatially and throughout the bloom season can aid in toxin concentration forecasts. We quantified microcystin production and biodegradation during two growth seasons in two western Lake Erie sites with different physicochemical properties commonly plagued by summer Microcystis blooms. Microcystin production rates were greater with elevated nutrients than under ambient conditions and were highest nearshore during the initial phases of the bloom, and production rates were lower in later bloom phases. We examined biodegradation rates of the most common and toxic microcystin by adding extracellular stable isotope‐labeled microcystin‐LR (1 μ g L −1 ), which remained stable in the abiotic treatment (without bacteria) with minimal adsorption onto sediment, but strongly decreased in all unaltered biotic treatments, suggesting biodegradation. Greatest biodegradation rates (highest of −8.76 d −1 , equivalent to the removal of 99.98% in 18 h) were observed during peak bloom conditions, while lower rates were observed with lower cyanobacteria biomass. Cell‐specific nitrogen incorporation from microcystin‐LR by nanoscale imaging mass spectrometry showed that a small percentage of the heterotrophic bacterial community actively degraded microcystin‐LR. Microcystin production and biodegradation rates, combined with the microcystin incorporation by single cells, suggest that microcystin predictive models could be improved by incorporating toxin production and biodegradation rates, which are influenced by cyanobacterial bloom stage (early vs. late bloom), nutrient availability, and bacterial community composition.

59 BASIC BIOLOGICAL SCIENCES↗

A cellular selection identifies elongated flavodoxins that support electron transfer to sulfite reductase

Flavodoxins (Flds) mediate the flux of electrons between oxidoreductases in diverse metabolic pathways. To investigate whether Flds can support electron transfer to a sulfite reductase (SIR) that evolved to couple with a ferredoxin, we evaluated the ability of Flds to transfer electrons from a ferredoxin-NADP reductase (FNR) to a ferredoxin-dependent SIR using growth complementation of an Escherichia coli strain with a sulfur metabolism defect. We show that Flds from cyanobacteria complement this growth defect when coexpressed with an FNR and an SIR that evolved to couple with a plant ferredoxin. When we evaluated the effect of peptide insertion on Fld-mediated electron transfer, we observed a sensitivity to insertions within regions predicted to be proximal to the cofactor and partner binding sites, while a high insertion tolerance was detected within loops distal from the cofactor and within regions of helices and sheets that are proximal to those loops. Bioinformatic analysis showed that natural Fld sequence variability predicts a large fraction of the motifs that tolerate insertion of the octapeptide SGRPGSLS. In conclusion, these results represent the first evidence that Flds can support electron transfer to assimilatory SIRs, and they suggest that the pattern of insertion tolerance is influenced by interactions with oxidoreductase partners.

59 BASIC BIOLOGICAL SCIENCES↗

Challenges, progress, and future perspectives for cyanobacterial polyhydroxyalkanoate production

Polyhydroxyalkanoates (PHA) are a promising bio-based alternative to traditional plastics derived from petroleum. Cyanobacteria are photosynthetic organisms that produce PHA from CO 2 and sunlight, which can potentially reduce production costs and environmental footprint in comparison to heterotrophic bacteria cultures because (1) they utilize inorganic carbon sources for growth and (2) they do not require intensive aeration for oxygenation. Moreover, supplementing precursors such as propionate, acetate, valerate, etc., can be used to obtain various copolymers with plastic customizable properties in comparison to the classical homopolymers, such as polyhydroxybutyrate, PHB. This critical review covers the latest advances in PHA production, including recent discoveries in the metabolism interplay between PHA and glycogen production, and new insights into cultivation strategies that enhance PHA accumulation, and purification processes. This review also addresses the challenges and suggests potential solutions for a viable industrial PHAs production process.

59 BASIC BIOLOGICAL SCIENCES↗

Microalgal lipids: biochemistry and biotechnology

Microalgae, including cyanobacteria, are a genetically diverse and biochemically diverse group of mostly photosynthetic organisms that can be found in nearly every ecosystem. They produce a broad range of compounds, including structural and bioactive lipids with various roles in the metabolism of the cell and potential applications in biotechnology. Here, the majority of lipids are synthesized in the chloroplast using acetate to produce fatty acids and terpenoids via the acetate pathway and methylerythritol phosphate deoxy-xylulose phosphate pathway, respectively. This review will highlight the major groups of microalgal lipids as well as their applications in food, fuels, and specialty chemicals. Future directions include the development of microalgal chassis organisms and molecular tools to manipulate lipid synthesis for the enhanced production of target metabolites.

cyanobacteria↗

Structure of a photosystem I-ferredoxin complex from a marine cyanobacterium provides insights into far-red light photoacclimation

Far-red light photoacclimation (FaRLiP) exhibited by some cyanobacteria allows these organisms to use light of the far-red region of the solar spectrum (700 to 800 nm) for photosynthesis. Part of this process includes the replacement of six photosystem I (PSI) subunits with isoforms that confer the binding of chlorophyll (Chl) f molecules that are synthesized by chlorophyll f synthase and that absorb far-red light. However, the exact sites at which Chl f molecules are bound are still challenging to determine. To aid in the identification of Chl fbinding sites, we solved the cryo-EM structure of PSI from far-red light-acclimated cells (FRLPSI) of the cyanobacterium Synechococcus sp. PCC 7335. We identified six sites that bind Chl f with high specificity, and three additional sites that are likely to bind Chl f at lower specificity. All of these binding sites are in the core antenna regions of PSI, and Chl f was not observed among the electron transport cofactors. This structural analysis also reveals both conserved and non-conserved Chl f-binding sites, the latter of which exemplify the diversity in FRL-PSI among species. We found that the FRL-PSI structure also contains a bound soluble ferredoxin, PetF1, at low occupancy, which suggests that ferredoxin binds less transiently than expected according to the canonical view of ferredoxin-binding to facilitate electron transfer. We suggest that this may result from structural changes in FRL-PSI that occur specifically during FaRLiP.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Structure of a dimeric photosystem II complex from a cyanobacterium acclimated to far-red light

Photosystem II (PSII) is the water-splitting enzyme central to oxygenic photosynthesis. To drive water oxidation, light is harvested by accessory pigments, mostly chlorophyll (Chl) a molecules, which absorb visible light (400–700 nm). Some cyanobacteria facultatively acclimate to shaded environments by altering their photosynthetic machinery to additionally absorb far-red light (FRL, 700–800 nm), a process termed far-red light photoacclimation or FaRLiP. During far-red light photoacclimation, FRL-PSII is assembled with FRL-specific isoforms of the subunits PsbA, PsbB, PsbC, PsbD, and PsbH, and some Chl-binding sites contain Chls d or f instead of the usual Chl a. The structure of an apo-FRL-PSII monomer lacking the FRL-specific PsbH subunit has previously been determined, but visualization of the dimeric complex has remained elusive. Here, we report the cryo-EM structure of a dimeric FRL–PSII complex. The site assignments for Chls d and f are consistent with those assigned in the previous apo-FRL-PSII monomeric structure. All sites that bind Chl d or Chl f at high occupancy exhibit a FRL-specific interaction of the formyl moiety of the Chl d or Chl f with the protein environment, which in some cases involves a phenylalanine sidechain. The structure retains the FRL-specific PsbH2 subunit, which appears to alter the energetic landscape of FRL-PSII, redirecting energy transfer from the phycobiliprotein complex to a Chl f molecule bound by PsbB2 that acts as a bridge for energy transfer to the electron transfer chain. Collectively, these observations extend our previous understanding of the structure-function relationship that allows PSII to function using lower energy FRL.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Structure of the antenna complex expressed during far-red light photoacclimation in Synechococcus sp. PCC 7335

Far-red light photoacclimation, or FaRLiP, is a facultative response exhibited by some cyanobacteria that allows them to absorb and utilize lower energy light (700–800 nm) than the wavelengths typically used for oxygenic photosynthesis (400–700 nm). During this process, three essential components of the photosynthetic apparatus are altered: photosystem I, photosystem II, and the phycobilisome. In all three cases, at least some of the chromophores found in these pigment–protein complexes are replaced by chromophores that have red-shifted absorbance relative to the analogous complexes produced in visible light. Recent structural and spectroscopic studies have elucidated important features of the two photosystems when altered to absorb and utilize far-red light, but much less is understood about the modified phycobiliproteins made during FaRLiP. We used single-particle, cryo-EM to determine the molecular structure of a phycobiliprotein core complex comprising allophycocyanin variants that absorb far-red light during FaRLiP in the marine cyanobacterium Synechococcus sp. PCC 7335. The structure reveals the arrangement of the numerous red-shifted allophycocyanin variants and the probable locations of the chromophores that serve as the terminal emitters in this complex. It also suggests how energy is transferred to the photosystem II complexes produced during FaRLiP. The structure additionally allows comparisons with other previously studied allophycocyanins to gain insights into how phycocyanobilin chromophores can be tuned to absorb far-red light. These studies provide new insights into how far-red light is harvested and utilized during FaRLiP, a widespread cyanobacterial photoacclimation mechanism.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Structural and Biophysical Properties of a [4Fe-4S] Ferredoxin-Like Protein from Synechocystis sp. PCC 6803 with a Unique Two Domain Structure

Electron carrier proteins (ECPs), binding iron-sulfur clusters, are vital components within the intricate network of metabolic and photosynthetic reactions. They play a crucial role in the distribution of reducing equivalents. In Synechocystis sp. PCC 6803, the ECP network includes at least nine ferredoxins. Previous research, including global expression analyses and protein binding studies, has offered initial insights into the functional roles of individual ferredoxins within this network. This study primarily focuses on Ferredoxin 9 (slr2059). Through sequence analysis and computational modeling, Ferredoxin 9 emerges as a unique ECP with a distinctive two-domain architecture. It consists of a C-terminal iron-sulfur binding domain and an N-terminal domain with homology to Nil-domain proteins, connected by a structurally rigid 4-amino acid linker. Notably, in contrast to canonical [2Fe-2S] ferredoxins exemplified by PetF (ssl0020), which feature highly acidic surfaces facilitating electron transfer with photosystem I reaction centers, models of Ferredoxin 9 reveal a more neutral to basic protein surface. Using a combination of electron paramagnetic resonance spectroscopy and square-wave voltammetry on heterologously produced Ferredoxin 9, this study demonstrates that the protein coordinates 2x[4Fe-4S]2+/1+ redox-active and magnetically interacting clusters, with measured redox potentials of -420 +/- 9 mV and -516 +/- 10 mV vs SHE. A more in-depth analysis of Fdx9's unique structure and protein sequence suggests that this type of Nil-2[4Fe-4S] multi-domain ferredoxin is well conserved in cyanobacteria, bearing structural similarities to proteins involved in homocysteine synthesis in methanogens.

cyanobacteria↗

Improving the Transformation Efficiency of Synechococcus sp. PCC 7002 via Methylome-Guided Premethylation of DNA

Cyanobacteria are promising microbial platforms for a diverse set of biotechnology applications, from living materials to photosynthetic chemical production, but are less well characterized than commonly engineered microbes such as Escherichia coli. This study facilitates genetic engineering in Synechococcus sp. PCC 7002, a fast-growing, halotolerant, and naturally competent strain, by identifying ten native methylation motifs and designing shuttle strains that mimic the native methylation state by expressing a subset of heterologous methyltransferases. DNA methylation in E. coli with as few as two active methyltransferases increased transformation efficiency up to 30-fold across four distinct integration sites in PCC 7002. This work provides an experimental framework to bypass native restriction-modification systems for efficient genome editing and metabolic engineering in nonmodel bacteria.

59 BASIC BIOLOGICAL SCIENCES↗

Biosynthesis of Minimal C-Phycocyanin Chromophore Assemblies in E. coli Provides a Platform to Dissect Protein-Mediated Tuning of Exciton Transfer

Cyanobacteria are arguably among the most evolutionarily successful organisms on Earth, inhabiting a wide range of ocean, freshwater, soil, and even desert environments on every continent. The cyanobacterial phycobilisome consists of stacks of disk-like light-collecting moieties, allophycocyanin (APC) and phycocyanin (CPC), with covalently bound phycocyanobilin (PCB) pigments. The ways in which the energies of the specific chromophores in these complexes are tuned by the protein to achieve its highly efficient and directional energy transfer are not fully understood, as complex combinations of decay pathways are occurring simultaneously and competitively through this elaborate light-harvesting system. This makes it difficult to extract information about isolated protein-pigment interactions. We provide herein a description of a useful new experimental platform in which we have recombinantly expressed a fully functioning CPC complex and selectively created minimal chromophore sets to study their individual contributions to the overall CPC spectra. Structural and computational analysis of this protein system have provided a greater understanding of how the protein environment serves to alter the photophysics of each of these chromophores. Introduction of a quencher into various positions within CPC confirmed the ability of the protein environment to tune the directionality of energy transport in this assembly. Further mutational analysis suggested the roles of key amino acids surrounding the chromophores, showcasing the utility of heterologous expression techniques for understanding the effects of structure on EET mechanisms in the phycobilisome.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Niche expansion for phototrophic sulfur bacteria at the Proterozoic–Phanerozoic transition

Significance Carotenoid pigments afford valuable clues about the chemistry and biology of both modern and ancient aquatic environments. This study reveals that fossil aromatic carotenoids—long considered biomarkers for anoxygenic, phototrophic sulfur bacteria and their physiological requirement for hydrogen sulfide and illumination—can also be biosynthesized by oxygen-producing cyanobacteria. Cyanobacterial aromatic carotenoids, which are distinct in their chemical structures and occurrence patterns, are the most commonly encountered compounds in Proterozoic marine settings as well as in lakes from more recent eras. In contrast, carotenoids diagnostic for green sulfur bacteria of the family Chlorobiaceae became both prevalent and abundant in marine paleoenvironments beginning in the Phanerozoic Eon. This expansion occurs as marine sulfate inventories increased toward the end of the Proterozoic Eon.

59 BASIC BIOLOGICAL SCIENCES↗

Carbon isotope fractionation by an ancestral rubisco suggests that biological proxies for CO 2 through geologic time should be reevaluated

The history of Earth’s carbon cycle reflects trends in atmospheric composition convolved with the evolution of photosynthesis. Fortunately, key parts of the carbon cycle have been recorded in the carbon isotope ratios of sedimentary rocks. The dominant model used to interpret this record as a proxy for ancient atmospheric CO 2 is based on carbon isotope fractionations of modern photoautotrophs, and longstanding questions remain about how their evolution might have impacted the record. Therefore, we measured both biomass (ε p ) and enzymatic (ε Rubisco ) carbon isotope fractionations of a cyanobacterial strain ( Synechococcus elongatus PCC 7942) solely expressing a putative ancestral Form 1B rubisco dating to ≫1 Ga. This strain, nicknamed ANC, grows in ambient pCO 2 and displays larger ε p values than WT, despite having a much smaller ε Rubisco (17.23 ± 0.61‰ vs. 25.18 ± 0.31‰, respectively). Surprisingly, ANC ε p exceeded ANC ε Rubisco in all conditions tested, contradicting prevailing models of cyanobacterial carbon isotope fractionation. Such models can be rectified by introducing additional isotopic fractionation associated with powered inorganic carbon uptake mechanisms present in Cyanobacteria, but this amendment hinders the ability to accurately estimate historical pCO 2 from geological data. Understanding the evolution of rubisco and the CO 2 concentrating mechanism is therefore critical for interpreting the carbon isotope record, and fluctuations in the record may reflect the evolving efficiency of carbon fixing metabolisms in addition to changes in atmospheric CO 2 .

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Natural diversity provides a broad spectrum of cyanobacteriochrome-based diguanylate cyclases

Cyanobacteriochromes (CBCRs) are spectrally diverse photosensors from cyanobacteria distantly related to phytochromes that exploit photoisomerization of linear tetrapyrrole (bilin) chromophores to regulate associated signaling output domains. Unlike phytochromes, a single CBCR domain is sufficient for photoperception. CBCR domains that regulate the production or degradation of cyclic nucleotide second messengers are becoming increasingly well characterized. Furthermore, cyclic di-guanosine monophosphate (c-di-GMP) is a widespread small-molecule regulator of bacterial motility, developmental transitions, and biofilm formation whose biosynthesis is regulated by CBCRs coupled to GGDEF (diguanylate cyclase) output domains. In this study, we compare the properties of diverse CBCR-GGDEF proteins with those of synthetic CBCR-GGDEF chimeras. Our investigation shows that natural diversity generates promising candidates for robust, broad spectrum optogenetic applications in live cells. Since light quality is constantly changing during plant development as upper leaves begin to shade lower leaves—affecting elongation growth, initiation of flowering, and responses to pathogens, these studies presage application of CBCR-GGDEF sensors to regulate orthogonal, c-di-GMP-regulated circuits in agronomically important plants for robust mitigation of such deleterious responses under natural growing conditions in the field.

59 BASIC BIOLOGICAL SCIENCES↗