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PubMed2026

Green genes from blue greens: challenges and solutions to unlocking the potential of cyanobacteria in drug discovery.

Cyanobacteria are prolific producers of biologically active compounds that are important in influencing ecology, behavior of interacting organisms, and as leads in drug discovery efforts. Here we discuss the challenges faced by all natural product researchers, especially those that focus on cyanobacteria, and then describe progress that has been made in these areas. We also propose some solutions, paths forward, and thoughts for consideration on these challenges.

Biological Products↗

PubMed2025

UAV-based multispectral image analysis revealed stay-green haplotypes in wheat specific for different soil nitrogen levels.

BACKGROUND: The so-called stay-green trait, a delay in onset and progression of leaf senescence, is associated with slower chlorophyll degradation and higher photosynthesis rates during maturation resulting in higher crop yields. Understanding the genetic and physiological basis of the stay-green trait and breeding cultivars with stable stay-green behaviour across a range of different nitrogen (N) conditions and specifically under low N availability can contribute to ensuring wheat yields and reducing N fertilizer application. The goal of this study was therefore to identify haplotypes associated with high stay-green capacity under different N availability conditions in wheat. A diverse set of 221 wheat cultivars was grown under three different N levels and phenotyped by uncrewed aerial vehicle (UAV)-based multispectral imaging to characterise genetic and environmental variation in stay-green. Haplotypes associated with stay-green were identified across N levels and specifically under low N availability. RESULTS: The plant senescence reflectance index (PSRI) calculated from multispectral images was identified as the most specific stay-green indicator allowing for differentiation of genotypic effects due to its greater sensitivity to senescence-related changes in pigment composition and its higher reliability. We found genetic variance for stay-green and a consistent genetic correlation between stay-green and grain yield at all imaging dates and N levels within the utilised diversity panel confirming its potential as a future breeding target. Haplotype analyses revealed two favourable major allele haplotypes present in 95% of the stay-green cultivars, i.e. the top 25% of the diversity set based on PSRI values, which significantly enhance stay-green performance and grain yield. In addition, we identified a favourable minor allele haplotype specifically associated with stay-green under low N availability and capable of further increasing stay-green and grain yield when stacked onto the two favourable major allele haplotypes. CONCLUSIONS: The newly identified stay-green haplotypes can be further used for fine-mapping and identifying the underlying genes as well as for selecting for higher stay-green and grain yield. Thereby our results can contribute to improving our understanding of the complex genetic regulation underlying stay-green in different environments and to breeding new cultivars with stable performance across N levels or specifically under low N availability.

Triticum↗

PubMed1978

A reappraisal of the reaction of butyryl-coenzyme A dehydrogenase with phenylmercuric acetate. Evidence that de-greening involves a reaction of the tightly bound thioester.

Phenylmercuric acetate reversibly de-greens butyryl-CoA dehydrogenase from Megasphaera elsdenii, abolishing the absorption band at 710nm. The view that this is a result of modification of a protein thiol group is re-examined in the light of the following new observations. (i) After treatment with phenylmercuric acetate, the enzyme's ability to be re-greened by addition of thiols was not decreased by gel filtration or precipitation with (NH(4))(2)SO(4). (ii) Phenylmercuric acetate caused the same extent of de-greening whether added in a few large amounts or many small ones. The overall time taken for de-greening was, however, greatly extended when many small additions were made. (iii) In Tris/acetate buffer, pH7.5, 3.5mol of phenylmercuric acetate/mol of enzyme subunit was required for complete de-greening, compared with only 2.5mol/mol in phosphate buffer, pH7. (iv) None of the groups that react with phenylmercuric acetate is accessible to iodoacetate or iodoacetamide. (v) On a molar basis dithiothreitol, mercaptoethanol and CoA are equally effective in re-greening the enzyme. (vi) Provided that phenylmercuric acetate is not present in excess, the de-greened enzyme forms normal and stable complexes with crotonyl-CoA and acetoacetyl-CoA. (vii) When a small excess of phenylmercuric acetate is present, full stable development of the enzyme-acetoacetyl-CoA complex requires addition of several mol of acetoacetyl-CoA/mol of enzyme subunit. (viii) The ability of de-greened enzyme to be immediately re-greened by an excess of thiol declines with time, more rapidly at pH6 than at pH7 or 8, but at all three pH values the instantaneous re-greening was followed by a slow phase of further increase in A(710). This further recovery was most extensive and most rapid at pH8. These findings are reminiscent of the previously described reversible decline in the re-greening capacity of a protein-free acid extract of green butyryl-CoA dehydrogenase. It is concluded that the likely cause of de-greening is chemical modification of the tightly bound thioester rather than a protein thiol group. The reversibility would be explained if the thioester exists on the surface of the enzyme in equilibrium with free CoA and a lactone, or if the acyl group is readily and reversibly transferred from the thiol of CoA to a protein side chain.

Acyl Coenzyme A↗

PubMed2025

Association between residential greenness and coronary heart disease: A proteomics and miRNA microarray analysis.

Greenness has been linked to cardiovascular disease. However, the specific biological mechanisms through which greenness impacts coronary heart disease (CHD) remain unclear. We aim to explore the underlying epigenetic mechanisms linking greenness and CHD by using proteomics and miRNA microarray. A total of 2387 participants were included in the population study, 816 of whom were diagnosed with CHD. Residential greenness exposure was characterized using the normalized difference vegetation index (NDVI). Generalized additive models and restricted cubic splines investigated the association between greenness and CHD. Mediation analysis examined whether cardiovascular metabolic risk factors (blood pressure, inflammation indicators, and glucose) mediated the association. After proteomics and miRNA microarray screening, Elisa and qRT-PCR validated selected proteins (THBS1, FCN3, and LTBP1) and miRNAs (miR-671-5p, miR-124-3p, and miR-379-5p) in CHD. Among these, LTBP1 and miR-379-5p showed significant differential expression (P < 0.05) and were examined as potential molecular mediators. Higher greenness exposure within a 1000-m area was associated with a lower risk of CHD (OR: 0.86, 95 % CI: 0.81, 0.92). Systolic blood pressure (6.32 % [95 % CI: 1.49 %, 13.12 %]), lymphocyte (10.98 % [95 % CI: 3.76 %, 22.00 %]), monocyte (9.94 % [95 % CI: 3.42 %, 20.87 %]), and fasting blood glucose (3.41 % [95 % CI: 0.56 %, 7.84 %]) mediated this association. LTBP1 and miR-379-5p were differentially expressed in CHD and mediated 7.19 % [95 % CI: 0.01 %, 23.37 %] and 20.03 % [95 % CI: 2.85 %, 69.71 %] of greenness effect on CHD, respectively. Combining the population study and experiments, we found that miR-379-5p and LTBP1 may jointly modulate vascular constriction and immune inflammation in the association between greenness and CHD.

Humans↗

PubMed1977

Somatic segregation and rate of greening after ultraviolet irradiation of Euglena gracilis.

1. During multiplication of irradiated cells, a segregation may take place between bleached cells, whose progeny is unable to green, and green ones. Some of the green cells give progenies exclusively made of green cells; the progeny of others is partly composed of bleached cells. 2. If one assumes that greening results from the activity of functional units endowed with genetic continuity (Plastidial Segregating Units = PSU), segregation of these units seems to occur according to a model involving random sorting out during the three first divisions. During the following divisions, functional units seem to multiply faster than those impaired by irradiation. 3. The greening rate of colonies issued from irradiated cells seems to be conditioned mostly by the number of functional PSU remaining in the mother cell of the colony.

Animals↗

PubMed1979

Screening of red-green defects of colour vision with pseudoisochromatic tests.

Fifty red-green defectives and 100 normal subjects were examined with the second edition of the Boström-Kugelberg (BK II, 1972) series and the Ishihara complete edition (1976) of pseudoisochromatic plates. The results are related to those obtained with the first edition of the Boström-Kugelberg test (BK I, 1944) and the Boström test (II B, 1950) and to the classification of defects obtained with the Nagel anomaloscope. The 50 red-green defectives were originally selected by using a combination of the BK I and II B tests. The normal subjects also, passed this preliminary test, as well as an examination with the Nagel anomaloscope. In the final examinations performed under standardized conditions, three red-green defectives passed both the BK II and the BK I test, while eight defectives passed the Ishihara test. Combination of BK II or BK I test with the Ishihara test does not improve the result. Only one defective (a borderline case of protanomaly) passed the separate II B test. Normal subjects were not classified as colour defectives with any of the four pseudoisochromatic tests used. All normal subjects passed both the BK II and the Ishihara test. Classified as suspected red-green defectives (one misreading made in standardized conditions) were five normal subjects with the II B test and one normal subject with the BK I test. In the second edition of Boström-Kugelberg series, the plates numbered 3, 5, 11, 16 and 18 are clearly less effective than respective plates of the first edition. Only the plates numbered 1 and 10 have markedly improved in the second edition. Red-green defectives made on average 0.54 misreadings per plate in the BK II test as compared with respective 0.62 in BK I, 0.69 in the Ishihara and 0.56 in the II B test.

Color Perception Tests↗

PubMed1977

The red and green cone visual pigments of deuternomalous trichromacy.

1. Three "simple" deuteranomalous trichromats match with abnormally low "red" tristimulus values throughout the spectrum and abnormally high "green" tristimulus values in the long wave end of the spectrum which become normal (and then low) in the yellow-green. The spectrum locus of this transition differs from one anomalous to the other. Differences in the matches of two of these cannot be due to differences in eye media transmissivities alone. Therefore these two deuteranomalous have different cone visual pigments. 2. The analytical anomaloscope was used in the confrontation of one deuteranomalous with six deuteranopes in turn. In each confrontation the deuteranope set the anomaloscope in his mode and adjusted the intensity of the monochromatic light for a match. Deuteranomalous matches were rejected by four of these six deuteranopes. 3. They were accepted by two of the six. These two rejected each other's matches in a way not attributable to differences in eye media transmissivity. 4. Three different psychophysical techniques were used to measure the action spectra of the long wave cones of these two deuternopes. All three methods reveal small but systematic differences in lambdamax and shape of the curve for the one deuteranope compared with that of the other. 5. In red-green spectral range, these spectra are accurately described by different linear combinations of the color matching functions of the same deuteranomalous whose matches the two deuteranopes accept. Linear combinations of those of a second deuteranomalous, with at least one different kind of cone, fit less well. 6. The wave length discrimination curve of the former deuteranomalous was measured with a new method. The curves of two normals were also obtained for comparison. Wave-length discrimination predictions from the Stiles (1946) line element theory were compared to the anomalous curve. The deuteranopic action spectra were used in the line element to compute this deuteranomalous' discrimination. There is reasonable first order correspondence between prediction and observation, but the prediction is sensitive to small changes in the derivatives of the logarithms of the action spectra. 7. Line element prediction of the deuteranomalous step-by-step luminous efficiency curve is insensitive to such uncertainties. The agreement with expectation from the above assumptions and the measured step-by-step deuteranomalous luminous efficiency curve in the red-green part of the spectrum is therefore good. 8. It is concluded that the erythrolabe in one deuternope's long wave cones has the action spectrum of this deuteranomalous' long and the erythrolabe in the other deuternope's long wave sensitive cones has that of this deuternomalous' medium wave cones. This leads to a general hypothesis about the nature of all forms of red-green colour vision defects transmitted recessively on the X chromosome.

Color Vision Defects↗

PubMed1978

Janus green resistance in Saccharomyces cerevisiae: interaction of nuclear and cytoplasmic factors.

Janus green B was found to be a specific inhibitor of mitochondrial function in yeast. This is consistent with the Janus green specificity in supravital staining of mitochondria. A mutant of S. Cerevisiae resistant to Janus green B was isolated. It shows cross resistance to oligomycin, ethidium bromide and a weak resistance to chlormaphenicol. The mutant was found to be sensitive to cycloheximide and erythromycin. Genetic analysis of this mutant showed that mitochondrial genes are not involved in the determination of Janus Green resistance. Tetrad analysis suggested that two more more nuclear genes are concerned, but many unusal genetic features suggestive of the involvement of a cytoplasmic element remain to be explained.

Azo Compounds↗

PubMed1976

Observations on brilliant green agar with H2S indicator.

Several formulations of brilliant green agar with an added H2S indicator were evaluated. Results were optimum with variations of a basic formula consisting of 40 g of tryptic soy agar (Difco), 8 g of lactose, 8 g of sucrose, 80 mg of phenol red, 1 g of sulfanilamide, 1.5 g of ferric ammonium citrate, 5 g of sodium thiosulfate pentahydrate, and 7 mg of brilliant green dye per liter. Brilliant green dye was added after sterilization of the other components This formulation supported good growth of all of 39 strains of Salmonella tested. Normal biochemical types formed pink colonies with black centers, and an H2S-negative S. choleraesuis formed pink colonies without black centers. Of other bacteria tested, only Enterobacter, Klebsiella, and a few Citrobacter strains showed significant growth in 24 h. When lactose was omitted from the formulation, a lactose-fermenting strain formed pink colonies with black centers, and differentiation of Salmonella from the Enterobacter-Klebsiella groups was equally good. Addition of xylose (4.0 g) and L-lysine hydrochloride (5.4 g) to the above formulation improved differentiation between Salmonella and the few Citrobacter strains that grew and produced more intense blackening in Salmonella colonies. Addition of an H2S indicator to brilliant green agar formulations aided in identification of Salmonella colonies, especially in mixtures with other bacteria. These media were judged to give better differentiation of salmonellae from other bacteria than Hektoen agar with added novobiocin (10 mg/liter).

Agar↗

PubMed1978

Malachite green: applications in electron microscopy.

Incorporation of malachite green into a glutaraldehyde fixative results in enhanced staining of a number of cellular elements. Ribosomes and myofilaments exhibit increased electron density, but cell membranes generally are not stained. In certain tissues, lipid inclusions are uniformly and heavily stained. Other populations of lipid droplets exhibit differential affinity for malachite green, facilitating their division into subclasses. In addition to its function as a dye, malachite green has previously been reported to stabilize lipid elements soluble in aqueous glutaraldehyde. Such a component was observed in the stroma of uterine endometrium. The variety of cell components which exhibit increased contrast after preparation with malachite green suggests that this technique may find widespread application in fine structure studies.

Adrenal Cortex↗

PubMedDate not supplied

[Several characteristics of new strains of diploid green marmoset fibroblasts].

Two strains of diploid fibroblasts were derived from green monkey fetal lungs and their main properties were studied. These strains were shown not to differ principally from the well studied human diploid fibroblasts. The derived cultures are free from contaminating viruses. The titers of attenuated poliomyelitis virus strains achieved in green monkey diploid cells are comparable to those in primary cultures of green monkey kidneys. The possiblity of using green monkey diploid fibroblasts in virological practice is discussed.

Animals↗

PubMed1978

Detection of red and green flashes: evidence for cancellation and facilitation.

Green and red flashes of light will differentially stimulate the middle- and long-wavelength sensitive cones. Interaction of cone signals was studied by measuring increment thresholds for combinations of green and red flashes on a yellow adapting field. When the yellow adapting field was at 10.000 trolands (td), green and red incremental flashes (1 degree, 200-msec duration) produced cancellation when presented simultaneously and facilitation when presented sequentially. A green incremental flash (1.15 degrees, 200 msec, 5000-td adaptation field) and red decremental flash, or vice versa, produced facilitation when presented simultaneously. The results can be explained by color-differencing, opponent-mechanisms. The cancellation effect for the simultaneous incremental flashes largely disappeared when the flashes were exposed briefly (10 msec) or reduced in size (0.04 degrees). It is unlikely that the stimuli were exclusively detected by achromatic, luminance channels, as suggested by previous work, since observers could partially distinguish the hue of threshold flashes of 570- and 590-nm light (0.04 degrees, 10 msec) on a bright yellow field.

Color Perception↗

PubMed2026

An AI-assisted Clinical Decision Support System for Green Classification of Cystocele on Dynamic Transperineal Ultrasound.

Green classification of cystocele on dynamic transperineal ultrasound (TPUS) remains operator-dependent because it requires manual frame selection and landmark-based assessment of the Valsalva maneuver. We developed a workflow-oriented AI-assisted clinical decision support system for automated urethrovesical junction localization and dynamic Green classification and prospectively evaluated its standalone and reader-support performance. This diagnostic accuracy and reader study included 881 patients from a tertiary referral hospital, comprising a retrospective development cohort (n = 688) and an independent prospective test cohort (n = 193). A nested subset of 67 prospective patients was used for a reader study involving two junior and two intermediate radiologists under unaided and AI-assisted conditions. In the complete prospective test cohort, Green-AttGRU achieved a macro-averaged AUC of 0.939 (95% CI, 0.897-0.971) and an overall accuracy of 0.902 (95% CI, 0.860-0.943). In the reader study, overall accuracy increased from 0.761 to 0.821 without AI to 0.851-0.881 with AI, while macro-F1 increased from 0.660 to 0.777 to 0.820-0.860. Overall inter-reader agreement increased from a Fleiss' κ of 0.453 to 0.786, and pooled median interpretation time decreased from 26.7 s to 9.9 s. These findings support the preliminary feasibility of the system as a workflow-oriented decision-support tool for dynamic TPUS interpretation.

Humans↗

PubMed1976

The use of methyl green-pyronin staining after glutaraldehyde fixation and paraffin or araldite embedding.

Methyl green-pyronin staining has been used for localization of RNA and DNA in chick and mouse embryonic tissues and in insect larval salivary glands. Glutaraldehyde or tricholoracetic acid-lanthanum acetate (TCA-LA) was used as fixative and paraffin wax or Araldite was used as embedding medium. For good results the following are specially desirable: fixation with 2.5% glutaraldehyde, dehydration in alcohols for short time, and the use of fresh staining solutions. After TCA-LA fixation the final results are much less specific. The digestion with RNAse appears essential for the detection of RNA because pyronin does not seem to be entirely specific to RNA. The results show that glutaraldehyde a common fixative for electron microscopic work, is particularly suitable fixative for light microscopic cytochemical investigations if followed by methyl green-pyronin staining; furthermore, methyl green-pyronin staining after glutaraldehyde fixation can be carried out on Araldite sections.

Animals↗

PubMed2026

Environmental coupling between metal resistance genes and bacterial communities in Beijing urban green-space soils.

Urban green spaces are intensively managed ecosystems exposed to chronic, multisource, low-intensity anthropogenic inputs. These inputs may alter soil microbial communities and influence the distribution of metal resistance genes (MRGs). However, MRG distributions and their relationships with environmental conditions and bacterial communities remain unclear under the complex, non-extreme pollution conditions typical of these ecosystems. We investigated Beijing urban green spaces as a representative system using metagenomic sequencing and metagenome-assembled genome (MAG) analysis. We characterized soil MRG composition, its environmental associations, and the distribution of potential hosts. MRG composition differed significantly among ecological conservation (EC), transitional urban (TU), and central urban (CU) zones. These differences were closely associated with soil physicochemical properties and bacterial community structure. Available phosphorus (AP) was significantly associated with variation in both bacterial community structure and MRG composition. MAG-based analysis identified distinct potential-host compositions across the three functional zones. Proteobacteria were more frequently represented among dereplicated MAGs from EC soils, whereas Actinobacteria were more frequent in TU and CU soils. Heavy metal concentrations correlated with MRG composition. However, variation partitioning analysis did not identify an independent contribution from heavy metals after accounting for soil physicochemical properties and bacterial community structure. These findings indicate that urban green-space soil monitoring should incorporate environmental conditions and microbial community characteristics rather than rely solely on total metal concentrations.

Soil Microbiology↗

PubMed2026

Genome analysis of the glycosphingolipid-producing green alga tetraselmis sp. NKG400013.

Microalgae are gaining attention as sustainable resources for the production of valuable compounds, including biofuels, pigments, and bioactive metabolites. To support metabolic engineering and genome editing approaches aimed at enhancing these traits, high-quality genome assemblies are essential; however, genomic information remains limited for many microalgal lineages. Tetraselmis sp. NKG400013 is a green alga known for high glycosphingolipid accumulation with distinctive structural features. Here, we report a draft genome assembly of this strain generated using PacBio HiFi sequencing and transcriptome-supported annotation. The assembled genome spans 423.7 Mbp, with 74.5% repetitive sequences and 15,322 predicted protein-coding genes. Comparative analyses across 11 green algal species revealed a positive correlation between genome sizes and repeat contents, indicating that transposable element expansion, particularly long terminal repeat retrotransposons, has substantially contributed to genome enlargement in Tetraselmis. Genome-wide functional annotation and ortholog inference identified core enzymes required for glycosylceramide biosynthesis. Both sphingolipid Δ4 and Δ8 desaturases were identified in Tetraselmis and their coexistence suggests an expanded capacity for long-chain base modification that may underlie its distinctive glycosphingolipid profile. These results establish a genomic framework for understanding the high glycosphingolipid-producing capacity of NKG400013 and provide insights into the evolutionary diversification of sphingolipid metabolism in green algae.

Chlorophyta↗

PubMed2025

Genome editing in the green alga Chlamydomonas: past, present practice and future prospects.

The green alga Chlamydomonas is an important and versatile model organism for research topics ranging from photosynthesis and metabolism, cilia, and basal bodies to cellular communication and the cellular cycle and is of significant interest for green bioengineering processes. The genome in this unicellular green alga is contained in 17 haploid chromosomes and codes for 16 883 protein coding genes. Functional genomics, as well as biotechnological applications, rely on the ability to remove, add, and change these genes in a controlled and efficient manner. In this review, the history of gene editing in Chlamydomonas is put in the context of the wider developments in genetics to demonstrate how many of the key developments to engineer these algae follow the global trends and the availability of technology. Building on this background, an overview of the state of the art in Chlamydomonas engineering is given, focusing primarily on the practical aspects while giving examples of recent applications. Commonly encountered Chlamydomonas-specific challenges, recent developments, and community resources are presented, and finally, a comprehensive discussion on the emergence and evolution of CRISPR/Cas-based precision gene editing is given. An outline of possible future paths for gene editing based on current global trends in genetic engineering and tools for gene editing is presented.

Gene Editing↗

PubMed1979

Electron microscopy of malachite green--glutaraldehyde fixed bacteria.

Malachite green combined with glutaraldehyde has been used recently as a fixative for preserving and revaling lipid complexes in thin sections of eukaryotic cells examined by electron microscopy. When bacteria were prefixed with the above mixture granular electron dense inclusions were revealed in all cultures tested. These inclusions were replaced by electron transparent areas in cells fixed with glutaraldehyde alone. The structures were frequently located near to or within the nucleoid and adjacent to the cell membrane in Gram-negative bacteria and were associated with the nucleoid and mesosomes in Gram-positive species. Polyhydroxybutyrate granules, generally poorly preserved in thin sections of Aquaspirillum serpens, were well preserved by the malachite green-glutaraldehyde fixative. Malachite green complexes were observed outside of the cells in all preparations. Capsules were neither preserved nor stained.

Aldehydes↗