Crassulacean Research Articles

Genome-wide mapping of DNase I hypersensitive sites in pineapple leaves.

Pineapple [Ananas comosus (L.) Merr.] is the most economically important crop possessing crassulacean acid metabolism (CAM) photosynthesis which has a higher water use efficiency by control of nocturnal opening and diurnal closure of stomata. To provide novel insights into the diel regulatory landscape in pineapple leaves, we performed genome-wide mapping of DNase I hypersensitive sites (DHSs) in pineapple leaves at day (2a.m.) and night (10a.m.) using a simplified DNase-seq method. As a result, totally 33340 and 28753 DHSs were found in green-tip tissue, and 29597 and 40068 were identified in white-base tissue at 2a.m. and 10a.m., respectively. We observed that majority of the pineapple genes occupied less than two DHSs with length shorter than 1kb, and the promotor DHSs showed a proximal trend to the transcription start site (>77% promotor DHSs within 1kb). In addition, more intergenic DHSs were identified around transcription factors or transcription co-regulators (TFs/TCs) than other functional genes, indicating complex regulatory contexts around TFs/TCs. Through combined analysis of tissue preferential DHSs and genes, we respectively found 839 and 888 coordinately changed genes in green-tip at 2a.m. and 10a.m. (AcG2 and AcG10). Furthermore, AcG2-specific, AcG10-specific and common accessible DHSs were dissected from the total photosynthetic preferential DHSs, and the regulatory networks indicated dynamic regulations with multiple cis-regulatory elements occurred to genes preferentially expressed in photosynthetic tissues. Interestingly, binding motifs of several cycling TFs were identified in the DHSs of key CAM genes, revealing a circadian regulation to CAM coordinately diurnal expression. Our results provide a chromatin regulatory landscape in pineapple leaves during the day and night. This will provide important information to assist with deciphering the circadian regulation of CAM photosynthesis.

Dauricine alleviates cognitive impairment in Alzheimer's disease mice induced by D-galactose and AlCl3 via the Ca2+/CaM pathway

Alzheimer's disease (AD) is a common neurodegenerative disease in the elderly. Dysregulation of intracellular Ca2+ homeostasis plays a critical role in the pathological development of AD. Dauricine (DAU) is a bisbenzylisoquinoline alkaloid isolated from Menispermum dauricum DC., which can prevent the influx of extracellular Ca2+ and inhibit the release of Ca2+ from the endoplasmic reticulum. DAU has a potential for anti-AD. However, it is unclear whether DAU can exert its anti-AD effect in vivo by regulating the Ca2+ related signaling pathways. Here, we investigated the effect and mechanism of DAU on D-galactose and AlCl3 combined-induced AD mice based on the Ca2+/CaM pathway. The results showed that DAU (1 mg/kg and 10 mg/kg for 30 days) treatment attenuated learning and memory deficits and improved the nesting ability of AD mice. The HE staining assay showed that DAU could inhibit the histopathological alterations and attenuate neuronal damage in the hippocampus and cortex of AD mice. Studies on the mechanism indicated that DAU decreased the phosphorylation of CaMKII and Tau and reduced the formation of NFTs in the hippocampus and cortex. DAU treatment also reduced the abnormally high expression of APP, BACE1, and Aβ1–42, which inhibited the deposition of Aβ plaques. Moreover, DAU could decrease Ca2+ levels and inhibit elevated CaM protein expression in the hippocampus and cortex of AD mice. The molecular docking results showed that DAU may have a high affinity with CaM or BACE1. DAU has a beneficial impact on pathological changes in AD mice induced by D-galactose and AlCl3 and may act by negative regulation of the Ca2+/CaM pathway and its downstream molecules such as CaMKII and BACE1.

The first released available genome of the common ice plant (Mesembryanthemum crystallinum L.) extended the research region on salt tolerance, C3-CAM photosynthetic conversion, and halophism

Background: The common ice plant (Mesembryanthemum crystallinum L.) is an annual herb belonging to the genus Mesembryanthemum of the family Aizoaceae, native to Southern Africa. Methods: We performed shotgun genome paired-end sequencing using the Illumina platform to determine the genome sequence of the ice plants. We assembled the whole genome sequences using the genome assembler “ALGA” and “Redundans”, then released them as available genomic information. Finally, we mainly estimated the potential genomic function by the homology search method. Results: A draft genome was generated with a total length of 286 Mb corresponding to 79.2% of the estimated genome size (361 Mb), consisting of 49,782 contigs. It encompassed 93.49% of the genes of terrestrial higher plants, 99.5% of the ice plant transcriptome, and 100% of known DNA sequences. In addition, 110.9 Mb (38.8%) of repetitive sequences and untranslated regions, 971 tRNA, and 100 miRNA loci were identified, and their effects on stress tolerance and photosynthesis were investigated. Molecular phylogenetic analysis based on ribosomal DNA among 26 kinds of plant species revealed genetic similarity between the ice plant and poplar, which have salt tolerance. Overall, 35,702 protein-coding regions were identified in the genome, of which 56.05% to 82.59% were annotated and submitted to domain searches and gene ontology (GO) analyses, which found that eighteen GO terms stood out among five plant species. These terms were related to biological defense, growth, reproduction, transcription, post-transcription, and intermembrane transportation, regarded as one of the fundamental results of using the utilized ice plant genome. Conclusions: The information that we characterized is useful for elucidation of the mechanism of growth promotion under salinity and reversible conversion of the photosynthetic type from C3 to Crassulacean Acid Metabolism (CAM).

Comparison of Growth and Leaf Color Quality of <italic>Mesembryanthemum cordifolium</italic> f. <italic>variegata</italic> as Affected by Shading Levels

Background and objective: Mesembryanthemum cordifolium is a plant belonging to the Aizoaceae family and is native to South Africa and Namibia. As a CAM plant, M. cordifolium exhibits strong resistance to drought stress and can be utilized for the removal of soil salinity and heavy metals. Furthermore, it is an important medicinal crop with anti-inflammatory and antidepressant effects. However, despite its potential benefits, research on the optimal growth environment for M. cordifolium remains scarce.Methods: Therefore, this study selected M. cordifolium f. variegata with high ornamental value as the experimental plant and compared the effects of different shading levels on the growth and leaf color quality of variegated baby sun rose (M. cordifolium f. variegata). Six shading levels (0, 35, 45, 60, 75, and 99%, respectively) were designed using polyethylene (PE) shading films.Results: The results showed that the shoot height, shoot width, dry weight, and chlorophyll content (SPAD units) were highest under the 60% shading level, while leaf length, leaf width, and fresh weight were highest under the 75% shading level. On the other hand, growth was relatively lower under the 0% shading level, which suggests that M. cordifolium f. variegata prefers shade over direct sunlight. On the contrary, M. cordifolium f. variegata showed growth even under the 99% shading level, proving to have strong shade tolerance, but the leaf color quality was lowerst.Conclusion: In conclusion, to significantly increase the size and biomass of plants and improve leaf color quality when cultivating M. cordifolium f. variegata under shading treatment, it is recommended to cultivate it under 60-75% shading levels.

A comprehensive analysis of transcriptomic data for comparison of plants with different photosynthetic pathways in response to drought stress.

The main factor leading to a decrease in crop productivity is abiotic stresses, particularly drought. Plants with C4 and CAM photosynthesis are better adapted to drought-prone areas than C3 plants. Therefore, it is beneficial to compare the stress response of plants with different photosynthetic pathways. Since most crops are C3 and C4 plants, this study focused on conducting an RNA-seq meta-analysis to investigate and compare how C3 and C4 plants respond to drought stress at the gene expression level in their leaves. Additionally, the accuracy of the meta-analysis results was confirmed with RT-qPCR. Based on the functional enrichment and network analysis, hub genes related to ribosomal proteins and photosynthesis were found to play a potential role in stress response. Moreover, our findings suggest that the low abundant amino acid degradation pathway, possibly through providing ATP source for the TCA cycle, in both groups of plants and the activation of the OPPP pathway in C4 plants, through providing the electron source required by this plant, can help to improve drought stress tolerance.

Metabolic Adjustment of High Intertidal Alga Pelvetia canaliculata to the Tidal Cycle Includes Oscillations of Soluble Carbohydrates, Phlorotannins, and Citric Acid Content.

The brown alga Pelvetia canaliculata is one of the species successfully adapted to intertidal conditions. Inhabiting the high intertidal zone, Pelvetia spends most of its life exposed to air, where it is subjected to desiccation, light, and temperature stresses. However, the physiological and biochemical mechanisms allowing this alga to tolerate such extreme conditions are still largely unknown. The objective of our study is to compare the biochemical composition of Pelvetia during the different phases of the tidal cycle. To our knowledge, this study is the first attempt to draft a detailed biochemical network underneath the complex physiological processes, conferring the successful survival of this organism in the harsh conditions of the high intertidal zone of the polar seas. We considered the tide-induced changes in relative water content, stress markers, titratable acidity, pigment, and phlorotannin content, as well as the low molecular weight metabolite profiles (GC-MS-based approach) in Pelvetia thalli. Thallus desiccation was not accompanied by considerable increase in reactive oxygen species content. Metabolic adjustment of P. canaliculata to emersion included accumulation of soluble carbohydrates, various phenolic compounds, including intracellular phlorotannins, and fatty acids. Changes in titratable acidity accompanied by the oscillations of citric acid content imply that some processes related to the crassulacean acid metabolism (CAM) may be involved in Pelvetia adaptation to the tidal cycle.

Paleodiets of hunter‐gatherers from the central Patagonian coast: Reviewing scopes and limitations of stable isotope analyses

AbstractThis work reviews paleodietary reconstructions of hunter‐gatherers from the northeast province of Chubut, Argentina. The isotopic record of human bone can complement information from zooarchaeological and technological studies with its particular resolution on the long‐term diet of individuals. Previous δ13C and δ15N studies found dietary differences between the coastal area and the lower basin of the Chubut River, both in the proportion of marine resources in the diet and the trophic positions of prey. However, it is challenging to test hypotheses concerning human diets in northeastern Patagonia, where C3 and CAM plants grow and terrestrial, riverine, and marine prey are available. Here, we focus on the local isotopic ecology to assess the scopes and limitations of δ13C and δ15N stable isotopes analyses. We provide new data on native faunas spanning different trophic levels of terrestrial, riverine, and marine environments. After establishing the basic patterns in the local isotope record, we reanalyze the available δ13Ccol, δ15N, and δ13Capa measurements from 23 individuals studied 15 years ago. That analysis reveals a relatively continuous distribution of several overlapping resources on the coast of northeastern Patagonia. Bayesian mixing models identified a combination of terrestrial and riverine resources aggregated as the main component of long‐term diets. However, it generated diffuse estimations. The Bayesian ellipses, describing isotopic niches, emphasized more the spatial variations in diet between coastal and valley groups.

Acclimation to a combination of water deficit and nutrient deprivation through simultaneous increases in abscisic acid and bioactive jasmonates in the succulent plant Sempervivum tectorum L

Activation of hormonal responses defines the drought acclimation ability of plants and may condition their survival. However, aside ABA, little is known about the possible contribution of other phytohormones, such as jasmonates and salicylates, in the response of CAM plants to water deficit. Here, we aimed to study the physiological mechanisms underlying the stress tolerance of house leek (Sempervivum tectorum L.), a CAM plant adapted to survive harsh environments, to a combination of water deficit and nutrient deprivation. We exposed plants to the combination of these two abiotic stresses by withholding nutrient solution for 10 weeks and monitored their physiological response every two weeks by measuring various stress makers together with the accumulation of stress-related phytohormones and photoprotective molecules, such as tocopherols (vitamin E). Results showed that ABA content increased by 4.2-fold after four weeks of water deficit to keep later constant up to 10 weeks of stress, variations that occurred concomitantly with reductions in the relative leaf water content, which decreased by up to 20% only. The bioactive jasmonate, jasmonoyl-isoleucine was the other stress-related phytohormone that simultaneously increased under stress together with ABA. While contents of salicylic acid and the jasmonoyl-isoleucine precursors, 12-oxo-phytodienoic acid and jasmonic acid decreased with water deficit, those of jasmonoyl-isoleucine increased 3.6-fold at four weeks of stress. The contents of ABA and jasmonoyl-isoleucine correlated positively between them and with the content of α-tocopherol per unit of chlorophyll, thus suggesting a photoprotective activation role. It is concluded that S. tectorum not only withstands a combination of water deficit and nutrient deprivation for 10 weeks without any symptom of damage but also activates effective defense strategies through the simultaneous accumulation of ABA and the bioactive jasmonate form, jasmonoyl-isoleucine.

Developmental dynamics of crassulacean acid metabolism (CAM) in Opuntia ficus-indica.

The relative contributions of C3 photosynthesis and crassulacean acid metabolism (CAM) during the earliest stages of development were investigated to assess how much each might contribute to cactus pear (Opuntia ficus-indica) productivity. The developmental progression of C3 photosynthesis and CAM was assessed in seedlings and daughter cladodes of mature plants by titratable acidity, δ13C isotopic values and diel gas exchange measurements. Nocturnal acidification was observed in seedling cladodes and cotyledons at the earliest stages of development and became highly significant by 75 days of development. Seedling cotyledons showed mean δ13C values of -21.4 and -17.1 ‰ at 30 and 100 days of age, respectively. Seedling cladodes showed mean δ13C values of -19.4 and -14.5 ‰ at 30 and 100 days of age, respectively. These values are typical of CAM plants. Net CO2 assimilation was negative, then occurred in both the day and the night, with nighttime fixation becoming predominant once the primary cladode reached 5 cm in size. Emergent daughter cladodes growing on mature plants showed nocturnal titratable acidity at the earliest stages of development, which became significant when daughter cladodes were >2.5-5 cm in height. Emergent daughter cladodes showed mean δ13C values of -14.5 to -15.6 ‰, typical of CAM plants. CO2 assimilation studies revealed that net CO2 uptake was negative in daughter cladodes <12 cm in length, but then exhibited net positive CO2 assimilation in both the day and the night, with net nocturnal CO2 assimilation predominating once the daughter cladode grew larger. Developing O. ficus-indica primary and daughter cladodes begin as respiring sink tissues that transition directly to performing CAM once net positive CO2 fixation is observed. Overall, these results demonstrate that CAM is the primary form of photosynthetic carbon assimilation for O. ficus-indica even at the earliest stages of seedling or daughter cladode development.

The Synergistic Mechanism of Photosynthesis and Antioxidant Metabolism between the Green and White Tissues of Ananas comosus var. bracteatus Chimeric Leaves.

Ananas comosus var. bracteatus (Ac. bracteatus) is a typical leaf-chimeric ornamental plant. The chimeric leaves are composed of central green photosynthetic tissue (GT) and marginal albino tissue (AT). The mosaic existence of GT and AT makes the chimeric leaves an ideal material for the study of the synergistic mechanism of photosynthesis and antioxidant metabolism. The daily changes in net photosynthetic rate (NPR) and stomatal conductance (SCT) of the leaves indicated the typical crassulacean acid metabolism (CAM) characteristic of Ac. bracteatus. Both the GT and AT of chimeric leaves fixed CO2 during the night and released CO2 from malic acid for photosynthesis during the daytime. The malic acid content and NADPH-ME activity of the AT during the night was significantly higher than that of GT, which suggests that the AT may work as a CO2 pool to store CO2 during the night and supply CO2 for photosynthesis in the GT during the daytime. Furthermore, the soluble sugar content (SSC) in the AT was significantly lower than that of GT, while the starch content (SC) of the AT was apparently higher than that of GT, indicating that AT was inefficient in photosynthesis but may function as a photosynthate sink to help the GT maintain high photosynthesis activity. Additionally, the AT maintained peroxide balance by enhancing the non-enzymatic antioxidant system and antioxidant enzyme system to avoid antioxidant damage. The enzyme activities of reductive ascorbic acid (AsA) and the glutathione (GSH) cycle (except DHAR) and superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were enhanced, apparently to make the AT grow normally. This study indicates that, although the AT of the chimeric leaves was inefficient at photosynthesis because of the lack of chlorophyll, it can cooperate with the GT by working as a CO2 supplier and photosynthate store to enhance the photosynthetic ability of GT to help chimeric plants grow well. Additionally, the AT can avoid peroxide damage caused by the lack of chlorophyll by enhancing the activity of the antioxidant system. The AT plays an active role in the normal growth of the chimeric leaves.

How does leaf succulence relate with plant drought resistance in woody shrubs?

Succulence describes the amount of water stored in cells or organs, regardless of plant life-form, including woody and herbaceous plants. In dry environments, plants with greater survival often have greater leaf succulence. However, it is unclear how leaf succulence relates with plant drought resistance strategies, including isohydry (closing stomata to maintain leaf water status) and anisohydry (adjusting cell turgor to tolerate low leaf water status), which exist on a continuum that can be quantified by hydroscape area (larger hydroscape area indicates more anisohydric). We evaluated 12 woody species with differing leaf succulence in a glasshouse dry-down experiment to determine relationships among leaf succulence (degree of leaf succulence, leaf succulent quotient, leaf thickness) and plant drought response (hydroscape area, plant water use, turgor loss point, pre-dawn leaf water potential when transpiration ceased). Hydroscape areas ranged from 0.72 MPa2 (Carpobrotus modestus; CAM plants) to 7.01 MPa2 (Rhagodia spinescens; C3 plants), suggesting that C. modestus was more isohydric and R. spinescens was more anisohydric. More isohydric species C. modestus, C. rossii and Disphyma crassifolium (CAM plants) had greater leaf succulence, lower root allocation, used stored water and ceased transpiration at higher pre-dawn leaf water potential, shortly after reaching their turgor loss point. The remaining nine species that are not CAM plants had larger hydroscape areas and ceased transpiration at lower pre-dawn leaf water potential. Greater leaf succulence was not related with cumulative water loss until transpiration ceased in drying soils. All 12 species had high turgor loss points (-1.32 MPa to -0.59 MPa), but turgor loss point was not related to hydroscape area or leaf succulence. Our data suggests that overall greater leaf succulence was related with isohydry but this may have been influenced by the fact that these species were also CAM plants.

A complex network regulating malate contents during fruit ripening in climacteric fruits.

This article is a Commentary on Wang et al. (2023), 239: 1014–1034.

Radon adsorption and air purification by Spanish moss (Tillandsia usneoides) and its metabolic response to radon exposure

The capacity of Spanish moss (Tillandsia usneoides), an aerial plant, to adsorb radon (Rn) and absorb CO2 was assessed to analyze its capacity to remove pollutants from indoor air and to determine its radon (Rn) tolerance mechanism. Transcriptomics and metabolomics techniques were used to analyze the response of the plant to Rn exposure. Spanish moss absorbed indoor CO2 at night using the type of photosynthesis termed crassulacean acid metabolism. The CO2 absorption efficiency of the plant was mainly affected by the light duration and diurnal temperature differences. The highest purification efficiency was 48.25%, and the scales on the Spanish moss leaf surface were the key sites for Rn adsorption. Metabolome analysis showed that Rn exposure induced differential metabolites significantly enriched in the metabolism of lipids, amino acids, nucleotides, and carbohydrates. Transcriptome analysis showed significantly upregulated expression levels of functional genes in Rn-exposed leaves. Rn had significant effects on respiratory metabolism, as indicated by upregulated expression of metabolites and functional genes related to the glycolysis pathway, pyruvate oxidation, tricarboxylic acid cycle, and oxidative phosphorylation pathway. These responses indicated that the internal mechanism by which Spanish moss alleviates Rn stress involves an enhancement of cellular energy supplies and regulation of respiratory metabolic pathways to allow adaptation to Rn pollution.

The first released available genome of the common ice plant (Mesembryanthemum crystallinum L.) extended the research region on salt tolerance, C3-CAM photosynthetic conversion, and halophism

Background: The common ice plant (Mesembryanthemum crystallinum L.) is an annual herb belonging to the genus Mesembryanthemum of the family Aizoaceae, native to Southern Africa. Methods: We performed shotgun genome paired-end sequencing using the Illumina platform to determine the genome sequence of the ice plants. We assembled the whole genome sequences using the genome assembler “ALGA” and “Redundans”, then released them as available genomic information. Finally, we mainly estimated the potential genomic function by the homology search method. Results: A draft genome was generated with a total length of 286 Mb corresponding to 79.2% of the estimated genome size (361 Mb), consisting of 49,782 contigs. It encompassed 93.49% of the genes of terrestrial higher plants, 99.5% of the ice plant transcriptome, and 100% of known DNA sequences. In addition, 110.9 Mb (38.8%) of repetitive sequences and untranslated regions, 971 tRNA, and 100 miRNA loci were identified, and their effects on stress tolerance and photosynthesis were investigated. Molecular phylogenetic analysis based on ribosomal DNA among 26 kinds of plant species revealed genetic similarity between the ice plant and poplar, which have salt tolerance. Overall, 35,702 protein-coding regions were identified in the genome, of which 56.05% to 82.59% were annotated and submitted to domain searches and gene ontology (GO) analyses, which found that eighteen GO terms stood out among five plant species. These terms were related to biological defense, growth, reproduction, transcription, post-transcription, and intermembrane transportation, regarded as one of the fundamental results of using the utilized ice plant genome. Conclusions: The information that we characterized is useful for elucidation of the mechanism of growth promotion under salinity and reversible conversion of the photosynthetic type from C3 to Crassulacean Acid Metabolism (CAM).

Simultaneous influence of light and CO2 on phytoremediation performance and physiological response of plants to formaldehyde

Phytoremediation technology is an effective method to remove formaldehyde indoors, but the purification capacity and physiological response of plants to formaldehyde under the simultaneous influence of light and CO2 have not been examined in previous studies. In this study, formaldehyde fumigation experiments were conducted on the C3 plants Epipremnum aureum A. and Chlorophytum comosum L., and the crassulacean acid metabolism (CAM) plant Dieffenbachia maculate A. The phytoremediation performance and physiological response of plants were studied. The initial concentration of formaldehyde was established at 11.950 ± 1.442 [Formula: see text]; the light intensities were 448 ± 7 [Formula: see text], 1628 ± 22 [Formula: see text], and 3259 ± 22 [Formula: see text], respectively; and the concentrations of CO2 were 455 ± 29 [Formula: see text], 978 ± 50 [Formula: see text], 2020 ± 66 [Formula: see text], and 3006 ± 95 [Formula: see text], respectively. The results indicated that the highest purification rates of formaldehyde by E. aureum, D. maculata, and C. comosum were 55.8%, 43.7%, and 53.2%, respectively. The light intensity had a positive effect on the formaldehyde purification rates of all three plants and positively stimulated peroxidase (POD) activity, while the CO2 concentration had no significant impact on the formaldehyde purification capacity and plants' physiological characteristics. Exposure to formaldehyde inhibited formaldehyde dehydrogenase (FADH) activity and positively stimulated catalase (CAT) activity. The superoxide dismutase (SOD) activity positively correlated with the formaldehyde purification capacity of plants.

The climate changes promoted the chloroplast genomic evolution of Dendrobium orchids among multiple photosynthetic pathways

Dendrobium orchids have multiple photosynthetic pathways, which can be used as a model system for studying the evolution of crassulacean acid metabolism (CAM). In this study, based on the results of the net photosynthetic rates (Pn), we classified Dendrobium species into three photosynthetic pathways, then employed and compared their chloroplast genomes. The Dendrobium chloroplast genomes have typical quartile structures, ranging from 150,841–153,038 bp. The apparent differences in GC content, sequence variability, and IR junctions of SSC/IRB junctions (JSBs) were measured within chloroplast genomes among different photosynthetic pathways. The phylogenetic analysis has revealed multiple independent CAM origins among the selected Dendrobium species. After counting insertions and deletions (InDels), we found that the occurrence rates and distribution densities among different photosynthetic pathways were inconsistent. Moreover, the evolution patterns of chloroplast genes in Dendrobium among three photosynthetic pathways were also diversified. Considering the diversified genome structure variations and the evolution patterns of protein-coding genes among Dendrobium species, we proposed that the evolution of the chloroplast genomes was disproportional among different photosynthetic pathways. Furthermore, climatic correlation revealed that temperature and precipitation have influenced the distribution among different photosynthetic pathways and promoted the foundation of CAM pathway in Dendrobium orchids. Based on our study, we provided not only new insights into the CAM evolution of Dendrobium but also provided beneficial genetic data resources for the further systematical study of Dendrobium.

Adapting from Low to High: An Update to CO2-Concentrating Mechanisms of Cyanobacteria and Microalgae.

The intracellular accumulation of inorganic carbon (Ci) by microalgae and cyanobacteria under ambient atmospheric CO2 levels was first documented in the 80s of the 20th Century. Hence, a third variety of the CO2-concentrating mechanism (CCM), acting in aquatic photoautotrophs with the C3 photosynthetic pathway, was revealed in addition to the then-known schemes of CCM, functioning in CAM and C4 higher plants. Despite the low affinity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) of microalgae and cyanobacteria for the CO2 substrate and low CO2/O2 specificity, CCM allows them to perform efficient CO2 fixation in the reductive pentose phosphate (RPP) cycle. CCM is based on the coordinated operation of strategically located carbonic anhydrases and CO2/HCO3- uptake systems. This cooperation enables the intracellular accumulation of HCO3-, which is then employed to generate a high concentration of CO2 molecules in the vicinity of Rubisco's active centers compensating up for the shortcomings of enzyme features. CCM functions as an add-on to the RPP cycle while also acting as an important regulatory link in the interaction of dark and light reactions of photosynthesis. This review summarizes recent advances in the study of CCM molecular and cellular organization in microalgae and cyanobacteria, as well as the fundamental principles of its functioning and regulation.

Klaus Winter - the indefatigable CAM experimentalist.

In January 1972, Klaus Winter submitted his first paper on crassulacean acid metabolism (CAM) whilst still an undergraduate student in Darmstadt. During the subsequent half-century, he passed his Staatsexamensarbeit, obtained his Dr. rer. nat. summa cum laude and Dr. rer. nat. habil., won a Heinz Maier-Leibnitz Prize and a Heisenberg Fellowship, and has occupied positions in Germany, Australia, the USA and Panama. Now a doyen in CAM circles, and a Senior Staff Scientist at the Smithsonian Tropical Research Institute (STRI), he has published over 300 articles, of which about 44 % are about CAM. I document Winter's career, attempting to place his CAM-related scientific output and evolution in the context of factors that have influenced him as he and his science progressed from the 1970s to the 2020s.

The Effect of Plant Metabolism on Some Physiological Immune Responses Under Biotic Stress

This work is part of a larger research project studying the immune responses of a variety of plants. Once image-based (unpublished data) and another time, as in this study, biochemical parameters based. Plant models included three metabolically different plants, Vica faba L. (C3), Zea mays L. (C4), Mesembryanthemum L. (CAM). Pathogen models had Alternaria alternate and Pseudomonas syringe. All of which, along with the control treatment, represent nine plant-pathogen systems applied with three replicates of each studied species. To investigate the extent to which the immune response is transferred from infected leaves to uninfected leaves on the same plant, several physiological and biochemical immune responses were examined, including: two plant hormones, abscisic acid and Salicylic acid, and two active enzymes, Catalase enzyme and Peroxidase enzyme, two free radicals, ROS and H2O2. Results show that The studied hormones were significantly affected by the type of plant metabolism and the type of pathogen, sometimes by an increase and sometimes by a decrease in which ABA and SA increased in C3 and CAM plants when infected by Alternaria and varied when infected by pseudomonas syringe. At the same time, both ABA and SA decrease in C4 plants. The studied enzymes did not follow a specific pattern of increasing and decreasing response to a bacterial or fungal infection. Still, they differed significantly between plants of different metabolism according to the type of pathogen causing the infection. Infection of all three studied plants by Alternaria alternate and Pseudomonas syringae did not cause any significant changes at p≤ 0.05 in the free radical production of the leaves for both ROS and H2O2.

Diel dynamics of multi-omics in elkhorn fern provide new insights into weak CAM photosynthesis

Crassulacean acid metabolism (CAM) has high water-use efficiency (WUE) and is widely recognized to have evolved from C3 photosynthesis. Different plant lineages have convergently evolved CAM, but the molecular mechanism that underlies C3-to-CAM evolution remains to be clarified. Platycerium bifurcatum (elkhorn fern) provides an opportunity to study the molecular changes underlying the transition from C3 to CAM photosynthesis because both modes of photosynthesis occur in this species, with sporotrophophyll leaves (SLs) and cover leaves (CLs) performing C3 and weak CAM photosynthesis, respectively. Here, we report that the physiological and biochemical attributes of CAM in weak CAM-performing CLs differed from those in strong CAM species. We investigated the diel dynamics of the metabolome, proteome, and transcriptome in these dimorphic leaves within the same genetic background and under identical environmental conditions. We found that multi-omic diel dynamics in P. bifurcatum exhibit both tissue and diel effects. Our analysis revealed temporal rewiring of biochemistry relevant to the energy-producing pathway (TCA cycle), CAM pathway, and stomatal movement in CLs compared with SLs. We also confirmed that PHOSPHOENOLPYRUVATE CARBOXYLASE KINASE (PPCK) exhibits convergence in gene expression among highly divergent CAM lineages. Gene regulatory network analysis identified candidate transcription factors regulating the CAM pathway and stomatal movement. Taken together, our results provide new insights into weak CAM photosynthesis and new avenues for CAM bioengineering.