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Raman spectral analysis of microbial pigment compositions in vegetative cells and heterocysts of multicellular cyanobacterium.

ABSTRACT: The one-dimensional multicellular cyanobacterium, Anabaena sp. PCC 7120, exhibits a simple topology consisting of two types of cells under the nitrogen-depleted conditions. Although the differentiated (heterocyst) and undifferentiated cells (vegetative cells) were distinguished by their cellular shapes, we found that their internal states, that is, microbial pigment compositions, were distinguished by using a Raman microscope. Almost of Raman bands of the cellular components were assigned to vibrations of the pigments; chlorophyll a, β-carotene, phycocyanin, and allophycocyanin. We found that the Raman spectral measurement can detect the decomposition of both phycocyanin and allophycocyanin, which are components of the light-harvesting phycobilisome complex in the photosystem II. We observed that the Raman bands of phycocyanin and allophycocyanin exhibited more remarkable decrease in the heterocysts when compared to those of chlorophyll a and β-carotene. This result indicated the prior decomposition of phycobilisome in the heterocysts. We show that the Raman measurement is useful to detect the change of pigment composition in the cell differentiation.

SUBMITTER: Ishihara JI

PROVIDER: S-EPMC10133670 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature

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Raman spectral analysis of microbial pigment compositions in vegetative cells and heterocysts of multicellular cyanobacterium.

Ishihara Jun-Ichi JI Takahashi Hiroki H

Biochemistry and biophysics reports 20230414

The one-dimensional multicellular cyanobacterium, <i>Anabaena</i> sp. PCC 7120, exhibits a simple topology consisting of two types of cells under the nitrogen-depleted conditions. Although the differentiated (heterocyst) and undifferentiated cells (vegetative cells) were distinguished by their cellular shapes, we found that their internal states, that is, microbial pigment compositions, were distinguished by using a Raman microscope. Almost of Raman bands of the cellular components were assigned ...[more]

PMID: 37125074

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Project description:IntroductionEnvironmental microparticle is becoming a global pollutant and the entire population is increasingly exposed to the microparticles from artificial materials. The accumulation of microparticles including microplastics and its subsequent effects need to be investigated timely to keep sustainable development of human society.ObjectivesThis study aimed to explore the accumulation of environmental particles in thrombus, the pathological structure in the blood circulation system.MethodsPatients receiving cardiovascular surgical operations were screened and twenty-six thrombi were collected, digested and filtered. Non-soluble microparticles were enriched on the filter membrane and then were analyzed and identified with Raman Spectrometer. The associations of particle status (presence or absence) or particle number in the thrombus and clinical indicators were examined. One strict quality control-particle detection system was designed to eliminate environmental contaminations.ResultsAmong twenty-six thrombi, sixteen contained eighty-seven identified particles ranging from 2.1 to 26.0 μm in size. The number of microparticles in each thrombus ranged from one to fifteen with the median reaching five. All the particles found in thrombi were irregularly block-shaped. Totally, twenty-one phthalocyanine particles, one Hostasol-Green particle, and one low-density polyethylene microplastic, which were from synthetic materials, were identified in thrombi. The rest microparticles included iron compounds and metallic oxides. After the adjustment for potential confounders, a significantly positive association between microparticle number and blood platelet levels was detected (P < 0.01).ConclusionThis study provides the first photograph and Raman spectrum evidence of microparticles in thrombi. A large number of non-soluble particles including synthetic material microparticles could accumulate in arteries, suggesting that the risk of microparticle exposure was under-estimated and the re-evaluation of its health effects is urgently needed. There will be a series of reports on assessing the health effects of microparticle exposure in humans in the future and this research provided clues for the subsequent research.

Project description:BackgroundColonial and multicellular volvocine green algae have been extensively studied recently in various fields of the biological sciences. However, only one species (Pandorina morum) has been cryopreserved in public culture collections.ResultsHere, we investigated conditions for cryopreservation of the multicellular volvocine alga Gonium pectorale using vegetative colonies or cells and zygotes. Rates of vegetative cell survival in a G. pectorale strain after two-step cooling and freezing in liquid nitrogen were compared between different concentrations (3% and 6%) of the cryoprotectant N,N-dimethylformamide (DMF) and two types of tubes (0.2-mL polymerase chain reaction tubes and 2-mL cryotubes) used for cryopreservation. Among the four conditions investigated, the highest rate of survival [2.7 ± 3.6% (0.54-10%) by the most probable number (MPN) method] was obtained when 2.0-mL cryotubes containing 1.0 mL of culture samples with 6% DMF were subjected to cryogenic treatment. Using these optimized cryopreservation conditions, survival rates after freezing in liquid nitrogen were examined for twelve other strains of G. pectorale and twelve strains of five other Gonium species. We obtained ≥ 0.1% MPN survival in nine of the twelve G. pectorale strains tested. However, < 0.1% MPN survival was detected in eleven of twelve strains of five other Gonium species. In total, ten cryopreserved strains of G. pectorale were newly established in the Microbial Culture Collection at the National Institute for Environmental Studies. Although the cryopreservation of zygotes of volvocine algae has not been previously reported, high rates (approximately 60%) of G. pectorale zygote germination were observed after thawing zygotes that had been cryopreserved with 5% or 10% methanol as the cryoprotectant during two-step cooling and freezing in liquid nitrogen.ConclusionsThe present study demonstrated that cryopreservation of G. pectorale is possible with 6% DMF as a cryoprotectant and 1.0-mL culture samples in 2.0-mL cryotubes subjected to two-step cooling in a programmable freezer.

Project description:BackgroundAn ancient cyanobacterial incorporation into a eukaryotic organism led to the evolution of plastids (chloroplasts) and subsequently to the origin of the plant kingdom. The underlying mechanism and the identities of the partners in this monophyletic event remain elusive.Methodology/principal findingsTo shed light on this evolutionary process, we sequenced the genome of a cyanobacterium residing extracellularly in an endosymbiosis with a plant, the water-fern Azolla filiculoides Lam. This symbiosis was selected as it has characters which make it unique among extant cyanobacterial plant symbioses: the cyanobacterium lacks autonomous growth and is vertically transmitted between plant generations. Our results reveal features of evolutionary significance. The genome is in an eroding state, evidenced by a large proportion of pseudogenes (31.2%) and a high frequency of transposable elements (approximately 600) scattered throughout the genome. Pseudogenization is found in genes such as the replication initiator dnaA and DNA repair genes, considered essential to free-living cyanobacteria. For some functional categories of genes pseudogenes are more prevalent than functional genes. Loss of function is apparent even within the 'core' gene categories of bacteria, such as genes involved in glycolysis and nutrient uptake. In contrast, serving as a critical source of nitrogen for the host, genes related to metabolic processes such as cell differentiation and nitrogen-fixation are well preserved.Conclusions/significanceThis is the first finding of genome degradation in a plant symbiont and phenotypically complex cyanobacterium and one of only a few extracellular endosymbionts described showing signs of reductive genome evolution. Our findings suggest an ongoing selective streamlining of this cyanobacterial genome which has resulted in an organism devoted to nitrogen fixation and devoid of autonomous growth. The cyanobacterial symbiont of Azolla can thus be considered at the initial phase of a transition from free-living organism to a nitrogen-fixing plant entity, a transition process which may mimic what drove the evolution of chloroplasts from a cyanobacterial ancestor.

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Raman spectral analysis of microbial pigment compositions in vegetative cells and heterocysts of multicellular cyanobacterium.

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Raman spectral analysis of microbial pigment compositions in vegetative cells and heterocysts of multicellular cyanobacterium.

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