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Ulva prolifera, a type of green algae that can be consumed, was utilized in the production of an angiotensin-I converting enzyme (ACE) inhibitory peptide. The protein from the algae was isolated and subsequently hydrolyzed using a neutral protease. The resulting hydrolysate underwent several processes including Sephadex-G100 filtration chromatography, ultrafiltration, HPLC-Q-TOF-MS analysis, ADMET screening, UV spectrum detection test, molecular docking, and molecular dynamic simulation. Then, the ACE inhibitory peptide named KAF (IC50, 0.63 ± 0.26 µM) was identified. The effectiveness of this peptide in inhibiting ACE can be primarily attributed to two conventional hydrogen bonds. Additionally, it could activate endothelial nitric oxide synthase (eNOS) activity to promote the generation of nitric oxide (NO). Additionally, KAF primarily increased the intracellular calcium (Ca2+) level by acting on L-type Ca2+ channel (LTCC) and the ryanodine receptor (RyR) in the endoplasmic reticulum, and completed the activation of eNOS under the mediation of protein kinase B (Akt) signaling pathway. Our study has confirmed that KAF has the potential to be processed into pharmaceutical candidate functions on vasoconstriction.
Assuntos
Inibidores da Enzima Conversora de Angiotensina , Simulação de Acoplamento Molecular , Peptídeos , Ulva , Ulva/química , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Inibidores da Enzima Conversora de Angiotensina/química , Peptídeos/farmacologia , Peptídeos/química , Peptídeos/isolamento & purificação , Óxido Nítrico Sintase Tipo III/metabolismo , Animais , Óxido Nítrico/metabolismo , Vasodilatação/efeitos dos fármacos , Cálcio/metabolismo , Peptidil Dipeptidase A/metabolismo , Vasodilatadores/farmacologia , Vasodilatadores/isolamento & purificação , Vasodilatadores/química , Humanos , Algas ComestíveisRESUMO
The xanthophyll cycle is a photoprotective mechanism in plants and algae, which protects the photosynthetic system from excess light damage under abiotic stress. Zeaxanthin is considered to play a pivotal role in this process. In this study, the relative content of xanthophylls was determined using HPLC-MS/MS in Ulva prolifera exposed to different salinities. The results showed that high-salt stress significantly increased the relative content of xanthophylls and led to the accumulation of zeaxanthin. It was speculated that the accumulated zeaxanthin may contribute to the response of U. prolifera to high-salt stress. Zeaxanthin epoxidase (ZEP) is a key enzyme in the xanthophyll cycle and is also involved in the synthesis of abscisic acid and carotenoids. In order to explore the biological function of ZEP, a ZEP gene was cloned and identified from U. prolifera. The CDS of UpZEP is 1122 bp and encodes 373 amino acids. Phylogenetic analysis showed that UpZEP clusters within a clade of green algae. The results of qRT-PCR showed that high-salt stress induced the expression of UpZEP. In addition, heterologous overexpression of the UpZEP gene in yeast and Chlamydomonas reinhardtii improved the salt tolerance of transgenic organisms. In conclusion, the UpZEP gene may be involved in the response of U. prolifera to high-salt stress and can improve the high-salt tolerance of transgenic organisms.
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BACKGROUND: Pyropia yezoensis a commercially important red seaweed species, is susceptible to various microorganisms infections, among which bacterial infections are the most prominent ones. Pyropia yezoensis is often affected by harmful bacterial communities under high temperatures that can lead to its degradation and economic losses. The current study aimed to explore Pyropia yezoensis-associated microbiota and further identify potential isolates, which can degrade Pyropia yezoensis under high-temperature conditions. METHODS AND RESULTS: The 16S rRNA gene sequencing was used to identify the agarolytic bacterial species. The results showed that Chromohalobacter sp. strain AZ6, Pseudoalteromonas sp. strain AZ, Psychrobacter sp. strain AZ3, Vibrio sp. strain AZ, and Halomonas sp. strain AZ07 exhibited algicidal properties as these strains were more abundant at high temperature (25 °C). Among the five isolated strains, the potential isolate Halomonas sp. strain AZ07 showed high production of agarolytic enzymes, including lipase, protease, cellulase, and amylase. This study confirmed that the isolated strain could produce these four different enzymes. The strain Halomonas AZ07 was co-treated with Pyropia yezoensis cells under two different temperature environments, including 13 °C and 25 °C. The degradation of Pyropia yezoensis occurred at the optimum temperature of 25 °C and effectively degraded their cell wall, proteins, lipids, and carbohydrates. CONCLUSION: The successful cultivation of Pyropia yezoensis in coastal farm environments is dependent on specific temperature and environmental factors, and lower temperatures have been observed to be particularly beneficial for the survival and growth of Pyropia yezoensis. The temperature below 13 °C was confirmed to be the best niche for the symbiotic relationship of microbiota associated with Pyropia yezoensis for its growth, development, and production.
Assuntos
Halomonas , RNA Ribossômico 16S , Halomonas/genética , Halomonas/metabolismo , Halomonas/enzimologia , RNA Ribossômico 16S/genética , Temperatura Alta , Rodófitas/genética , Filogenia , Microbiota/genética , Alga Marinha/metabolismo , Alga Marinha/microbiologia , Temperatura , Algas Comestíveis , PorphyraRESUMO
In the present study, Ulva prolifera, an edible alga, was used to prepare angiotensin-I converting enzyme (ACE) inhibitory peptide. The algae protein was isolated and later hydrolyzed by five commercial enzymes (alcalase, papain, pepsin, trypsin, neutral protease), either individually or in combination. Hydrolysate, with the highest in vitro ACE inhibitory activity, was processed using the Sephadex-G100, ultrafiltration, HPLC-Q-TOF-MS, ADMET screening and molecular docking, respectively. The ACE inhibitory peptide DIGGL with a IC50 value of 10.32 ± 0.96 µM was then identified. The peptide against ACE by a non-competitive mode and mainly attributable to the three Conventional Hydrogen Bonds. It could activate Endothelial nitric oxide synthase activity in NO generation and reduce Endothelin-1 secretion induced by Angiotensin II in Human umbilical vein endothelial cells. Meanwhile, DIGGL could promote mice splenocytes proliferation, which was also effective when co-incubated with Con A or LPS, respectively. Besides, the anti-ACE peptide could remain active during the digestion of gastrointestinal proteases (pepsin-trypsin) in vitro.
Assuntos
Peptidil Dipeptidase A , Ulva , Animais , Humanos , Camundongos , Peptidil Dipeptidase A/metabolismo , Ulva/metabolismo , Inibidores da Enzima Conversora de Angiotensina/química , Hidrolisados de Proteína/química , Óxido Nítrico Sintase Tipo III , Tripsina/metabolismo , Pepsina A/metabolismo , Simulação de Acoplamento Molecular , Endotelina-1 , Angiotensina II , Papaína , Células Endoteliais/metabolismo , Lipopolissacarídeos , Hidrólise , Peptídeos/química , Peptídeo Hidrolases/metabolismo , SubtilisinasRESUMO
The influence of the social environment on healthy investment behavior is a vital research topic. This paper focuses on foreign direct investment (FDI) as an important part of its broad impact in improving the level of capital circulation and diversifying the non-systemic risk of a single country portfolio. Using data from 35 countries on direct investment in China, we find that the impact of the social environment on healthy investment behavior is mainly reflected in investors' resistance to cultural distance and their benefit compensation across institutional distance. In addition, their joint influence is still negative, dominated by cultural distance, which can still verify that institutional distance mitigates the negative effect of cultural distance on FDI. Therefore, in order to promote international healthy investment behavior, it is feasible to improve both the mitigation effect of the institution in the short term and promote the level of cultural exchange in the long term, according to the research results of this paper.
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Internacionalidade , Investimentos em Saúde , China , Desenvolvimento Econômico , Instalações de Saúde , Meio SocialRESUMO
Four new purified polysaccharides (PAP) were isolated and purified from the Enteromorpha prolifera by alkali extraction, and further characterization was investigated. Their average molecular weights of PAP-1, PAP-2, PAP-3, and PAP-4 were estimated as 3.44 × 104, 6.42 × 104, 1.20 × 105, and 4.82 × 104 Da, respectively. The results from monosaccharide analysis indicated that PAP-1, PAP-2, PAP-3 were acidic polysaccharides and PAP-4 was a neutral polysaccharide. PAP-1 and PAP-2 mainly consist of galacturonic acid, while PAP-3 and PAP-4 mainly contained rhamnose. Congo red test showed that no triple helical structure was detected in the four polysaccharides. The antioxidant activities were investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH), Superoxide, and 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical assay. In vitro antitumor activities were evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. PAP-1 exhibited relatively stronger antioxidant activities among the four polysaccharides in a dose-dependent manner. At a concentration of 1.00 mg/mL, the antioxidant activities of PAP-1 on the DPPH radical scavenging rate, superoxide anion radical scavenging rate, and ABTS radical rate at 1.00 mg/mL were 56.40%, 54.27%, and 42.07%, respectively. They also showed no significant inhibitory activity against MGC-803, HepG2, T24, and Bel-7402 cells. These investigations of polysaccharides provide a scientific basis for the use of E. prolifera as an ingredient in functional foods and medicines.
Assuntos
Antioxidantes/farmacologia , Clorófitas/metabolismo , Polissacarídeos/farmacologia , Álcalis/química , Animais , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Antioxidantes/isolamento & purificação , Fracionamento Químico , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Eritrócitos/patologia , Hemólise/efeitos dos fármacos , Células Hep G2 , Humanos , Peróxido de Hidrogênio/toxicidade , Camundongos Endogâmicos ICR , Estrutura Molecular , Polissacarídeos/isolamento & purificação , Relação Estrutura-AtividadeRESUMO
[This corrects the article DOI: 10.1371/journal.pone.0099741.].
RESUMO
Large-scale green tides caused by Ulva prolifera, occurred for 12 consecutive years in the Yellow Sea of China. To resolve the abrupt shift in species composition between attached and floating macroalgal assemblages, field experiments were conducted from May to July 2017 to quantify the net buoyancy force and compare the floating potential of the common green macroalgae from the red algal seaweed Pyropia yezoensis rafts. At the same time, U. prolifera from different sampling locations were tested to study variable buoyancy of this species and the associated influencing factors. Our results illustrated a stronger positive buoyant force and a proportionally greater buoyancy capacity of U. prolifera, compared to the other co-occurring species. Buoyancy is a dynamic trait and is closely correlated with light intensity, morphology and physiological status. The positive buoyancy of U. prolifera is an important factor that helps explain its predominance in the Yellow Sea's large-scale green tides.
Assuntos
Monitoramento Ambiental/métodos , Eutrofização/fisiologia , Ulva/fisiologia , China , Oceanos e Mares , Estações do Ano , Água do Mar/química , Alga Marinha/fisiologia , Ulva/crescimento & desenvolvimentoRESUMO
BACKGROUND: Carotenoids are widely distributed in plants and algae, and their biosynthesis has attracted widespread interest. Carotenoid-related research has mostly focused on model species, and there is a lack of data on the carotenoid biosynthetic pathway in U. prolifera that is the main species leading to green tide, a harmful plague of floating green algae. RESULTS: The carotenoid content of U. prolifera samples, that is the main species leading to green tide, a harmful plague of floating green algae at different temperatures revealed that its terpenoid was highest in the samples subjected to high temperature at 28 °C (H), followed by the samples subjected to low temperature at 12 °C (L). Its terpenoid was lowest in the samples subjected to medium temperature at 20 °C (M). We conducted transcriptome sequencing (148.5 million raw reads and 49,676 unigenes in total) of samples that were subjected to different temperatures to study the carotenoid biosynthesis of U. prolifera. There were 1125-3164 significant differentially expressed genes between L, M and H incubation temperatures, of which 11-672 genes were upregulated and 453-3102 genes were downregulated. A total of 3164 genes were significantly differentially expressed between H and M, of which 62 genes were upregulated and 3102 genes were downregulated. A total of 2669 significant differentially expressed genes were observed between L and H, of which 11 genes were upregulated and 2658 genes were downregulated. A total of 13 genes were identified to be involved in carotenoid biosynthesis in U. prolifera, and the expression levels of the majority were highest at H and lowest at M of incubation temperature. Both the carotenoid concentrations and the expression of the analysed genes were lowest in the normal temperature group, while low temperature and high temperature seemed to activate the biosynthesis of carotenoids in U. prolifera. CONCLUSIONS: In this study, transcriptome sequencing provided critical information for understanding the accumulation of carotenoids and will serve as an important reference for the study of other metabolic pathways in U. prolifera.
Assuntos
Carotenoides/biossíntese , Perfilação da Expressão Gênica , Genes de Plantas , Ulva/genética , Carotenoides/análise , Clorofila/análise , Clorofila A/análise , Regulação da Expressão Gênica de Plantas , Filogenia , RNA de Plantas/química , RNA de Plantas/isolamento & purificação , RNA de Plantas/metabolismo , Análise de Sequência de RNA , Temperatura , Ulva/classificaçãoRESUMO
The 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase (HDS) gene and the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) gene are two important genes in the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. In this study, we reported the isolation and characterization of full-length HDS (MF101802) and HDR (MF159558) from Pyropia haitanensis. Characteristics of 3-D structures of the PhHDS and PhHDR proteins were analysed respectively. The results showed that the full-length cDNA of PhHDS, which is 1801 bp long, contained a 1455 bp open reading frame (ORF) encoding a putative 484 amino acid residue protein with a predicted molecular mass of 51.60 kDa. Meanwhile, the full-length cDNA of PhHDR was 1668 bp and contained a 1434 bp ORF encoding a putative 477 amino acid 2 residue protein with a predicted molecular mass of 51.49 kDa. The expression levels of the two genes were higher in conchocelis than that in leafy thallus. Additionally, the expression levels could be influenced by light, temperature and salinity and induced by methyl jasmonate (MJ) and salicylic acid (SA). This study contributed to our in-depth understanding of the roles of PhHDS and PhHDR in terpenoid biosynthesis in Pyropia haitanensis and the regulation of the two genes by external environments.
Assuntos
Proteínas de Algas/metabolismo , Rodófitas/metabolismo , Acetatos/administração & dosagem , Acetatos/metabolismo , Proteínas de Algas/genética , Sequência de Aminoácidos , Clonagem Molecular , Ciclopentanos/administração & dosagem , Ciclopentanos/metabolismo , Regulação da Expressão Gênica de Plantas , Luz , Modelos Moleculares , Oxilipinas/administração & dosagem , Oxilipinas/metabolismo , Filogenia , Conformação Proteica , Rodófitas/genética , Ácido Salicílico/administração & dosagem , Ácido Salicílico/metabolismo , Salinidade , Água do Mar , Alinhamento de Sequência , TemperaturaRESUMO
Cyanobacteria blooms possess serious threats to water resources. Herein, we report the synthesis of polyanionic membranes (PA-M) by in situ photo-crosslinking of a sulfate-based anionic monomer followed by cation-exchange with metal cations, Fe3+ (PA-Fe), Cu2+ (PA-Cu), or Zn2+ (PA-Zn). The effect of cations on the anti-cyanobacterial activities against both Microcystis aeruginosa (M.â aeruginosa) and Anabaena flos-aquae (A.â flos-aquae) was investigated. All the prepared metal-containing membranes (PA-Fe, PA-Cu, PA-Zn) exhibit high anti-cyanobacterial activities and long-term anti-cyanobacterial stability, demonstrating that the synthesized PA-M membranes can be used as an effective and safe inhibitor to control cyanobacterial blooms.
Assuntos
Antibacterianos/farmacologia , Reagentes de Ligações Cruzadas/farmacologia , Dolichospermum flosaquae/efeitos dos fármacos , Microcystis/efeitos dos fármacos , Polímeros/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Reagentes de Ligações Cruzadas/síntese química , Reagentes de Ligações Cruzadas/química , Testes de Sensibilidade Microbiana , Estrutura Molecular , Polieletrólitos , Polímeros/síntese química , Polímeros/químicaRESUMO
The filamentous Cyanobacterium Arthrospira is commercially produced and is a functional, high-value, health food. We identified 5 low temperature and low light intensity tolerant strains of Arthrospira sp. (GMPA1, GMPA7, GMPB1, GMPC1, and GMPC3) using ethyl methanesulfonate mutagenesis and low temperature screening. The 5 Arthrospira strains grew rapidly below 14 °C, 43.75 µmol photons m-2 s-1 and performed breed conservation at 2.5 °C, 8.75 µmol photons m-2 s-1. We used morphological identification and molecular genetic analysis to identify GMPA1, GMPA7, GMPB1 and GMPC1 as Arthrospira platensis, while GMPC3 was identified as Arthrospira maxima. Growth at different culture temperatures was determined at regular intervals using dry biomass. At 16 °C and 43.75 µmol photons m-2 s-1, the maximum dry biomass production and the mean dry biomass productivity of GMPA1, GMPB1, and GMPC1 were 2057 ± 80 mg l-1, 68.7 ± 2.5 mg l-1 day-1, 1839 ± 44 mg l-1, 60.6 ± 1.8 mg l-1 day-1, and 2113 ± 64 mg l-1, 77.7 ± 2.5 mg l-1 day-1 respectively. GMPB1 was chosen for additional low temperature tolerance studies and growth temperature preference. In winter, GMPB1 grew well at mean temperatures <10 °C, achieving 3258 mg dry biomass from a starting 68 mg. In summer, GMPB1 grew rapidly at mean temperatures more than 28 °C, achieving 1140 mg l-1 dry biomass from a starting 240 mg. Phytonutrient analysis of GMPB1 showed high levels of C-phycocyanin and carotenoids. Arthrospira metabolism relates to terpenoids, and the methyl-D-erythritol 4-phosphate pathway is the only terpenoid biosynthetic pathway in Cyanobacteria. The 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) gene from GMPB1 was cloned and phylogenetic analysis showed that GMPB1 is closest to the Cyanobacterium Oscillatoria nigro-viridis PCC711. Low temperature tolerant Arthrospira strains could broaden the areas suitable for cultivation, extend the seasonal cultivation time, and lower production costs.
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Biomassa , Oscillatoria/crescimento & desenvolvimento , Oscillatoria/metabolismo , Terpenos/metabolismo , Temperatura Baixa , Luz , Mutagênese , Oscillatoria/genética , Filogenia , Alinhamento de SequênciaRESUMO
Chloroplast formation is associated with embryo development and seedling growth. However, the relationship between chloroplast differentiation and embryo development remains unclear. Five FtsHi genes that encode proteins with high similarity to FtsH proteins, but lack Zn2+-binding motifs, are present in the Arabidopsis genome. In this study, we showed that T-DNA insertion mutations in the Arabidopsis FtsHi4 gene resulted in embryo arrest at the globular-to-heart-shaped transition stage. Transmission electron microscopic analyses revealed abnormal plastid differentiation with a severe defect in thylakoid formation in the mutant embryos. Immunocytological studies demonstrated that FtsHi4 localized in chloroplasts as a thylakoid membrane-associated protein, supporting its essential role in thylakoid membrane formation. We further showed that FtsHi4 forms protein complexes, and that there was a significant reduction in the accumulation of D2 and PsbO (two photosystem II proteins) in mutant ovules. The role of FtsHi4 in chloroplast development was confirmed using an RNA-interfering approach. Additionally, mutations in other FtsHi genes including FtsHi1, FtsHi2, and FtsHi5 caused phenotypic abnormalities similar to ftshi4 with respect to plastid differentiation during embryogenesis. Taken together, our data suggest that FtsHi4, together with FtsHi1, FtsHi2, and FtsHi5 are essential for chloroplast development in Arabidopsis.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/embriologia , Proteínas das Membranas dos Tilacoides/fisiologia , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/análise , Proteínas de Arabidopsis/genética , Cloroplastos/metabolismo , Cloroplastos/fisiologia , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Mutagênese Sítio-Dirigida , Interferência de RNA , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Proteínas das Membranas dos Tilacoides/análise , Proteínas das Membranas dos Tilacoides/genética , Tilacoides/metabolismoRESUMO
The xanthophyll cycle (Xc), which involves violaxanthin de-epoxidase (VDE) and the zeaxanthin epoxidase (ZEP), is one of the most rapid and efficient responses of plant and algae to high irradiance. High light intensity can activate VDE to convert violaxanthin (Vx) to zeaxanthin (Zx) via antheraxanthin (Ax). However, it remains unclear whether VDE remains active under low light or dark conditions when there is no significant accumulation of Ax and Zx, and if so, how the ΔpH required for activation of VDE is built. In this study, we used salicylaldoxime (SA) to inhibit ZEP activity in the intertidal macro-algae Ulva sp. (Ulvales, Chlorophyta) and then characterized VDE under low light and dark conditions with various metabolic inhibitors. With inhibition of ZEP by SA, VDE remained active under low light and dark conditions, as indicated by large accumulations of Ax and Zx at the expense of Vx. When PSII-mediated linear electron transport systems were completely inhibited by SA and DCMU, alternative electron transport systems (i.e., cyclic electron transport and chlororespiration) could maintain VDE activity. Furthermore, accumulations of Ax and Zx decreased significantly when SA, DCMU, or DBMIB together with an inhibitor of chlororespiration (i.e., propyl gallate (PG)) were applied to Ulva sp. This result suggests that chlororespiration not only participates in the build-up of the necessary ΔpH, but that it also possibly influences VDE activity indirectly by diminishing the oxygen level in the chloroplast.
Assuntos
Cloroplastos/fisiologia , Transporte de Elétrons/fisiologia , Oxirredução , Fotoperíodo , Clorofila/metabolismo , Cloroplastos/metabolismo , Luz , Oxirredutases/metabolismo , Prótons , Espécies Reativas de Oxigênio/metabolismo , Ulva/metabolismo , Ulva/fisiologia , Xantofilas/metabolismo , Zeaxantinas/metabolismoRESUMO
Porphyra yezoensis Ueda is an intertidal marine red algae that has received increasing attention as a model organism owing to its important role in biological research and the agronomic industry. The two generations of Porphyra yezoensis, the sporophyte and the gametophyte, have the same genome but show great differences in many aspects, including structural features, habitat, and gene expression. To identify miRNAs and their probable roles in P. yezoensis development, we constructed and sequenced libraries of small RNA from P. yezoensis sporophytes and gametophytes. The sequencing data were analyzed, and 14 miRNAs were identified, with only one common to these two samples. Our results show that P. yezoensis has a complex small RNA processing system containing novel miRNAs that have no identifiable homolog in other organisms. These miRNAs might have important regulatory roles in development of the different generations of P. yezoensis.
RESUMO
The endosperm is an essential part of the seed which sustains embryo development and contains storage reserves. Endosperm development involves a series of nuclear divisions in the absence of cytokinesis. In this study, it is shown that a mutant Arabidopsis plant carrying the disrupted At4g24900 gene exhibits severe defects during seed development. At4g24900 encodes a nuclear-localized C2H2-containing protein and the transcripts of this gene are alternatively spliced, consequently producing at least nine differentially spliced isoforms. The heterozygous T-DNA insertion mutation in this gene resulted in abortion of 25% of seeds, and the homozygous mutant allele displayed embryo lethality. Differential interfernce contrast (DIC) analyses revealed that the mutant has endosperm nuclei with more than one nucleolus and an embryo arrested at the globular to heart stage transition. Because this mutant exhibits a titan-like phenotype, it was designated ttl. The TTL gene is preferentially expressed in tissues with quickly dividing cells as revealed in P(TTL)::GUS (ß-glucuronidase) transgenic plants. It is proposed that TTL is likely to function as a key regulator of endosperm nuclear division.
Assuntos
Processamento Alternativo , Arabidopsis/genética , Endosperma/embriologia , Sequência de Aminoácidos , Arabidopsis/química , Arabidopsis/embriologia , Arabidopsis/metabolismo , Endosperma/química , Endosperma/genética , Endosperma/metabolismo , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Alinhamento de Sequência , Dedos de ZincoRESUMO
Porphyra yezoensis has a macroscopic foliage gametophyte phase with only a single cell layer, and is ideally suited for the study of the sexual differentiation process, from the vegetative cell to the spermatia. Firstly, we compared variations in the responses of the vegetative and male sectors to desiccation. Later, cell tracking experiments were carried out during the formation of spermatia from vegetative cells. The two sectors showed similar tolerance to desiccation, and the formation of spermatia from vegetative cells was independent of the degree of desiccation. Both light and scanning electron microscopy (SEM) observations of the differentiation process showed that the formation of spermatia could be divided into six phases: the one-cell, two-cell, four-cell, eight-cell, pre-release and spermatia phases. Photomicrographs of Fluorescent Brightener staining showed that the released spermatia had no cell walls. Photosynthetic data showed that there was a significant rise in Y(II) in the four-cell phase, indicating an increase in photosynthetic efficiency of PSII during this phase. We propose that this photosynthetic rise may be substantial and provide the increased energy needed for the formation and release of spermatia in P. yezoensis.
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Aclimatação/fisiologia , Células Germinativas Vegetais/citologia , Células Germinativas Vegetais/crescimento & desenvolvimento , Fotossíntese/fisiologia , Células Vegetais/fisiologia , Porphyra/citologia , Porphyra/fisiologia , Organismos Aquáticos/fisiologia , Diferenciação Celular , China , Dessecação , Secas , Diferenciação Sexual/fisiologia , Água/metabolismoRESUMO
BACKGROUND: Porphyra yezoensis Ueda is one of the most important edible seaweed, with a dimorphic life cycle which consists of gametophyte as macroscopical blade and sporophyte as microscopic filamentous. Conspicuous differences exist in the two generations, such as morphology, cell structure, biochemistry, physiology, and so on. The developmental process of Porphyra yezoensis has been studied thoroughly, but the mechanism is still ambiguous and few studies on genetic expression have been carried out.In this study, the suppression subtractive hybridization (SSH) method conducted to generate large-scale expressed sequence tags (EST) is designed to identify gene candidates related to the morphological and physiological differences between the gametophytic and sporophytic generations of Porphyra yezoensis Ueda. FINDINGS: Each 300 clones of sporophyte and gametophyte cells were dipped onto the membrane for hybridization. The result of dot-blot suggested there were 222 positive clones in gametophyte library and 236 positive clones in sporophyte library. 383 positive clones of strongest signals had been sequenced, and 191 EST sequences of gametophyte and 192 of sporophyte were obtained.A total of 196 genes were obtained, within which 104 genes were identified from the gametophyte and 92 from the sporophyte. Thirty-nine genes of the gametophyte and 62 genes of the sporophyte showed sequence similarity to those genes with known or putative functions which were classified according to their putative biological roles and molecular functions. The GO annotation showed about 58% of the cellular component of sporophyte and gametophyte cells were mainly located in cytoplasm and nucleus. The special genes were located in Golgi apparatus, and high expression in plastid, ribosome and endoplasmic reticulum. The main biological functions of gametophyte cells contributed to DNA repair/replication, carbohydrate metabolism, transport and transcription, especially in response to heat and oxidative stress. The sporophyte cell expresses more genes in transcription, transport, carbohydrate metabolism, particularly in signal transduction, DNA and protein modification, protein and nucleotide metabolism. Four genes are expressed on both gametophyte and sporophyte cells and eighteen genes have not been annotated. CONCLUSION: According to the information of GO annotation, the gametophyte tends to growth and self- protection while the sporophyte tends to be more active in development. Interpretation of the differentially expressed genes revealed new insights into the molecular processes of the generation alternation of Porphyra yezoensis. Further investigation are needed due to insufficiency of functional genes research and indeterminancy of the functions of many sequences.
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Although natural sources have long been exploited for astaxanthin production, it is still uncertain if natural astaxanthin can be produced at lower cost than that of synthetic astaxanthin or not. In order to give a comprehensive cost analysis of astaxanthin production from Haematococcus, a pilot plant with two large scale outdoor photobioreactors and a raceway pond was established and operated for 2 years to develop processes for astaxanthin production from Haematococcus. The developed processes were scaled up to a hypothetical plant with a production capacity about 900 kg astaxanthin per year, and the process economics was preliminarily assessed. Based on the analysis, the production cost of astaxanthin and microalgae biomass can be as low as $718/kg and $18/kg respectively. The results are very encouraging because the estimated cost might be lower than that of chemically synthesized astaxanthin.
Assuntos
Biotecnologia/métodos , Clorófitas/metabolismo , Microalgas/metabolismo , Fotobiorreatores/economia , Biocombustíveis , Biomassa , Biotecnologia/economia , Biotecnologia/instrumentação , Xantofilas/biossínteseRESUMO
Ulva sp. (Chlorophyta) is a representative species of the intertidal macro-algae responsible for the green tides that occurred along the shores of Qingdao in 2008 and had detrimental effects on the preparation for the 2008 Beijing Olympic Games sailing competition. In view of its significance, we have investigated the photosynthetic performance of the photosystems and the changes in photosynthetic electron transport that occur during desiccation and rehydration of Ulva sp. The PSII activity in Ulva sp. declined gradually during the course of desiccation, which was reflected by the decreased maximum quantum yield and effective quantum yield, whereas the PSI activity fluctuated significantly. In contrast, the electron transport rates of PSII approached zero at severe levels of desiccation, but the electron transport of PSI, which still operated, could be suppressed effectively by a specific inhibitor. Furthermore, the electron transport of PSI during rehydration of desiccated thalli was recovered faster than that of PSII. All these results implied that the linear electron flow was abolished in desiccated Ulva sp., whereas the cyclic PSI activity was significantly elevated, was still active at severe levels of desiccation and could be restored faster than PSII activity. Based on these results, we concluded the PSI-driven cyclic electron flow might provide desiccation tolerance and additional flexibility for the cell physiology of Ulva sp. under desiccation conditions, which might be one of the most important factors that make Ulva sp. well suited to experience daily cycles of desiccation at low tide and rehydration at high tide.