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Underground personnel behavior recognition is an important measure to ensure safe production in coal mines. The existing research on behavior recognition of underground personnel lacks research and analysis on the perception mechanism, and the feature extraction method is simple. In order to solve the above problems, a behavior recognition method for underground personnel based on fusion networks is proposed. The method mainly includes three parts: data preprocessing, feature construction, and recognition network construction. Data preprocessing: the collected channel status information (CSI) data is processed through CSI quotient models, subcarrier denoising, and discrete wavelet denoising to reduce the impact of environmental noise and equipment noise. Feature construction: the processed data is transformed into images using the Gramian angular summation/difference fields (GASF/GADF) to preserve the spatial and temporal features of the data. Recognition network construction: according to the features of personnel actions, a fusion network composed of a gate recurrent unit (GRU) based encoding and decoding network and a multiscale convolutional neural network (CNN) is proposed. GRU is used to preserve the correlation between pre and post data. The weight allocation strategy of the attention mechanism is used to effectively extract key features to improve the accuracy of behavior recognition. The experimental results show that the average recognition accuracy of this method for eight movements, namely walking, taking off a hat, throwing things, sitting, smoking, waving, running, and sleeping, is 97.37%. The recognition accuracy for sleeping and sitting is the highest, and the most prone to misjudgment are walking and running. Using accuracy, precision, recall, and F1 score as evaluation indicators, it is concluded that the performance of the fusion network is superior to CNN and GRU. The accuracy of personnel behavior recognition is higher than the HAR system, WiWave system and Wi-Sense system. The average recognition accuracy of walking and taking off a hat at normal speed is 95.6%, which is higher than 93.6% for fast motion and 92.7% for slow motion. When the distance between transceiver devices is 2 m and 2.5 m, the recognition accuracy is higher.

In the application of mobile edge computing(MEC) in intelligent mine, the mobile users unload tasks to non-optimal edge servers due to unreasonable resource allocation, which leads to extra transmission time and execution delay, thus resulting in the decrease of the total task completion rate. In order to solve the above problem, a two-dimensional dynamic matching algorithm based on preference is proposed to optimize the resource allocation decision in MEC system. The data of the position of a mobile user in MEC system and the calculation amount required by a task in one time slot is sent to the edge server. The preference table of the edge server for the mobile user is formed according to the set preference value. At the same time, the preference table for all the edge servers is formed by the mobile user according to different physical distances. The two preference tables are combined to form a two-dimensional dynamic preference table, which is abstracted into a two-dimensional matrix. The two-dimensional matrix is processed by a two-dimensional dynamic matching algorithm based on preference, and the matching optimization results of mobile users and edge servers are obtained. The simulation results show that compared with the conventional MEC scene unloading algorithm, the preference-based two-dimensional dynamic matching algorithm can effectively alleviate the problem of the decrease of the total task completion rate in a large number of sudden task scenes, and can achieve the total task completion rate of more than 60% in extreme cases.

The difficulty in fault diagnosis of mine hoist inverters lies in extracting the features that characterize faults. At present, signal processing methods are mainly used to obtain fault statistical features, or the fault depth features are extracted by neural networks. In the actual working environment, the hoist inverter is affected by factors such as background noise and load changes. Therefore, it is difficult to obtain features that can characterize the faults effectively by using a single feature extraction method, resulting in low fault diagnosis accuracy of the hoist inverter. In order to solve the above problems, a fault diagnosis method of hoist inverter based on the fusion of statistical features and depth features is proposed. Firstly, the Hilbert-Huang transform(HHT) is used to conduct modified ensemble empirical mode decomposition(MEEMD) of the inverter output current signal so as to obtain the fault statistical features. At the same time, the squeeze and excitation with densely connected convolutional network(SE-DenseNet) is used to extract the depth features of the output current signal. Secondly, the local fisher discriminant analysis(LFDA) is used to perform fusion and dimensionality reduction processing on the combination of the two features to obtain low-dimensional fusion features of statistical features and depth features. Finally, the low-dimensional fusion features are input to the extreme learning machine to obtain inverter fault classification. Experiments are conducted for a single IGBT open-circuit fault in the hoist inverter. The results show that the low-dimensional fusion features obtained by this method are more capable of fault characterization than single features, which improves the fault recognition accuracy effectively.

In coal mine working face, there are problems such as uneven energy distribution of wireless monitoring network, the difficulty of manual maintenance and node battery replacement and energy limitation of some monitoring nodes. In order to solve the above problems, an energy optimization method of wireless monitoring network in coal mine working face based on mobile nodes is proposed by using the characteristics of periodic movement of the shearer. This method installs mobile nodes on the shearer to forward data that collected by monitoring nodes in the network to reduce multi-hop data transmission, and reduces redundant data transmission by setting transmission thresholds to reduce node energy consumption. The mobile nodes are used to radiate radio frequency energy to the monitoring nodes, which provides targeted energy supplement to the monitoring nodes so as to achieve energy balance of the network nodes. The experimental results show that compared with LEACH protocol, this method can make the wireless monitoring network energy distribution more balanced, reduce the network energy consumption and monitoring node mortality rate effectively under the same operation round, and extend the effective life of the network by 2.8 times.

Simple Summary: This review highlights the intricate interactions between two major constituents of marine ecosystems: dinoflagellates and heterotrophic bacteria. Dinoflagellates typically associate with a diverse array of heterotrophic bacteria, which are not limited to but predominantly consist of phyla such as Alphaproteobacteria, Gammaproteobacteria, and the Cytophaga–Flavobacterium–Bacteroides group. These bacterial communities engage with dinoflagellates in a multifaceted manner, including nutrient exchange, the secretion of pathogenic substances, and involvement in the synthesis of chemical entities. Crucially, the process of horizontal gene transfer from bacteria is a significant force sculpting the genomic architecture of dinoflagellates. The objective of this review is to shed light on the dynamic interactions between dinoflagellates and their bacterial partners, examining the relationship through a physiological, ecological, and evolutionary perspective. Dinoflagellates and heterotrophic bacteria are two major micro-organism groups within marine ecosystems. Their coexistence has led to a co-evolutionary relationship characterized by intricate interactions that not only alter their individual behaviors but also exert a significant influence on the broader biogeochemical cycles. Our review commenced with an analysis of bacterial populations, both free-living and adherent to dinoflagellate surfaces. Members of Alphaproteobacteria, Gammaproteobacteria, and the Cytophaga–Flavobacterium–Bacteroides group are repeatedly found to be associated with dinoflagellates, with representation by relatively few genera, such as Methylophaga, Marinobacter, and Alteromonas. These bacterial taxa engage with dinoflagellates in a limited capacity, involving nutrient exchange, the secretion of pathogenic substances, or participation in chemical production. Furthermore, the genomic evolution of dinoflagellates has been profoundly impacted by the horizontal gene transfer from bacteria. The integration of bacterial genes into dinoflagellates has been instrumental in defining their biological characteristics and nutritional strategies. This review aims to elucidate the nuanced interactions between dinoflagellates and their associated bacteria, offering a detailed perspective on their complex relationship. [ABSTRACT FROM AUTHOR]

Global climate change intensifies the water cycle and makes freshest waters become fresher and vice‐versa. But how this change impacts phytoplankton in coastal, particularly harmful algal blooms (HABs), remains poorly understood. Here, we monitored a coastal bay for a decade and found a significant correlation between salinity decline and the increase of Karenia mikimotoi blooms. To examine the physiological linkage between salinity decreases and K. mikimotoi blooms, we compare chemical, physiological and multi‐omic profiles of this species in laboratory cultures under high (33) and low (25) salinities. Under low salinity, photosynthetic efficiency and capacity as well as growth rate and cellular protein content were significantly higher than that under high salinity. More strikingly, the omics data show that low salinity activated the glyoxylate shunt to bypass the decarboxylation reaction in the tricarboxylic acid cycle, hence redirecting carbon from CO2 release to biosynthesis. Furthermore, the enhanced glyoxylate cycle could promote hydrogen peroxide metabolism, consistent with the detected decrease in reactive oxygen species. These findings suggest that salinity declines can reprogram metabolism to enhance cell proliferation, thus promoting bloom formation in HAB species like K. mikimotoi, which has important ecological implications for future climate‐driven salinity declines in the coastal ocean with respect to HAB outbreaks. [ABSTRACT FROM AUTHOR]

Simple Summary: Recently, the occurrence of harmful algal blooms (HABs) in coastal areas has increased rapidly, negatively impacting fishing resources, public health, and marine ecosystems. Certain microorganisms play a significant role in the termination of HABs. To exploit these algicidal bacteria effectively, it is imperative to amplify their algicidal ratio and devise efficient large-scale cultivation methods; however, research in these areas is underdeveloped. We previously identified Pseudoalteromonas sp. strain FDHY-MZ2, which exhibited significant algicidal activity against Karenia mikimotoi, a species globally acknowledged for forming HAB. To improve the algicidal efficacy of FDHY-MZ2, cultures were progressed from shaking flask conditions to small-scale (5 L) and pilot-scale fermentation (50 L). The optimal fermentation medium and conditions at a pilot-scale level were established, resulting in a substantial improvement in algicidal properties. This enhancement might be due to a rise in algal H2O2 production accompanied by increased cell membrane disintegration, pronounced damage to cell chlorophyll and algal photosynthesis, and severe protein degradation. These findings provide new insights into the previously unknown potential of systematically applied microbial agents for the treatment of HABs. The role of microorganisms in effectively terminating harmful algal blooms (HABs) is crucial for maintaining environmental stability. Recent studies have placed increased emphasis on bio-agents capable of inhibiting HABs. The bacterium Pseudoalteromonas sp. strain FDHY-MZ2 has exhibited impressive algicidal abilities against Karenia mikimotoi, a notorious global HAB-forming species. To augment this capability, cultures were progressively scaled from shake flask conditions to small-scale (5 L) and pilot-scale (50 L) fermentation. By employing a specifically tailored culture medium (2216E basal medium with 1.5% soluble starch and 0.5% peptone), under precise conditions (66 h, 20 °C, 450 rpm, 30 L/min ventilation, 3% seeding, and constant starch flow), a notable increase in algicidal bacterial biomass was observed; the bacterial dosage required to entirely wipe out K. mikimotoi within a day decreased from 1% to 0.025%. Compared to an unoptimized shake flask group, the optimized fermentation culture caused significant reductions in algal chlorophyll and protein levels (21.85% and 78.3%, respectively). Co-culturing induced increases in algal malondialdehyde and H2O2 by 5.98 and 5.38 times, respectively, leading to further disruption of algal photosynthesis. This study underscores the unexplored potential of systematically utilized microbial agents in mitigating HABs, providing a pathway for their wider application. [ABSTRACT FROM AUTHOR]

Prorocentrum donghaiense is a harmful-algal-bloom-forming species of planktonic dinoflagellates widely distributed around the world, which threatens the marine environment and human health. Bacteria are promising biological agents to control algal growth in HABs. Previously, we isolated an Alteromonas sp. FDHY-03, a P. donghaiense-lysing bacteria strain, from Xiapu Sea area of China. In order to improve the algicidal activity of this strain, we optimized the medium composition and culture conditions. Based on single-factor method experiment design, the optimum medium component of algicidal effect for strain FDHY-03 was sucrose and peptone. The result of Plackett–Burman design indicated that three significant factors (sucrose, peptone, and rotational speed) appeared. Finally, the concentrations of key factors were confirmed by central composite design (CCD) and response surface methodology (RSM). Under the optimized medium, the algicidal rate of strain FDHY-03 against P. donghaiense improved by 67.15%, and the OD600 value increased by 2.86-fold. The optimal source and condition were sucrose 0.46% (w/v), peptone 4.25% (w/v) addition, and rotation speed 255 rpm. Overall, this study provides an optimized method and optimum medium for improving the algicidal activity against P. donghaiense, and has a positive influence on algae-lysing bacteria for controlling the blooms of the algae in the environment. [ABSTRACT FROM AUTHOR]

In the actual industrial scenarios, most existing fault diagnosis approaches are faced with two challenges, insufficient labeled training data and distribution divergences between training and testing datasets. For the above issues, a new transferable fault diagnosis approach of rotating machinery based on deep autoencoder and dominant features selection is proposed in this article. First, maximal overlap discrete wavelet packet transform is applied for signals processing and mix-domains statistical feature extraction. Second, dominant features selection by importance score and differences between domains is proposed to select dominant features with high fault-discriminative ability and domain invariance. Then, selected dominant features are used for pretraining deep autoencoder (source model), which helps in enhancing the fault representative ability of deep features. The parameters of the source model are transferred to the target model, and normal state features from target domain are adopted for fine-tuning the target model. Finally, the target model is applied for fault patterns classification. Motor and bearing fault datasets are used for a series of experiments, and the results verify that the proposed methods have better cross-domain diagnosis performance than comparative models. [ABSTRACT FROM AUTHOR]

Diatoms and dinoflagellates dominate coastal marine phytoplankton communities as major players of marine biogeochemical cycles and their seasonal succession often leads to harmful algal blooms (HABs). What regulates their respective dominances and the development of the HABs remains elusive. Here we conducted time-sequential metatranscriptomic profiling on a natural assemblage that evolved from diatom dominance to a dinoflagellate bloom to interrogate the underlying major metabolic and ecological drivers. Data reveals similarity between diatoms and dinoflagellates in exhibiting high capacities of energy production, nutrient acquisition, and stress protection in their respective dominance stages. The diatom-to-dinoflagellate succession coincided with an increase in turbidity and sharp declines in silicate and phosphate availability, concomitant with the transcriptomic shift from expression of silicate uptake and urea utilization genes in diatoms to that of genes for light harvesting, diversified phosphorus acquisition and autophagy-based internal nutrient recycling in dinoflagellates. Furthermore, the diatom-dominant community featured strong potential to carbohydrate metabolism and a strikingly high expression of trypsin potentially promoting frustule building. In contrast, the dinoflagellate bloom featured elevated expression of xanthorhodopsin, and antimicrobial defensin genes, indicating potential importance of energy harnessing and microbial defense in bloom development. This study sheds light on mechanisms potentially governing diatom- and dinoflagellate-dominance and regulating bloom development in the natural environment and raises new questions to be addressed in future studies. [ABSTRACT FROM AUTHOR]

Proliferating cell nuclear antigen (PCNA) plays critical roles in eukaryotic DNA replication and replication‐associated processes. It is typically encoded by one or two gene copies (pcna) in eukaryotic genomes. Recently reported higher copy numbers of pcna in some dinoflagellates raised a question of how this gene has uniquely evolved in this phylum. Through real‐time PCR quantification, we found a wide range of pcna copy number (2–287 copies) in 11 dinoflagellate species (n = 38), and a strong positive correlation between pcna copy number and genome size (log10–log10 transformed). Intraspecific pcna diverged up to 21% and are dominated by nonsynonymous substitutions, indicating strong purifying selection pressure on and hence functional necessity of this gene. By surveying pcna copy numbers in eukaryotes, we observed a genome size threshold at 4 pg DNA, above which more than two pcna copies are found. To examine whether retrotransposition is a mechanism of pcna duplication, we measured the copy number of retroposed pcna, taking advantage of the 22‐nt dinoflagellate‐specific spliced leader (DinoSL) capping the 5′ end of dinoflagellate nuclear‐encoded mRNAs, which would exist in the upstream region of a retroposed gene copy. We found that retroposed pcna copy number increased with total pcna copy number and genome size. These results indicate co‐evolution of dinoflagellate pcna copy number with genome size, and retroposition as a major mechanism of pcna duplication in dinoflagellates. Furthermore, we posit that the demand of faithful replication and maintenance of the large dinoflagellate genomes might have favored the preservation of the retroposed pcna as functional genes. [ABSTRACT FROM AUTHOR]

Summary: Despite numerous laboratory studies on physiologies of harmful algal bloom (HAB) species, physiologies of these algae during a natural bloom are understudied. Here, we investigated a bloom of the raphidophyte Heterosigma akashiwo in the East China Sea in 2014 using metabarcode (18S rDNA) and metatranscriptome sequencing. Based on 18S rDNA analyses, the phytoplankton community shifted from high diversity in the pre‐bloom stage to H. akashiwo predominance during the bloom. A sharp decrease in ambient dissolved inorganic phosphate and strong up‐regulation of phosphate and dissolved organic phosphorus (DOP) uptake genes, including the rarely documented (ppGpp)ase, in H. akashiwo from pre‐bloom to bloom was indicative of rapid phosphorus uptake and efficient utilization of DOP that might be a driver of the H. akashiwo bloom. Furthermore, observed up‐regulated expression of mixotrophy‐related genes suggests potential contribution of mixotrophy to the bloom. Accelerating photosynthetic carbon fixation was also implied by the up‐regulation of carbonic anhydrase genes during the bloom. Notably, we also observed a strong morning‐to‐afternoon shift in the expression of many genes. Our findings provide insights into metabolic processes likely important for H. akashiwo bloom formation, and suggest the need to consider timing of sampling in field studies on this alga. [ABSTRACT FROM AUTHOR]

Although gene regulation can occur at both transcriptional and epigenetic (microRNA) levels, combined transcriptomic and microRNAomic responses to environmental stress are still largely unexplored for marine plankton. Here, we conducted transcriptome and microRNAome sequencing for Prorocentrum donghaiense to understand the molecular mechanisms by which this dinoflagellate copes with phosphorus (P) deficiency. Under P-depleted conditions, G1/S specific cyclin gene was markedly downregulated, consistent with growth inhibition, and genes related to dissolved organic phosphorus (DOP) hydrolysis, carbon fixation, nitrate assimilation, glycolysis, and cellular motility were upregulated. The elevated expression of ATP-generating genes (for example, rhodopsin) and ATP-consuming genes suggests some metabolic reconfiguration towards accelerated ATP recycling under P deficiency. MicroRNAome sequencing revealed 17 microRNAs, potentially regulating 3268 protein-coding genes. Functional enrichment analysis of these microRNA-targeted genes predicted decreases in sulfatide (sulfolipid) catabolism under P deficiency. Strikingly, we detected a significant increase in sulfolipid sulfatide content (but not in sulphoquinovosyldiacylglycerol content) and its biosynthesis gene expression, indicating a different sulfolipid-substituting-phospholipid mechanism in this dinoflagellate than other phytoplankters studied previously. Taken together, our integrative transcriptomic and microRNAomic analyses show that enhanced DOP utilization, accelerated ATP cycling and repressed sulfolipid degradation constitute a comprehensive strategy to cope with P deficiency in a model dinoflagellate. [ABSTRACT FROM AUTHOR]

Gene structure and expression regulation of form II RuBisCO (rbcII) in dinoflagellates are still poorly understood. Here we isolated this gene (Pdrbc) and investigated its diel expression pattern in a harmful algal bloom forming dinoflagellate Prorocentrum donghaiense. We obtained cDNA sequences with triple tandem repeats of the coding unit (CU); the 5′ region has the sequence of a typical dinoflagellate plastid gene, encoding an N-terminus with two transmembrane regions separated by a plastid transit peptide. The CUs (1,455 bp except 1464 bp in last CU) are connected through a 63 bp spacer. Phylogenetic analysis showed that rbcII CUs within species formed monophyletic clusters, indicative of intraspecific gene duplication or purifying evolution. Using quantitative PCR (qPCR) we estimated 117±40 CUs of Pdrbc in the P. donghaiense genome. Although it is commonly believed that most dinoflagellate genes lack transcriptional regulation, our RT-qPCR analysis on synchronized cultures revealed remarkable diel rhythm of Pdrbc expression, showing significant correlations of transcript abundance with the timing of the dark-to-light transition and cell cycle G2M-phase. When the cultures were shifted to continuous light, Pdrbc expression remained significantly correlated with the G2M-phase. Under continuous darkness the cell cycle was arrested at the G1 phase, and the rhythm of Pdrbc transcription disappeared. Our results suggest that dinoflagellate rbcII 1) undergoes duplication or sequence purification within species, 2) is organized in tandem arrays in most species probably to facilitate efficient translation and import of the encoded enzyme, and 3) is regulated transcriptionally in a cell cycle-dependent fashion at least in some dinoflagellates. [ABSTRACT FROM AUTHOR]

The toxic dinoflagellate Karenia mikimotoi is a harmful algal bloom-forming species in coastal areas around the world. It produces ichthyotoxins and hemolytic toxins, with deleterious effects on marine ecosystems. In this study, the bacterium Pseudoalteromonas sp. FDHY-MZ2, with high algicidal efficiency against K. mikimotoi , was isolated from a bloom event. Based on the results, it completely lysed K. mikimotoi cells within 24 h 0.5% (vol/vol), with the algicidal activity of the supernatant of the bacterium culture. Algal cell wall fragmentation occurred, leading to cell death. There was a marked decline in various photochemical traits. When treated with the supernatant, cellulase, pheophorbide a oxygenase (PAO) and cyclin B genes were significantly increased, suggesting induced cell wall deterioration, chloroplast degradation and cell cycle regulation of K. mikimotoi cells. In addition, the expression levels of reactive oxygen species (ROS) scavenging gene was significantly inhibited, indicating that the ROS removal system was damaged. The bacterial culture was dried to obtain the spray-dried powder, which showed algicidal activity rates of 92.2 and 100% against a laboratory K. mikimotoi culture and a field microcosm of Karlodinium sp. bloom within 24 h with the addition of 0.04% mass fraction powder. Our results demonstrate that FDHY-MZ2 is a suitable strain for K. mikimotoi and Karlodinium sp. blooms management. In addition, this study provides a new strategy for the anthropogenic control of harmful algal bloom-forming species in situ . IMPORTANCE K. mikimotoi is a noxious algal bloom-forming species that cause damaging of the aquaculture industry and great financial losses. Bacterium with algicidal activity is an ideal agency to inhibit the growth of harmful algae. In this approach application, the bacterium with high algicidal activity is required and the final management material is ideal for easy-to-use. The algicidal characteristics are also needed to understand the effects of the bacterium for managing strategy exploration. In this study, we isolated a novel algicidal bacterium with extremely high lysis efficiency for K. mikimotoi . The algicidal characteristics of the bacterium as well as the chemical and molecular response of K. mikimotoi with the strain challenge were examined. Finally, the algicidal powder was explored for application. The results demonstrate that FDHY-MZ2 is suitable for K. mikimotoi and Karlodinium sp. blooms controlling, and this study provides a new strategy for algicidal bacterium application.

Marine microalgae are regarded as potential feedstock because of their multiple valuable compounds, including lipids, pigments, carbohydrates, and proteins. Some of these compounds exhibit attractive bioactivities, such as carotenoids, ω-3 polyunsaturated fatty acids, polysaccharides, and peptides. However, the production cost of bioactive compounds is quite high, due to the low contents in marine microalgae. Comprehensive utilization of marine microalgae for multiple compounds production instead of the sole product can be an efficient way to increase the economic feasibility of bioactive compounds production and improve the production efficiency. This paper discusses the metabolic network of marine microalgal compounds, and indicates their interaction in biosynthesis pathways. Furthermore, potential applications of co-production of multiple compounds under various cultivation conditions by shifting metabolic flux are discussed, and cultivation strategies based on environmental and/or nutrient conditions are proposed to improve the co-production. Moreover, biorefinery techniques for the integral use of microalgal biomass are summarized. These techniques include the co-extraction of multiple bioactive compounds from marine microalgae by conventional methods, super/subcritical fluids, and ionic liquids, as well as direct utilization and biochemical or thermochemical conversion of microalgal residues. Overall, this review sheds light on the potential of the comprehensive utilization of marine microalgae for improving bioeconomy in practical industrial application.

Despite numerous laboratory studies on physiologies of harmful algal bloom (HAB) species, physiologies of these algae during a natural bloom are understudied. Here, we investigated a bloom of the raphidophyte Heterosigma akashiwo in the East China Sea in 2014 using metabarcode (18S rDNA) and metatranscriptome sequencing. Based on 18S rDNA analyses, the phytoplankton community shifted from high diversity in the pre-bloom stage to H. akashiwo predominance during the bloom. A sharp decrease in ambient dissolved inorganic phosphate and strong up-regulation of phosphate and dissolved organic phosphorus (DOP) uptake genes, including the rarely documented (ppGpp)ase, in H. akashiwo from pre-bloom to bloom was indicative of rapid phosphorus uptake and efficient utilization of DOP that might be a driver of the H. akashiwo bloom. Furthermore, observed up-regulated expression of mixotrophy-related genes suggests potential contribution of mixotrophy to the bloom. Accelerating photosynthetic carbon fixation was also implied by the up-regulation of carbonic anhydrase genes during the bloom. Notably, we also observed a strong morning-to-afternoon shift in the expression of many genes. Our findings provide insights into metabolic processes likely important for H. akashiwo bloom formation, and suggest the need to consider timing of sampling in field studies on this alga., (© 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Dinoflagellates undergo a typical eukaryotic cell cycle consisting of G1, S, G2, and M phases and some of the typical cell cycle related genes have been computationally identified. However, very few of these genes have been experimentally linked to the cell cycle phases. Besides, although thecate dinoflagellates are known to possess theca composed of cellulose, information on cellulose synthesis and degradation associated with the cell cycle is also limited. In this study, we isolated G1/S cyclin, cellulose synthase and cellulase encoding genes in dinoflagellate Prorocentrum donghaiense . Further, using reverse transcription quantitative PCR (RT-qPCR), we characterized the expression profiles of the three genes throughout the cell cycle. All three showed clear expression dynamics throughout the cell cycle, with fold changes of 26, 2.4 and 9.3 for G1/S cyclin, cellulose synthase and cellulase gene, respectively. The transcript abundance of G1/S cyclin increased in late G1 phase and dropped in early S phase, indicating that this protein is involved in the G1/S transition. Throughout the cell cycle, the average transcript level of cellulose synthase was 4.5-fold higher than that of cellulase. Cellulose synthase and cellulase gene expressions showed peak transcript abundances at middle G1 phase and G2M phase, respectively, indicating the respective roles of these enzymes in the growth of newly divided cells and in cytokinesis. Our results suggest that G1/S cyclin, cellulase, and cellulose synthase genes associated with G1/S transition, G2M, and G1 phases of the cell cycle and are candidates of biomarkers for assessing growth status of P. donghaiense .

Phosphorus (P) is an essential nutrient element for the growth of phytoplankton. How P deficiency affects population growth and the cell division cycle in dinoflagellates has only been studied in some species, and how it affects photosynthesis and cell growth remains poorly understood. In the present study, we investigated the impact of P deficiency on the cell division cycle, the abundance of the carbon-fixing enzyme Rubisco, and other cellular characteristics in the Gymnodiniales peridinin-plastid species Amphidinium carterae. We found that under P-replete condition, the cell cycle actively progressed in the culture in a 24-h diel cycle with daily growth rates markedly higher than the P-deficient cultures, in which cells were arrested in the G1 phase and cell size significantly enlarged. The results suggest that, as in previously studied dinoflagellates, P deficiency likely disenables A. carterae to complete DNA duplication or check-point protein phosphorylation. We further found that under P-deficient condition, overall photosystem II quantum efficiency (Fv/Fm ratio) and Rubisco abundance decreased but not significantly, while cellular contents of carbon, nitrogen, and proteins increased significantly. These observations indicated that under P-deficiency, this dinoflagellate was able to continue photosynthesis and carbon fixation, such that proteins and photosynthetically fixed carbon could accumulate resulting in continued cell growth in the absence of division. This is likely an adaptive strategy thereby P-limited cells can be ready to resume the cell division cycle upon resupply of phosphorus.

Alkaline phosphatase (AP) is a key enzyme that enables marine phytoplankton to scavenge phosphorus (P) from dissolved organic phosphorus (DOP) when inorganic phosphate is scarce in the ocean. Yet how the AP gene has evolved in phytoplankton, particularly dinoflagellates, is poorly understood. We sequenced full-length AP genes and corresponding complementary DNA (cDNA) from 15 strains (10 species), representing four classes of the core dinoflagellate lineage, Gymnodiniales, Prorocentrales, Suessiales, and Gonyaulacales. Dinoflagellate AP gene sequences exhibited high variability, containing variable introns, pseudogenes, single nucleotide polymorphisms and consequent variations in amino acid sequence, indicative of gene duplication events and consistent with the "birth-and-death" model of gene evolution. Further sequence comparison showed that dinoflagellate APs likely belong to an atypical type AP (PhoA(aty)), which shares conserved motifs with counterparts in marine bacteria, cyanobacteria, green algae, haptophytes, and stramenopiles. Phylogenetic analysis suggested that PhoA(aty) probably originated from an ancestral gene in bacteria and evolved divergently in marine phytoplankton. Because variations in AP amino acid sequences may lead to differential subcellular localization and potentially different metal ion requirements, the multiple types of APs in algae may have resulted from selection for diversifying strategies to utilize DOP in the P variable marine environment.

The proton pump rhodopsin is widely found in marine bacteria and archaea, where it functions to capture light energy and convert it to ATP. While found in several lineages of dinoflagellates, this gene has not been studied in Prorocentrales species and whether it functionally tunes to light spectra and intensities as in bacteria remains unclear. Here we identified and characterized this gene in the bloom-forming Prorocentrum donghaiense. It is a 7-helix transmembrane polypeptide containing conserved domains and critical amino acid residues of PPR. This gene is phylogenetically affiliated to the xanthorhodopsin clade, but seems to have a distinct evolutionary origin. Quantitative reverse transcription PCR showed that in regular cultures, the transcript abundance of the gene exhibited a clear diel pattern, high abundance in the light period and low in the dark. The same diel pattern was observed for protein abundance with a Western blot using specific antiserum. The rhythm was dampened when the cultures were shifted to continuous dark or light condition, suggesting that this gene is not under circadian clock control. Rhodopsin transcript and protein abundances varied with light intensity, both being highest at a moderate illumination level. Furthermore, the expression of this gene responded to different light spectra, with slightly higher transcript abundance under green than blue light, and lowest abundance under red light. Transformed Escherichia coli over-expressing this rhodopsin gene also exhibited an absorption maximum in the blue-green region with slightly higher absorption in the green. These rhodopsin-promoting light conditions are similar to the relatively turbid marine habitat where the species forms blooms, suggesting that this gene may function to compensate for the light-limited photosynthesis in the dim environment.

Gene structure and expression regulation of form II RuBisCO (rbcII) in dinoflagellates are still poorly understood. Here we isolated this gene (Pdrbc) and investigated its diel expression pattern in a harmful algal bloom forming dinoflagellate Prorocentrum donghaiense. We obtained cDNA sequences with triple tandem repeats of the coding unit (CU); the 5' region has the sequence of a typical dinoflagellate plastid gene, encoding an N-terminus with two transmembrane regions separated by a plastid transit peptide. The CUs (1,455 bp except 1464 bp in last CU) are connected through a 63 bp spacer. Phylogenetic analysis showed that rbcII CUs within species formed monophyletic clusters, indicative of intraspecific gene duplication or purifying evolution. Using quantitative PCR (qPCR) we estimated 117±40 CUs of Pdrbc in the P. donghaiense genome. Although it is commonly believed that most dinoflagellate genes lack transcriptional regulation, our RT-qPCR analysis on synchronized cultures revealed remarkable diel rhythm of Pdrbc expression, showing significant correlations of transcript abundance with the timing of the dark-to-light transition and cell cycle G2M-phase. When the cultures were shifted to continuous light, Pdrbc expression remained significantly correlated with the G2M-phase. Under continuous darkness the cell cycle was arrested at the G1 phase, and the rhythm of Pdrbc transcription disappeared. Our results suggest that dinoflagellate rbcII 1) undergoes duplication or sequence purification within species, 2) is organized in tandem arrays in most species probably to facilitate efficient translation and import of the encoded enzyme, and 3) is regulated transcriptionally in a cell cycle-dependent fashion at least in some dinoflagellates.