Effects of different hydrological conditions on the taxonomic structure and functional traits of mollusk communities in a large floodplain wetland.

Floodplain wetlands are critical to the conservation of aquatic biodiversity and the ecological integrity of river networks. However, increasing drought severity and frequency caused by climate change can reduce floodplain wetlands' resistance and recovery capacities. Mollusks, which are common inhabitants of floodplain wetlands, are among the most vulnerable species to drought. However, the response of mollusk communities to drought has received little attention. Here, we investigated how the structure and functional traits of mollusk communities changed in response to varying hydrological conditions, including a flash drought (FD) in the Poyang Lake floodplain wetland. Our findings showed that FD strongly reduced mollusk abundance and biomass, decreased both α- and ß-diversity, and resulted in the extinction of bivalve taxa. A sudden shift in community trait structure was discovered due to the extinction of many species. These traits, which include deposit feeding, crawling, scraping, aerial respiration, and dormancy, help mollusks survive in FD and tolerate completely dry out of their Changhuchi habitat. Finally, we discovered that dissolved oxygen was an important controlling variable for mollusk communities during drought. Our findings provide a scientific basis for the management and conservation of floodplain wetland biodiversity in the context of increasing drought frequency and intensity.

Geographic origin shapes the adaptive divergences of Rotaria rotatoria (Rotifera, Bdelloidea) to thermal stress: Insights from ecology and transcriptomics.

Global warming has raised concerns regarding the potential impact on aquatic biosafety and health. To illuminate the adaptive mechanisms of bdelloid rotifers in response to global warming, the ecological and transcriptomic characteristics of two strains (HX and ZJ) of Rotaria rotatoria were investigated at 25°C and 35°C. Our results showed an obvious genetic divergence between the two geographic populations. Thermal stress significantly reduced the average lifespan of R. rotatoria in both strains, but increased the offspring production in the ZJ strain. Furthermore, the expression levels of genes Hsp70 were significantly upregulated in the HX strain, while GSTo1 and Cu/Zn-SOD were on the contrary. In the ZJ strain, the expression levels of genes Hsp70, CAT2, and GSTo1 were upregulated under thermal stress. Conversely, a significant decrease in the expression level of the Mn-SOD gene was observed in the ZJ strain under thermal stress. Transcriptomic profiling analysis revealed a total of 105 and 5288 differentially expressed genes (DEGs) in the HX and ZJ strains under thermal stress, respectively. The PCA results showed clear differences in gene expression pattern between HX and ZJ strains under thermal stress. Interestingly, compared to the HX strain, numerous downregulated DEGs in the ZJ strain were enriched into pathways related to metabolism under thermal stress, suggesting that rotifers from the ZJ strain prioritize resource allocation to reproduction by suppressing costly metabolic processes. This finding is consistent with the life table results. This study provides new insights into the adaptive evolution of aquatic animals in the context of global climate change.

Temporal Distribution Patterns of Cryptic Brachionus calyciflorus (Rotifera) Species in Relation to Biogeographical Gradient Associated with Latitude.

Sympatric distribution and temporal overlap of cryptic zooplankton species pose a challenge to the framework of the niche differentiation theory and the mechanisms allowing competitor coexistence. We applied the methods of phylogenetic analysis, DNA taxonomy, and statistical analysis to study the temporal distribution patterns of the cryptic B. calyciflorus species, an excellent model, in three lakes, and to explore the putative mechanisms for their seasonal succession and temporal overlap. The results showed that in the warm-temperate Lake Yunlong, B. fernandoi and B. calyciflorus s.s. underwent a seasonal succession, which was largely attributed to their differential adaptation to water temperature. In the subtropical Lake Jinghu, B. fernandoi, B. calyciflorus s.s., and B. dorcas exhibited both seasonal succession and temporal overlap. Seasonal successions were largely attributed to their differential adaptation to temperature, and temporal overlap resulted from their differential responses to algal food concentration. In the tropical Lake Jinniu, B. calyciflorus s.s. persisted throughout the year and overlapped with B. dorcas for 5 months. The temporal overlap resulted from their differential responses to copepod predation. These results indicated that the temporal distribution pattern of the cryptic B. calyciforus species and the mechanism that allows competitor coexistence vary with different climate zones.

The Multifaceted Effects of Short-Term Acute Hypoxia Stress: Insights into the Tolerance Mechanism of Propsilocerus akamusi (Diptera: Chironomidae).

Plenty of freshwater species, especially macroinvertebrates that are essential to the provision of numerous ecosystem functions, encounter higher mortality due to acute hypoxia. However, within the family Chironomidae, a wide range of tolerance to hypoxia/anoxia is displayed. Propsilocerus akamusi depends on this great tolerance to become a dominant species in eutrophic lakes. To further understand how P. akamusi responds to acute hypoxic stress, we used multi-omics analysis in combination with histomorphological characteristics and physiological indicators. Thus, we set up two groups-a control group (DO 8.4 mg/L) and a hypoxic group (DO 0.39 mg/L)-to evaluate enzyme activity and the transcriptome, metabolome, and histomorphological characteristics. With blue-black chromatin, cell tightness, cell membrane invagination, and the production of apoptotic vesicles, tissue cells displayed typical apoptotic features in the hypoxic group. Although lactate dehydrogenase (LDH), alcohol dehydrogenase (ADH), catalase (CAT), and Na+/K+ -ATPase (NKA) activities were dramatically enhanced under hypoxic stress, glycogen content, and superoxide dismutase (SOD) activities were significantly reduced compared to the control group. The combined analysis of the transcriptome and metabolome, which further demonstrated, in addition to carbohydrates, including glycogen, the involvement of energy metabolism pathways, including fatty acid, protein, trehalose, and glyoxylate cycles, provided additional support for the aforementioned findings. Lactate is the end product of glycogen degradation, and HIF-1 plays an important role in promoting glycogenolysis in acute hypoxic conditions. However, we discovered that the ethanol tested under hypoxic stress likely originates from the symbiodinium of P. akamusi. These results imply that some parameters related to energy metabolism, antioxidant enzyme activities, and histomorphological features may be used as biomarkers of eutrophic lakes in Chironomus riparius larvae. The study also provides a scientific reference for assessing toxicity and favoring policies to reduce their impact on the environment.

The elevated toxicity of the biodegradation product (guanylurea) from metformin and the antagonistic pattern recognition of combined toxicity: Insight from the pharmaceutical risk assessment and the simulated wastewater treatment.

The emerging contaminants metformin (MET) and its degradation product guanylurea (GUA) are released into aquatic environments through wastewater treatment plants (WWTPs). Thus, the environmental risks of wastewater with more treatments may be underestimated due to the lower effect concentration of GUA and the higher detected concentration of GUA in treated wastewater in comparison with MET. In this study, we aimed to investigate the combined toxicity mode of MET and GUA to Brachionus calyciflorus by simulating the degrees of wastewater treatments through adjustments to the ratio of MET and GUA in medium. The results showed that the 24 h-LC50 of MET, GUA, their mixtures of equal concentrations and the mixtures of equal toxic units to B. calyciflorus were 907.44, 544.53, 1185.82 and 940.52 mg/L, respectively, demonstrating GUA is significantly more toxic than MET. An antagonistic interaction between MET and GUA was found in mixture toxicity assessments. Compared with the control, MET treatments only significantly affected the intrinsic rate of population increase of rotifers (rm), while all life-table parameters were significantly affected by GUA. In addition, at medium and high concentrations (120 and 600 µmol/L), the net reproductive rate (R0) and rm of rotifers under GUA were significantly lower than those under MET. Notably, increased proportion of GUA relative to MET in binary-mixture treatments resulted in increased survival risk and reduced fecundity of rotifers. Moreover, the responses of population dynamics to exposures of MET and GUA were mainly attributed to the reproduction of rotifer, indicating that an improved wastewater treatment process is necessary to protect aquatic ecosystems. The study highlights the importance of considering the combined toxicity of emerging contaminants and degradation product in environmental risk assessment, especially the unintentional transformations of parent compound in treated wastewater.

Inhibitory Effects of Antipsychotic Chlorpromazine on the Survival, Reproduction and Population Growth Other Than Neurotransmitters of Zooplankton in Light of Global Warming.

Global warming and environmental pollution have created a unique combination of abiotic and biotic stresses to zooplankton. However, little information is available on the effects of antipsychotic drugs commonly used to treat psychosis, such as chlorpromazine (CPZ), on non-target aquatic organisms in light of global warming. This study investigated how dopamine concentrations (DAC), acute toxicity and chronic toxicity of Brachionus calyciflorus changed in response to CPZ and gradually increasing temperatures. The results showed that the concentration range of rotifer DAC was 1.06~2.51 ng/g. At 18, 25 and 32 °C, the 24 h LC50 was 1.795, 1.242 and 0.833 mg/L, respectively. Compared to the control, exposure to CPZ significantly decreased life expectancy at hatching, the net reproduction rate, generation time, population growth rate and dopamine concentration of B. calyciflorus in all three temperatures (p < 0.05). The toxicity of CPZ to rotifers was increased by high temperature. These findings indicated that CPZ is highly toxic to rotifers, displaying high ecological risks to aquatic ecosystems.

The Relationship between Brachionus calyciflorus-Associated Bacterial and Bacterioplankton Communities in a Subtropical Freshwater Lake.

Zooplankton bodies are organic-rich micro-environments that support fast bacterial growth. Therefore, the abundance of zooplankton-associated bacteria is much higher than that of free-living bacteria, which has profound effects on the nutrient cycling of freshwater ecosystems. However, a detailed analysis of associated bacteria is still less known, especially the relationship between those bacteria and bacterioplankton. In this study, we analyzed the relationships between Brachionus calyciflorus-associated bacterial and bacterioplankton communities in freshwater using high-throughput sequencing. The results indicated that there were significant differences between the two bacterial communities, with only 29.47% sharing OTUs. The alpha diversity of the bacterioplankton community was significantly higher than that of B. calyciflorus-associated bacteria. PCoA analysis showed that the bacterioplankton community gathered deeply, while the B. calyciflorus-associated bacterial community was far away from the whole bacterioplankton community, and the distribution was relatively discrete. CCA analysis suggested that many environmental factors (T, DO, pH, TP, PO43-, NH4+, and NO3-) regulated the community composition of B. calyciflorus-associated bacteria, but the explanatory degree of variability was only 37.80%. High-throughput sequencing revealed that Raoultella and Delftia in Proteobacteria were the dominant genus in the B. calyciflorus-associated bacterial community, and closely related to the biodegradation function. Moreover, several abundant bacterial members participating in carbon and nitrogen cycles were found in the associated bacterial community by network analysis. Predictive results from FAPROTAX showed that the predominant biogeochemical cycle functions of the B. calyciflorus-associated bacterial community were plastic degradation, chemoheterotrophy, and aerobic chemoheterotrophy. Overall, our study expands the current understanding of zooplankton-bacteria interaction and promotes the combination of two different research fields.

Bioaccumulation and biomagnification effects of nano-TiO2 in the aquatic food chain.

The increasing production of nano-TiO2 has attracted extensive concerns about the ecological consequence and health risk of these compounds in natural ecosystem. However, little is known about its toxicity on zooplankton, especially its possibility to access to the food chain via dietary exposure. To address this concern, the toxic and cumulative effects of nano-TiO2 on an aquatic food chain were explored through two trophic levels independently or jointly including producer and consumer. The results revealed that exposure to suspensions of nanomaterials had negative effects on both producers and consumers. Specifically, nanoparticles reduced the density of algal cells in a concentration-dependent way, and hatching life expectancy, average lifespan, net reproductive rate, and population intrinsic growth rate of rotifers decreased significantly with the concentration of nanomaterials increased (P < 0.05). Notably, nanoparticles accumulated in algal cells and were transferred to consumers through dietary exposure. Biomagnification of nano-TiO2 was observed in this simplified food chain, as many of the biomagnification factor (BMF) values in this study were >1. Exposure concentration, exposure time and their interactions play a strong part in the accumulation of nanoparticles in algae and rotifers. Overall, the present findings confirmed that nano-TiO2 was deleterious to plankton, posing a significant environmental threat to aquatic ecosystems. Graphical abstract.

Population dynamics of Brachionus calyciflorus driven by the associated natural bacterioplankton.

Zooplankton provides bacteria with a complex microhabitat richen in organic and inorganic nutrients, and the bacteria community also changes the physiochemical conditions for zooplankton, where the symbiotic relationship between them plays an important role in the nutrient cycle. However, there are few studies on the effect of associated bacteria on the population dynamics of rotifers. In order to make clear their relationships, we reconstructed the associated bacterial community in Brachionus calyciflorus culture, and examined the life history and population growth parameters, and analyzed the diversity and community composition of the associated bacteria at different growth stages of B. calyciflorus. The results showed that the addition of bacteria from natural water can promote the population growth and asexual reproduction of B. calyciflorus, but has no significant effect on sexual reproduction, exhibited by the improvement of its life expectancy at hatching, net reproduction rates and intrinsic growth rate, no significant effects on the generation time and mixis ratio of offspring. It was found that the B. calyciflorus-associated bacterial community was mainly composed of Proteobacteria, Bacteroidota, Actinobacteriota, Cyanobacteria and Firmicutes. Through correlation network analysis, the members of Burkholderiales, Pseudomonadales, Micrococcales, Caulobacterales and Bifidobacteriales were the keystone taxa of B. calyciflorus-associated bacteria. In addition, the relative abundance of some specific bacteria strains increased as the population density of B. calyciflorus increased, such as Hydrogenophaga, Acidovorax, Flavobacterium, Rheinheimera, Novosphingobium and Limnobacter, and their relative abundance increased obviously during the slow and exponential phases of population growth. Meanwhile, the relative abundance of adverse taxa (such as Elizabethkingia and Rickettsiales) decreased significantly with the increase in rotifer population density. In conclusion, the closely associated bacteria are not sufficient for the best growth of B. calyciflorus, and external bacterioplankton is necessary. Furthermore, the function of keystone and rare taxa is necessary for further exploration. The investigation of the symbiotic relationship between zooplankton-associated bacterial and bacterioplankton communities will contribute to monitoring their roles in freshwater ecosystems, and regulate the population dynamics of the micro-food web.

[Spatio-temporal Distribution Characteristics and Driving Factors of Zooplankton in Hongze Lake].

Hongze Lake is the fourth largest freshwater lake in China and is an important source of water for surrounding industrial and agricultural processes and fishery resources. Analyzing the changes in the zooplankton community structure in Hongze Lake can provide scientific support for the scientific management of its ecology and environment. A one-year monthly monitoring study was conducted from March 2017 to February 2018 to analyze the temporal and spatial changes in species composition, density, and biomass of zooplankton in Hongze Lake, as well as the seasonal changes in community diversity and dominant species. Canonical correspondence analysis was employed to explore the relationships between the temporal and spatial changes in zooplankton and the environmental factors of Hongze Lake. The results showed that the average annual density of zooplankton in Hongze Lake was 383.87 ind ·L-1, and the average annual biomass was 1.36 mg ·L-1. The community structure of zooplankton in Hongze Lake varied greatly across time and space. Community structure varied greatly in summer, and zooplankton density and biomass reached a maximum in autumn. The community structure of the zooplankton was the simplest in winter. Chengzi Bay and Lihewa Bay exhibited an abundance of many different zooplankton species with limited spatial differentiation, whereas the zooplankton in the overflow area comprised fewer species but exhibited greater spatial variation. In summer, water level and temperature are the dominant factors, whereas in autumn and winter, the dominant factors are water temperature, nutrients, and chlorophyll. Canonical correspondence analysis showed that the temporal and spatial changes in zooplankton community structure in Hongze Lake were mainly determined by water level, total phosphorus, water temperature, and total nitrogen content. Water level fluctuation has the greatest direct impact on zooplankton community structure, and water quality regulation has indirect impact.

Combined Effects of Temperature and the Microcystin MC-LR on the Feeding Behavior of the Rotifer Brachionus calyciflorus.

The aim of this study was to investigate the responses in filtration and grazing rates of five rotifer strains of the species Brachionus calyciflorus under different temperatures and MC-LR concentrations. The results showed that strain identity, MC-LR concentration, temperature, and the interactions of these factors significantly affected both response variables, with the exception of the interaction of strain and MC-LR on the grazing rates. At low MC-LR concentrations and for the control group, the filtration and grazing rates increased with increasing temperature. The filtering and grazing rates of B. calyciflorus exposed to higher MC-LR concentrations, however, showed no evident enhancement with increasing of temperature. At high temperatures, the filtration and grazing rates of all rotifer strains decreased significantly with increasing concentration of MC-LR, however B. calyciflorus exhibited a refractory stability in the presence of increased MC-LR levels at lower temperatures.

Effects of Multi-metal (Cu, Zn, Cd, Cr, and Mn) Mixtures on the Reproduction of Freshwater Rotifer Brachionus calyciflorus.

In the field, organisms are usually exposed to mixtures of various metals. However, the effects of multi-metal mixtures on growth and reproduction of rotifers remain unknown. In the present study, effects of multi-metal mixtures (Cu, Zn, Cd, Cr, and Mn) on reproduction of the freshwater rotifer Brachionus calyciflorus were assessed by determining various endpoints, including the ratio of ovigerous females to nonovigerous females, the ratio of mictic to amictic females, the mictic rate, the fertilization rate, the population growth rate, and the resting eggs production. The results demonstrated that reproduction of rotifers was significantly affected by all multi-metal mixtures assessed. Moreover, the ratio of mictic to amictic females was the most sensitive endpoint and might be suitable to evaluate effects of multi-metal mixtures to rotifers.

Coal fly ash effluent affects the distributions of Brachionus calyciflorus sibling species.

Fly ash, a coal combustion residue of thermal power plants and a source of multiple pollutants, has been recognized as an environmental hazard all over the world. Although it is known that fly ash effluent affects density, diversity and distribution of rotifers in drainage systems and receiving water bodies, the effect of fly ash effluent on the distributions of highly similar rotifer species remains unknown. In this study, the mtDNA COI genes of 90 individuals in Brachionus calyciflorus complex from Lake Hui (as a fly ash discharge water pond) and other two neighboring lakes (Lake Fengming and Lake Tingtang) were sequenced and analyzed, and the responses in selected life table demographic parameters (life expectancy at hatching, net reproductive rate, intrinsic rate of population increase and proportion of sexual offspring) of different rotifer populations to fly ash effluent were investigated. Overall, 72 mtDNA haplotypes were defined, and were split into two clades by the phylogenetic trees. The divergence of COI gene sequences between the two clades ranged from 11.8% to17.8%, indicating the occurrence of two sibling species (sibling species I and sibling species II). Sibling species I distributed in all the three lakes, showing strong capabilities for dispersal and colonization, which were supported by its higher level of gene flow (2.60-4.04) between the populations from Lake Hui and each of the other two lakes, longer life expectancy at hatching (101.6-148.2 h), and higher net reproductive rate (4.4-16.4 offspring/female) and intrinsic rate of population increase (0.60-0.98/d) when cultured in aerated tap water and fly ash effluent. Sibling species II distributed in both Lake Tingtang and Lake Fengming, showing that its dispersal existed between the two lakes. Considering that the distance between Lake Hui and Lake Fengming is shorter than that between Lake Tingtang and Lake Fengming, sibling species II is able to disperse at least from Lake Fengming to Lake Hui. The restricted distribution of sibling species II in Lake Hui might be attributed to its lower intrinsic rate of population increase (0.34-0.39/d) when cultured in aerated tap water and fiy ash effluent, which might be further lowered by the lower algal food level and quality in Lake Hui.

Effects of DDT and dicofol on population growth of Brachionus calyciflorus under different algal (Scenedesmus obliquus) densities.

A number of organochlorine pesticides, including DDT and dicofol, used to be important in crop protection and management. Their residues may reach water bodies and eventually affect the non-target organisms such as rotifers. In the present study, we evaluated the effects of DDT (0.05, 0.1, 0.2 and 0.4 mg l(-1)) and dicofol (0.1, 0.2, 0.4 and 0.8 mg l(-1)) on the population growth of rotifer Brachionus calyciflorus under two levels of Scenedesmus obliquus (1.0 x 10(6) and 3.0 x 10(6) cell ml(-1)). Regardless of the food level, DDT was more toxic than dicofol to B. calyciflorus. Under low food level, DDT at 0.1 and 0.2 mg l(-1) decreased the population growth rate (r), and DDT at 0.05-0.4 mg l(-1) decreased the maximum population density (K). Dicofol at 0.4 and 0.8 mg l(-1) decreased r and K, but dicofol at 0.2 mg l(-1) increased K. Under high food level, DDT at 0.05-0.2 mg l(-1) increased K, whereas DDT at 0.4 mg l(-1) as well as dicofol at 0.4 and 0.8 mg l(-1) decreased r and K. Increase in food level increased r exposed to DDT at 0.05-0.2 mg l(-1) as well as dicofol at 0.8 mg l(-1), and Kexposed to DDTat 0.05-0.2 mg l(-1) as well as dicofol at 0.1 and 0.2 mg l(-1). DDT concentration, algal density and their interaction affected r and K of B. calyciflorus. Both dicofol concentration and algal density affected r. Dicofol concentration, algal density and their interaction affected K. Both r and K were suitable endpoints for assessing the effects of DDT and dicofol on the rotifers population dynamics under two algal densities, and the latter was more sensitive.

The life-table demographic response of freshwater rotifer Brachionus calyciflorus to multi-metal (Cu, Zn, Cd, Cr, and Mn) mixture interaction.

The effects of multi-metal mixtures on the life-table demography of rotifers are not well known. In this study, the freshwater rotifer Brachionus calyciflorus was exposed to mixture of Cu, Zn, Cd, Cr, and Mn, and the life-table demographic parameters including net reproductive rate, generation time, life expectancy at hatching, and intrinsic rate of population increase were calculated. The results showed that interactions between a given element concentration, except Mn, and the other four elements mixture concentration affected the intrinsic rate of population increase (p < 0.01). Interactions between Zn concentration, as well as Mn, and the other four elements mixture concentration affected the net reproductive rate and the life expectancy at hatching, respectively (p < 0.05). The variation of parameters with the rise of the other four elements mixture concentrations from 0 to high was mainly attributed to the difference of interaction among the five metals mixture with different ratio of concentrations.

Features of Pro-σK important for cleavage by SpoIVFB, an intramembrane metalloprotease.

Intramembrane proteases regulate diverse processes by cleaving substrates within a transmembrane segment or near the membrane surface. Bacillus subtilis SpoIVFB is an intramembrane metalloprotease that cleaves Pro-σ(K) during sporulation. To elucidate features of Pro-σ(K) important for cleavage by SpoIVFB, coexpression of the two proteins in Escherichia coli was used along with cell fractionation. In the absence of SpoIVFB, a portion of the Pro-σ(K) was peripherally membrane associated. This portion was not observed in the presence of SpoIVFB, suggesting that it serves as the substrate. Deletion of Pro-σ(K) residues 2 to 8, addition of residues at its N terminus, or certain single-residue substitutions near the cleavage site impaired cleavage. Certain multiresidue substitutions near the cleavage site changed the position of cleavage, revealing preferences for a small residue preceding the cleavage site N-terminally (i.e., at the P1 position) and a hydrophobic residue at the second position following the cleavage site C-terminally (i.e., P2'). These features appear to be conserved among Pro-σ(K) orthologs. SpoIVFB did not tolerate an aromatic residue at P1 or P2' of Pro-σ(K). A Lys residue at P3' of Pro-σ(K) could not be replaced with Ala unless a Lys was provided farther C-terminally (e.g., at P9'). α-Helix-destabilizing residues near the cleavage site were not crucial for SpoIVFB to cleave Pro-σ(K). The preferences and tolerances of SpoIVFB are somewhat different from those of other intramembrane metalloproteases, perhaps reflecting differences in the interaction of the substrate with the membrane and the enzyme.

Identification and characterization of five intramembrane metalloproteases in Anabaena variabilis.

Regulated intramembrane proteolysis (RIP) involves cleavage of a transmembrane segment of a protein, releasing the active form of a membrane-anchored transcription factor (MTF) or a membrane-tethered signaling protein in response to an extracellular or intracellular signal. RIP is conserved from bacteria to humans and governs many important signaling pathways in both prokaryotes and eukaryotes. Proteases that carry out these cleavages are named intramembrane cleaving proteases (I-CLips). To date, little is known about I-CLips in cyanobacteria. In this study, five putative site-2 type I-Clips (Ava_1070, Ava_1730, Ava_1797, Ava_3438, and Ava_4785) were identified through a genome-wide survey in Anabaena variabilis. Biochemical analysis demonstrated that these five putative A. variabilis site-2 proteases (S2Ps(Av)) have authentic protease activities toward an artificial substrate pro-σ(K), a Bacillus subtilis MTF, in our reconstituted Escherichia coli system. The enzymatic activities of processing pro-σ(K) differ among these five S2Ps(Av). Substitution of glutamic acid (E) by glutamine (Q) in the conserved HEXXH zinc-coordinated motif caused the loss of protease activities in these five S2Ps(Av), suggesting that they belonged to the metalloprotease family. Further mapping of the cleaved peptides of pro-σ(K) by Ava_4785 and Ava_1797 revealed that Ava_4785 and Ava_1797 recognized the same cleavage site in pro-σ(K) as SpoIVFB, a cognate S2P of pro-σ(K) from B. subtilis. Taking these results together, we report here for the first time the identification of five metallo-intramembrane cleaving proteases in Anabaena variabilis. The experimental system described herein should be applicable to studies of other RIP events and amenable to developing in vitro assays for I-CLips.

Genetic differentiation and phylogeographical structure of the Brachionus calyciflorus complex in eastern China.

Spatio-temporal patterns and processes of genetic differentiation in passively dispersing zooplankton are drawing much attention from both ecologists and evolutionary biologists. Two opposite phylogeographical scenarios have already been demonstrated in rotifers, which consist of high levels of genetic differentiation among populations even on small geographical scales on the one hand and the traditionally known cosmopolitanism that is associated with high levels of gene flow and long-distance dispersal via diapausing stages on the other hand. Here, we analysed the population genetic structure and the phylogeography of the Brachionus calyciflorus species complex in eastern China. By screening a total of 318 individuals from ten locations along a 2320-km gradient and analysing samples from two growing seasons, we aimed at focusing on both small- and large-scale patterns. We identified eight cryptic species and verified species status of two of these by sexual reproduction tests. Samples in summer and winter yielded different cryptic species. The distribution patterns of these genetically distinct cryptic species were diverse across eastern China, from full cosmopolitanism to local endemism. The two most abundant cryptic species BcWIII and BcSW showed a pattern of strong genetic differentiation among populations and no significant isolation by distance. Long-distance colonization, secondary contact and recent range expansion are probably responsible for the indistinct pattern of isolation by distance. Our results suggest that geographical distance is more important than temporal segregation across seasons in explaining population differentiation and the occurrence of cryptic species. We explain the current phylogeographical structure in the B. calyciflorus species complex by a combination of recent population expansion, restricted gene flow, priority effects and long-distance colonization.

Patterns and processes in the genetic differentiation of the Brachionus calyciflorus complex, a passively dispersing freshwater zooplankton.

Elucidating the evolutionary patterns and processes of extant species is an important objective of any research program that seeks to understand population divergence and, ultimately, speciation. The island-like nature and temporal fluctuation of limnetic habitats create opportunities for genetic differentiation in rotifers through space and time. To gain further understanding of spatio-temporal patterns of genetic differentiation in rotifers other than the well-studied Brachionus plicatilis complex in brackish water, a total of 318 nrDNA ITS sequences from the B. calyciflorus complex in freshwater were analysed using phylogenetic and phylogeographic methods. DNA taxonomy conducted by both the sequence divergence and the GMYC model suggested the occurrence of six potential cryptic species, supported also by reproductive isolation among the tested lineages. The significant genetic differentiation and non-significant correlation between geographic and genetic distances existed in the most abundant cryptic species, BcI-W and Bc-SW. The large proportion of genetic variability for cryptic species Bc-SW was due to differences between sampling localities within seasons, rather than between different seasons. Nested Clade Analysis suggested allopatric or past fragmentation, contiguous range expansion and long-distance colonization possibly coupled with subsequent fragmentation as the probable main forces shaping the present-day phylogeographic structure of the B. calyciflorus species complex.

[Spatial patterns of genetic differentiation in Brachionus calyciflorus species complex].

To understand spatial patterns of genetic differentiation in the Brachionus calyciflorus species complex, the rDNA ITS regions of 124 rotifer clones collected from eight geographic regions in East China were sequenced and analyzed. A total of 73 haplotypes were defined, and were grouped into three clades by the phylogenetic trees. The divergences of ITS sequence among the three clades ranged from 4.2% to 25.3%, indicating the occurrence of three cryptic species (cryptic species I, cryptic species II and cryptic species III). The sympatric co-occurrence of these cryptic species was observed at Guangzhou, Danzhou and Wuhu. The nucleotide diversity in B. calyciflorus complex was higher, and so was haplotype diversity. Within cryptic species III, the higher genetic differentiation might be attributed to fragmentation and restricted gene flow. In addition, the rapid increase of colonizers after an ambiguous glacial bottleneck might interfere with effective gene flow and accelerate genetic differentiation of geographic populations. The nested clade analysis suggested to some extent the geographic structure, and the non-significant correlation existed between the geographic distance and the average pairwise difference between populations (Da) as well as Fst. The Younger Dryas(YD) Event after the last full glacial period might has considerable effect on the patterns of geographic structure and distribution of cryptic B. calyciflorus species. During the YD event, the three cryptic species probably retreated and co-occurred in multiple relict refugia to the south of Nanling area. Long-distance colonization possibly coupled with subsequent fragmentation might be the main reason of the current patterns of geographic structure. The geographic distribution of B. calyciflorus complex on a global scale might be attributed to the motion of continental plates.