C-Phycocyanin improves the quality of goat oocytes after in vitro maturation and vitrification.

In vitro maturation (IVM) and cryopreservation of goat oocytes are important for establishing a valuable genetic bank for domesticated female animals and improving livestock reproductive efficiency. C-Phycocyanin (PC) is a Spirulina extract with antioxidant, antiinflammatory, and radical scavenging properties. However, whether PC has positive effect on goat oocytes IVM or developmental competence after vitrification is still unknown. In this study, we found that first polar body extrusion (n = 293), cumulus expansion index (n = 269), and parthenogenetic blastocyst formation (n = 281) were facilitated by adding 30 µg/mL PC to the oocyte maturation medium when compared with the control groups and that supplemented with 3, 10, 100 or 300 µg/mL PC (P < 0.05). Although PC supplementation did not affect spindle formation or chromosome alignment (n = 115), it facilitated or improved cortical granules migration (n = 46, P < 0.05), mitochondria distribution (n = 39, P < 0.05), and mitochondrial membrane potential (n = 46, P < 10-4). Meanwhile, supplementation with 30 µg/mL PC in the maturation medium could significantly inhibit the reactive oxygen species accumulation (n = 65, P < 10-4), and cell apoptosis (n = 42, P < 0.05). In addition, PC increased the oocyte mRNA levels of GPX4 (P < 0.01), and decreased the mRNA and protein levels of BAX (P < 0.01). Next, we investigated the effect of PC supplementation in the vitrification solution on oocyte cryopreservation. When compared with the those equilibrate in the vitrification solution without PC, recovered oocytes in the 30 µg/mL PC group showed higher ratios of normal morphology (n = 85, P < 0.05), survival (n = 85, P < 0.05), first polar body extrusion (n = 62, P < 0.05), and parthenogenetic blastocyst formation (n = 107, P < 0.05). Meanwhile, PC supplementation of the vitrification solution increased oocyte mitochondrial membrane potential (n = 53, P < 0.05), decreased the reactive oxygen species accumulation (n = 73, P < 0.05), promoted mitochondria distribution (n = 58, P < 0.05), and inhibited apoptosis (n = 46, P < 10-3). Collectively, our findings suggest that PC improves goat oocyte IVM and vitrification by reducing oxidative stress and early apoptosis, which providing a novel strategy for livestock gamete preservation and utilization.

C-phycocyanin improves the developmental potential of cryopreserved human oocytes by minimizing ROS production and cell apoptosis.

PURPOSE: The cryopreservation process damages oocytes and impairs development potential. As a potent antioxidant, C-phycocyanin (PC) regulates reproductive performance. However, its beneficial effects on vitrified human oocytes remain unknown. METHODS: In this study, human GV-stage oocytes obtained from controlled ovarian hyperstimulation (COH) cycles were randomly allocated to three groups: fresh oocyte without freezing (F group), vitrification in medium supplemented with PC (P group), and vitrification in medium without PC as control group (C group). After warming, viable oocytes underwent in vitro maturation. RESULTS: Our results showed that 3 µg/mL PC treatment increased the oocyte maturation rate after cryopreservation. We also found that PC treatment maintains the regular morphological features of oocytes. After PC treatment, confocal fluorescence staining showed a significant increase in the mitochondrial membrane potential of the vitrified oocytes, along with a notable decrease in intracellular reactive oxygen species and the early apoptosis rate. Finally, after in vitro maturation and parthenogenetic activation, vitrified oocytes had a higher potential for cleavage and blastocyst formation after PC treatment. CONCLUSION: Our results suggest that PC improves the developmental potential of cryopreserved human GV-stage oocytes by attenuating oxidative stress and early apoptosis and increasing the mitochondrial membrane potential.

Population structure and genetic diversity of Tamarix chinensis as revealed with microsatellite markers in two estuarine flats.

Background: Tamarix chinensis Lour. is a 3-6-meter-tall small tree with high salt- and alkali- tolerance and aggressive invasiveness, mainly distributed in the eastern part of China in warm-temperate and subtropical climate zones, yet there is little information available regarding genetic diversity and population structure. Methods: A total of 204 individuals of nine T. chinensis populations were investigated for genetic diversity and population structure using a set of 12 highly polymorphic microsatellite markers. Results: The total number of alleles detected was 162, the average number of effective allele was 4.607, the average polymorphism information content (PIC) value of the 12 loci was 0.685, and the mean observed heterozygosity (Ho) and the mean expected heterozygosity (He) was 0.653 and 0.711, respectively. Analysis of molecular variance (AMOVA) showed a 5.32% genetic variation among T. chinensis populations. Despite a low population differentiation, Bayesian clustering analysis, discriminant analysis of principal components (DAPC) and the unweighted pair group method with arithmetic mean (UPGMA) clearly identified three genetic clusters correlated to the populations' geographic origin: the northern populations including those from Yellow River Delta, the Fangshan (FS) population from Beijing, the Changyi (CY) population from Bohai Bay, the Huanjiabu (HHJ) population from Hangzhou Bay, and the remaining two populations from Hangzhou Bay. There was a significant relationship between the genetic distance and geographical distance of the paired populations. Gene flow (Nm) was 4.254 estimated from FST. Conclusion: T. chinensis possessed high genetic diversity comparable to tree species, and although the population differentiation is shallow, our results classified the sampled populations according to sampling localities, suggesting the different origins of the study populations.

Cumulus Cells Accelerate Postovulatory Oocyte Aging through IL1-IL1R1 Interaction in Mice.

The oocytes of female mammals will undergo aging after ovulation, also known as postovulatory oocyte aging (POA). Until now, the mechanisms of POA have not been fully understood. Although studies have shown that cumulus cells accelerate POA over time, the exact relationship between the two is still unclear. In the study, by employing the methods of mouse cumulus cells and oocytes transcriptome sequencing and experimental verification, we revealed the unique characteristics of cumulus cells and oocytes through ligand-receptor interactions. The results indicate that cumulus cells activated NF-κB signaling in oocytes through the IL1-IL1R1 interaction. Furthermore, it promoted mitochondrial dysfunction, excessive ROS accumulation, and increased early apoptosis, ultimately leading to a decline in the oocyte quality and the appearance of POA. Our results indicate that cumulus cells have a role in accelerating POA, and this result lays a foundation for an in-depth understanding of the molecular mechanism of POA. Moreover, it provides clues for exploring the relationship between cumulus cells and oocytes.

Bacterial community variations in the South China Sea driven by different chemical conditions.

In this study, Illumina MiSeq sequencing of the 16 S rRNA gene was used to describe the bacterial communities in the South China Sea (SCS) during the southwest monsoon period. We targeted different regions in the SCS and showed that bacterial community was driven by the effects of the river, upwelling, and mesoscale eddy through changing the environmental factors (salinity, temperature, and nutrients). Distinct bacterial communities were observed among different chemical conditions, especially between the estuary and the open sea. The abundance of Burkholderiales, Frankiales, Flavobacteriales, and Rhodobacterales dominated the estuary and its adjacent waters. Bacteria in cyclonic eddy were dominated by Methylophilales and Pseudomonadales, whereas Prochlorococcus, SAR11 clade, and Oceanospirillales had relatively high abundance in the anticyclonic eddy. Overall, the abundance of specific phylotypes significantly varied among samples with different chemical conditions. Chemical conditions probably act as a driver that shapes and controls the diversity of bacteria in the SCS. This study suggests that the interaction between microbial and environmental conditions needs to be further considered to fully understand the diversity and function of marine microbes.

Effect of mangrove restoration on sediment properties and bacterial community.

Mangrove reconstruction is an efficient approach for mangrove conservation and restoration. The present study aimed to explore the effects of mangrove reconstruction on sediment properties and bacterial community. The results showed that mangrove restoration greatly promoted sediment fertility, whereas the improvements were more obvious induced by Kandelia obovata when compared to Avicennia marina. In all the samples, the dominant top5 bacterial group were Proteobacteria (48.31-54.52%), Planctomycetes (5.98-8.48%), Bacteroidetes (4.49-11.14%) and Acidobacteria (5.69-8.16%). As for the differences among the groups, the relative abundance of Chloroflexi was higher in the sediments of K. obovata, while Bacteroidetes was more abundant in A. marina group. Furthermore, the two bacterial genera (Rhodoplanes and Novosphingobium) were more dominant in the sediments of K. obovata, while the sediments of A. marina contained higher abundance of Vibrio and Marinobacterium. Besides, bacterial community was highly correlated with mangrove species and sediment property and nutrient status. The results of this study would provide a better understanding of the ecological benefits of mangroves and highlighted the information on biogeochemical processes driven by mangrove restoration and microorganisms.

Radial oxygen loss is correlated with nitrogen nutrition in mangroves.

The present study aimed to explore the possible functions of radial oxygen loss (ROL) on mangrove nutrition. A field survey was conducted to explore the relations among ROL, root anatomy and leaf N in different mangrove species along a continuous tidal gradient. Three mangroves with different ROL (Avicennia marina [A. marina], Kandelia obovata and Rhizophora stylosa) were then selected to further explore the dynamics of N at the root-soil interface. The results showed that seaward pioneer mangrove species such as A. marina appeared to exhibit higher leaf N despite growing under poorer nutrient conditions. Greater leaf N in pioneer mangroves coincided with their special root structure (e.g., high porosity together with a thin lignified/suberized exodermis) and powerful ROL. An interesting positive relation was observed between ROL and leaf N in mangroves. Moreover, rhizo-box data further showed that soil nitrification was also strongly correlated with ROL. A. marina, which had the highest ROL among the three mangrove species studied, consistently possessed the highest levels of NO3-, nitrification and ammonia-oxidizing bacteria and archaea gene copies in the rhizosphere. Besides, both NO3- and NH4+ influxes were found to be higher in the roots of A. marina when compared to those of K. obovata and R. stylosa. In summary, greater N acquisition by pioneer mangroves such as A. marina was strongly correlated with ROL which would regulate N transformation and translocation at the root-soil interface. The implications of this study may be significant in mangrove nutrition and the mechanisms involved in mangrove zonation.

Nitrogen dynamics in the mangrove sediments affected by crabs in the intertidal regions.

Crab is an important benthonic animal in mangrove ecosystem, however, the potential function of crabs on nitrogen (N) transformation in mangrove ecosystems is still poorly understood. The present study aimed to explore the potential effect of crab burrows on nitrification/denitrification within the sediments. The results showed that the presence of crab burrows could directly promote soil nitrification, the regions within more crab burrows appeared to possess higher nitrification. Higher AOA and AOB gene copies were also observed in the sediments surrounding crab burrows than those in the sediments without crab burrow. On the contrary, lower nirS copies, a denitrification related gene, were detected in the sediments surrounding crab burrows. In summary, the present study proposed new evidences of nitrification enhancement deriving by crabs, the presence of crabs might be significant in alleviating nitrification inhibition and benefits the growth of mangroves under tidal flooding.

Distribution and sources of the polycyclic aromatic hydrocarbons in the sediments of the Pearl River estuary, China.

The Pearl River delta, one of the most prosperous economically region in China, has experienced significant contaminant inputs. However, the dynamics of pollutants in the Pearl River estuary and the adjacent coastal areas are still unclear at present. In the paper, distribution and sources of polycyclic aromatic hydrocarbons (PAHs) were investigated in the surface sediments of the Pearl River estuary. The total PAHs concentrations ranged from 126.08 to 3828.58 ng/g with a mean value of 563.52 ng/g, whereas the highest PAHs were observed in Guangzhou channel. Among the U.S. Environmental Protection Agency's 16 priority PAHs, PAHs with 3-4 rings exhibited relative higher levels. A positive relationship was found between PAHs and total organic carbon. The source analysis further showed that the major sources of PAHs in the Pearl River estuary were originated from the pyrolytic inputs, reflecting a mixed energy structure such as wood, coal and petroleum combustion. In summary, although PAHs in Lingding Bay and the adjacent coastal areas of the Pearl River estuary exhibited a relatively low pollution level, the relatively high pollution level of PAHs in Guangzhou channel will be attended.

Identification of cold tolerance genes from leaves of mangrove plant Kandelia obovata by suppression subtractive hybridization.

Low temperature is a major abiotic stress that seriously limits mangrove productivity and distribution, the molecular mechanisms of cold tolerance involved in mangroves are still poorly understood at present. It was used to identify the potential cold-related genes in Kandelia obovata (K. obovata) by suppression subtractive hybridization. 334 cold-related expressed sequence tags (ESTs) out of 670 clones were isolated and sequenced. Among these ESTs, 143 unique cDNAs were identified and classified into ten groups, such as metabolism, energy, cell rescue and defense, transcription and photosynthesis according to NCBI blast. Based on bioinformatics analysis, these ESTs were mainly related to response to stimulus and metabolic process, and were included to 72 KEGG pathways. Two selected genes (e.g., aquaporin gene and zinc family protein gene) from the library were further analyzed by quantitative real-time PCR analysis. Both the two genes were found to be transcriptionally up-regulated under cold stress, which partly approve the construction of the subtractive cDNA library. The diversity of the putative functions of these genes indicated that cold stress resulted in a complex response in K. obovata. Further investigation on the functions and potential pathways of these genes will facilitate the understanding of the molecular adaptations to cold tolerance in mangrove plants.

Variation of phytoplankton community structure from the Pearl River estuary to South China Sea.

The Pearl River is located in the northern part of South China Sea. The environment of the Pearl River estuary (PRE) is significantly impacted by nutrients from anthropogenic activities. Along the anthropogenic pollution gradient from the PRE to South China Sea, the phylogenetic diversity and biomass of phytoplankton was examined in relation to physic-chemical variables. The richness of rbcL gene was higher in the open sea than the estuary, while the concentration of chlorophyll a (Chl a) was higher in the estuary than in the open sea. The cluster analysis of the sequences data resulted in seven phytoplankton community types and the dominant species of phytoplankton changed from Cryptophytes and Diatoms to Prymnesiophytes and Diatoms along the gradient. The community structure of phytoplankton was shaped by nutrients and salinity. The phytoplankton biomass was significantly positively affected by phosphorus, nitrite and ammonium (P < 0.01) but negatively by salinity (P < 0.05); the phytoplankton diversity was highly positively affected by salinity (P < 0.05) but negatively by silicate and nitrate (P < 0.01; P < 0.05, respectively). Anthropogenic activities played a critical role in the phytoplankton distribution and biomass of the study area. Further research is necessary to reveal the influence mechanism of environmental factors on the phytoplankton.

Physiological and biochemical response to drought stress in the leaves of Aegiceras corniculatum and Kandelia obovata.

Drought stress is one of the major abiotic stresses that affects plant growth and metabolism adversely around the world. According to this research, the effect of drought stress on the activity of antioxidative enzymes, soluble sugar, protein and lipid peroxidation were studied in leaves of two mangrove plants, Kandelia obovata and Aegiceras corniculatum. The result showed that superoxide dismutase (SOD) and peroxidase (POD) varied significantly between the leaves and roots studied. The activities increased in different stress levels. The production rate of O 2 (-·) changed with the activity of SOD and POD. Lipid peroxidation was enhanced and Glycine betaine (GB) could decrease the level of malonaldehyde in order to reduce the damage of membrane system. The content of soluble sugar and protein also increased under drought stress and GB helped to eliminate the accumulation of them which somehow enhance the ability of defensing the plants under drought stress. These results indicated that antioxidative activity may play an important role in A. corniculatum and K. obovata and that cell membrane in leaves of K. obovata had greater stability than those of A. corniculatum. Exogenous application of GB had positive effects on A. corniculatum and K. obovata under drought stress which could be products exogenously applied to mangrove plants in order to alleviates the adverse effects.

Spatial variation of phytoplankton community structure in Daya Bay, China.

Daya Bay is one of the largest and most important gulfs in the southern coast of China, in the northern part of the South China Sea. The phylogenetic diversity and spatial distribution of phytoplankton from the Daya Bay surface water and the relationship with the in situ water environment were investigated by the clone library of the large subunit of ribulose-1, 5-bisphosphate carboxylase (rbcL) gene. The dominant species of phytoplankton were diatoms and eustigmatophytes, which accounted for 81.9 % of all the clones of the rbcL genes. Prymnesiophytes were widely spread and wide varieties lived in Daya Bay, whereas the quantity was limited. The community structure of phytoplankton was shaped by pH and salinity and the concentration of silicate, phosphorus and nitrite. The phytoplankton biomass was significantly positively affected by phosphorus and nitrite but negatively by salinity and pH. Therefore, the phytoplankton distribution and biomass from Daya Bay were doubly affected by anthropic activities and natural factors.

Differences in root aeration, iron plaque formation and waterlogging tolerance in six mangroves along a continues tidal gradient.

Mangrove is a special coastal forest along tropical and subtropical intertidal shores. However, how mangroves adapt to tidal flooding and the mechanisms involved in mangrove zonation are still poorly understood. In this study, a pot trial with different tide treatments was conducted to investigate the differences in root anatomy, porosity, radial oxygen loss, iron plaque formation and waterlogging tolerance among six mangroves along a continuous tidal gradient. The index of waterlogging tolerance illustrated that Sonneratia apetala possessed the highest index, followed by Aeguceras corniculatum/Kandelia, Rhizophora stylosa, Heritiera littorlis and Thespesia populnea. Waterlogging tolerances of the mangroves were found to be positively correlated with their root porosity, radial oxygen loss and iron plaque formation. Waterlogging-sensitive species such as landward semi-mangroves exhibited small root porosity and ROL, while waterlogging-tolerant species such as seaward pioneer and rhizophoraceous mangroves exhibited extensive porosity, ROL and iron plaque formation. Nevertheless, grater root porosity and iron plaque formation were detected in permanent waterlogged plants when compared to drained plants. In conclusion, The present study proposes a structural adaptive strategy to tidal flooding in mangroves, such that the mangroves with higher root porosity, ROL and iron plaque appeared to exhibit higher waterlogging tolerance and adaptability in anaerobic foreshores.

Genetic diversity of bacterial communities and gene transfer agents in northern South China Sea.

Pyrosequencing of the 16S ribosomal RNA gene (rDNA) amplicons was performed to investigate the unique distribution of bacterial communities in northern South China Sea (nSCS) and evaluate community structure and spatial differences of bacterial diversity. Cyanobacteria, Proteobacteria, Actinobacteria, and Bacteroidetes constitute the majority of bacteria. The taxonomic description of bacterial communities revealed that more Chroococcales, SAR11 clade, Acidimicrobiales, Rhodobacterales, and Flavobacteriales are present in the nSCS waters than other bacterial groups. Rhodobacterales were less abundant in tropical water (nSCS) than in temperate and cold waters. Furthermore, the diversity of Rhodobacterales based on the gene transfer agent (GTA) major capsid gene (g5) was investigated. Four g5 gene clone libraries were constructed from samples representing different regions and yielded diverse sequences. Fourteen g5 clusters could be identified among 197 nSCS clones. These clusters were also related to known g5 sequences derived from genome-sequenced Rhodobacterales. The composition of g5 sequences in surface water varied with the g5 sequences in the sampling sites; this result indicated that the Rhodobacterales population could be highly diverse in nSCS. Phylogenetic tree analysis result indicated distinguishable diversity patterns among tropical (nSCS), temperate, and cold waters, thereby supporting the niche adaptation of specific Rhodobacterales members in unique environments.

Metal (Pb, Zn and Cu) uptake and tolerance by mangroves in relation to root anatomy and lignification/suberization.

Metal pollution has been widely reported in mangrove wetlands; however, the mechanisms involved in metal detoxification by mangroves are still poorly understood. This study aimed to investigate the possible function of root anatomy and lignification/suberization on metal uptake and tolerance in seedlings of six species of mangroves. The results revealed that the three rhizophoraceous species (Bruguiera gymnorrhiza (L.) Poir, Kandelia obovata Sheue, Liu & Yong and Rhizophora stylosa Griff) consistently exhibited higher metal tolerances than the three pioneer species (Aegiceras corniculatum (Linn.) Blanco, Acanthus ilicifolius L. and Avicennia marina (Forsk.) Viern.). Moreover, metal-tolerant species often exhibited a thick exodermis with high lignification and suberization. The tolerance indices of the mangroves were found to be positively correlated with the amounts of lignin and suberin deposition within the exodermal cell walls. The observed metal uptake by the excised roots further illustrated that a lignified/suberized exodermis directly delayed the entry of metals into the roots, and thereby contributed to a higher tolerance to heavy metals. In summary, the present study proposes a barrier property of the lignified/suberized exodermis in dealing with the stresses of heavy metals, such that the mangroves which possessed more extensive lignification/suberization within the exodermis appeared to exhibit higher metal tolerance.