Overexpression of Zostera japonica J protein gene ZjDjB1 in Arabidopsis enhanced the tolerance to lead stress.

BACKGROUND: Among the heavy metal pollution in soil, lead pollution is particularly prominent. The lead in contaminated soil will not only cause damage to plants, animals and microorganisms, but also seriously affect the progress of the entire ecosystem. Under lead stress, the abundance of DnaJ protein in plants will increase. However, little is known about the role of DnaJ in lead stress. METHODS AND RESULTS: We used transgenic Arabidopsis that overexpressed DnaJ gene ZjDjB1 of Zostera japonica as material to study the role of DnaJ in the mechanism of lead induced stress response. Under lead stress, the seedlings and adult plants of transgenic ZjDjB1 Arabidopsis have higher tolerance to lead stress than wild type. Under lead stress, the content of NO and O2·- free radicals in transgenic ZjDjB1 Arabidopsis was lower than that of wild type. The negative effect of catalase in transgenic ZjDjB1 Arabidopsis under lead stress was weaker than that of wild type. The expression of ABC transporter of mitochondrion 3 (ATM3; systematic name: ABCB25) in transgenic ZjDjB1 Arabidopsis under lead stress was higher than that in wild type. CONCLUSIONS: These results confirmed that ZjDjB1, the DnaJ gene of Z. japonica, was involved in the reaction mechanism to lead pollution, which might improve the tolerance of plants to lead stress by maintaining catalase activity and increasing the expression level of ATM3 under lead stress.

A review of metal contamination in seagrasses with an emphasis on metal kinetics and detoxification.

Seagrasses are important foundation species in coastal ecosystems, and they provide food and habitat that supports high biodiversity. However, seagrasses are increasingly subjected to anthropogenic disturbances such as metal pollution, which has been implicated as a significant factor driving seagrass losses. There have been several reviews synthesizing the metal concentrations in seagrasses and evaluating their utility as biomonitors for metal pollution in the coastal environment at the local scale. However, the interpretation of metal data in seagrass biomonitors requires a more mechanistic understanding of the processes governing metal bioaccumulation and detoxification. In this review, the progress and trends in metal studies in seagrasses between 1973 and 2022 were analyzed to identify frontier topics in this field. In addition, we tried to (1) analyze and assess the current status of metal contamination in seagrasses on a global scale by incorporating more metal data from tropical and Indo-Pacific seagrasses, (2) summarize the geochemical and biological factors governing metal uptake and loss in seagrasses, and (3) provide an up-to-date understanding of metals' effects on seagrasses and their physiological responses to metal challenges. This review improves our understanding of the highly variable metal concentrations observed in the field.

Overexpression of Zostera japonica 14-3-3 gene ZjGRF1 enhances the resistance of transgenic Arabidopsis to copper stress.

BACKGROUND: Copper is both a nutrient essential for plant growth and a pollutant. In recent decades, with the rapid development of industrial and agricultural production, copper has been used more and more widely, and its consumption has also increased rapidly. Excessive soil copper contents induce phytotoxicity, affecting plant growth, development and yields. Moreover, copper can accumulate in crops and enter the food chain through enrichment, harming human health. METHODS AND RESULTS: In this study, Arabidopsis wild-type (WT) and Zostera japonica 14-3-3 gene ZjGRF1 overexpression lines were used to explore the physiological function and molecular mechanism of ZjGRF1 in Arabidopsis in the copper stress response. Under copper stress, compared with WT plants, transgenic ZjGRF1 Arabidopsis plants exhibited less inhibition of root growth and development and had higher fresh weights. Under copper stress, the soluble sugar and soluble protein contents in transgenic ZjGRF1 Arabidopsis plants were significantly higher than those in WT plants, while the superoxide dismutase (SOD), peroxidase and catalase (CAT) activities were significantly higher than those in WT plants. Additionally, the malonaldehyde content of transgenic plants was significantly lower than that of WT plants. Furthermore, qRT-PCR results showed that under copper stress, the SOD, CAT1 and HMA5 expression levels in transgenic ZjGRF1 Arabidopsis plants were significantly higher than those in WT plants, while COPT1 expression was significantly lower than that in WT plants. CONCLUSIONS: ZjGRF1 enhanced the copper stress resistance of Arabidopsis by maintaining high antioxidant enzyme activity, increasing copper efflux and reducing copper uptake under copper stress.

Single-molecule real-time sequencing of the full-length transcriptome of Halophila beccarii.

Ecologically, Halophila beccarii Asch. is considered as a colonizing or a pioneer seagrass species and a "tiny but mighty" seagrass species, since it may recover quickly from disturbance generally. The use of transcriptome technology can provide a better understanding of the physiological processes of seagrasses. To date, little is known about the genome and transcriptome information of H. beccarii. In this study, we used single molecule real-time (SMRT) sequencing to obtain full-length transcriptome data and characterize the transcriptome structure. A total of 11,773 of the 15,348 transcripts were successfully annotated in seven databases. In addition, 1573 long non-coding RNAs, 8402 simple sequence repeats and 2567 transcription factors were predicted in all the transcripts. A GO analysis showed that 5843 transcripts were divided into three categories, including biological process (BP), cellular component (CC) and molecular function (MF). In these three categories, metabolic process (1603 transcripts), protein-containing complex (515 transcripts) and binding (3233 transcripts) were the primary terms in BP, CC, and MF, respectively. The major types of transcription factors were involved in MYB-related and NF-YB families. To the best of our knowledge, this is the first report of the transcriptome of H. beccarii using SMRT sequencing technology.

14-3-3 gene of Zostera japonica ZjGRF1 participates in gibberellin signaling pathway.

BACKGROUND: Zostera japonica is a unique seagrass species in Asia with important ecological value. Gibberellin (GA) is a plant hormone crucial in the regulation of plant growth and development, including seed longevity, seed germination, anti-aging, hypocotyl elongation, leaf development, reproductive organ development, and abiotic stress responses. However, the role of 14-3-3 gene of Z. japonica in GA signaling pathway remains unclear. METHODS AND RESULTS: Herein, gibberellin content and expression of GA synthesis genes were lower in Arabidopsis overexpressing ZjGRF1, 14-3-3 gene of Z. japonica, than in wild type (WT). Moreover, the expression level of GA receptors was lower in transgenic ZjGRF1 Arabidopsis than in WT. The expression level of GA response gene expansin8 (EXP8) was lower in transgenic ZjGRF1 Arabidopsis than in WT. In contrast, the expression levels of PACLOBUTRAZOL RESISTANT1 (PRE1), PRE5 and SCARECROW-LIKE 3 (SCL3) were higher in the transgenic ZjGRF1 Arabidopsis than in WT. Transgenic ZjGRF1 Arabidopsis has decreased sensitivity to paclobutrazol, an inhibitor of GA synthesis. The expression level of flowering regulatory genes was lower in transgenic ZjGRF1 Arabidopsis than in WT. CONCLUSION: This is the first study to report the function of 14-3-3 gene family in seagrass. These findings can be used in future studies on the regulation role of 14-3-3 gene on plant development.

Overexpression of Zostera japonica heat shock protein gene ZjHsp70 enhances the thermotolerance of transgenic Arabidopsis.

BACKGROUND: Heat shock protein 70s (Hsp70s) are major members of the heat shock protein family and play a variety of roles to protect plants against stress. Plant Hsp70s are a conserved and widely expressed family of heat shock proteins. They have two main functional regions: N-terminal nucleic acid binding region and C-terminal substrate binding region. METHODS AND RESULTS: In this study, we cloned the Hsp70 gene of Zostera japonica (ZjHsp70) based on the sequence obtained by transcriptome sequencing. The transcriptional levels of ZjHsp70 increased significantly at 1 h after heat treatment. ZjHsp70 was located in the cytoplasm and nucleus. The overexpression of ZjHsp70 in Arabidopsis resulted in increased heat tolerance, lower contents of malondialdehyde and higher antioxidant enzyme activity than in the wild type. ZjHsp70 may achieve this goal by maintaining highly active antioxidant enzymes. CONCLUSIONS: We show that ZjHsp70 can improve plant heat tolerance by maintaining high antioxidant enzyme activity under high temperature stress. This study provided a basis to study the role of ZjHsp70 in thermotolerance in more detail.

[Latitudinal variations of seed functional traits of Phragmites australis in Chinese coastal marsh]. / 中国滨海沼泽湿地芦苇种子功能性状的纬度梯度变异.

Functional traits of seeds reflect plant reproductive strategies adapting to environmental changes, which is an evolutionary behavior in natural selection and genetics. Study on seed functional traits is of great significance to deeply understand the long-term adaptive evolution of plants and seeds. We measured seed functional traits of a main indigenous species Phragmites australis, including seed size, seed weight, seed set, and seed production, in nine coastal marshes of the six provinces/cities along the coastal zone of China (21°29'-40°57' N), and analyzed latitudinal variations of functional traits. The results showed that seed functional traits of P. australis in Chinese coastal marshes varied significantly with latitude and that there were significant correlations among different traits. Seed size (including seed length, seed width, seed shape index, aspect ratio, and seed surface area), and 100-seed weight showed significant quadratic function relation with latitude, which firstly decreased and then increased with the increases of latitude, while seed setting rate firstly increased and then reduced. There was a trade-off between the number and size of P. australis seeds. Seed production per unit area significantly increased with latitude. Results of stepwise regression analysis showed that climatic factors were the main driver resulting in the difference of seed functional traits of P. australis between latitudes, followed by pH and salinity of soil porewater.

Overexpression of seagrass DnaJ gene ZjDjB1 enhances the thermotolerance of transgenic arabidopsis thaliana.

Seagrass meadows are one of the most important marine resources that grow along the coast. They provide habitat and a food source for animals. They also protect the coast, fix sediment and purify seawater. In the current period of global climate change, anomalies in coastal water temperatures are increasing. A sudden increase in water temperature owing to a heat wave can have a profound effect on seagrass. Zostera japonica is a type of intertidal seagrasses, which is exposed to the air at low tide. High temperatures in the summer often lead to a decline in seagrass meadows. DnaJ proteins, also known as J proteins, are a family of conserved chaperone proteins. They are designated as J proteins because they contain a highly conserved J domain. They function as chaperones of heat shock proteins in organisms. In this study, the role of DnaJ protein (ZjDjB1) of Z. japonica under heat stress was studied. ZjDjB1 was localized to the cytoplasm and nucleus. The overexpression of ZjDjB1 in Arabidopsis thaliana results in an increase in thermotolerance and a decrease in the accumulation of reactive oxygen species and also a reduction in membrane damage. ZjDjB1 may achieve this goal by maintaining a low activity of proteolytic enzymes.

Seagrass beds store less carbon but support more macrobenthos than mangrove forests.

Seagrass beds (SG) have powerful ecosystem functions, but some seagrasses, e.g., Halophila beccarii, have been neglected because of their tiny plant structure and narrow and restricted distribution in the intertidal zone. In this study, we compared ecosystem structure and function between H. beccarii SG and adjacent coastal habitats through: (i) vegetation composition, (ii) carbon storage, and (iii) macrobenthic community. The vegetation carbon storage was only 0.1% of that in mangrove forests (MG). The sediment carbon storage (0-60 cm) of SG (67.4 ± 20.2 MgCorg·ha-1) was 74.31% of MG (90.7 ± 12.9 MgCorg·ha-1) and 13.51% higher than unvegetated flats (UnV) (55.1 ± 17.7 MgCorg·ha-1). The macrobenthic abundance, biomass and secondary productivity in SG were significantly higher than those in MG, indicating that SG support more macrobenthos than MG. However, SG experienced more disturbances than MG and UnV. Therefore, it is necessary to strengthen the protection of H. beccarii SG to preserve their essential functions and services.

Cloning and activity analysis of the promoter of nucleotide exchange factor gene ZjFes1 from the seagrasses Zostera japonica.

After HSP70 binds to the J domain of the substrate and co-chaperone protein, ATP is hydrolyzed to ADP, and the nucleotide exchange factors (NEFs) promote the release of ADP. Under physiological conditions, the nucleotide exchange step is the rate-limiting step, which is accelerated by NEFs. In this study, the promoter of nucleotide exchange factor ZjFes1 was cloned, and its expression in tissues and under heat stress was studied to understand the regulatory mechanism of ZjFes1 and provide the molecular basis to study heat tolerance mechanism of seagrass. It was found that the promoter has common cis-acting elements in promoter and enhancer regions CAAT-box, as well as light response elements AE-box, Box 4 and TCCC-motif, a cis-acting regulatory element essential for the anaerobic induction of ARE, hormone response elements CGTCA-motif and TGACG-motif (MeJA response element), GARE-motif (gibberellin response element), TGA-element (auxin response element), a cis-acting regulatory element related to meristem expression CAT-box, and a cis-acting element involved in defense and stress responsiveness of TC-rich repeats. Two-week-old seedlings exhibited weak GUS activities in their cotyledons. In addition, the AtFes1A promoter was constitutively active in the anthers. After exposure to 38 °C for 2 h, the root tips of two-week-old seedlings were stained a strong blue. Heat-inducible activities of GUS were also observed in the cotyledons, roots, leaves, anthers, sepals and siliques.

Heat-stress induced expression of stress-inducible nucleotide exchange factor Fes1 in seagrass Zostera japonica.

Seagrass meadows are among the four most productive marine natural ecosystems in the world. Zostera japonica (Z. japonica) is the most widely distributed species of seagrass in China. Nucleotide exchange factors (NEFs) promote the release of ADP during heat stress, accelerating the rate-limiting step of Heat shock protein 70 (Hsp70). Although NEFs play an important role in abiotic stress tolerance of plants, NEFs in seagrass have not been studied. In this study, we cloned Fes1 from Z. japonica (ZjFes1) by rapid amplification of the cDNA ends using RACE, and full length ZjFes1 was 1171 bp. It contained an 81 bp 5'-terminal untranslated region (UTR), 109 bp 3'-UTR and 981 bp open reading frame (ORF). The ORF (ZjFes1) was predicted to encode a polypeptide of 326 amino acids with theoretical molecular weight (MW) of 36.10 kDa and pI of 5.22. ZjFes1 shared 89% amino acid identity with Fes1 from Zostera marina (Z. marina). The transcriptional levels of ZjFes1 increased significantly 1 h after heat treatment. ZjFes1 was localized to the cytoplasm. Taken together, we found that ZjFes1 was a stress-inducible gene that may be involved in heat stress response. This study lays the foundation for further studies on the role of ZjFes1 in heat resistance.

Overexpression of Seagrass Nucleotide Exchange Factor Gene ZjFes1 Enhances Heat Tolerance in Transgenic Arabidopsis.

Nucleotide exchange factors (NEFs) play an important role in plant abiotic stress response, but their characteristics and functions in seagrass have not been studied. Zostera japonica (Z. japonica) is one of the most widely distributed seagrass species in China and are distributed in subtropical and temperate coastal areas. Z. japonica is intertidal seagrass, which often undergoes heat stress during summer when the tide ebbs. Overexpression of ZjFes1 in Arabidopsis results in an increase in heat tolerance. We found that ZjFes1 associates with ZjHsp70 in vivo by yeast two-hybrid and bimolecular fluorescence complementarity (BiFC). Overexpression of ZjFes1 leads to selective reduction of Hsp70 transcription and an increase in Hsp101. In conclusion, our results suggest that ZjFes1 may be an active regulator of heat tolerance.

Changes in carbon storage and macrobenthic communities in a mangrove-seagrass ecosystem after the invasion of smooth cordgrass in southern China.

The exotic smooth cordgrass (Spartina alterniflora) has invaded intertidal wetlands near the Ronggenshan tidal flats of Beihai, Guangxi, China, where historically seagrasses and mangroves coexisted. We investigated sediment organic carbon (SOC) storage and macrobenthic community structure in the existing mangroves (MG), S. alterniflora (SA), seagrass bed (SG), and unvegetated flat (UnV) habitats following the S. alterniflora invasion. SOC storage increased after S. alternifolia invasion in the SG and UnV habitats. Spartina alterniflora invasion changed the dominant species of the macrobenthos in the original habitats and reduced the diversity of macrobenthos in SG and UnV habitats. Clearly, S. alternifolia invasion can change the ecological functioning of south China's coastal ecosystems by altering carbon sequestration and affecting biodiversity.

SMRT sequencing of full-length transcriptome of seagrasses Zostera japonica.

Seagrass meadows are among the four most productive marine ecosystems in the world. Zostera japonica (Z. japonica) is the most widely distributed species of seagrass in China. However, there is no reference genome or transcriptome available for Z. japonica, impeding progress in functional genomic and molecular ecology studies in this species. Temperature is the main factor that controls the distribution and growth of seagrass around the world, yet how seagrass responds to heat stress remains poorly understood due to the lack of genomic and transcriptomic data. In this study, we applied a combination of second- and third-generation sequencing technologies to sequence full-length transcriptomes of Z. japonica. In total, we obtained 58,134 uniform transcripts, which included 46,070 high-quality full-length transcript sequences. We identified 15,411 simple sequence repeats, 258 long non-coding RNAs and 28,038 open reading frames. Exposure to heat elicited a complex transcriptional response in genes involved in posttranslational modification, protein turnover and chaperones. Overall, our study provides the first large-scale full-length trascriptome in Zostera japonica, allowing for structural, functional and comparative genomics studies in this important seagrass species. Although previous studies have focused specifically on heat shock proteins, we found that examination of other heat stress related genes is important for studying response to heat stress in seagrass. This study provides a genetic resource for the discovery of genes related to heat stress tolerance in this species. Our transcriptome can be further utilized in future studies to understand the molecular adaptation to heat stress in Zostera japonica.

[Seagrass ecosystems: contributions to and mechanisms of carbon sequestration].

The ocean's vegetated habitats, in particular seagrasses, mangroves and salt marshes, each capture and store a comparable amount of carbon per year, forming the Earth's blue carbon sinks, the most intense carbon sinks on the planet. Seagrass meadows, characterized by high primary productivity, efficient water column filtration and sediment stability, have a pronounced capacity for carbon sequestration. This is enhanced by low decomposition rates in anaerobic seagrass sediments. The carbon captured by seagrass meadows contributes significantly to the total blue carbon. At a global scale, seagrass ecosystems are carbon sink hot spots and have profound influences on the global carbon cycle. This importance combined with the many other functions of seagrass meadows places them among the most valuable ecosystems in the world. Unfortunately, seagrasses are declining globally at an alarming rate owing to anthropogenic disturbances and climate change, making them also among the most threatened ecosystems on the Earth. The role of coastal systems in carbon sequestration has received far too little attention and thus there are still many uncertainties in evaluating carbon sequestration of global seagrass meadows accurately. To better assess the carbon sequestration of global seagrass ecosystems, a number of scientific issues should be considered with high priorities: 1) more accurate measurements of seagrass coverage at national and global levels; 2) more comprehensive research into species- and location-specific carbon sequestration efficiencies; 3) in-depth exploration of the effects of human disturbance and global climate change on carbon capture and storage by seagrass ecosystems.

Natural and human influences on nutrient transport through a small subtropical Chinese estuary.

Global understanding of land-ocean nutrient fluxes increasingly recognizes the disproportionate importance of small rivers. We studied nutrient fluxes from a small catchment in fast developing southern China to uncover effects of land-use. Water was sampled in the macro-tidal estuary of Nanliu River and adjacent Lianzhou Bay in spring and summer of investigate spatial and temporal variations of dissolved nutrients. High riverine concentrations of nitrate (NO3; up to 220 µM) and phosphate (PO4; up to 3.7 µM) mainly originated from agricultural fertilizer input. Riverine dissolved silica (Si; up to 47 µM) increased in the oligosaline part of the estuary through human disturbance of bottom sediments. Dissolved organic nitrogen (DON; up to 194 µM) and ammonium (NH4; up to 40 µM) concentrations increased within the estuary due to inputs from livestock and mussel beds, respectively. Aquaculture ponds contained high concentrations of NH4 (up to 355 µM) and DON (up to 151 µM) but are not an important source to the estuary due to rare wastewater discharge and low absolute nutrient amounts relative to river export. Nutrient concentrations in Lianzhou Bay were low because tidal currents disperse land-derived nutrients offshore into the adjacent Beibu Gulf. A high proportion of regenerated nitrogen in the bay suggests that primary production is sustained by rapid in situ nutrient cycling between primary producers and benthic consumers. High nutrient export makes the Nanliu River an important nutrient source for the north-western South China Sea, despite its proportionately small size. Macro-tide induced short-term concentration changes exceed variability on seasonal and sub-seasonal scales. All nutrients vary inter-annually and between seasons, depending on precipitation-driven river runoff. Total nutrient export to Beibu Gulf coastal waters is stronger during the high discharge period in summer and autumn. In recent years changing nitrogen to phosphorus ratios have alleviated phosphorus limitation in Lianzhou Bay, permitting increased primary productivity.