The WRKY gene family in the halophyte Limonium bicolor: identification, expression analysis, and regulation of salt stress tolerance.

KEY MESSAGE: 63 L. bicolor WRKY genes were identified and their informatics was analyzed. The results suggested that the LbWRKY genes involved in the development and salt secretion of salt glands in L. bicolor. Salt stress, as a universal abiotic stress, severely inhibits the growth and development of plants. WRKY transcription factors play a vital role in plant growth and development, as well as in response to various stresses. Nevertheless, little is known of systematic genome-wide analysis of the WRKY genes in Limonium bicolor, a model recretohalophyte. In this study, 63 L. bicolor WRKY genes were identified (LbWRKY1-63), which were unevenly distributed across seven chromosomes and one scaffold. Based on the structural and phylogenetic characteristics, 63 LbWRKYs are divided into three main groups. Cis-elements in the LbWRKY promoters were related to growth and development, phytohormone responses, and stress responses. Colinearity analysis showed strong colinearity between LbWRKYs and GmWRKYs from soybean (Glycine max). Therefore, LbWRKY genes maybe have similar functions to GmWRKY genes. Expression analysis showed that 28 LbWRKY genes are highly expressed in roots, 9 in stems, 26 in leaves, and 12 in flowers and most LbWRKY genes responded to NaCl, ABA, and PEG6000. Silencing LbWRKY10 reduced salt gland density and salt secretion ability of leaves, and the salt tolerance of the species. Consistent with this, genes associated with salt gland development were markedly down-regulated in the LbWRKY10-silenced lines. Our findings suggested that the LbWRKY genes involved in the development and salt secretion of salt glands in L. bicolor. Our research provides new insights into the functions of the WRKY family in halophytes.

Melatonin increases growth and salt tolerance of Limonium bicolor by improving photosynthetic and antioxidant capacity.

BACKGROUND: Soil salinization is becoming an increasingly serious problem worldwide, resulting in cultivated land loss and desertification, as well as having a serious impact on agriculture and the economy. The indoleamine melatonin (N-acetyl-5-methoxytryptamine) has a wide array of biological roles in plants, including acting as an auxin analog and an antioxidant. Previous studies have shown that exogenous melatonin application alleviates the salt-induced growth inhibition in non-halophyte plants; however, to our knowledge, melatonin effects have not been examined on halophytes, and it is unclear whether melatonin provides similar protection to salt-exposed halophytic plants. RESULTS: We exposed the halophyte Limonium bicolor to salt stress (300 mM) and concomitantly treated the plants with 5 µM melatonin to examine the effect of melatonin on salt tolerance. Exogenous melatonin treatment promoted the growth of L. bicolor under salt stress, as reflected by increasing its fresh weight and leaf area. This increased growth was caused by an increase in net photosynthetic rate and water use efficiency. Treatment of salt-stressed L. bicolor seedlings with 5 µM melatonin also enhanced the activities of antioxidants (superoxide dismutase [SOD], peroxidase [POD], catalase [CAT], and ascorbate peroxidase [APX]), while significantly decreasing the contents of hydrogen peroxide (H2O2), superoxide anion (O2•-), and malondialdehyde (MDA). To screen for L. bicolor genes involved in the above physiological processes, high-throughput RNA sequencing was conducted. A gene ontology enrichment analysis indicated that genes related to photosynthesis, reactive oxygen species scavenging, the auxin-dependent signaling pathway and mitogen-activated protein kinase (MAPK) were highly expressed under melatonin treatment. These data indicated that melatonin improved photosynthesis, decreased reactive oxygen species (ROS) and activated MAPK-mediated antioxidant responses, triggering a downstream MAPK cascade that upregulated the expression of antioxidant-related genes. Thus, melatonin improves the salt tolerance of L. bicolor by increasing photosynthesis and improving cellular redox homeostasis under salt stress. CONCLUSIONS: Our results showed that melatonin can upregulate the expression of genes related to photosynthesis, reactive oxygen species scavenging and mitogen-activated protein kinase (MAPK) of L. bicolor under salt stress, which can improve photosynthesis and antioxidant enzyme activities. Thus melatonin can promote the growth of the species and maintain the homeostasis of reactive oxygen species to alleviate salt stress.

A meta-analysis of the bacterial and archaeal diversity observed in wetland soils.

This study examined the bacterial and archaeal diversity from a worldwide range of wetlands soils and sediments using a meta-analysis approach. All available 16S rRNA gene sequences recovered from wetlands in public databases were retrieved. In November 2012, a total of 12677 bacterial and 1747 archaeal sequences were collected in GenBank. All the bacterial sequences were assigned into 6383 operational taxonomic units (OTUs 0.03), representing 31 known bacterial phyla, predominant with Proteobacteria (2791 OTUs), Bacteroidetes (868 OTUs), Acidobacteria (731 OTUs), Firmicutes (540 OTUs), and Actinobacteria (418 OTUs). The genus Flavobacterium (11.6% of bacterial sequences) was the dominate bacteria in wetlands, followed by Gp1, Nitrosospira, and Nitrosomonas. Archaeal sequences were assigned to 521 OTUs from phyla Euryarchaeota and Crenarchaeota. The dominating archaeal genera were Fervidicoccus and Methanosaeta. Rarefaction analysis indicated that approximately 40% of bacterial and 83% of archaeal diversity in wetland soils and sediments have been presented. Our results should be significant for well-understanding the microbial diversity involved in worldwide wetlands.

Ecological effects of roads on the plant diversity of coastal wetland in the Yellow River Delta.

The 26 sample sites in 7 study plots adjacent to asphalt road and earth road in coastal wetland in the Yellow River Delta were selected to quantify plant diversity using quadrat sampling method in plant bloom phase of July and August 2012. The indice of ß T and Jaccard's coefficient were applied to evaluate the species diversity. The results showed that the plant diversities and alien plants were high in the range of 0-20 m to the road verge. There were more exotics and halophytes in plots of asphalt roadside than that of earth roadside. However, proportion of halophytes in habitats of asphalt roadsides was lower than that of earth roadside. By comparing ß-diversity, there were more common species in the asphalt roadsides than that in the earth roadsides. The similarity of plant communities in studied plots of asphalt roadsides and earth roadsides increased with increasing the distance to road verge. The effect range of roads for plant diversity in study region was about 20 m to road verge. Our results indicate that the construction and maintenance of roads in wetland could increase the plant species diversities of communities and risk of alien species invasion.

Wet and dry atmospheric depositions of inorganic nitrogen during plant growing season in the coastal zone of Yellow River Delta.

The ecological problems caused by dry and wet deposition of atmospheric nitrogen have been widespread concern in the world. In this study, wet and dry atmospheric depositions were monitored in plant growing season in the coastal zone of the Yellow River Delta (YRD) using automatic sampling equipment. The results showed that SO4 (2-) and Na(+) were the predominant anion and cation, respectively, in both wet and dry atmospheric depositions. The total atmospheric nitrogen deposition was ~2264.24 mg m(-2), in which dry atmospheric nitrogen deposition was about 32.02%. The highest values of dry and wet atmospheric nitrogen deposition appeared in May and August, respectively. In the studied area, NO3 (-)-N was the main nitrogen form in dry deposition, while the predominant nitrogen in wet atmospheric deposition was NH4 (+)-N with ~56.51% of total wet atmospheric nitrogen deposition. The average monthly attribution rate of atmospheric deposition of NO3 (-)-N and NH4 (+)-N was ~31.38% and ~20.50% for the contents of NO3 (-)-N and NH4 (+)-N in 0-10 cm soil layer, respectively, suggested that the atmospheric nitrogen was one of main sources for soil nitrogen in coastal zone of the YRD.

Effects of returning paddy field to wetland on composition and stability of soil aggregates in the Yellow River Delta.

The composition and stability of soil aggregates are important indicators for measuring soil quality, which would be affected by land use changes. Taking wetlands with different returning years (2 and 15 years) in the Yellow River Delta as the research object, paddy fields and natural wetlands as control, we analyzed the changes in soil physicochemical properties and soil aggregate composition. The results showed that soil water content, total organic carbon, dissolved organic carbon and total phosphorus of the returning soil (0-40 cm) showed an overall increasing trend with returning period, while soil pH and bulk density was in adverse. There was no significant change in clay content, electrical conductivity, and total nitrogen content. The contents of macro-aggregates and micro-aggregates showed overall increasing and decreasing trend with returning period, respectively. The stability of aggregates in the topsoil (0-10 cm) increased with returning years. Geometric mean diameter and mean weight diameter increased by 8.9% and 40.4% in the 15th year of returning, respectively, while the mass proportion of >2.5 mm fraction decreased by 10.5%. There was no effect of returning on aggregates in subsoil (10-40 cm). Our results indicated that returning paddy field to wetland in the Yellow River Delta would play a positive role in improving soil structure and aggregate stability.

Two birds, one stone: Enhancement of flame retardancy and antibacterial property of viscose fabric using an aminoazole-based cyclotriphosphazene.

Enhancing the fire-retardant and antibacterial properties of viscose fabric through a simple strategy is crucial and urgent. In this study, an aminoazole-based cyclotriphosphazene (HATA) was designed and synthesized through nucleophilic substitution between hexachlorocyclotriphosphazene and 3-amino-1,2,4-triazole. The application of a rapid dipping strategy and the use of 10 wt% HATA aqueous solution significantly increased the limiting oxygen index of the viscose fabric from 19.3 % to 28.4 %. In addition, the HATA-treated fabric exhibited self-extinguishing properties in vertical flame testing. The peak heat release rate of HATA-treated fabric, according to pyrolysis combustion flow calorimetry, significantly decreased by over 83 %. The scanning electron microscope images revealed the original woven fabric structure after burning. The thermogravimetric infrared spectroscopy and X-ray photoelectron spectroscopy results confirmed that the introduction HATA in viscose hindered the release of combustible gas and facilitated the formation of a protective char layer. In addition, 10 % HATA-viscose exhibited remarkable antimicrobial properties, achieving 99.96 % and 99.84 % antibacterial rates against Staphylococcus aureus and Escherichia coli, respectively. Furthermore, HATA-treated viscose fabric exhibited favorable mechanical performance, whiteness, and air permeability. This research provides a simple and effective flame-retardant and antibacterial treatment strategy for viscose fabric.

Corrosion Behavior of High-Nitrogen Steel Hybrid Welded Joints Fabricated by Hybrid Laser-Arc Welding.

To study the corrosion mechanism of high-nitrogen steel welds, this study investigated the effects of laser outputs on the corrosion behavior of high-nitrogen steel hybrid welded joints in hybrid laser-arc welding. The relationship between the ferrite content and laser output was characterized. The ferrite content increased with the increase in the laser power. The corrosion phenomenon first occurred at the two-phase interface, thereby forming corrosion pits. Ferritic dendrites were first corroded to form dendritic corrosion channels. Furthermore, first-principles calculations were performed to investigate the properties of the austenite and ferrite content. The work function and surface energy indicated that solid-solution nitrogen austenite exhibits a higher surface structural stability than austenite and ferrite. This study provides useful information for high-nitrogen steel weld corrosion.

Effect of electron acceptor addition on the temperature sensitivity of soil anaerobic carbon mineralization in the Yellow River Estuary wetland, China.

The temperature sensitivity of soil carbon mineralization (Q10) is an important index to evaluate the responses of ecosystem carbon cycling to climate change. We examined the effects of three electron acceptors [SO42-, NO3- and Fe(Ⅲ)] addition on the Q10 value of anaerobic carbon mineralization of Phragmites australis community soil (0-10 cm) in the Yellow River Estuary wetland with the closed culture-gas chromatography method. The results showed that the three electron acceptors addition inhibited the production of CO2 and CH4 during the 48-day culture period, with a decrease of 17.3%-20.8% for CO2 and 29.2%-36.2% for CH4. Generally, the CO2 production differed with the concentrations of electron acceptors, while CH4 production differed with the type of electron acceptors. The CO2:CH4 ratios were significantly different with temperature, indicating an obvious temperature dependence for the anaerobic carbon mineralization pathway. The Q10 values of CO2 and CH4 production under three electron acceptor additions ranged from 1.08 to 1.11 and from 1.19 to 1.37, respectively, showing an increasing trend compared with the control. The type and concentration of electron acceptors affected the temperature dependence of CO2 production, while electron acceptors affected that of CH4 production. It is suggested that the input of reducing salts would retard the mineralization loss of organic carbon in estuary freshwater wetlands under the background of climate change, but enhance the sensitivity of carbon mineralization to increasing temperature.

Association of COL5A1 gene polymorphisms and musculoskeletal soft tissue injuries: a meta-analysis based on 21 observational studies.

OBJECTIVE: Inconsistent findings existed on the correlation of collagen type V α1 (COL5A1) gene polymorphisms and musculoskeletal soft tissue injuries (MSTIs). The purpose of this study was to collect and combine the current evidences by a meta-analysis approach. METHODS: Six online databases were searched up to August, 2021. The methodological quality of each individual study was evaluated based upon Newcastle-Ottawa Scale (NOS). The strength of the effect size was presented by odds ratio (OR) with 95% confidence interval (95%CI) in five genetic models. The data were analyzed using Review Manager 5.3. RESULTS: Twenty-one studies were eligible to this meta-analysis. The study quality was deemed fair to excellent according to NOS. In the overall analyses, the merged data suggested that rs12722, rs71746744, and rs3196378 polymorphisms were correlated to an increased susceptibility to MSTIs. But the association was not established in rs13946 or rs11103544 polymorphism. For rs12722 polymorphism, stratified analyses by injury type and ethnicity identified the association mainly existed in ligament injury and among Caucasian population. For rs13946 polymorphism, subgroup analysis suggested the association existed in tendon and ligament injuries. CONCLUSION: This study supports that rs12722 is associated with an elevated susceptibility to ligament injury, especially in the Caucasian population. Rs13946 polymorphism appears to increase the risk to tendon and ligament injuries. Rs71746744 and rs3196378 polymorphisms have a tendency to confer an elevated risk to MSTIs. However, no relevance is found between rs11103544 polymorphism and MSTIs.

Association between matrix metalloproteinase-3 gene polymorphisms and tendon-ligament injuries: evidence from a meta-analysis.

BACKGROUND: Tendon-ligament injuries (TLIs), including Achilles tendinopathy, cruciate ligament injury, tennis elbow, rotator cuff injury, patellar tendinopathy, and tibial tendinopathy, are common musculoskeletal soft injuries during physical activity. Matrix metalloproteinase-3 (MMP-3) gene polymorphisms have been implicated in the etiology of TLIs in several genetic association studies with inconsistent results. The purpose of this study was to collect and synthesize the current evidences on the association of MMP-3 polymorphisms and TLIs. METHODS: The search was conducted using PubMed, Web of Science, EMBASE, Cochrane Library, CNKI and Wanfang databases, prior to July, 2021. Newcastle Ottawa Scale was used to appraise the study quality. Strengths of association were represented by odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULTS: Thirteen studies with 2871 cases and 4497 controls met the eligibility criteria, and each study was in high quality. The overall analyzes suggested rs3025058 was associated with an increased TLIs risk (5A vs. 6A, OR = 1.20, 95% CI 1.03-1.40, P = 0.020). However, the association was not found for rs679620, rs591058, and rs650108 polymorphisms. Subgroup analysis by injury type suggested that rs679620 polymorphism was associated with a reduced risk to Achilles tendon rupture (AA + AG vs. GG, OR = 0.46, 95% CI 0.25-0.87, P = 0.020), and rs3025058 was associated with an elevated risk to anterior cruciate ligament injury (5A5A + 5A6A vs. 6A6A, OR = 1.46, 95% CI 1.03-2.06, P = 0.030). When stratified by ethnicity, the findings indicated that rs3025058 polymorphism was associated with an increased TLIs risk among Caucasians (5A6A vs. 6A6A, OR = 1.55, 95% CI 1.09-2.42, P = 0.020) and Brazilians (5A5A vs. 5A6A + 6A6A, OR = 2.80, 95% CI 1.44-5.45, P = 0.002). CONCLUSION: Findings of this study suggest that rs679620 polymorphism is associated with a reduced Achilles tendon rupture risk, and rs3025058 polymorphism contributes to an increased TLIs risk in Caucasians and Brazilians. However, rs591058 and rs650108 polymorphisms do not show any association with TLIs.

Quaternary stratigraphy and paleoenvironmental evolution of the Ningbo Coastal Plain revealed by core Z02.

The Quaternary sediment in the Ningbo Coastal Plain was the deposit due to sea-land interaction, which recorded information of past climate changes. The region is therefore an ideal area to study paleoclimate changes and sedimentary characteristics. We determined the stratigraphic division and paleoenvironmental evolution based on 14C and paleomagnetic dating, along with detailed analyses of lithology, pollen assemblage, foraminifera and ostracodes assemblage, and grain size of sediment in core Z02 located in the southeastern Ningbo Coastal Plain. The results showed that the boundary between the Holocene and Upper Pleistocene in the core Z02 record was at 30.5 m, the boundary between the Upper and Middle Pleistocene was at 82.65 m, and the boundary between the Quaternary and Lower Cretaceous was at 90 m. The Middle Pleistocene section of the core contained few sediments, while the Lower Pleistocene section was completely missed. During the late Pleistocene, the hydrodynamic conditions experienced energy levels of medium to medium low to medium, and sedimentary facies changed from alluvial lake to overbank to river to lake to alluvial lake to lake to overbank. During the Holocene, the hydrodynamic changes experienced energy levels of medium low to low to medium, and the sedimentary facies changed from shoreland to shallow sea to shoreland lake. The Ningbo Coastal Plain had experienced tectonic uplift, weathering and erosion stage in the Early and Middle Pleistocene, from warm and humid to dry in the Late Pleistocene, and from warm and humid to dry and cool in the Holocene, as revealed by the core Z02 record. This study provided useful information in investigating past environmental changes in the subtropical coastal region of eastern China.

Prognostic implications of Kindlin proteins in human osteosarcoma.

The Kindlin protein family, comprising Kindlin-1, Kindlin-2 and Kindlin-3, play important roles in various human cancers. Here, to explore the clinical significance of Kindlins in human osteosarcomas, quantitative real-time PCR and Western blot analyses were performed to detect the expression of Kindlin-1, Kindlin-2 and Kindlin-3 mRNAs and proteins in 20 self-pairs of osteosarcoma and adjacent noncancerous tissues. Then, immunohistochemistry was performed to examine subcellular localizations and expression patterns of Kindlin proteins in 100 osteosarcoma and matched adjacent noncancerous tissues. Kindlin-1, Kindlin-2 and Kindlin-3 protein immunostainings were localized in the cytoplasm, nucleus and cytoplasm, respectively, of tumor cells in primary osteosarcoma tissues. Statistically, the expression levels of Kindlin-1 and Kindlin-2 mRNAs and proteins in osteosarcoma tissues were all significantly higher (both P<0.01), but those of Kindlin-3 mRNA and protein were both dramatically lower (both P<0.05), than in matched adjacent noncancerous tissues. In addition, the overexpressions of Kindlin-1 and Kindlin-2 proteins were both significantly associated with high tumor grade (both P=0.01), presence of metastasis (both P=0.006), recurrence (both P=0.006) and poor response to chemotherapy (both P=0.02). Moreover, Kindlin-1 and Kindlin-2 expressions were both identified as independent prognostic factors for overall (both P=0.01) and disease-free (P=0.02 and 0.01, respectively) survivals of osteosarcoma patients. However, no associations were observed between Kindlin-3 expression and various clinicopathologic features and patients' prognosis. In conclusion, aberrant expression of Kindlin-1 and Kindlin-2 may function as reliable markers for progression and prognosis in osteosarcoma patients, especially for tumor recurrence.

[Study of determination method for zinc in semen and liquor folliculi of artificial fecundation].

Zinc one of the key elements that influence human genetic health, was selectively determined in this paper. In the area of metal elements determination in body fluid, most samples used in the study are blood, plasma and saliva. No literature about genetic fluid determination has been found so far. In this paper the preparation of samples, volume of samples and determination parameters were investigated. An easy reliable method of determining zinc in semen and liquor folliculi was established. The recoveries are 92.0%-107.6%, the detection limit is 0.002 mg x L(-1) and the relative standard deviations (RSD) (n = 5) are 1.3%-10.0%.

Prognostic value of microRNA-223/epithelial cell transforming sequence 2 signaling in patients with osteosarcoma.

MicroRNA-223 (miR-223) has been demonstrated to be implicated in cell proliferation and cell cycle progression of osteosarcoma cell lines by regulating its target gene epithelial cell transforming sequence 2 (ECT2). However, the clinical significance of the deregulation of the miR-223/Ect2 axis in human osteosarcoma has not been fully elucidated. To address this problem, we firstly showed that the expression levels of miR-223 and Ect2 messenger RNA were, respectively, down-regulated and up-regulated in osteosarcoma tissues compared with those in noncancerous bone tissues significantly (both P < .001), according to the results of quantitative real-time reverse transcription-polymerase chain reaction. Notably, miR-223 down-regulation was negatively correlated with Ect2 messenger RNA up-regulation in osteosarcoma tissues (r = -0.68, P = .01). Then, the combined low miR-223 expression and high Ect2 expression (miR-223-low/Ect2-high) was significantly associated with high tumor grade (P = .01), poor response to chemotherapy (P = .01), positive metastasis (P < .001), and recurrence (P < .001) of osteosarcomas. Moreover, patients with miR-223-low/Ect2-high expression had the shortest overall survival (P < .001) and disease-free survival (P < .001) compared with patients in the other 3 groups (miR-223-low/Ect2-low, miR-223-high/Ect2-high, and miR-223-high/Ect2-low). Furthermore, the multivariate analysis identified miR-223/Ect2 expression and the status of metastasis as independent prognostic factors for overall survival and disease-free survival. In conclusion, our data offer convincing evidence that the deregulation of miR-223 and its target gene ECT2 may be associated with the aggressive tumor progression of human osteosarcoma. Of note, the combined miR-223 down-regulation and Ect2 up-regulation may be a possible marker of poor prognosis in this malignancy.