The family of chloride channel proteins that mediate Cl- transportation play vital roles in plant nutrient supply, cellular action potential and turgor pressure adjustment, stomatal movement, hormone signal recognition and transduction, Cl- homeostasis, and abiotic and biotic stress tolerance. The anionic toxicity, mainly caused by chloride ions (Cl-), on plants under salt stress remains poorly understood. In this work, we investigated the function of soybean Cl-/H+ antiporter GmCLC1 under salt stress in transgenic Arabidopsis thaliana, soybean, and yeast. We found that GmCLC1 enhanced salt tolerance in transgenic A. thaliana by reducing the Cl- accumulation in shoots and hence released the negative impact of salt stress on plant growth. Overexpression of GmCLC1 in the hairy roots of soybean sequestered more Cl- in their roots and transferred less Cl- to their shoots, leading to lower relative electrolyte leakage values in the roots and leaves. When either the soybean GmCLC1 or the yeast chloride transporter gene, GEF1, was transformed into the yeast gef1 mutant, and then treated with different chloride salts (MnCl2, KCl, NaCl), enhanced survival rate was observed. The result indicates that GmCLC1 and GEF1 exerted similar effects on alleviating the stress of diverse chloride salts on the yeast gef1 mutant. Together, this work suggests a protective function of GmCLC1 under Cl- stress.

Background The anionic toxicity of plants under salt stress is mainly caused by chloride (Cl − ). Thus Cl − influx, transport and their regulatory mechanisms should be one of the most important aspects of plant salt tolerance studies, but are often sidelined by the focus on sodium (Na + ) toxicity and its associated adaptations. Plant chloride channels (CLCs) are transport proteins for anions including Cl − and nitrate (NO 3 − ), and are critical for nutrition uptake and transport, adjustment of cellular turgor, stomatal movement, signal transduction, and Cl − and NO 3 − homeostasis under salt stress. Results Among the eight soybean CLC genes, the tonoplast-localized c2 has uniquely different transcriptional patterns between cultivated soybean N23674 and wild soybean BB52. Using soybean hairy root transformation, we found that GsCLC-c2 over-expression contributed to Cl − and NO 3 − homeostasis, and therefore conferred salt tolerance, through increasing the accumulation of Cl − in the roots, thereby reducing their transportation to the shoots where most of the cellular damages occur. Also, by keeping relatively high levels of NO 3 − in the aerial part of the plant, GsCLC-c2 could reduce the Cl − /NO 3 − ratio. Wild type GsCLC-c2, but not its mutants ( S184P , E227V and E294G ) with mutations in the conserved domains, is able to complement Saccharomyces cerevisiae △gef1 Cl − sensitive phenotype. Using two-electrode voltage clamp on Xenopus laevis oocytes injected with GsCLC-c2 cRNA, we found that GsCLC-c2 transports both Cl − and NO 3 − with slightly different affinity, and the affinity toward Cl − was pH-independent. Conclusion This study revealed that the expression of GsCLC-c2 is induced by NaCl-stress in the root of wild soybean. The tonoplast localized GsCLC-c2 transports Cl − with a higher affinity than NO 3 − in a pH-independent fashion. GsCLC-c2 probably alleviates salt stress in planta through the sequestration of excess Cl − into the vacuoles of root cells and thus preventing Cl ...

Water is essential for plant growth and development. Water deficiency leads to loss of yield and decreased crop quality. To understand water transport mechanisms in plants, we cloned and characterized a novel tonoplast intrinsic protein (TIP) gene from soybean with the highest similarity to TIP2-type from other plants, and thus designated GmTIP2;3. The protein sequence contains two conserved NPA motifs and six transmembrane domains. The expression analysis indicated that this gene was constitutively expressed in all detected tissues, with higher levels in the root, stem and pod, and the accumulation of GmTIP2;3 transcript showed a significant response to osmotic stresses, including 20% PEG6000 (polyethylene glycol) and 100 μM ABA (abscisic acid) treatments. The promoter-GUS (glucuronidase) activity analysis suggested that GmTIP2;3 was also expressed in the root, stem, and leaf, and preferentially expressed in the stele of root and stem, and the core promoter region was 1000 bp in length, located upstream of the ATG start codon. The GUS tissue and induced expression observations were consistent with the findings in soybean. In addition, subcellular localization showed that GmTIP2;3 was a plasma membrane-localized protein. Yeast heterologous expression revealed that GmTIP2;3 could improve tolerance to osmotic stress in yeast cells. Integrating these results, GmTIP2;3 might play an important role in response to osmotic stress in plants.

Plant response to environmental stresses is regulated by a complicated network of regulatory and functional genes. In this study, we isolated the putative stress-associated gene GmZFP3 (a C2H2-type Zinc finger protein gene) based on the previous finding that it was one of two genes located in the QTL region between the Satt590 and Satt567 markers related to soybean tolerance to drought. Temporal and spatial expression analysis using quantitative real-time PCR indicated that GmZFP3 was primarily expressed in roots, stems and leaf organs and was expressed at low levels in flowers and soybean pods. Moreover, GmZFP3 expression increased in response to polyethylene glycol (PEG) and Abscisic acid (ABA) treatments. In addition, subcellular localization analysis indicated that GmZFP3 was ubiquitously distributed in plant cells. Transgenic experiments indicated that GmZFP3 played a negative role in plant tolerance to drought. Analysis of ABA-related marker gene expression in Arabidopsis suggested that GmZFP3 might be involved in the ABA-dependent pathway during the drought stress response. Taken together, these results suggest that soybean GmZFP3 negatively regulates the drought response.

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Purpose: No final conclusion has yet been reached on characteristics of postoperative pain and pain-related factors after video-assisted thoracoscopic surgery (VATS). This study was designed to explore features of acute severe pain and chronic post-surgical pain (CPSP), and the pain-related factors of VATS. Patients and Methods: Data of patients who underwent VATS for lung cancer in Cancer Hospital, Chinese Academy of Medical Sciences between March 2017 and January 2019 were reviewed in this retrospective study. A numerical rating scale (NRS) was used for evaluating the intensity of postoperative pain including no pain (NRS=0), mild pain (NRS=1-3), moderate pain (NRS=4-6), and severe pain (NRS=7-10). Pain intensity was assessed daily within a week after operation, and also evaluated at 3 months postoperatively. Results: One hundred and five (3.4%) of the 3072 patients enrolled experienced severe pain (NRS=7-10) on the 1st day after operation, and 17 (0.6%) on the 2nd day. Smoking history, three-port VATS, prolonged operation time, and without patient-controlled analgesia (PCA) were correlated to increased incidence of severe pain. Among all patients, 237 (7.7%) cases generated CPSP, and VATS type, operation time, duration of drainage, and severe pain on the 1st day were four independent risk factors related to CPSP. Conclusion: Patients seemed to experience a lower incidence of acute severe pain and CPSP after VATS than traditional open surgery. Acute severe pain was correlated with smoking history, VATS type, operation time, and PCA; VATS type, operation time, duration of drainage, and severe pain on the 1st day postoperatively were four independent risk factors of CPSP.

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Dendrobium huoshanense is used to treat various diseases in traditional Chinese medicine. Recent studies have identified active components. However, the lack of genomic data limits research on the biosynthesis and application of these therapeutic ingredients. To address this issue, we generated the first chromosome-level genome assembly and annotation of D. huoshanense. We integrated PacBio sequencing data, Illumina paired-end sequencing data, and Hi-C sequencing data to assemble a 1.285 Gb genome, with contig and scaffold N50 lengths of 598 kb and 71.79Mb, respectively. We annotated 21,070 protein-coding genes and 0.96 Gb transposable elements, constituting 74.92% of the whole assembly. In addition, we identified 252 genes responsible for polysaccharide biosynthesis by Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation. Our data provide a basis for further functional studies, particularly those focused on genes related to glycan biosynthesis and metabolism, and have implications for both conservation and medicine.