ABSTRACT
Legumes have evolved a nitrogen-fixing symbiotic interaction with rhizobia, and this association helps them to cope with the limited nitrogen conditions in soil. The compatible interaction between the host plant and rhizobia leads to the formation of root nodules, wherein internalization and transition of rhizobia into their symbiotic form, termed bacteroids, occur. Rhizobia in the nodules of the Inverted Repeat-Lacking Clade legumes, including Medicago truncatula, undergo terminal differentiation, resulting in elongated and endoreduplicated bacteroids. This transition of endocytosed rhizobia is mediated by a large gene family of host-produced nodule-specific cysteine-rich (NCR) peptides in M. truncatula. Few NCRs have been recently found to be essential for complete differentiation and persistence of bacteroids. Here, we show that a M. truncatula symbiotic mutant FN9285, defective in the complete transition of rhizobia, is deficient in a cluster of NCR genes. More specifically, we show that the loss of the duplicated genes NCR086 and NCR314 in the A17 genotype, found in a single copy in Medicago littoralis R108, is responsible for the ineffective symbiotic phenotype of FN9285. The NCR086 and NCR314 gene pair encodes the same mature peptide but their transcriptional activity varies considerably. Nevertheless, both genes can restore the effective symbiosis in FN9285 indicating that their complementation ability does not depend on the strength of their expression activity. The identification of the NCR086/NCR314 peptide, essential for complete bacteroid differentiation, has extended the list of peptides, from a gene family of several hundred members, that are essential for effective nitrogen-fixing symbiosis in M. truncatula.
Subject(s)
Medicago truncatula , Multigene Family , Plant Proteins , Root Nodules, Plant , Symbiosis , Medicago truncatula/microbiology , Medicago truncatula/genetics , Medicago truncatula/physiology , Root Nodules, Plant/microbiology , Root Nodules, Plant/genetics , Symbiosis/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Regulation, Plant , Rhizobium/physiology , Rhizobium/genetics , Nitrogen Fixation/genetics , Peptides/metabolism , Peptides/genetics , Sinorhizobium meliloti/physiology , Sinorhizobium meliloti/genetics , Cysteine/metabolismABSTRACT
The bio-based coatings of cellulose fabrics (cotton) had attracted increasing attention for multifunction and sustainability but suffered from poor durability and low efficiency. Here, the aldehyde-free and durable coatings for cotton fabrics (CPZ@CF) with satisfactory flame retardancy, antibacteria as well as wearing performance were prepared through the interfacial coordination effect where the well-organized zinc phytate complex were in situ grew on the pre-treated surface of cotton fabrics with chitosan (CS) and Zn2+. The CZP@CF exhibited excellent antibacterial activity for Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) with 99.99 % antibacterial rates benefiting from the synergistic effect between Zn2+ and CS. Meanwhile, even the CPZ coatings loading was only 1.5 wt%, the fire safety of CZP@CF remarkably enhanced owing to the excellent synergistic catalytic charring and free radical capture. More importantly, the antibacterial rates of CZP@CF for S. aureus and E. coli still reached 99.99 % and 91.67 % after 50 washing cycles. Additionally, this treatment method did not deteriorate the fabrics properties, including mechanical and breathability as well as wearing performance, which provided the approach to fabricate the flame retardant and antibacterial textiles with well durability and wearing performance.
Subject(s)
Cellulose , Chitosan , Cotton Fiber , Escherichia coli , Staphylococcus aureus , Aldehydes , Anti-Bacterial AgentsABSTRACT
Medicago truncatula has been selected as one of the model legume species for gene functional studies. To elucidate the functions of the very large number of genes present in plant genomes, genetic mutant resources are very useful and necessary tools. Fast Neutron (FN) mutagenesis is effective in inducing deletion mutations in genomes of diverse species. Through this method, we have generated a large mutant resource in M. truncatula. This mutant resources have been used to screen for different mutant using a forward genetics methods. We have isolated and identified a large amount of symbiotic nitrogen fixation (SNF) deficiency mutants. Here, we describe the detail procedures that are being used to characterize symbiotic mutants in M. truncatula. In recent years, whole genome sequencing has been used to speed up and scale up the deletion identification in the mutant. Using this method, we have successfully isolated a SNF defective mutant FN007 and identified that it has a large segment deletion on chromosome 3. The causal deletion in the mutant was confirmed by tail PCR amplication and sequencing. Our results illustrate the utility of whole genome sequencing analysis in the characterization of FN induced deletion mutants for gene discovery and functional studies in the M. truncatula. It is expected to improve our understanding of molecular mechanisms underlying symbiotic nitrogen fixation in legume plants to a great extent.
ABSTRACT
In the "treat all" era, the high rate of late HIV diagnosis (LHD) worldwide remains an impediment to ending the HIV epidemic. In this study, we analyzed LHD in newly diagnosed people living with HIV (PLWH) and its impact on HIV transmission in Northeast China. Sociodemographic information, baseline clinical data, and plasma samples obtained from all newly diagnosed PLWH in Shenyang, the largest city in Northeast China, between 2016 and 2019 were evaluated. Multivariate logistic regression analysis was performed to identify risk factors associated with LHD. A molecular network based on the HIV pol gene was constructed to assess the risk of HIV transmission with LHD. A total of 2882 PLWH, including 882 (30.6%) patients with LHD and 1390 (48.2%) patients with non-LHD, were enrolled. The risk factors for LHD were older age (≥ 30 years: p < .01) and diagnosis in the general population through physical examination (p < .0001). Moreover, the molecular network analysis revealed that the clustering rate (p < .0001), the fraction of individuals with ≥ 4 links (p = .0847), and the fraction of individuals linked to recent HIV infection (p < .0001) for LHD were significantly or marginally significantly lower than those recorded for non-LHD. Our study indicates the major risk factors associated with LHD in Shenyang and their limited contribution to HIV transmission, revealing that the peak of HIV transmission of LHD at diagnosis may have been missed. Early detection, diagnosis, and timely intervention for LHD may prevent HIV transmission.
Subject(s)
HIV Infections , Humans , HIV Infections/diagnosis , HIV Infections/epidemiology , HIV Infections/drug therapy , Risk Factors , Early Diagnosis , China/epidemiologyABSTRACT
Universal antiretroviral therapy (ART, "treat all") was recommended by the World Health Organization in 2015; however, HIV-1 transmission is still ongoing. This study characterizes the drivers of HIV transmission in the "treat all" era. Demographic and clinical information and HIV pol gene were collected from all newly diagnosed cases in Shenyang, the largest city in Northeast China, during 2016 to 2019. Molecular networks were constructed based on genetic distance and logistic regression analysis was used to assess potential transmission source characteristics. The cumulative ART coverage in Shenyang increased significantly from 77.0% (485/630) in 2016 to 93.0% (2598/2794) in 2019 (p < 0.001). Molecular networks showed that recent HIV infections linked to untreated individuals decreased from 61.6% in 2017 to 28.9% in 2019, while linking to individuals with viral suppression (VS) increased from 9.0% to 49.0% during the same time frame (p < 0.001). Undiagnosed people living with HIV (PLWH) hidden behind the links between index cases and individuals with VS were likely to be male, younger than 25 years of age, with Manchu nationality (p < 0.05). HIV transmission has declined significantly in the era of "treat all". Undiagnosed PLWH may drive HIV transmission and should be the target for early detection and intervention.
Subject(s)
HIV Infections , HIV-1 , China/epidemiology , Female , Genes, pol , HIV Infections/diagnosis , HIV Infections/drug therapy , HIV Infections/epidemiology , HIV-1/genetics , Humans , Male , Specimen HandlingABSTRACT
Objective: To explore the effects of pamidronate disodium combined with calcium on BMD values and the severity of pain in elderly patients with osteoporosis based on the mobile terminal platform for the Internet of Things. Methods: The data of 120 patients admitted to our hospital from January 2019 to December 2020 were retrospectively analyzed. According to the patients' condition and medication wills, they were divided into the experimental group (n = 68) and the control group (n = 52). All patients were given chronic disease management based on the mobile terminals for the Internet of Things, and they received the treatment of bisphosphonates and calcium, with the supplement of calcium at a daily dose of 1000 mg. The control group was given alendronate sodium once a week, and the experimental group was given pamidronate disodium by intravenous infusion three times a month, with the treatment cycle as 1 year. The patients' bone mineral density (BMD) values and the pain indexes were compared after treatment. Results: There was no statistical difference in general information between the two groups (p > 0.05). The BMD values of the lumbar vertebrae L2-4, total hip, and femur neck at 6 months and 1 year after treatment in the experimental group were significantly higher than those in the control group (p < 0.001). The pain scores at 6 months and 1 year after treatment in the experimental group were significantly lower than those in the control group (p < 0.001). Conclusion: The treatment of pamidronate disodium combined with calcium based on the mobile terminal platform for the Internet of Things can reduce the severity of pain in elderly patients with osteoporosis and improve the BMD, which has a generalization value.
Subject(s)
Internet of Things , Osteoporosis , Aged , Bone Density , Calcium , Diphosphonates/therapeutic use , Humans , Lumbar Vertebrae , Osteoporosis/drug therapy , Pain/drug therapy , Pamidronate/pharmacology , Pamidronate/therapeutic use , Retrospective StudiesABSTRACT
We estimated the optimum time to initiate antiretroviral therapy (ART) in a retrospective observational cohort. We observed that ART initiation 7âdays or less ( n â=â817) and 8-30âdays ( n â=â1009) were the most important factors with viral suppression, and had similar viral suppression rate, CD4 + T-cell count increase and fractions of individuals with links at least 4 and individuals linked to recent HIV infection in HIV molecular networks. This study provides real-world evidence on the benefits of rapid ART initiation in resource-limited setting.
Subject(s)
Anti-HIV Agents , HIV Infections , Anti-HIV Agents/therapeutic use , CD4 Lymphocyte Count , Cohort Studies , HIV Infections/diagnosis , HIV Infections/drug therapy , Humans , Retrospective Studies , Viral LoadABSTRACT
The formation and development of legumes nodules requires a lot of energy. Legumes must strictly control the number and activity of nodules to ensure efficient energy distribution. The AON system can limit the number of rhizobia infections and nodule numbers through the systemic signal pathway network that the aboveground and belowground parts participate in together. It can also promote the formation of nodules when plants are deficient in nitrogen. The currently known AON pathway includes four parts: soil NO3- signal and Rhizobium signal recognition and transmission, CLE-SUNN is the negative regulation pathway, CEP-CRA2 is the positive regulation pathway and the miR2111/TML module regulates nodule formation and development. In order to ensure the biological function of this important approach, plants use a variety of plant hormones, polypeptides, receptor kinases, transcription factors and miRNAs for signal transmission and transcriptional regulation. This review summarizes and discusses the research progress of the AON pathway in Legume nodule development.
Subject(s)
Fabaceae , Rhizobium , Self-Control , Fabaceae/genetics , Fabaceae/metabolism , Gene Expression Regulation, Plant , Homeostasis , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Root Nodulation/genetics , Rhizobium/metabolism , Root Nodules, Plant/metabolism , Symbiosis/physiologyABSTRACT
Postmenopausal osteoporosis (PMOP) is known as one of the prevalent diseases among middle-aged and elderly women. This paper revolves around the alteration of miR-211-5p in PMOP patients and its function in osteogenic differentiation. Quantitative real-time polymerase chain reaction (qRT-PCR) was implemented to check the miR-211-5p level in the plasma of PMOP patients. Knockdown and overexpression experiments were done to verify the influence of miR-211-5p on human-derived mesenchymal stem cell (hMSC) osteogenic differentiation and osteogenesis. The alkaline phosphatase (ALP) assay kit was taken to test ALP activity. Alizarin red staining monitored osteogenic differentiation, while oil red O staining examined adipogenesis. Western blot confirmed the profiles of osteoclastogenesis-concerned factors (TRAP, NFAT2, c-FOS, Runx2, OCN, CTSK), dual specific phosphatase 6 (DUSP6), ERK, SMAD, and ß-catenin. Dual-luciferase reporter and RNA immunoprecipitation assays were implemented to identify the association between miR-211-5p and DUSP6. Our data displayed that miR-211-5p was down-regulated in the PMOP patients' plasma (in contrast with the healthy controls), and it was positively correlated with Vit-D and BMD levels. miR-211-5p overexpression vigorously facilitated hMSC osteogenic differentiation, while miR-211-5p inhibition contributed to the opposite situation. miR-211-5p initiated the ERK/SMAD/ß-catenin pathway and repressed DUSP6's expression. Overexpression of DUSP6 counteracted the miR-211-5p-mediated function to a great extent and inactivated ERK/SMAD/ß-catenin, whereas enhancing ERK phosphorylation weakened the DUSP6 overexpression-induced function. Consequently, this research unveiled that miR-211-5p promotes osteogenic differentiation by interfering with the DUSP6-mediated ERK/SMAD/ß-catenin pathway.
Subject(s)
Dual Specificity Phosphatase 6 , Mesenchymal Stem Cells , MicroRNAs , Osteoporosis, Postmenopausal , Aged , Cell Differentiation/genetics , Cells, Cultured , Dual Specificity Phosphatase 6/metabolism , Female , Humans , Mesenchymal Stem Cells/metabolism , MicroRNAs/genetics , Middle Aged , Osteogenesis/genetics , Osteoporosis, Postmenopausal/genetics , Osteoporosis, Postmenopausal/metabolism , beta Catenin/genetics , beta Catenin/metabolismABSTRACT
Circular RNA (circRNA) is a crucial regulator in multiple human diseases, including osteoporosis (OP). However, the function of numerous circRNAs remains unclear. This study aimed to explore the role and mechanism of circ_0019693 in bone marrow mesenchymal stem cell (BMSC) osteogenic differentiation and osteogenesis-coupled angiogenesis. The expression of circ_0019693, miR-942-5p and purkinje cell protein 4 (PCP4) was measured using quantitative real-time PCR (qPCR) or Western blot. Osteogenic differentiation was monitored according to the protein levels of RUNX family transcription factor 2 (RUNX2), osteopontin (OPN) and osteocalcin (OCN) by Western blot analysis, and the activity of alkaline phosphatase (ALP). Angiogenesis was evaluated by tube formation assay. The targeting relationship between miR-942-5p and circ_0019693 or PCP4 was identified using pull-down, dual-luciferase reporter, and RNA immunoprecipitation assays. Circ_0019693 was downregulated in serum samples and bone tissues from OP patients relative to normal subjects. Circ_0019693 expression was enhanced in the stages of BMSC osteogenic differentiation. Circ_0019693 overexpression enhanced the activity of ALP and the expression of RUNX2, OPN and OCN, and its overexpression also promoted angiogenesis. However, circ_0019693 knockdown played the opposite effects. MiR-942-5p was ensured to be a target of circ_0019693, and miR-942-5p enrichment reversed the effects of circ_0019693. In addition, PCP4 was a target of miR-942-5p, and miR-942-5p inhibitor-promoted BMSC osteogenic differentiation and angiogenesis were partly repressed by PCP4 knockdown. In conclusion, circ_0019693 promotes BMSC osteogenic differentiation osteogenesis-coupled angiogenesis via regulating miR-942-5p-targeted PCP4, thus blocking the development of OP.
Subject(s)
Bone Marrow Cells/metabolism , Cell Differentiation , Mesenchymal Stem Cells/metabolism , MicroRNAs/metabolism , Neovascularization, Pathologic/metabolism , Nerve Tissue Proteins/metabolism , Osteogenesis , Osteoporosis/metabolism , RNA, Circular/metabolism , Aged , Aged, 80 and over , Female , Humans , Male , MicroRNAs/genetics , Middle Aged , Neovascularization, Pathologic/genetics , Nerve Tissue Proteins/genetics , Osteoporosis/genetics , RNA, Circular/geneticsABSTRACT
Ischemic stroke is a kind of disease with high mortality and high disability, which brings a huge burden to the public health system (Hu et al. (2017)), and it poses a serious threat to the quality of life of patients. Cerebral ischemia/reperfusion injury is an important pathophysiological mechanism. This study aims to assess the mechanism of SNHG15 in the occurrence and development of cerebral ischemia/reperfusion injury of nerve cells and to investigate its potential value for diagnosis and treatment. SNHG15 targeted miRNA molecules and target genes were predicted with bioinformatics tools such as StarBase and TargetScan. The process of ischemic reperfusion in cerebral apoplexy in normal cultured and oxygen-glucose-deprived and reoxygenated neurons was simulated with RT-PCR and western blot technique. The expressions of SNHG15 and miR-141 were detected with qPCR, and the expressions of SIRT1 and p65, TNF-α, ROS, iNOS, and IL-6 were detected with western blot. Meanwhile, SNHG15 siRNAs and miR-141 mimics were transfected for SH-SY5Y, with western blot testing. And the expressions of miR-141, SIRT1, and p65, TNF-α, ROS, iNOS, and IL-6 were tested. According to the prediction with bioinformatics tools of StarBase and TargetScan, miR-141 is the target of lncSNHG15. In the luciferase reporter plasmid double-luciferase assay, miR-141 and SIRT1 were defined as the target relationship. In the oxygen-glucose-deprived reoxygenation model group, SNHG15 expression increased, miR-141 expression decreased, SIRT1 expression increased, and the expressions of p65, TNF-α, ROS, iNOS, and IL-6 decreased. In the SNHG15-siRNA-transfected oxygen-glucose-deprived reoxygenation cell model group, miR-141 expression increased, SIRT1 expression decreased, and the expressions of p65, TNF-α, and IL-6 increased compared with the si-NC group. In the miR-141-mimic-transfected oxygen-glucose-deprived reoxygenation cell model, SNHG15 expression decreased, SIRT1 expression decreased, and the expressions of p65, TNF-α, IL-1ß, and IL-6 increased. In conclusion, SNHG15 expression increased during the process of oxygen-glucose-deprived reoxygenation, and the oxidative stress process was reduced by miR-141/SIRT1.
Subject(s)
Brain Ischemia , MicroRNAs , RNA, Long Noncoding , Reperfusion Injury , Apoptosis/genetics , Brain Ischemia/genetics , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , Oxidative Stress/genetics , Quality of Life , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Reperfusion Injury/genetics , Reperfusion Injury/metabolism , Sirtuin 1/genetics , Sirtuin 1/metabolismABSTRACT
BACKGROUND: Circular RNAs (circRNAs) can act as vital players in osteoarthritis (OA). However, the roles of circRNAs in OA remain obscure. Herein, we explored the roles of exosomal circRNA bromodomain and WD repeat domain containing 1(circ-BRWD1) in OA pathology. METHODS: In vitro model of OA was constructed by treating CHON-001 cells with interleukin-1ß (IL-1ß). Quantitative real-time polymerase chain reaction (qRT-PCR) assay was used for circ-BRWD1, BRWD, miR-1277, and TNF receptor-associated factor 6 (TRAF6) levels. RNase R assay was conducted for the feature of circ-BRWD1. Transmission electron microscopy (TEM) was employed to analyze the morphology of exosomes. Western blot assay was performed for protein levels. Cell Counting Kit-8 (CCK-8) assay, flow cytometry analysis, and 5-Ethynyl-2'-deoxyuridine (EDU) assay were adopted for cell viability, apoptosis, and proliferation, respectively. Enzyme-linked immunosorbent assay (ELISA) was carried out for the concentrations of interleukin-6 (IL-6) and interleukin-8 (IL-8). Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were used to analyze the interaction between miR-1277 and circ-BRWD1 or TRAF6. RESULTS: Circ-BRWD1 was increased in OA cartilage tissues, IL-1ß-treated CHON-001 cells, and the exosomes derived from IL-1ß-treated CHON-001 cells. Exosome treatment elevated circ-BRWD1 level, while exosome blocker reduced circ-BRWD1 level in IL-1ß-treated CHON-001 cells. Silencing of circ-BRWD1 promoted cell viability and proliferation and repressed apoptosis, inflammation, and extracellular matrix (ECM) degradation in IL-1ß-stimulated CHON-001 cells. For mechanism analysis, circ-BRWD1 could serve as the sponge for miR-1277 to positively regulate TRAF6 expression. Moreover, miR-1277 inhibition ameliorated the effects of circ-BRWD1 knockdown on IL-1ß-mediated CHON-001 cell damage. Additionally, miR-1277 overexpression relieved IL-1ß-induced CHON-001 cell injury, while TRAF6 elevation restored the impact. CONCLUSION: Exosomal circ-BRWD1 promoted IL-1ß-induced CHON-001 cell progression by regulating miR-1277/TRAF6 axis.
Subject(s)
MicroRNAs , Osteoarthritis , RNA, Circular , Chondrocytes , Humans , Intracellular Signaling Peptides and Proteins , TNF Receptor-Associated Factor 6ABSTRACT
Earthworms (Annelida: Crassiclitellata) are widely distributed around the world due to their ancient origination as well as adaptation and invasion after introduction into new habitats over the past few centuries. Herein, we report a 1.2 Gb complete genome assembly of the earthworm Amynthas corticis based on a strategy combining third-generation long-read sequencing and Hi-C mapping. A total of 29,256 protein-coding genes are annotated in this genome. Analysis of resequencing data indicates that this earthworm is a triploid species. Furthermore, gene family evolution analysis shows that comprehensive expansion of gene families in the Amynthas corticis genome has produced more defensive functions compared with other species in Annelida. Quantitative proteomic iTRAQ analysis shows that expression of 147 proteins changed in the body of Amynthas corticis and 16 S rDNA sequencing shows that abundance of 28 microorganisms changed in the gut of Amynthas corticis when the earthworm was incubated with pathogenic Escherichia coli O157:H7. Our genome assembly provides abundant and valuable resources for the earthworm research community, serving as a first step toward uncovering the mysteries of this species, and may provide molecular level indicators of its powerful defensive functions, adaptation to complex environments and invasion ability.
Subject(s)
Evolution, Molecular , Genome , Oligochaeta/genetics , Animals , Escherichia coli/pathogenicity , Gene Expression Regulation , Host-Pathogen Interactions , Oligochaeta/metabolism , Oligochaeta/microbiology , Phylogeny , Proteomics , TranscriptomeABSTRACT
Since the implementation of the "treat all" policy in China in 2016, there have been few data on the prevalence of transmitted drug resistance (TDR) in China. In this study, we describe TDR in patients newly diagnosed with human immunodeficiency virus (HIV) infection between 2016 and 2019 in Shenyang city, China. Demographic information and plasma samples from all newly reported HIV-infected individuals in Shenyang from 2016 to 2019 were collected. The HIV pol gene was amplified and sequenced for subtyping and TDR. The spread of TDR was analyzed by inferring an HIV molecular network based on pairwise genetic distance. In total, 2,882 sequences including CRF01_AE (2019/2,882, 70.0%), CRF07_BC (526/2,882, 18.3%), subtype B (132/2,882, 4.6%), and other subtypes (205/2,882, 7.1%) were obtained. The overall prevalence of TDR was 9.1% [95% confidence interval (CI): 8.1-10.2%]; the prevalence of TDR in each subtype in descending order was CRF07_BC [14.6% (95% CI: 11.7-18.0%)], subtype B [9.1% (95% CI: 4.8-15.3%)], CRF01_AE [7.9% (95% CI: 6.7-9.1%)], and other sequences [7.3% (95% CI: 4.2-11.8%)]. TDR mutations detected in more than 10 cases were Q58E (n = 51), M46ILV (n = 46), K103N (n = 26), E138AGKQ (n = 25), K103R/V179D (n = 20), and A98G (n = 12). Molecular network analysis revealed three CRF07_BC clusters with TDR [two with Q58E (29/29) and one with K103N (10/19)]; and five CRF01_AE clusters with TDR [two with M46L (6/6), one with A98G (4/4), one with E138A (3/3), and one with K103R/V179D (3/3)]. In the TDR clusters, 96.4% (53/55) of individuals were men who have sex with men (MSM). These results indicate that TDR is moderately prevalent in Shenyang (5-15%) and that TDR strains are mainly transmitted among MSM, providing precise targets for interventions in China.
ABSTRACT
To disclose how phosphorus deficiency influence phytoremediation of Cd contamination using poplars, root architecture, Cd absorption, Cd translocation and antioxidant defense in poplar roots were investigated using a clone of Populus × euramericana. Root growth was unaltered by Cd exposure regardless of P conditions, while the degree of root proliferation upon P deficiency was changed by high level of Cd exposure. The concentration and content of Cd accumulation in roots were increased by P deficiency. This can be partially explained by the increased expression of genes encoding PM H + -ATPase under the combined conditions of P deficiency and high Cd exposure, which enhanced Cd2+-H+ exchanges and led to an increment of Cd uptake under P deficiency. Despite of the increasing Cd accumulation in roots, the translocation of Cd from roots to aerial tissues sharply decreased upon P deficiency. The relative expression of genes responsible for Cd translocation (HMA4) decreased upon P deficiency and thus inhibited Cd translocation via xylem. GR activity was decreased by P deficiency, which can inhibit the form of GSH and GSH-Cd complexes and decrease Cd translocation via GSH-Cd complexes. The transportation of PC-Cd complexes into vacuole decreased under P deficiency as a result of the low expression of PCS and ABCC1, and thus suppressed Cd tolerance and Cd detoxification in roots. Moreover, P deficiency decreased the levels of antioxidase (GR and CAT) and phytohormones including JA, ABA and GA3, which synchronously reduced antioxidant capacity in roots.
Subject(s)
Cadmium/metabolism , Phosphorus/metabolism , Populus/physiology , Adaptation, Physiological , Antioxidants/metabolism , Biodegradation, Environmental , Biological Transport , Cadmium/toxicity , Cell Proliferation , Plant Growth Regulators/metabolism , Plant Roots/metabolism , Populus/metabolism , Xylem/metabolismABSTRACT
BACKGROUND: MicroRNAs (miRNAs) are approximately 20-22 nt non-coding RNAs that play key roles in many biological processes in both animals and plants. Although a number of miRNAs were identified in maize, the function of miRNA in seed development was merely discussed. RESULTS: In this study, two small RNA libraries were sequenced, and a total reads of 9,705,761 and 9,005,563 were generated from developing seeds and growing leaves, respectively. Further analysis identified 125 known miRNAs in seeds and 127 known miRNAs in leaves. 54 novel miRNAs were identified and they were not reported in other plants. Additionally, some miRNA*s of these novel miRNAs were detected. Potential targets of all novel miRNAs were predicted based on our strict criteria. In addition to deep-sequencing, miRNA microarray study confirmed the higher expression of several miRNAs in seeds. In summary, our results indicated the distinct expression of miRNAs during seed development. CONCLUSIONS: We had identified 125 and 127 known miRNAs from seeds and leaves in maize, and a total of 54 novel miRNAs were discovered. The different miRNA expression profile in developing seeds were revealed by both sequencing and microarray studies.
Subject(s)
MicroRNAs/genetics , RNA, Plant/genetics , RNA, Untranslated/genetics , Zea mays/genetics , Oligonucleotide Array Sequence Analysis/methods , Plant Leaves/genetics , Seeds/geneticsABSTRACT
Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.