Mitochondria fulfil essential functions in respiration and metabolism as well as regulating stress responses and apoptosis. Most native mitochondrial proteins are encoded by nuclear genes and are imported into mitochondria via one of several receptors that recognize N-terminal signal peptides. The targeting of recombinant proteins to mitochondria therefore requires the presence of an appropriate N-terminal peptide, but little is known about mitochondrial import in monocotyledonous plants such as rice (Oryza sativa). To gain insight into this phenomenon, we targeted nuclear-encoded enhanced green fluorescent protein (eGFP) to rice mitochondria using six mitochondrial pre-sequences with diverse phylogenetic origins, and investigated their effectiveness by immunoblot analysis as well as confocal and electron microscopy. We found that the ATPA and COX4 (Saccharomyces cerevisiae), SU9 (Neurospora crassa), pFA (Arabidopsis thaliana) and OsSCSb (Oryza sativa) peptides successfully directed most of the eGFP to the mitochondria, whereas the MTS2 peptide (Nicotiana plumbaginifolia) showed little or no evidence of targeting ability even though it is a native plant sequence. Our data therefore indicate that the presence of particular recognition motifs may be required for mitochondrial targeting, whereas the phylogenetic origin of the pre-sequences probably does not play a key role in the success of mitochondrial targeting in dedifferentiated rice callus and plants.
Mammalian cells were grown to high density in a 3,000 L culture using perfusion with hollow fibers operated in a tangential flow filtration mode. The high-density culture was used to inoculate the production stage of a biomanufacturing process. At constant permeate flux operation, increased transmembrane pressures (TMPs) were observed on the final day of the manufacturing batches. Small scale studies suggested that the filters were not irreversibly fouled, but rather exposed to membrane concentration polarization that could be relieved by tangential sweeping of the hollow fibers. Studies were undertaken to analyze parameters that influence the hydrodynamic profile within hollow fibers; including filter area, cell density, recirculation flow rate, and permeate flow rate. Results indicated that permeate flow rate had the greatest influence on modulating TMP. Further evaluation showed a significant decrease in TMP when permeate flow was reduced, and this occurred without any negative effect on cell growth or viability. Hence, a 30% reduction of permeate flow rate was implemented at manufacturing scale. A stable operation was achieved as TMP was successfully reduced by 75% while preserving all critical factors for performance in the perfusion bioreactor. 相似文献
Identifying seasonal shifts in community assembly for multiple biological groups is important to help enhance our understanding of their ecological dynamics. However, such knowledge on lotic assemblages is still limited. In this study, we used biological traits and functional diversity indices in association with null model analyses to detect seasonal shifts in the community assembly mechanisms of lotic macroinvertebrates and diatoms in an unregulated subtropical river in China. We found that functional composition and functional diversity (FRic, FEve, FDis, MNN, and SDNN) showed seasonal variation for macroinvertebrate and diatom assemblages. Null models suggested that environmental filtering, competitive exclusion, and neutral process were all important community assembly mechanisms for both biological groups. However, environmental filtering had a stronger effect on spring macroinvertebrate assemblages than autumn assemblages, but the effect on diatom assemblages was the same in both seasons. Moreover, macroinvertebrate and diatom assemblages were shaped by different environmental factors. Macroinvertebrates were filtered mainly by substrate types, velocity, and CODMn, while diatoms were mainly shaped by altitude, substrate types, and water quality. Therefore, our study showed (a) that different biological assemblages in a river system presented similarities and differences in community assembly mechanisms, (b) that multiple processes play important roles in maintaining benthic community structure, and (c) that these patterns and underlying mechanisms are seasonally variable. Thus, we highlight the importance of exploring the community assembly mechanisms of multiple biological groups, especially in different seasons, as this is crucial to improve the understanding of river community changes and their responses to environmental degradation. 相似文献
Plant and Soil - Phosphorus (P) mobility depends on the availability of water in soil, both of which are limited resources for crop production. We studied the mechanisms governing tolerance to... 相似文献
Chrysanthemum is one of the most important ornamental flowers in the world, and temperature has a significant influence on its field production. In the present study, differentially expressed proteins were investigated in the leaves of Dendranthema grandiflorum ‘Jinba’ under high temperature stress using label-free quantitative proteomics techniques. The expressed proteins were comparatively identified and analyzed. A total of 1,463 heat-related, differentially expressed proteins were successfully identified by Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS), and 1,463 heat-related, differentially expressed proteins were successfully identified by mass spectrometry after a high temperature treatment. Among these, 701 proteins were upregulated and 762 proteins were downregulated. The in-depth bioinformatics analysis of these differentially expressed proteins revealed that these were involved in energy metabolism pathways, protein metabolism, and heat shock. In the present study, the investigators determined the changes in the levels of some proteins, and their expression at the protein and molecular levels in chrysanthemum to help reveal the mechanism of heat resistance in chrysanthemum. Furthermore, the present study elucidated some of the proteins correlated to heat resistance in chrysanthemum, and their expression changes at the protein and molecular levels to help reveal the mechanism of heat resistance in this flower species. These results provide a theoretical basis for the selection of new heat resistant varieties of chrysanthemum in the field. 相似文献
Arbuscular mycorrhizal fungi (AMF) can improve plant tolerance to several abiotic stresses, including heavy
metals, drought or salinity exposure. However, the role of AMF in alleviation of soil cadmium (Cd)-induced toxicity to plants is still largely unknown. In this study, Cd speciation in soil and subcellular distribution of Cd were
used to characterize the roles of application AM fungi in the alleviation of Cd toxicity in alfalfa plants. Our results
showed that the addition of Glomus mosseae in Cd contaminated soil (10 mg/Kg) significantly increased soil pH,
cation exchange capacity (CEC) and organic matter in rhizosphere soil with Medicago truncatula L., and then
account for significantly decreased contents of exchangeable and carbonate-bounded Cd speciation in rhizosphere
soil, indicating alleviation of plant toxicity by reduction of bioavailable fractions of Cd. Although there is no significant difference found in Cd accumulation by roots and shoots respectively between Cd and AM-Cd treatments, more portion of Cd was recorded compartmentalization in cell wall fraction of both root and shoot in
treatment of Cd with AM application, indicating alleviation of Cd toxicity to plant cell. Herein, application of
AM fungi in Cd treatments performed to inhibit the appearance of Cd toxicity symptoms, including the improvement of leaf electrolyte leakage, root elongation, seedling growth and biomass. This information provides a clearer
understanding of detoxification strategy of AM fungi on Cd behavior with development and stabilization of soil
structure and subcellular distribution of plant. 相似文献
Purpose: Cervical cancer (CC) is one of the most general gynecological malignancies and is associated with high morbidity and mortality. We aimed to select candidate genes related to the diagnosis and prognosis of CC.Methods: The mRNA expression profile datasets were downloaded. We also downloaded RNA-sequencing gene expression data and related clinical materials from TCGA, which included 307 CC samples and 3 normal samples. Differentially expressed genes (DEGs) were obtained by R software. GO function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs were performed in the DAVID dataset. Using machine learning, the optimal diagnostic mRNA biomarkers for CC were identified. We used qRT-PCR and Human Protein Atlas (HPA) database to exhibit the differences in gene and protein levels of candidate genes.Results: A total of 313 DEGs were screened from the microarray expression profile datasets. DNA methyltransferase 1 (DNMT1), Chromatin Assembly Factor 1, subunit B (CHAF1B), Chromatin Assembly Factor 1, subunit A (CHAF1A), MCM2, CDKN2A were identified as optimal diagnostic mRNA biomarkers for CC. Additionally, the GEPIA database showed that the DNMT1, CHAF1B, CHAF1A, MCM2 and CDKN2A were associated with the poor survival of CC patients. HPA database and qRT-PCR confirmed that these genes were highly expressed in CC tissues.Conclusion: The present study identified five DEmRNAs, including DNMT1, CHAF1B, CHAF1A, MCM2 and Kinetochore-related protein 1 (KNTC1), as potential diagnostic and prognostic biomarkers of CC. 相似文献
Excessive pulmonary inflammatory response is critical in the development of acute lung injury (ALI). Previously, microRNAs (miRNAs) have been recognized as an important regulator of inflammation in various diseases. However, the effects and mechanisms of miRNAs on inflammatory response in ALI remain unclear. Herein, we tried to screen miRNAs in the processes of ALI and elucidate the potential mechanism. Using a microarray assay, microRNA let-7e (let-7e) was chose as our target for its reported suppressive roles in several inflammatory diseases. Down-regulation of let-7e by antagomiR-let-7e injection attenuated LPS-induced acute lung injury. We also found that antagomiR-let-7e could obviously improve the survival rate in ALI mice. Moreover, antagomiR-let-7e treatment reduced the production of proinflammatory cytokines (i.e., TNF-α, IL-1β and IL-6) in bronchoalveolar lavage fluid (BALF) of LPS-induced ALI mice. Luciferase reporter assays confirmed that suppressor of cytokine signaling 1 (SOCS1), a powerful attenuator of nuclear factor kappa B (NF-κB) signaling pathway, was directly targeted and suppressed by let-7e in RAW264.7 cells. In addition, it was further observed that SOCS1 was down-regulated, and inversely correlated with let-7e expression levels in lung tissues of ALI mice. Finally, down-regulation of let-7e suppressed the activation of NF-κB pathway, as evidenced by the reduction of p-IκBα, and nuclear p-p65 expressions in ALI mice. Collectively, our findings indicate that let-7e antagomir protects mice against LPS-induced lung injury via repressing the pulmonary inflammation though regulation of SOCS1/NF-κB pathway, and let-7e may act as a potential therapeutic target for ALI. 相似文献