Magnesium (Mg(2+)) is an essential macronutrient for plant growth and development, and the CorA/MRS2/MGT-type Mg(2+) transporters play important roles in maintaining Mg(2+) homeostasis in plants. Although the MRS2/MGT genes have been identified in two model plant species, Arabidopsis and rice, a comprehensive analysis of the MRS2/MGT gene family in other plants is lacking. In this work, 12 putative MRS2/MGT genes (ZmMGT1- ZmMGT12) were identified in maize and all of them were classified into five distinct subfamilies by phylogenetic analysis. A complementation assay in the Salmonella typhimurium MM281 strain showed that five representatives of the 12 members possess Mg(2+) transport abilities. Inhibition of ZmMGT protein activity using the hexaamminecobalt (III) (Co-Hex) inhibitor indicated that the ZmMGT protein mediated both low-affinity and high-affinity Mg(2+) transport in maize. A semi-quantitative reverse transcription-PCR (RT-PCR) analysis revealed that eight genes were constitutively expressed in all of the detected tissues, with one being specifically expressed in roots and three having no detectable expression signals. A quantitative RT-PCR analysis showed that some ZmMGT members displayed differential responses to Mg(2+) deficiency and aluminum (Al) stress. Furthermore, root growth inhibition and Mg(2+) accumulation analyses in two maize inbred lines, which conferred different levels of Al tolerance, revealed that ZmMGT proteins contributed to the Al resistance of the Al tolerance genotype. We hypothesize that ZmMGT family members function as Mg(2+) transporters and may play a role in linking Mg(2+) deficiency and Al stress responses. Our results will be valuable in a further analysis of the important biological functions of ZmMGT members in maize.
Aim: Proteins with legume lectin domains are known to possess a wide range of biological functions. Here, the antitumor effects of two representative legume lectins, concanavalin A (ConA) and Sophora flavescens lectin (SFL), on human breast carcinoma cells were investigated in vitro and in vivo. Methods: Human breast carcinoma MCF-7 cells and human normal mammary epithelial MCF-10A cells were examined. Cell viability was detected using WST-1 and CCK-8 assays. Cell apoptosis was analyzed with Hoechst 33258 staining. Cell cycle was investigated using flow cytometry. The expression of relevant proteins was measured using Western blotting. Breast carcinoma MCF-7 bearing nude mice were used to study the antitumor effects in vivo. The mice were injected with ConA (40 mg/kg, ip) and SFL (55 mg/kg, ip) daily for 14 d. Results: ConA and SFL inhibited the growth of MCF-7 cells in dose-and time-dependent manners (IC 50 values were 15 and 20 μg/mL, respectively). Both ConA and SFL induced apoptotic morphology in MCF-7 cells without affecting MCF-10A cells. ConA and SFL dosedependently increased the sub-G 1 proportion in MCF-7 cells, while SFL also triggered the G 2 /M phase cell cycle arrest. Both ConA and SFL dose-dependently increased the activities of caspase-3 and caspase-9 and release of cytochrome c from mitochondria into cytoplasm, up-regulated Bax and Bid, and down-regulated Bcl-2 and Bcl-X L in MCF-7 cells. ConA reduced NF-κB, ERK, and JNK levels, and increased p53 and p21 levels, while SFL caused similar changes in NF-κB, ERK, p53, and p21 levels, but did not affect JNK expression. Administration of ConA and SFL significantly decreased the subcutaneous tumor mass volume and weight in MCF-7 bearing nude mice. Conclusion: ConA and SFL exert anti-tumor actions against human breast carcinoma MCF-7 cells both in vitro and in vivo.
Common buckwheat is a health-care crop, and continuous cropping is one of the main factors restricting its high-yielding. In order to clarify the mechanism of continuous cropping of buckwheat, 4, 5and 6 years continuous cropping treatments were set up. We obtained the following results. The available nutrients content in rhizosphere soil, soil enzyme activity, leaf area, chlorophyll and soluble protein content, the peroxidase(POD) activity of leaves, agronomic traits, root index (except average diameter) of Fengtian1(FT1) decreased significantly with the increase of continuous cropping years. While, soil pH and the number of fungi in rhizosphere soil increased significantly, the number of bacteria in rhizosphere soil and the activity of catalase (CAT) and superoxide dismutase(SOD) in leaves first increased and then decreased. The yields of continuous cropping for 6 years were 12 times lower than the control crop. In conclusion, continuous cropping has certain effects on growth and yields of buckwheat.
Antibiotics have been described to modulate bacterial virulence gene expression. This study aimed to assess the changes caused by anti-Staphylococcus agents in the transcription of leucocidin ED (lukED) gene of Staphylococcus aureus strain Newman in vitro and in vivo and to determine whether the altered expression is agr dependent. The bacteria were exposed to subinhibitory concentrations [1/2, 1/4, or 1/8 minimal inhibitory concentration (MIC)] of 11 antibiotics, and the expression of lukE and agr-effector RNAIII was determined using qRT-PCR. In vivo experiments were performed to evaluate the impact exerted by six representative antibiotics on the transcription of both genes. Molecular analysis showed that in vitro lukE transcription was dramatically promoted in the Newman strain exposed to sub-MICs of vancomycin, trimethoprim-sulfamethoxazole, clindamycin, gentamicin, daptomycin, and ciprofloxacin and considerably reduced when stimulated by cefazolin, erythromycin, rifampicin, tigecycline, and linezolid. In the murine abscess model, tigecycline significantly decreased the transcription of lukE and the bacterial numbers, whereas vancomycin increased them; although cefazolin increased the lukE expression (contrary to the in vitro effect), it had a remarkable role in reducing bacterial load. The correspondence analysis shows that RNAIII expression varied under seven of 11 antibiotics in vitro, and six drugs in vivo were consistent with lukE transcripts. In conclusion, our data show that anti-Staphylococcus antibiotics exert modulatory effects on lukE expression in vitro and/or in vivo, and the changed expression caused by some drugs may be involved with agr activity, thus providing a guide to choose appropriate agents to avoid promoting bacterial virulence in lukED-positive S. aureus infections.
Aluminum (Al) toxicity is a major factor limiting crop production and plant growth in acid soils. The complex inheritance of Al toxicity and tolerance mechanisms in maize has uncharacterized yet. In thsi study, the maize inbred line 178 seedlings were treated with 200 μmol L-1 CaCl 2 +0 μmol L-1 AlCl 3 (control) and 200 μmol L-1 CaCl 2 +60 μmol L-1 AlCl 3 (Al treatment) for 1 and 6 h, respectively. The experiment was repeated three times. Then a detailed temporal analysis of root gene expression was performed using an Agilent GeneChip with 34 715 genes, only the genes showing more than 2.0-fold difference (P<0.01) between the control and the Al treatment maize seedlings was analyzed further. Thus, a total of 832 different expression genes, 689 significantly up-regulated and 143 down-regulated, were identified after the seedlings were treated with Al for 6 h. And 60 genes, 59 up-regulated and 1 down-regulated, were also detected after the seedlings were treated for 1 h. Replicated transcriptome analyses further showed that about 61% of total significantly genes could be annotated based on plant genome resources. Quantitative real-time PCR (qRT-PCT) of some selected candidate genes was used to demonstrate the microarray data, indicating significant differences between the control and Al treated seedlings. Exposure to Al for 6 h triggered changes in the transcript levels for several genes, which were primarily related to cell wall structure and metabolism, oxidative stress response, membrane transporters, organic acid metabolism, signaling and hormones, and transcription factors, etc. After Al treated for 1 h, differential abundance of transcripts for several transporters, kinase, and transcription factors were specifically induced. In this study, the diversity of the putative functions of these genes indicates that Al stress for a short stage induced a complex transcriptome changes in maize. These results would further help us to understand rapid and early mechanisms of Al toxicity and tolerance in maize regulated at the transcriptional level.
ZmMGT10 was specifically expressed in maize roots and induced by a deficiency of magnesium. Overexpression of ZmMGT10 restored growth deficiency of the Salmonella typhimurium MM281 strain and enhanced the tolerance in Arabidopsis to stress induced by low magnesium levels by increasing uptake of Mg via roots. CorA/MRS2/MGT-type Mg transporters play a significant role in maintaining magnesium (Mg) homeostasis in plants. Although the maize CorA/MRS2/MGT family comprises of 12 members, currently no member has been functionally characterized. Here, we report the isolation and functional characterization of ZmMGT10 from the maize MRS2/MGT gene family. ZmMGT10 has a typical structure feature which includes two conserved TMs near the C-terminal end and an altered AMN tripeptide motif. The high sequence similarity and close phylogenetic relationship indicates that ZmMGT10 is probably the counterpart of Arabidopsis AtMGT6. The complementation of the Salmonella typhimurium mutated MM281 strain indicates that ZmMGT10 possesses the ability to transport Mg. ZmMGT10 was specifically expressed in the plant roots and it can be stimulated by a deficiency of Mg. Transgenic Arabidopsis plants which overexpressed ZmMGT10 grew more vigorously than wild-type plants under low Mg conditions, exhibited by longer root length, higher plant fresh weight and chlorophyll content, suggesting ZmMGT10 was essential for plant growth and development under low Mg conditions. Further investigations found that high accumulation of Mg occurred in transgenic plants attributed to improved Mg uptake and thereby enhanced tolerance to Mg deficiency. Results from this investigation illustrate that ZmMGT10 is a Mg transporter of maize which can enhance the tolerance to Mg deficient conditions by improving Mg uptake in the transgenic plants of Arabidopsis.
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