MrERF039 transcription factor plays an active role in the cold response of Medicago ruthenica as a sugar molecular switch.

Cold stress severely restricts plant development, causing significant agricultural losses. We found a critical transcription factor network in Medicago ruthenica was involved in plant adaptation to low-temperature. APETALA2/ethylene responsive factor (AP2/ERF) transcription factor MrERF039 was transcriptionally induced by cold stress in M. ruthenica. Overexpression of MrERF039 significantly increased the glucose and maltose content, thereby improving the tolerance of M. ruthenica. MrERF039 could bind to the DRE cis-acting element in the MrCAS15A promoter. Additionally, the methyl group of the 14th amino acid in MrERF039 was required for binding. Transcriptome analysis showed that MrERF039 acted as a sugar molecular switch, regulating numerous sugar transporters and sugar metabolism-related genes. In addition, we found that MrERF039 could directly regulate ß-amylase gene, UDP glycosyltransferase gene, and C2H2 zinc finger protein gene expression. In conclusion, these findings suggest that high expression of MrERF039 can significantly improve the cold tolerance of M. ruthenica root tissues during cold acclimation. Our results provide a new theoretical basis and candidate genes for breeding new legume forage varieties with high resistance.

Study of a Si-based light initiated multi-gate semiconductor switch for high temperatures.

Light initiated multi-gate semiconductor switch (LIMS) is a kind of power electronic device which has many differences from traditional thyristor triggered by electric pulse. LIMS is triggered by laser, the turn-on time is smaller, and the anti-electromagnetic interferences is strong. The opening mode of LIMS is obviously different to traditional thyristor. After the laser into the gate area, a large number of electrons and holes will appear in P-base region, holes gather in the area of P-base in PN junction J2, and electrons gather in N-drift region around the PN junction J2. PN junction J2 will open first, then PN junction J3 opens. The delay time of the NPN and PNP thyristors is close to zero when the laser pulse is narrow and the peak power is high, so the turn-on velocity is fast. To optimize the characteristics of the LIMS at high temperatures, we propose a new structure of the LIMS with the optimization of the n+ layer, circular light gate, and the new-style edge termination. The diameter of the LIMS is 23 mm. The experiment results show that the leakage current of the proposed LIMS has been decreased from more than 1 mA to 500 µA at 125 °C, the output current of the LIMS is 10.2 kA with a voltage of 4 kV at 85 °C, and the output current of the LIMS is 12.1 kA with a voltage of 4 kV at - 55 °C. Additionally, di/dt is larger than 30 kA/µs.

Melatonin Enhances Seed Germination and Seedling Growth of Medicago sativa Under Salinity via a Putative Melatonin Receptor MsPMTR1.

Alfalfa (Medicago sativa L.) is an important forage crop, and salt stress is a major limiting factor in its yield. Melatonin (MT) is a multi-regulatory molecule in plants. We showed that basal MT content was positively correlated with the salt tolerance degree of different alfalfa varieties. MT and its precursor 5-HT fully recovered seed germination while partially ameliorated seedling growth of salt-stressed alfalfa. The 5-HT showed some divergent effects from MT with regards to growth amelioration under salinity. Salt stress caused stunted plant growth in soil culture, while MT ameliorated it by elevating plant height, fresh weight, branching number, and chlorophyll content. Silencing of a putative MT receptor, MsPMTR1, which was shown to be membrane-localized, abolished the ameliorative effects of MT on salt-stressed alfalfa seedling growth, while overexpression of MsPMTR1 improved plant growth under salt stress. The RNA sequencing analysis showed that nine pathway genes were specifically induced by MT treatment compared with salt stress. These MT-responsive differentially expressed genes include basal metabolic pathway genes, such as "ribosome, elongation factor," "sugar and lipid metabolism," and "photosynthesis" and stress-related genes encoding "membrane integrity" related proteins, heat shock protein, peroxidase/oxidoreductase, and protease. Several abiotic stress response-related genes, such as DRE, ARF, HD-ZF, MYB, and REM were repressed by NaCl treatment while induced by MT treatment. In summary, we demonstrated the importance of MsPMTR1 in MT-mediated salt tolerance in alfalfa, and we also analyzed the regulatory mechanism of MT during alfalfa seed germination under salt stress.

Acidic pH promotes nucleus pulposus cell senescence through activating the p38 MAPK pathway.

BACKGROUND: Nucleus pulposus (NP) cell senescence is an important cellular feature within the degenerative disc. It is known that a very acidic niche exists in the degenerative disc, which participates in regulating disc cell viability and matrix metabolism. OBJECTIVE: The present study was aimed to investigate the role and potential signaling transduction pathway of an acidic pH in regulating NP cell senescence. METHODS: Rat NP cells were cultured in an acidic pH of 7.2 close to that in a healthy disc (Control group) or in an acidic pH of 6.2 close to that in a severe degenerative disc (Experiment group) for 10 days. Additionally, the experimental NP cells were incubated along with the inhibitor SB203580 to analyze the role of p38 MAPK pathway in this process. RESULTS: Compared with the control NP cells, experimental NP cells showed a suppressed cell proliferation potency, an increased G0/G1 phase fraction whereas a decreased S-phase fraction and a declined telomerase activity, an up-regulated expression of senescence-related molecules (p16 and p53), and a down-regulated expression of matrix-related moleucles (aggrecan and collagen II). Further analysis showed that inhibition of the p38 MAPK pathway partly reversed effects of acidic pH of 6.2 on the experimental NP cells. CONCLUSION: The very acidic niche identified in a severe degenerative disc promotes NP cell senescence through regulating the p38 MAPK pathway. The present study provides a new mechanism that drives NP cell senescence during disc degeneration.

Osteogenic protein-1 inhibits nucleus pulposus cell apoptosis through regulating the NF-κB/ROS pathway in an inflammation environment.

BACKGROUND: Intervertebral disc degeneration is a pathological process that involves an inflammation response. As a classical cellular feature, several studies have demonstrated that inflammation can promote nucleus pulposus (NP) cell apoptosis. Therefore, attenuation of NP cell apoptosis may be a potential way to retard disc degeneration. OBJECTIVE: The present study was aimed to investigate the protective effects of osteogenic protein-1 (OP-1) against NP cell apoptosis in an inflammation environment, and the potential signaling transduction pathway. METHODS: Rat NP cells were cultured in medium with or without inflammatory cytokine tumor necrosis factor (TNF)-α for 6 days. The exogenous TNF-α was added into the medium to investigate its protective effects. NP cell apoptosis was evaluated by cell apoptosis ratio, caspase-3 activity, gene/protein expression of apoptosis-related molecules (Bcl-2, Bax, and caspase-3). Additionally, the intracellular reactive oxygen species (ROS) content and activity of the NF-κB pathway were also analyzed. RESULTS: Compared with the control NP cells, TNF-α significantly increased cell apoptosis ratio, caspase-3 activity, gene/protein expression of Bcl-2, Bax and caspase-3, ROS content, and activity of the NF-κB pathway. However, OP-1 partly attenuated these effects in NP cells treated with TNF-α. CONCLUSION: OP-1 is effective in attenuating TNF-α-caused NP cell apoptosis, and the ROS/NF-κB pathway may be the potential signaling transduction pathway. The present study indicates that OP-1 may be helpful to inhibit inflammation-mediated disc degeneration.

SALL4 promotes osteoblast differentiation by deactivating NOTCH2 signaling.

BACKGROUND: The Spalt-Like Transcription Factor 4 (SALL4) is reported to regulate cell proliferation, migration and invasion. However, the roles of SALL4 in osteoblast differentiation are unclear. This study was aimed to explore the underlying mechanism of SALL4 in osteoporosis. METHODS: Firstly, the expression of SALL4 was assessed in vivo and in vitro at various stages of development of rats (E14, E20, postnatal 2, 4, and 9 day) or different incubation time (0, 6, 9, 12 and 15 day) of C2C12 and MC3T3-E1 cells. Then, alkaline phosphatase (ALP) activities and positive cells percentages were respectively detected after oeSall4 or siSall4 transfection. Cell differentiation related markers and chondrogenesis-related genes expressions in C2C12 cells were tested by western blot assay and qRT-PCR. Finally, the connection and interaction between SALL4 and NOTCH2 were studied. RESULTS: The results showed that SALL4 expression was increased in vivo and in vitro with the growth of rats or the incubation of cells. SALL4 overexpression promoted osteoblast differentiation; on the contrary, SALL4 knockdown inhibited osteoblast differentiation. Moreover, SALL4 participated in the middle and late stages of cell differentiation. Then, SALL4 and NOTCH2 interacted with each other. NOTCH2 expression was decreased both in vivo and in vitro, and negatively regulated by SALL4. Besides, SALL4 overexpression suppressed NOTCH2 target genes expressions and nuclear entry, while deactivated NOTCH2 signaling. CONCLUSIONS: Our study found that SALL4 played very important roles in the process of osteoblast differentiation by deactivating NOTCH2 signaling. These findings might provide a new insight for treatment of osteoporosis.