WRKY transcription factors are reported to be involved in defense regulation, stress response and plant growth and development. However, the precise role of WRKY transcription factors in abiotic stress tolerance is not completely understood, especially in crops. In this study, we identified and cloned 10 WRKY genes from genome of wheat (Triticum aestivum L.). TaWRKY10, a gene induced by multiple stresses, was selected for further investigation. TaWRKY10 was upregulated by treatment with polyethylene glycol, NaCl, cold and H2O2. Result of Southern blot indicates that the wheat genome contains three copies of TaWRKY10. The TaWRKY10 protein is localized in the nucleus and functions as a transcriptional activator. Overexpression of TaWRKY10 in tobacco (Nicotiana tabacum L.) resulted in enhanced drought and salt stress tolerance, mainly demonstrated by the transgenic plants exhibiting of increased germination rate, root length, survival rate, and relative water content under these stress conditions. Further investigation showed that transgenic plants also retained higher proline and soluble sugar contents, and lower reactive oxygen species and malonaldehyde contents. Moreover, overexpression of the TaWRKY10 regulated the expression of a series of stress related genes. Taken together, our results indicate that TaWRKY10 functions as a positive factor under drought and salt stresses by regulating the osmotic balance, ROS scavenging and transcription of stress related genes.
Atmospheric pressure room temperature plasma jets (APRTP-Js) that can emit a mixture of different active species have recently found entry in various medical applications. Apoptosis is a key event in APRTP-Js-induced cellular toxicity, but the exact biological mechanisms underlying remain elusive. Here, we explored the role of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in APRTP-Js-induced apoptosis using in vitro model of HepG2 cells. We found that APRTP-Js facilitated the accumulation of ROS and RNS in cells, which resulted in the compromised cellular antioxidant defense system, as evidenced by the inactivation of cellular antioxidants including glutathione (GSH), superoxide dismutase (SOD) and catalase. Nitrotyrosine and protein carbonyl content analysis indicated that APRTP-Js treatment caused nitrative and oxidative injury of cells. Meanwhile, intracellular calcium homeostasis was disturbed along with the alteration in the expressions of GRP78, CHOP and pro-caspase12. These effects accumulated and eventually culminated into the cellular dysfunction and endoplasmic reticulum stress (ER stress)-mediated apoptosis. The apoptosis could be markedly attenuated by N-acetylcysteine (NAC, a free radical scavenger), which confirmed the involvement of oxidative and nitrative stress in the process leading to HepG2 cell apoptosis by APRTP-Js treatment.
SummaryCo-expression of CrtB and CrtI enhanced carotenoid in endosperm through upregulation of the endogenous carotenogenic genes. Our results also indicate important roles of LCYB and HYD in wheat carotenoid biosynthesis.
A cold atmospheric-pressure plasma, which has been widely used for biomedical applications, may potentially affect the cell cycle and cause cell apoptosis. In this paper, a human hepatocellular carcinoma cell (HepG2) is treated by a singleelectrode plasma jet device. Further investigation by using flow cytometric analysis demonstrates that plasma treatment increases the percentage of apoptotic cells being associated with cell cycle arrest at the G2/M phase. Moreover, the reverse transcription polymerase chain reaction assay shows that the cyclin B1 and Cdc2 are decreased at the transcription level after plasma treatment, while the expression of the p21 Cdk inhibitor, as well as that of tumor suppressor p53, is enhanced. On the other hand, the levels of certain pro-or antiapoptotic genes are checked, and the experimental results suggest that the plasma induces apoptosis by shifting the Bax/Bcl-2 ratio to trigger HepG2 cell apoptosis. The results of the present investigation indicate that the plasma jet device may have potential therapeutic activities, such as sterilization of living tissue and tumor therapy of the postoperative treatment process.
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