茶树小分子量热激蛋白基因CsHSP17.2的克隆与表达分析

[目的]进一步了解茶树小分子量热激蛋白基因CsHSP17.2在逆境胁迫条件下的分子生物学功能.[方法]利用RT-PCR技术从茶树‘迎霜’中克隆得到CsHSP1 7.2基因,运用生物信息学软件分析其核苷酸和编码蛋白,通过Real-timePCR分析其表达模式.[结果]该基因开放阅读框长度为453 bp,编码150个氨基酸,蛋白质相对分子质量为17.2×10a,理论等电点5.56;无信号肽位点,属于非分泌型蛋白;被定位于细胞质中.系统发育树分析表明,茶树CsHSP1 7.2与水稻(GenBank登录号:P27777)和花生(ABC41131)的进化关系较近,属于小分子量热激蛋白基因家族第Ⅰ亚族.qRT-PCR分析发现,茶树CsHSP17.2属于组成型基因;高温(38℃)处理1h能显著提高CsHSP17.2 mRNA的相对表达量(P＜0.05);干旱(100 g·L-1 PEG 6000)、高盐(200 mmol· L-1 NaCl)和外源脱落酸(200 mg· L-1 ABA)处理条件下,该基因的转录水平均出现不同程度的上调.[结论]克隆得到茶树‘迎霜’小分子量热激蛋白基因CsHSP17.2,其在花中表达量最高,且响应高温、干旱、高盐和外源脱落酸胁迫.     

Over-expression of Hsp75 in neural stem cells reduced Aβ-mediated neurotoxicity

AIM: To investigate the effect of heat shock protein 75 (Hsp75) over-expression on Aβ-induced neurotoxicity in the neural stem cells and to explore its mechanism. METHODS: An adenovirus-mediated Hsp75 over-expression vector was used in vitro. The mouse neural stem cell C17.2 was cultured in vitro and divided into control group, Aβ group, negative adenovirus vector transfection group and Hsp75 over-expression adenovirus vector transfection group. The transfection and cellular immune identification were detected by fluorescence microscopy. The cell morphology was observed under inverted phase-contrast microscope. The cell viability and apoptosis were detected by MTT assay and flow cytometry, respectively. Hsp75 over-expression and cleaved caspase-3 protein level were measured by Western blot. RESULTS: Observation by fluorescence microscopy indicated that C17.2 cells were successfully transfected and Hsp75 gene was effectively expressed in the neural stem cells after transfection. In addition, the morphology and viability of the cells did not change and these cells did not differentiate after transfection. As compared with control group, the cell viability in Aβ group and negative adenovirus vector transfection group was significantly decreased (P < 0.05), and the cell apoptotic rate and cleaved caspase-3 level (P < 0.05) were increased. As compared with Aβ group and negative adenovirus vector transfection group, Hsp75 over-expression significantly increased the cell viability, and decreased the cell apoptosis and cleaved caspase-3 level (P < 0.05). CONCLUSION: Hsp75 over-expression protects the neural stem cells against Aβ-induced injury. The mechanism may be related to inhibiting caspase-3 pathway-dependent apoptosis.     

17.2％吡唑醚菌酯·氟环唑悬乳剂对玉米的防病保健作用

[目的]了解17.2％吡唑醚菌酯·氟环唑悬乳剂在防治玉米大斑病中的应用效果.[方法]通过2012、2013年两年的田间示范试验研究17.2％吡唑醚菌酯·氟环唑悬乳剂对玉米大斑病的防治效果及其对玉米植株的保健作用.[结果]17.2％吡唑醚菌酯·氟环唑悬乳剂对玉米大斑病的防治效果达到90％以上,该药剂处理后的玉米穗长明显增加、叶绿素含量增加、秃头率降低、产量增加.[结论]17.2％吡唑醚菌酯·氟环唑悬乳剂对玉米大斑病有很好的防治效果,并且对玉米有保健增产作用.