玉米热激蛋白ZmHSP70-12基因的克隆及表达分析

热激蛋白70(HSP70)是原核和真核细胞中普遍存在的一种高度保守的分子伴侣。从玉米中克隆1个HSP70家族成员。该基因cDNA序列全长为1 992 bp,开放阅读框为2 352 bp,编码663个氨基酸,蛋白质分子量约75.0 kD。蛋白结构预测及同源比对分析表明,该基因编码蛋白含ATPase位点和HSP70保守结构域,与拟南芥AtHSP70-12序列高度相似,命名为ZmHSP70-12。蛋白亚细胞定位显示,ZmHSP70-12蛋白在内质网中表达。实时荧光定量PCR分析表明,ZmHSP70-12对非生物胁迫高温、干旱均具有明显的应答反应,推测ZmHSP70-12是玉米中与胁迫逆境相关的基因。     

Application of urease inhibitor improves protein composition and bread‐baking quality of urea fertilized winter wheat

Background: Nitrogen losses is an economic problem for wheat production and a high risk to the environment. Therefore, improved N fertilizer management is a key to increasing the N efficiency and minimizing N losses. To increase N efficiency, enhanced fertilizers such as urea combined with urease inhibitor can be used. Aims: The aim of present study was to evaluate the effects of different N forms on grain storage protein subunits in winter wheat and to examine whether the observed changes correlate with parameters of baking quality. Methods: The investigation was performed over two consecutive years at two locations in Germany. Protein subunits were analyzed by SDS‐PAGE. Results: Protein concentrations were similarly increased after fertilization with ammonium nitrate and urea + urease inhibitor. Analysis of the individual storage protein fractions indicated that both fertilizers specifically enhanced ω‐gliadins and HMW glutenins, but the effect was more pronounced in the ammonium nitrate treatment. Application of urea + urease inhibitor had greater influence on the protein composition and resulted in higher specific baking volume as well as the best fresh keeping ability, in comparison with urea treatment. Conclusion: Considering that the urea + urease inhibitor treatment resulted in almost comparable improvements of NUE and baking quality, with the additional benefit of reduced N losses in combination with easy handling, urea + urease inhibitor can be recommended as a viable alternative to both urea alone and ammonium nitrate treatments. This opens up an opportunity for the reduction of N loss in wheat production when use of urea is preferred.     

Effect of CHOP expression knock-down on apoptosis of renal tubular epithelial cells

AIM:To study the effect of C/EBP homologous protein (CHOP) on the apoptosis of renal tubular epithelial HK2 cells. METHODS:The serum mRNA levels of CHOP in the patients with acute kidney injury and healthy controls were detected by qPCR. In vitro, renal tubular epithelial HK2 cells were divided into control group, negative group (transfected with negative control siRNA), si-CHOP group (transfected with CHOP siRNA), and induced by transforming growth factor-β1 (TGF-β1). The viability of the cells was measured by MTT assay, and the apoptotic rate was analyzed by flow cytometry. The protein levels of nuclear antigen Ki-67, proliferating cell nuclear antigen (PCNA), caspase-3 and cleaved caspase-3 were determined by Western blot. RESULTS:Compared with the healthy controls, the serum mRNA levels of CHOP in the patients with acute kidney injury were increased significantly (P<0.05). Transfection with CHOP siRNA significantly decreased the expression of CHOP in the renal tubular epithelial HK2 cells (P<0.05). Knock-down of CHOP expression by siRNA significantly increased the viability of renal tubular epithelial HK2 cells (P<0.05), decreased the apoptotic rate (P<0.05), increased the expression of Ki-67 and PCNA (P<0.05), and down-regulated the protein level of cleaved caspase-3 (P<0.05). CONCLUSION:The serum mRNA levels of CHOP were increased in the patients with acute kidney injury. Knock-down of CHOP expression inhibits the apoptosis of renal tubular epithelial cells by regulating the expression of proliferation-and apoptosis-related proteins.     

miR-204 inhibits proliferation of Hodgkin lymphoma cells by targeting Sirt1/p53 signaling pathway

AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53.     

利用荧光观测明确载球孢白僵菌松毛虫赤眼蜂对亚洲玉米螟的防控增效作用

载球孢白僵菌松毛虫赤眼蜂能够显著提高对亚洲玉米螟的防控效果，为了明确其对亚洲玉米螟卵期及幼虫期可持续防控的作用机理，采用松毛虫赤眼蜂吸附绿色荧光蛋白（GFP）标记的球孢白僵菌分生孢子，进行赤眼蜂载菌情况，以及亚洲玉米螟卵和幼虫的寄生、侵染过程观察。结果表明，赤眼蜂羽化后能够吸附柞蚕卵表面粘附的白僵菌分生孢子，并将其携带至亚洲玉米螟卵块表面，并吸附于未被赤眼蜂寄生的亚洲玉米螟卵孵化的幼虫体表，实现侵染并致死，幼虫带菌率达60.00%，网室内杀虫生物测定僵虫率达27.00%。本研究表明，载菌赤眼蜂在提高杀虫效率的同时实现害虫可持续防控，该方法为其他载菌天敌的应用提供了依据。     

异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。     

生物炭对Bt Cry1Ac蛋白抗紫外能力影响

为了研究生物炭对紫外线的防护作用，以豆壳烧制的生物炭作为载体，研究生物炭对Bt Cry1Ac蛋白的吸附行为以及生物炭对Cry1Ac蛋白的紫外保护作用。使用扫描电子显微镜、透射电子显微镜、X-射线粉末衍射以及傅立叶红外光谱等手段对生物炭的形貌和结构进行表征。结果表明，生物炭是典型的多孔结构材料，表面具有丰富的官能团。Cry1Ac蛋白与生物炭吸附平衡时间为50 min，最合适的吸附浓度比（生物炭：蛋白）为1：100，二者吸附符合准二级动力学模型和Langmuir模型。在UVB紫外照射4 h后，生物炭与Cry1Ac蛋白复合物对棉铃虫的生物活性是单纯蛋白的4.93倍，显示生物炭具有较好的紫外抵抗效果。研究结果初步表明，制备得到的生物炭能够显著提高Cry1Ac蛋白的抗紫外能力，为后续研发耐受紫外线的农药剂型提供新材料。     

电子鼻和电子舌单独与联合检测掺大豆蛋白或淀粉的鸡肉糜

为实现掺杂掺假鸡肉的快速、客观评价，该研究利用电子鼻和电子舌联合检测技术对掺杂鸡肉糜进行快速检测，通过对采集的数据进行主成分分析和偏最小二乘法分析，所得结果表明，采用主成分分析，电子鼻和电子舌联合检测掺大豆蛋白鸡肉糜和掺淀粉鸡肉糜的主成分总贡献率分别为99.8%和99.1%；采用偏最小二乘法分析，电子鼻和电子舌联合检测鸡肉糜中掺杂大豆蛋白含量的预测值与真实值之间的决定系数为0.992，均方根误差为2.8%；联合检测鸡肉糜中掺杂淀粉含量的预测值与真实值之间的决定系数为0.996，均方根误差为2.4%。表明电子鼻和电子舌联合检测对鸡肉糜的掺杂情况具有良好的区分和预测能力，并且是一种有效、高精度的肉类掺假检测方法。     

产漆酶真菌对高寒草甸土壤有机质矿化特性的研究

为了了解产漆酶微生物真菌对高寒草甸土壤有机质的矿化作用及其特性，实验采用室内密闭静置培养法，对添加产漆酶真菌Marasmius tricolor 310b发酵粗酶液对高寒草甸土壤生物活性有机碳库的影响进行了研究。结果表明：与对照组相比，添加产漆酶真菌Marasmius tricolor 310b发酵粗酶液后，在整个培养期内土壤CO₂释放量和微生物生物量碳含量随培养时间的延长，呈现不断下降的趋势，35 d后趋于稳定；在整个培养周期，微生物呼吸熵的变化总体呈对数降低趋势，处理组符合方程y=-9.964ln （x）+32.281（R²=0.9744），对照组符合方程y=-9.788ln （x）+28.683（R²=0.95），处理组土壤CO₂的累计释放量始终高于对照组（P<0.05），说明菌株310b促进了土壤有机质矿化作用。土壤有机碳含量在培养前期开始降低，但差异不显著，15 d后处理组的土壤有机碳含量低于对照组（P<0.05），说明不同阶段产漆酶真菌对有机质的周转速率不同，推测产漆酶真菌对有机质的转化作用与土壤环境条件有关。     

羟丙基二淀粉磷酸酯对虾糜凝胶特性及其蛋白结构的影响

为考察羟丙基二淀粉磷酸酯（hydroxypropyl distarch phosphate, HPDSP）对中华管鞭虾Solenocera melantho虾糜凝胶性能的改善效果，在虾糜中分别添加1%、3%、5%、7%、9%的HPDSP，以凝胶强度、质地剖面分析、持水性、凝胶微观结构等为指标，并提取虾糜溶胶、凝胶化虾糜、虾糜凝胶中的肌原纤维蛋白，通过圆二色光谱分析蛋白分子二级结构，探究HPDSP对虾糜凝胶性能的影响及其机理。结果表明：添加HPDSP能提高虾糜凝胶的凝胶强度和持水性，并在添加量为7%时达到最大值；HPDSP使虾糜凝胶a*值和b*值下降，但L*值和白度除9%添加量组外，均没有显著变化；适量HPDSP能提高虾糜凝胶硬度、弹性、粘聚性和咀嚼性，促进虾糜形成更加致密稳定的凝胶网状结构。在虾糜凝胶化过程中，加热引起肌原纤维蛋白的α-螺旋含量降低，β-折叠和无规卷曲含量升高，同时HPDSP会对肌原纤维蛋白热变性和聚集产生影响，从而改变其二级结构，影响虾糜凝胶的形成。结论：在中华管鞭虾虾糜中，添加7%的HPDSP能显著改善虾糜凝胶性能。