茶树蛋白激酶基因CsCIPK的克隆与表达特性分析

以茶树品种‘龙井43’作为材料,利用RT-PCR方法,从茶树的cDNA中克隆得到1个编码蛋白激酶的基因,命名为CsCIPK。序列分析表明,CsCIPK开放阅读框长度为1 341 bp,编码446个氨基酸,蛋白质分子量为414234。蛋白功能域预测和多重对比显示,CsCIPK蛋白含有1个保守的N端激酶结构域和1个相对不保守的C端调节结构域,即丝氨酸/苏氨酸激酶结构域和NAF结构域。理化性质、亲/疏水性、无序化分析显示,CsCIPK属于疏水性蛋白,理论等电点为7.04,有4段无序化区域,其二级结构分析显示主要由α螺旋、不规则卷曲组成。通过实时荧光定量PCR对‘龙井43’和‘安吉白茶’中的CsCIPK表达特性进行分析。结果显示‘龙井43’中CsCIPK的相对表达量在高温、干旱及盐处理4 h、低温处理24 h时达到最高。‘安吉白茶’中CsCIPK的相对表达量在高温及盐处理4 h、低温及干旱处理1h时达到最高。CsCIPK在‘龙井43’的根中,‘安吉白茶’茎中表达量最高。不同浓度的GA和IBA处理‘龙井43’茶苗,结果显示0.2 mmol·L-1 GA处理后,CsCIPK表达量先升高后下降,6 d时处理组为对照组的62倍;0.6 mmol·L-1 IBA处理后,CsCIPK的表达量在3 d时显著高于对照组;不同浓度GA和IBA处理后,9 d时CsCIPK表达量均显著低于对照。     

CUDC-907 induces DNA damage and autophagy in glioma cells

AIM:To investigate the effect of CUDC-907, a dual histone deacetylase (HDAC) and phosphatidylinositol 3-kinase (PI3K) inhibitor, on the DNA damage, cell cycle distribution and autophagy in human glioma U251 cells. METHODS:U251 cells were treated with CUDC-907 of different concentrations, and the cell viability was detected by MTT assay. The quantitative γ-H2AX foci were determined by laser scanning confocal microscopy. The cell cycle distribution of U251 cells was examined by flow cytometry. The protein expression was determined by Western blot analysis. RESULTS:CUDC-907 inhibited the cell viability and the phosphorylation of Akt and p70 ribosomal protein S6 kinase (p70s6K) in the U251 cells (P<0.05). In CUDC-907-treated cells, the number of γ-H2AX foci and protein expression of γ-H2AX were increased significantly (P<0.05). CUDC-907 also induced cell arrest in the G₂/M phase by up-regulating the expression of p21, and inhibiting the protein level of cyclin B1 and the phosphorylation of cell division cycle protein 2 (Cdc2). In addition, CUDC-907 triggered cell autophagy, and inhibition of autophagy increased CUDC-907-induced DNA damage of U251 cells. CONCLUSION:CUDC-907 significantly inhibits PI3K/Akt signaling pathway, induces DNA damage and arrests cell cycle in G₂/M phase. Blockage of autophagy promotes CUDC-907-induced DNA damage of U251 cells.     

NRF2 attenuates oxidative stress and lysosomal dysfunction in doxorubicin-induced H9C2 cells

AIM:To study the effect of nuclear factor E2-related factor 2 (NRF2) on oxidative stress injury and lysosomal dysfunction in doxorubicin (DOX)-induced rat myocardial H9C2 cells. METHODS:The H9C2 cells were treated with DOX. The expression of NRF2 at mRNA and protein levels was determined by real-time PCR and Western blot. The H9C2 cells stably over-expressing NRF2 were established by lentiviral infection. Real-time PCR and Western blot were used to identify the efficiency of over-expression. After DOX treatment, the cell viability was measured by CCK-8 assay, the activity of lactate dehydrogenase (LDH), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), and the content of malondialdehyde (MDA) in the cell supernatant were detected. FITC-dextran was used to analyze lysosomal pH, and the protein expression of lysosomal-associated membrane protein 1 (LAMP1) and cathepsin B was determined by Western blot.RESULTS:The expression of NRF2 at mRNA and protein levels in DOX-treated H9C2 cells was significantly decreased (P<0.05). Over-expression of NRF2 significantly up-regulated the mRNA and protein expression of NRF2 in DOX-treated H9C2 cells (P<0.05). After DOX treatment, the cell viability was decreased, and LDH activity was increased. The activity of SOD, GSH-Px and CAT was decreased, and the content of MDA was increased (P<0.05). The lysosomal pH was increased, and the protein expression of LAMP1 and cathepsin B decreased (P<0.05). Over-expression of NRF2 increased the cell viability, decreased LDH activity, increased the activity of SOD, GSH-Px and CAT, and decreased the content of MDA in cell supernatant (P<0.05). Over-expression of NRF2 also decreased the lysosomal pH, and increased the protein expression of LAMP1 and cathepsin B (P<0.05). CONCLUSION:DOX inhibits the expression of NRF2 in the myocardial H9C2 cells. Over-expression of NRF2 attenuates oxidative stress and lysosomal dysfunction in the H9C2 cells induced by DOX.     

Effect of Jieyuwan on depression-like behaviors and expression of BDNF in hippocampus and prefrontal cortex of WKY rats

AIM:To investigate the mechanism of depression and its development, and to study the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus and prefrontal cortex of Wistar-Kyoto (WKY) rats treated with Jieyuwan. METHODS:Adult male WKY rats were used as an animal model of endogenous depression. Wistar rats of the same strain were selected as control group. WKY rats were randomly divided into model group, citalopram group and Jieyuwan group. After intragastric administration for 21 d, the changes of depression-like behaviors were observed by sucrose preference test and forced swimming test. Immunofluorescence and Western blot were used to detect the expression of BDNF in the hippocampus and prefrontal cortex. RESULTS:WKY rats showed significant depression-like behaviors, and the expression of BDNF was significantly decreased in the hippocampus and prefrontal cortex (P<0.01). The reduction of neuronal axons in hippocampus was also observed. After drug treatment, the depression-like behaviors of WKY rats were significantly attenuated, and the expression of BDNF and the number of axons were increased (P<0.01). CONCLUSION:Jieyuwan effectively attenuates the depression-like behaviors of WKY rats, and BDNF is a key factor in its antidepressant effect. Our findings further confirm the involvement of BDNF in the development of depression.     

百香果品种‘紫香1号’果实糖、酸和维生素成分分析

为深入了解‘紫香1号’百香果成熟果实中的可溶性糖、有机酸和维生素组分及含量,采用高效液相色谱法测定各组分的含量。结果表明:‘紫香1号’百香果中的可溶性糖主要为葡萄糖、果糖和蔗糖,各糖分之间差异显著,其中蔗糖含量最高,达38.52mg/g,属蔗糖积累型;有机酸包括苹果酸、丙二酸、乳酸、柠檬酸和琥珀酸,各有机酸之间差异显著,以柠檬酸含量最高,达0.995 mg/g,属柠檬酸优势型;维生素包括维生素A、维生素B1、维生素C和维生素E,各种维生素之间差异达显著水平,其中维生素E含量最高,为430.61 mg/kg。可见,‘紫香1号’百香果含有丰富的可溶性糖、有机酸和维生素,具有较高的甜度和酸度值,故果实成熟时鲜食口感酸甜,风味浓郁。     

紫花苜蓿不同种植布局对苹果害虫及其天敌的影响

为探究紫花苜蓿不同种植布局对苹果园害虫及天敌的影响,在山东省沂水市苹果园设置了紫花苜蓿逐行、隔1行和隔2行种植的布局方式,以自然生草区为对照,系统调查了4种生草方式苹果园果树上和生草上害虫及天敌的发生动态。结果表明:害虫的调查总量,紫花苜蓿逐行种植区最少,比隔1行种植区少20.29%,比隔2行种植区少48.53%,比自然生草区少49.91%;苹果树上天敌的调查总量,紫花苜蓿隔1行种植区最多,是逐行种植区的1.13倍、隔2行种植区的1.29倍、自然生草区的1.78倍;生草上天敌的调查总量,紫花苜蓿逐行种植区最多,是隔1行种植区的1.05倍、隔2行种植区的1.19倍、自然生草区的1.66倍;益害比方面,紫花苜蓿逐行种植区和隔1行种植区的益害比明显高于其他生草区。综合防治效果和种植成本,紫花苜蓿隔1行种植的布局方式更合理。     

两段式花生摘果特性试验研究

为探明两段式收获条件下花生摘果特性,提高花生联合收获机的收获质量,以江苏宿迁地区天府9号为试验对象,应用室内与田间试验相结合的研究方法,研究收获期影响摘果特性的主要因素,揭示花生荚果含水率、晾晒时间、果柄力学特性和摘果作业质量的内在关系。结果表明:基于不同的试验条件与方法,果柄拉断力有所差异,但随晾晒时间(含水率)变化趋势一致,在晾晒2~3天时,果柄拉断力达到峰值,且秧柄节点拉断力大于果柄节点;确定最佳摘果时间为晾晒5~6天,并经捡拾联合收获机摘果试验验证。研究结果为摘果装置的结构参数和运动参数提供了直接依据,并为两段式收获模式下最佳捡拾摘果时间提供了参考,同时为深入研究摘果效率与摘果损失提供了参考。     

收获期花椒的力学特性与形态特征测定与分析

我国花椒的收获主要以人工收获为主。随着社会的发展,花椒市场需求不断扩大,迫切需要解决花椒收获效率低下的问题。因此,花椒收获机械化已成为椒农的心声,研究分析收获期花椒的力学特性可以为花椒收获机械的设计者提供必须的基础理论数据。因花椒在品种、气候条件、土壤及其他方面的差异,其力学特性会有较大的差别。为此,通过陕西省韩城市收获期花椒的实验室测定,采用图像解析和数理统计方法对收获期花椒的力学特性进行分析,得出了花椒主要几何特性指标值的变化区间,并统计分析了花椒果实与果柄及果柄与树枝之间的分离力以及花椒果实的颜色特征,为花椒的收获和加工机器系统的开发提供科学依据。     

我国农用无人机发展概况与展望

近年来,随着我国农业结构的转变与调整,农业生产对农业机械的需要也从普遍的耕作机械、收获机械、农副产品加工机械等方面扩展到了农用无人机。无人机在有动力源的情况下,能通过操控来执行多种作业任务,可广泛应用于农业生产,主要用途体现在植保作业、林业监测、作物授粉及牧群定位等方面。为此,介绍了农用无人机的分类、国内外农用无人机的技术发展概况和现状,以及其在农业领域的应用,阐明了其在我国的发展前景,指出了其在发展过程中存在的一些问题,并提出了相应的发展建议。