菊花花瓣伸长相关基因CmXTHs的克隆与功能验证

从杭白菊(Chrysanthemum morifolium‘Hangbaiju’)中克隆了4个细胞壁松弛和伸展相关的木葡聚糖内转糖苷酶/水解酶基因,分别命名为CmXTH1、CmXTH2、CmXTH3和CmXTH4,序列分析表明其编码的氨基酸序列N端都含有23~30个氨基酸组成的信号肽区域和保守的催化活性位点DE(I/L)DEFLG以及紧邻的N(R/A)T组成的N端糖基化位点,CmXTH1/2/3的C端均含有4个保守的半胱氨酸残基,CmXTH1/2/3/4蛋白的相似度为66.2%。系统进化分析结果表明,CmXTH1/2/3属于XTH家族Ⅰ组,CmXTH4属于Ⅱ组。实时荧光定量结果表明:(1)菊花CmXTH1/2/3/4在根、茎、叶和花蕾中均有表达。CmXTH1在根中表达量较高,CmXTH2/3在茎中表达量较高,CmXTH4在花蕾中表达量最高。(2)CmXTH1/2/3/4在花序不同部位表达量有差异,CmXTH1/4在舌状花中表达量最高,CmXTH2/3在筒状花中表达量较高。(3)CmXTH1/2/3/4在舌状花不同发育阶段表达量差异显著,CmXTH1/2在盛花期表达量最高,CmXTH3在衰老期表达量最高,CmXTH4在初开期表达量最高。病毒诱导基因沉默结果表明,CmXTH1/2/3/4沉默系的花序直径和舌状花长度与对照相比均有所减小,其中CmXTH4沉默系减小最大,分别减小了25.67%和10.42%,舌状花花瓣表皮细胞与对照相比明显变小,影响了舌状花花瓣的伸长;CmXTH2沉默系的筒状花雄蕊长度和CmXTH4沉默系的花萼直径分别与对照相比显著减小。这表明菊花CmXTH1/2/3/4基因参与了花序的开放,促进了舌状花花瓣的伸长,对于菊花的花序增大具有重要作用。     

支管射流三通结构优化与试验

为了提高支管射流三通水力性能,改善滴灌的灌水均匀性,基于CFX数值模拟技术,对进口宽度为15 mm的支管射流三通进行结构优化.选取位差、劈距、劈尖半径和侧壁倾角为影响因素,通过四因素三水平正交设计了9组模型,边界条件设定为进口压力100 kPa.选取支管射流三通出口设计流量为评价标准,支管射流三通最优结构尺寸为位差5.5 mm、劈距113 mm、劈尖半径13 mm、侧壁倾角10°.此结构尺寸参数下的支管射流三通水力性能试验结果表明:在进口水压为100 kPa时,支管射流三通脉冲频率为148次/min,水头压力振幅为37.9 kPa,水头压力损失为16.7 kPa,出口流量为0.698 L/s;支管射流三通所接滴灌带长度为60 m时,与普通支管三通相比,支管射流三通的灌水均匀系数提高了2.78%,流量偏差率降低了4.72%.该研究可为射流技术在脉冲滴灌系统的研究、开发与应用提供理论依据.     

柑橘胼胝质合成酶基因家族的表达分析

胼胝质由胼胝质合成酶合成,胼胝质沉积是植物对入侵病原的防御反应。为了探索胼胝质合成酶(Callose synthase,CalS)基因在柑橘中对黄龙病的应答情况,从甜橙基因组中筛选并利用生物信息学分析了柑橘胼胝质合成酶(CsCalS)家族的12个基因。这些基因编码的氨基酸序列与拟南芥中的胼胝质合成酶具有较高的同源性。荧光实时定量PCR结果显示,CsCalS5和CsCalS12在柑橘健康叶片中的表达量高于其他胼胝质合成酶基因。比较黄龙病感病材料和健康对照材料,发现CsCalS5、CsCalS7和CsCalS8在感病的‘砂糖橘’和‘强德勒’柚中均上调表达,可能参与柑橘对黄龙病病原的应答反应。     

不同外植体及冷藏处理对华中枸骨组织培养的影响

[目的]研究不同外植体及冷藏处理对华中枸骨组织培养的影响。[方法]以华中枸骨带芽新梢茎段为外植体,考察外植体的取材时间、部位以及4℃冷藏处理对其初代培养的影响。[结果]华中枸骨以带芽的新梢茎段作为外植体进行组织培养时,以春季采取植株中上部枝条为宜,其中春季萌条的芽增殖倍数可达到4,中上部萌条的芽萌动率可达到45%。适当的冷藏处理对外植体的消毒以及芽的萌动有利,4℃冷藏处理14 h后的消毒成功率最高,可达到92%;冷藏处理22 h的芽萌动率最高,可达到18%。[结论]华中枸骨组织培养中外植体最好于春季选取植株中上部萌发的新梢顶芽;华中枸骨芽的萌动率随冷藏时间的增加呈线性增长,可能与低温打破休眠有关。     

Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

Lycopene protects primary mouse cerebrocortical neurons against t-BHP-induced damage in vitro

AIM: To investigate the protective effect of lycopene on primary mouse cerebrocortical neurons exposed to tert-butyl hydroperoxide (t-BHP) and its mechanisms of in vitro.METHODS: Primary cerebrocortical neurons of newborn C57 mice were extracted and divided into normal group, t-BHP group, lycopene+t-BHP group and lycopene group. The neuronal damage was induced by t-BHP exposure for 24 h, and the cell viability was examined by MTT assay. ROS content was measured by flow cytometry, and the protein levels of Bax, Bcl-2, caspase-3, cleaved caspase-3 and cytochrome C were examined by Western blot.RESULTS: The primary mouse cortical neurons expressed MAP-2 protein. Lycopene at concentration of 4 μmol/L reversed the decrease in cell viability. Flow cytometry revealed that lycopene treatment attenuated ROS content under the condition of t-BHP exposure. In addition, the protein level of Bcl-2 was increased, and the expression of Bax, cleaved caspase-3 and cytochrome-C was suppressed in lycopene+t-BHP group.CONCLUSION: The protective effect of lycopene on cortical neurons with t-BHP-induced injury may be involved in the mechanism of neuronal antioxidative response by down-regulating caspase-3 and Bax/Bcl-2 through the mitochondrial apoptotic pathway.     

绿洲灌区不同密度玉米群体的耗水特性研究

针对水资源不足严重制约绿洲灌区玉米生产,密植对玉米耗水特性影响研究薄弱,以及生产实践中缺乏调控种植密度以同步提高产量和水分利用效率的理论依据等问题,2012—2015年,以先玉335为参试品种,在相同施肥、灌水制度下,设75 000株hm–2(低,D1)、87 000株hm–2(中,D2)、99 000株hm–2(高,D3)3个密度水平,探讨密度对玉米耗水的时间动态、耗水结构以及利用效率的影响,以期为优化密植增产节水技术提供理论依据。结果表明,随密度增大,玉米全生育期总耗水量(ET)增大,大喇叭口期至吐丝期是耗水量增大的主要时期,高、中密度处理与低密度处理相比,全生育期总耗水量4年平均分别高22.8%、14.4%,大喇叭口期至吐丝期平均分别高28.4%、18.2%,其他生育时期不同处理的耗水量差异不显著。增加密度可降低玉米的无效耗水,提高水分利用率,高、中密度处理较低密度处理全生育期的棵间蒸发量(E)4年平均分别减少56.5 mm、27.6 mm,密植降低棵间蒸发量的主要时期在拔节至灌浆期,其中大喇叭口期至吐丝期的日均棵间蒸发量分别减少0.51 mm、0.27 mm;高、中密度与低密度处理相比,全生育期的E/ET分别降低15.8%、6.2%,其中拔节至大喇叭口期减小幅度最大,分别为22.1%、10.7%。与低密度处理相比,高、中密度处理的籽粒产量分别提高了17.9%、14.8%,但高、中密度处理间的产量差异不显著;高、中、低密度处理的水分利用效率(WUE)分别为18.2、19.3和16.8 kg hm–2 mm–1,中密度处理的WUE显著高于低密度处理;与低密度处理相比,高、中密度处理4年平均灌溉水利用效率(IWUE)分别提高了34.5%、19.6%。本研究说明,在传统供水覆膜条件下,进一步增加种植密度是干旱绿洲灌区提高玉米产量和灌溉水利用效率的可行措施。     

离子阱液质联用仪应用于植物叶片游离氨基酸快速分析方法的建立

建立一种液相色谱-离子阱串联质谱(LC-MS-MS)同时分析植物组织中23种游离氨基酸的方法。植物样品经盐酸-乙醇法一步提取后直接进样,无需进行衍生和固相萃取等其他前处理步骤。液相色谱采用Intertsil OSD-4C18(250mm×3.0mm)色谱柱,采用甲醇-甲酸体系进行洗脱,对23种氨基酸获得比较理想的分离效果。结果表明,该方法通用性良好、样品制备简单(一步提取,无需衍生)、灵敏度高、检出范围宽、高效快速,可以在30min内完成整个分析过程。     

不同供磷水平对辣椒根系形态和根际特征的影响

以辣椒品种线椒王为试验材料，采用盆栽方式，通过分析不同施磷水平下辣椒根系形态和根系生理的变化，探究辣 椒磷高效利用的主要根系及根际过程。结果表明，辣椒地上部和根系生物量随供磷水平的增加呈现先增加后下降的趋势，整 株磷浓度以及整株磷累积量呈增加趋势。随供磷水平增加，根系总根长呈先增加后下降趋势，根系平均直径、根冠比呈逐渐 降低趋势；辣椒根系菌根侵染率和土壤酸性磷酸酶活性呈现先增加后下降的趋势。相关分析结果表明，土壤有效磷浓度和整 株磷浓度与根系平均直径、菌根侵染率呈极显著负相关，与总根长可用一元二次方程拟合，表明辣椒根系适应低磷的根际特 征可能是通过增加根系生物量及总根长，同时降低根系平均直径来实现的。     

Cigarette dust particles induced lung function injury of chronic obstructive pulmonary disease in mice

AIM: To investigate the pathogenesis of chronic obstructive disease (COPD) in mice by using nasal drip of cigarette dust particles (DSP) induced pulmonary function damage model.METHODS: BALB/c mice were randomly divided into control group, LPS 100 mg/L group, DSP 0.75 mL/L group, DSP 1.5 mL/L group and DSP 3 mL/L group for 30 days. The method of nasal drip was used for 30 days to establish the COPD model. Rrs, Ers, Crs, Est, Cst, P-3/8Rn, P-3/8G and P-3/8H were measured for evaluating lung function of the mice in each group by the method of FlexiVent. The effect on the increase of airway resistance induced by methacholine (Mach) was determined using main bronchial rings by Myograph method. The HE, Masson and Resorcinol fuchsin staining of mouse tracheas and lung tissues were conducted. RESULTS: Continuous nasal drip with DSP for 30 days increased Rrs, Ers, Est, P-3/8Rn and decreased Crs, Cst, P-3/8G and P-3/8H in the mice. DSP significantly shifted the dose-effect curve of tracheal contraction induced by Mach to the left, increased the sensitivity of the airway to Mach, and significantly increased the maximal contractile airway effect of Mach. Exposure to DSP caused fibrosis of airway subepithelial, deposition of collagen in the airway basement membrane under the reticular plate, induced reticular plate thickening, pulmonary bronchial lumen serious deformation, and the inflammatory cell infiltration in the lung of mice. Significantly increased alveolar wall muscle fibers and collagen fibers were also observed. CONCLUSION: The lung function and pathomorphological changes of COPD mice induced by 30 days nasal drip of cigarette dust particles were similar to those of human COPD.