Importance of Cell Wall Degrading Enzymes Produced by Fusarium graminearum during Infection of Wheat Heads

Cytological studies were carried out to elucidate the importance of cell wall degrading enzymes (CWDE) during infection of wheat spikes by Fusarium graminearum. Scanning electron micrographs revealed that at 6–24 hours after inoculation (hai) of single spikelets with macroconidia of F. graminearum, the fungus germinated by forming several germ tubes and developed a dense hyphal network in the cavity of the spikelet. At 24–36hai, the fungus formed infection hyphae which invaded the ovary and inner surface of the lemma and palea. Transmission electron microscopical studies revealed that the fungus extended inter- and intracellularly in the ovary, lemma and rachis and caused considerable damage and alterations to the host cell walls. In different tissues of healthy and F. graminearum-infected wheat spikes the cell wall components cellulose, xylan and pectin were localized by means of enzyme-gold and immuno-gold labelling techniques. Localization of cellulose, xylan and pectin showed that host cell walls which were in direct contact with the pathogen surface had reduced gold labelling compared to considerable higher labelling densities of walls distant from the pathogen–host interface or in non-colonized tissues. The reduced gold labelling densities in the infected host cell walls indicate that these polysaccharide degrading enzymes might be important pathogenicity factors of F. graminearum during infection of wheat spikes. The results revealed that, infection and colonization of wheat spikes by F. graminearum and reactions of infected host tissue were similar to those reported for F. culmorum.     

Expression and subcellular localization of urocortin in syncytiotrophoblast of human term placenta

AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine.     

巴戟天MoDXS基因及其启动子的克隆与分析

从药用植物巴戟天根部组织中克隆了3个1–脱氧–D–木酮糖5–磷酸合成酶基因MoDXS1、MoDXS2-1和MoDXS2-2,其全长cDNA分别为2 676、2 667和2 610 bp,编码区序列长度为2 154、2 121和2 190 bp,编码717、706和729个氨基酸。BlastP序列比对分析表明MoDXS蛋白与其他植物的DXS蛋白高度同源;系统进化发育树分析显示,MoDXS蛋白与中粒咖啡和小粒咖啡DXS蛋白亲缘关系最近。MoDXS1、MoDXS2-1和MoDXS2-2蛋白被分别归为clade 1、clade 3和clade 2。MoDXS1在巴戟天根中表达量最高;MoDXS2-1在巴戟天根、茎、叶中的相对表达量差异显著,叶中最高;MoDXS2-2在根中表达量最高,而在茎和叶中最低。MoDXS1、MoDXS2-1和MoDXS2-2基因的5′端上游启动子序列长度分别为2 538、732和1 744 bp。亚细胞定位预测3个MoDXS均在叶绿体上。     

新麦草YUCCA基因克隆及其表达特性分析

【目的】克隆新麦草分蘖关键基因YUCCA并明确其表达特性,为该基因功能验证和育种利用奠定基础。【方法】以不同分蘖类型的新麦草分蘖节为材料,采用PCR法克隆新麦草YUCCA基因并进行同源序列比对,构建1300-cYFP-YUCCA过表达载体,利用烟草瞬时转染及蛋白质印迹法分析新麦草YUCCA蛋白的表达位置及表达情况,通过RNA-Seq数据及qRT-PCR分析验证YUCCA基因的相对表达量。【结果】成功克隆了新麦草YUCCA基因全长,大小为1 336 bp,最大阅读框为1 236 bp,并成功构建了1300-cYFP-YUCCA过表达载体。多序列比对结果显示,新麦草YUCCA主要与麦类作物的亲缘关系较近。表达特性分析发现,YUCCA基因在多分蘖型新麦草中的相对表达量远低于少分蘖型新麦草;与叶、根等组织相比,YUCCA基因在分蘖节中的相对表达量最高。烟草瞬时转化和蛋白质印迹法分析均显示,YUCCA蛋白可以正常表达,其亚细胞定位于细胞质中。【结论】YUCCA基因在调控新麦草分蘖中起下调作用,可用于后续YUCCA基因功能的研究。     

茶树CsSPMS基因全长cDNA克隆和时空表达分析

为探究精胺合成酶（spermine synthase,SPMS）与茶树（Camellia sinensis）耐寒性的关系,以茶树品种‘迎霜’为试验材料,利用简并引物PCR扩增技术结合5′/3′RACE方法,获得与低温胁迫相关的CsSPMS片段cDNA全长序列（GenBank登录号为KJ580429）。该序列全长1 374 bp,包含1 113 bp的完整开放阅读框（ORF）,编码371个氨基酸,预测分子量41.28 kD;构建亚细胞定位载体pJIT166-GFP/SPMS,亚细胞定位结果显示CsSPMS定位在细胞核上。荧光定量PCR分析表明CsSPMS在根、茎、叶、芽、花和果中都有表达,在根、茎中表达量较高;低温胁迫处理下不同组织CsSPMS的表达量,在叶片中2 h达到峰值,而在根中12 h达到高峰;在低温条件下4个茶树品种的耐寒性与CsSPMS表达量密切相关。     

植物脱水素对多种逆境的响应

综述了目前关于脱水素（Dehydrin,DHN）的研究进展,包括脱水素的蛋白结构特征、细胞定位及作用机制,脱水素在生物及非生物逆境下的转基因研究,脱水素的磷酸化修饰,脱水素的结合金属离子、清除活性氧、作为分子伴侣 保护酶活性等特性。展望了利用现代生物技术将脱水素用于提高植物对逆境的耐受力的应用前景,提出今后还需在脱水素作用机制、基因家族功能与生物胁迫相关性等几个方面进行深入研究。     

昆虫病原线虫共生菌胞内外蛋白的提取及杀虫活性比较

通过超声波破碎、硫酸铵盐析等方法从10株昆虫病原线虫共生菌的发酵液中分离出各菌株的胞内和胞外蛋白,测定各蛋白样品含量及其对棉铃虫初孵幼虫的生长抑制毒性,并用Native-PAGE和SDS-PAGE对各粗蛋白组分进行初步分析。结果表明有7个菌株的胞内和胞外蛋白对棉铃虫初孵幼虫生长均显示出不同程度的抑制生长活性。其中以嗜线虫致病杆菌3个菌株CB6、All和Bw活性最强,其胞内蛋白的活性分别达97.1%、97.9%、97.2%,胞外蛋白分别达95.8%9、6.3%、88.6%。不同菌株胞内胞外蛋白产量各不相同。电泳图谱表明,不同菌株间及同一菌株胞内胞外蛋白有差异,但近缘菌株相似。     

水稻糙米粒宽性状的QTL定位研究

利用由怀香B和筒恢211配制的F2群体进行糙米粒宽基因的QTL定位,定位到了2个粒宽基因的QTLs,其中有1个新的QTLs.有1个增效基因为主效基因,解释的表型变异高达20.2%.研究表明,所利用的F2群体的糙米粒宽可能主要是由1个主效QTL所控制.     

基于全质心WLS-HDS-TWR算法的无人驾驶农业机械协同定位算法研究

针对室内全球导航卫星系统（Global navigation satellite system,GNSS）信号受遮挡时,农用车辆协同定位精度低、稳定性差、信号丢包等问题,本文开展面向超宽带（Ultra wideband,UWB）调频技术的室内外农用车辆协同定位算法研究。首先,搭建三基站多边测距定位模型,实现主基站绝对位置标定以及辅助基站绝对位置坐标的变换求解;其次,提出全质心加权最小二乘的高速双边双向（Weighted least squares high double sided two-way ranging,WLS-HDS-TWR）农机协同定位算法,基于泰勒级数展开的WLS估计算法,求解主车位置。同时,提出面向室内环境的多状态基站组合的UWB定位模块布设模式,并验证其可行性;通过飞行时间法（Time of flight,TOF）获取主从车距离信息,融合GNSS标定位置信息、主车坐标信息以及测距信息,实现主从车协同定位。最后,基于Prescan/Simulink搭建联合仿真平台,验证提出算法的可靠性;通过农用履带车辆开展室内及室外协同定位实车试验,试验结果表明：全质心WLS-HDS-TWR协同定位算法可有效解决室内GNSS信号缺失问题,室内环境下,定位精度较HDS-TWR及全质心LS-HDS-TWR算法分别提高26.98%和22.03%,满足智能农机协同定位作业需求。