家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

家蚕质型多角体病毒苏州核包涵体小种的纯化

从BmCPV_t内发现和分离出一株新的核包涵体小种,初步命名为家蚕质型多角体病毒—苏州核包涵体小种(Cytoplasmic Polyhedrosis Virus——Suzhou nuclear polyhedra forming strain,简称CPV——Suzhou N strain),家蚕感梁CPV——Suzhou N Strain引起的多角体病,与国内外已知的CPV各变异株不同。本病蚕的多角体只在中肠组织圆筒形细胞核内检出,多角体内有许多病毒粒子,病毒粒子球状,具有核蛋白紫外吸收特征性光谱和较强的感染活性,病毒核酸初步鉴定为dsRNA。     

苹果退绿叶斑病毒组织印迹的免疫法检测与定位

用组织培养技术将富士苹果茎尖外植体建立在ＭＳ培养基中,形成试管苗。取４—６周龄的茎与愈伤组织,徒手切取约１ｍｍ厚的横切面,印迹在硝化纤维膜上。印迹组织经封闭后,与ＣＬＳＶ碱性磷酸酶标抗体反应。对反应结果进行显色。感染ＣＬＳＶ的印迹组织呈紫色反应,正常组织无显色反应。结果表明,该方法十分容易检测印迹组织中的ＣＬＳＶ。带毒愈伤组织较茎的显色反应敏感,显色反应时间仅为茎的一半（１５ｍｉｎ）。ＣＬＳＶ在愈伤组织中有两种分布情况：一是所有组织均有显色反应;二是呈不均匀分布。ＣＬＳＶ在茎组织中有三种分布情况：第一,除髓部外,表皮、皮层及维管系统中均有分布;第二,不均匀分布,ＣＬＳＶ集中分布于表度组织中,在皮层及韧皮部仅有少量分布;第三,“嵌合”分布,即同一种组织中部分组织带毒,部分为正常组织。     

苜蓿银纹夜蛾核多角体病毒在家蚕蛹体内的复制及重组病毒外源基因的表达

家蚕蛹在复眼着色期,经4℃冷藏24小时后,可被Ac NPV(苜蓿银纹夜蛾核多角体病毒)感染。在蛹体内复制出的多角体大小差异较大,且比在昆虫Sf—21细胞中繁殖的Ac NPV和在蚕蛹体内形成的Bm NPV多角体小。在蛹体内繁殖的Ac NPV的游离病毒可以回返感染Sf—21细胞。由蚕蛹内分离的Ac NPV基因组DNA的限制性内切酶图谱与野生型的Ac NPV相同。以上结果证明Ac NPV可以在蚕蛹体内复制。含HBsAg基因(人乙肝表面抗原)的重组Ac NPV亦可感染家蚕蛹,并能正确表达外源基因。此外,还发现化蛹后第2天的家蚕蛹被注射约5×10~5PFU的Ac NPV,可诱导“人工滞育蛹”现象的发生,在25℃经过30天后蛹仍存活,发育停滞在复眼着色前阶段。     

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.     

‘新红星’苹果果实蔗糖合酶的活性及亚细胞定位

 在‘新红星’苹果果实发育过程中, 伴随果糖、葡萄糖和蔗糖的积累, 蔗糖合酶的分解活性逐渐下降, 而合成活性逐渐升高。苹果果实的蔗糖合酶由分子量为90 kD 的亚基组成, 其免疫信号强度随发育进程而增加。蔗糖合酶主要定位于果肉细胞的细胞质中, 发育中后期该酶的分布密度有一定增加。综合结果认为, 蔗糖合酶调控发育早期蔗糖的降解和发育后期蔗糖的积累。     

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.     

基于GIS的农村居民点空间分布特征研究----以扶余市为例

科学认识乡村地域结构是全面推进农业农村现代化和乡村振兴的基础。本文以扶余市第二次土地调查调数据为基础，借助ArcGIS空间分析软件，运用规模分析、核密度估算、缓冲区分析，从用地规模、分布格局与土地利用条件方面探讨扶余市农村居民点地域分异特征。研究表明：扶余市以微、小型居民点居多，居民点数量随着规模的增大逐渐减少，居民点之间的距离差异较大，总体呈现大散居、小聚居，不均匀分散分布；农村居民点的平均核密度为0.4个/km2，空间布局呈东南部高密度集聚成片，向西逐渐密度降低，西部零散错落分布；居民点分布呈现较强的城镇指向性，交通用地对居民点有集聚的作用，河流对微型居民点和大型居民点有集聚吸引的功能。     

苦瓜转录因子McEIL2的克隆及其表达分析

本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。     

鸡Perilipin1基因的克隆及亚细胞定位

本研究旨在克隆鸡(Gallus gallus)脂滴包被蛋白基因(Perilipin1)的cDNA序列,并检测其在鸡前脂肪细胞分化过程中的亚细胞定位.本研究以7周龄肉鸡腹部脂肪组织为材料,采用RT-PCR和RACE的方法扩增并克隆了鸡Perilipin1基因的5'UTR、CDS和3'UTR片段,分析了该基因的结构;以鸡原代前脂肪细胞为材料,利用免疫荧光及激光共聚焦技术,在诱导分化的不同时间点(12～120 h),检测了鸡Perilipin1在前脂肪细胞中的表达位置.结果表明,本研究获得的鸡Perilipin11基因5'UTR、CDS和YUTR序列总长度为2 379 bp,该基因由9个外最子、8个内含子组成(ATG位于第二外显子上);在鸡原代前脂肪细胞诱导分化过程中,Perilipin1始终包被在脂滴周围.综上,本研究成功克隆了鸡Perilipinl基因的完整cDNA序列并确定了Perilipin1在鸡前脂肪细胞分化过程中与脂滴的空间位置关系,该结果为深入开展鸡Perilipinl基因的功能研究,揭示鸡脂肪代谢的分子遗传机理提供了重要的理论依据.