细胞凋亡的线粒体途径调控

细胞凋亡是近年来研究的一个热点问题，涉及到细胞内许多复杂的生化过程．线粒体是真核生物能量和代谢的中心，也是细胞凋亡信号传导途径中起关键调节作用的细胞器，对细胞凋亡的线粒体途径调控机制进行研究具有重要意义．本文综述了细胞凋亡过程中线粒体途径的信号传导及其调控机制、线粒体对细胞凋亡信号的反应、细胞凋亡过程中线粒体功能丧失几方面的研究进展．图1参43     

棉铃虫核多角体病毒基因组限制酶酶切分析及其多角体基因的定位

应用１１种限制性内酶ＢａｍＨⅠ、ＢｓｔＥⅡ、ＥｃｏＲⅠ、ＥｃｏＲⅤ、ＨｉｎｄⅢ、ＫｐｎⅠ、ＰｓｔⅠ、ＳａｌⅠ、ＳｓｔⅠ、ＸｂａⅠａⅠ和ＸⅠⅠ和ＸｈｏⅠ分别将中国棉铃虫核多角体病毒（单粒包埋ＨａＳＮＰＶ）湖北株基因组Ｄ组ＤＮＡ酶切为１０、１２、２２、２１、１３、６、６、４０、６、２１、６个片段，并求得基因组大小平均为１什什Ｍｒ≈７９．１×１０６．以家委核多角体病毒ＢｍＮＰＶ多角体基因为探针，利用Ｓｏｕｔｈｅｒｎ杂交技术将病毒多角体基因定位在ＳａｌⅠ４．２×１０３ｂ左右的片段上．与棉铃虫核多角体病毒其它株系酶切图谱比较结果表明，本株病毒与上海等株系及美洲棉铃虫核多角体病毒ＨａＳＮＰＶＥｌｋａｒ株系酶切图谱相似，它们之间的条带数和大小差异较小，而与已发现的所有多粒包埋型病毒ＨａＳＮＰＶ酶切图主谱差异较大．据此认为ＨａＳＮＰＶ和ＨａＭＮＰＶ属于基因型不同的两类病毒，而ＨａＳＮＰＶ不同分离株的同源性很高     

龟足精子发生和成熟精子的超微结构

对龟足(Pollicipes metella）精子发生和成熟精子的超微结构进行了研究．精子发生属于变态型．细胞由多边形变为椭圆形到长梨形直至弹开成线形；胞核由椭圆形逐渐变长弯曲成U形，最后变为细长形；中期精子细胞出现的椭圆形附属小滴也弯曲成U形，最终成长柱状．早期、中期、后期精子细胞出现核周微管及核囊，核周微管及核囊在成熟精子中消失．精子为前中部带有一个长柱状附属小滴的线状结构(长约36μm)，可分为顶体、颈部、中段和尾部四部分．顶体为锥状(约0．6μm)，顶体后的基体发出轴丝，轴丝纵贯末端．核细长线状(约16μm)．附属小滴长柱状(约8μm)，内有棒状致密斑．核、附属小滴与轴丝并行．鞭毛为典型的“9 2”结构。     

水稻抗虫转基因光温敏核不育系植株的获得

以质粒pCUBAC-HPT作抗虫基因供体，优良光温敏核不育系612S为受体，采用基因枪转化法，获得了转抗虫基因sck和sbk的植株．分子证据表明，外源基因己整合到受体基因组中．蛋白活性测定和Western blotting结果显示，转基因在受体植株中得到表达．田间抗虫性测定表明，外源基因的转入提高了受体的抗虫性．本文还讨论了转基因技术在水稻育种中的作用．图5表2参12     

桃ACC合酶基因组DNA（pACSG01）的序列分析及其表达

以桃基因组DNA为模板，用套式PCR技术扩增并克隆了桃ACC合酶基因片段，将其克隆（定名为pACSG01）并进行序列测定，表明该自然全长1320bp，并富含HindⅢ和EcoRⅠ位点，与其它ACC合酶基因结构序列有一定的相似性，内含有两个内含子，编码的氨基酸序列与已克隆桃ACC合酶cDNA推导的氨基酸序列的同源性分别为57．4％和56．8％，pACSG01与我们已克隆的两个桃ACC合酶cDNA基因表达有所不同，RNA点杂交和RT－PCR结合Southern杂交分析表明，该基因在成熟和乙烯处理的果实均不表达，伤处理、LiCl和生长素处理也不能诱导核基因的表达，在衰老花瓣中也不表达，图3参18。     

基于叶绿体基因组的浮萍亚科系统进化

由于浮萍形态高度退化,其系统进化研究较为困难,基于叶绿体基因组研究浮萍亚科系统进化对解决该问题具有重要意义.通过过滤全DNA数据(即包含细胞内的核DNA、叶绿体DNA和线粒体DNA的测序数据)和直接提取叶绿体DNA测序两种方法组装叶绿体基因组,然后分别基于rbc L基因和叶绿体全基因组序列构建浮萍亚科系统发育树,并比较浮萍叶绿体基因组大小.经比对发现,通过两种方法得到的Landoltia punctata ZH0051叶绿体基因组完全一致,全长均为171 013 bp,序列相似度100%.基于叶绿体全基因组序列构建浮萍亚科系统发育树的节点置信值都达到了1,确认了浮萍亚科的进化次序为多根紫萍属、少根紫萍属、绿萍属、扁平无根萍属、芜萍属.浮萍亚科中叶绿体基因组最大的为少根紫萍属(171 kb),最小的为绿萍属(166 kb).本研究表明过滤全DNA数据组装出的浮萍叶绿体基因组是可靠的;浮萍叶绿体基因组大小虽有所差异,但随进化没有明确的扩张或收缩的趋势.     

榨菜线粒体DNA雄性不育相关片段克隆及序列分析

研究了榨菜(Brassica juncea var．tumida Tsen and Lee)细胞质雄性不育性(CMS)与线粒体基因组的关系,以榨菜细胞质雄性不育系及其保持系线粒体DNA(mtDNA)为模板,设计合成引物进行PCR扩增,从榨菜胞质雄性不育系mtDNA上扩增得到与雄性不育相关的特异片段T1170,点杂交分析表明,T1170仅与榨菜胞质雄性不育系mtDNA有杂交信号,而保持系的mtDNA则无,序列测定该片段全长1173bp,编码389个氨基酸,其中疏水氨基酸占35％。图4参9。     

nano-TiO_2与1,2,4-TCB联合作用对斑马鱼鳃组织结构及染色体损伤研究

采用组织病理切片法、微核及彗星试验研究nano-TiO_2和1,2,4-TCB联合染毒对斑马鱼鳃组织结构、细胞微核率及DNA损伤的毒性效应。结果表明,单独nano-TiO_2暴露不会导致斑马鱼鳃组织结构损伤,也不会引起明显的微核及染色体损伤。1,2,4-TCB(≥10 mg·L~(-1))会引起鳃组织和细胞染色体的损伤。nano-TiO_2与1,2,4-TCB联合作用会加重斑马鱼的鳃组织结构损伤,出现鳃小片排列严重紊乱,表皮细胞断失严重,鳃小片上皮细胞坏死脱落,鳃丝结构模糊等症状。两者联合作用会引起鳃细胞微核率增加和彗星细胞率的增加,呈现一定的剂量-效应关系,且两者间的联合作用具有协同作用。对彗星试验的尾部DNA百分比、尾长及尾距的分析发现,与对照组和单独作用相比,两者联合作用对彗星指标均有显著增强作用,这表明两者联合作用对染色体的损伤属于非特定片段损伤。     

玉米多胞质系线粒体数量和结构特征的比较分析

以玉米多胞质系为材料,采用透射电镜的观察方法,比较观察了１１种材料中黄化幼芽细胞的线粒体结构特征的变化．观察表明,１１种材料黄化幼芽线粒体形态及外膜、内膜、泡状嵴和基质颗粒等没有什么显著的差异,而线粒体大小和每个细胞内的线粒体数量却是明显不同的．Ｆ检验说明,线粒体大小和每个细胞线粒体数量上有极显著的差异,也即是说,１１种不同细胞质类型的材料细胞质作用和核质（基因）互作是明显的．试验还观察到线粒体、分裂现象和线粒体、淀粉粒聚集在胞间连丝附近的有趣现象．     

广州白云山次生常绿阔叶林的群落结构动态

根据对广州白云山次生常绿阔叶林群落相隔６ａ的两次植被调查，分析了广州白云山次生常绿阔叶林在保护条件下，自然演变过程中的群落结构动态．群落的垂直结构变化主要以大量幼树进入第一层成树为特点；水平结构则以密度和盖度的增长为特点．在种类组成方面，６ａ前群落中居多的阳性树种现已逐渐为耐阴性较强的种类所代替．在组织结构水平上，乔木层的物种多样性指数、群落均匀度指标趋于增加，而生态优势度指标趋于降低，反映了群落向着组成结构复杂、物种多样性增高的方面演变，且前期变化速度较快，后期较慢并趋于稳定．     

Spermatozoa and spermatogenesis in a monoplacophoran mollusc,Laevipilina antarctica: ultrastructure and comparison with other Mollusca

Ultrastructural features of spermatozoa and spermatogenesis are described for the first time in a monoplacophoran and compared with data for other conchiferan molluses. Spermatozoa of Laevipilina antarctica Warén and Hain, 1992, are of the structurally simple, aquasperm type, featuring a conical acrosome, a compact nucleus with lacunae, a short midpiece and a single flagellum. The acrosomal vesicle shows an electron-dense inner zone, and a basal invagination (subacrosomal space) contains granular material but no axial rod. The nucleus exhibits a shallow indentation apically which contains subacrosomal material, and five (sometimes four) indentations posteriorly which partially accommodate the five (rarely four) midpiece mitochondria. Two centrioles are present, the distal connected to the annulus by satellite fibres and acting as a basal body for the flagellum (axoneme probably 9+2 structure). Spermatogonia, characterized by an oblong nucleus and one or two nucleoli, line the basal membrane of the testis wall; spermatids of varying stages of maturity occupy the remainder of the testis. Acrosome and flagellum production is already well advanced in spermatids and probably commences at the spermatocyte stage. Cytoplasmic bridges occur in all developmental stages, most visibly in spermatids. The spermatid chromatin condenses in large tracts, leaving electron-lucent lacunae. Mitochondria collect posteriorly and form, presumably by fusion, the five (or four) larger, spherical mitochondria which gather around the centrioles.     

玉兰油细胞的发育及其与器官结构发育的关系

利用石蜡切片法及半薄切片法对玉兰茎、叶、花和种子中油细胞的发育进行了研究，结果表明：在所研究的各器官中油细胞以相同的方式发生。油细胞原始细胞因其细胞质染色较深、细胞核大明显及液泡化程度低而易与周围组织细胞分辨，以后，原始细胞逐渐液泡化，最终形成一个中央大液泡，其细胞质及细胞核呈一薄层，当油细胞成熟时，其细胞质和细胞核解体，而大液泡充满整个油细胞腔，成为储油的囊，在一些油细胞中，可观察到怀形构造，但     

Spermatozoan ultrastructure and spermatogenesis in aplousobranch ascidians,with some phylogenetic considerations

Spermatozoa and some stages of spermatogenesis were studied for four species of aplousobranch ascidians. Spermatozoa of Clavelina lepadiformis (Müller) (family Clavelinidae) are plesiomorphous in that they have apical acrosomal vesicles and a moderately elongated, cylindrical nucleus. The elongated mitochondrion is twisted ea. 11/2 times around the nucleus. In their ultrastructural morphology, C. lepadiformis sperm conform in some respects to the least-derived ascidian sperm, those of the phlebobranch ascidian Ciona intestinalis L.; however, the sperm of Clavelina lepadiformis have two apomorphies not shared with those of Ciona sp.: (1) the mitochondrion of Clavelina lepadiformis is long and spiralled along the entire nucleus rather than being comparatively compact and not at all helical; (2) the mitochondrial cristae are elongated parallel to the long axis of the nucleus, whereas in Ciona sp. sperm the cristae are unmodified. In Distaplia sp., Aplidium sp. and Synoicum pulmonaria (Ellis and Solander) the spermatozoa are more derived and consist of a proximal cylindrical and a distal corkscrew-like part. The mitochondrion in Distaplia sp. and Aplidium sp. contains electron-dense material and extends in a long thread around the nucleus. In S. pulmonaria the mitochondrion surrounds the anterior part of the nucleus in mature spermatozoa, and an elongated, dense structure displaying fine striation is enclosed in the mitochondrion in late spermatids. The sperm ultrastructural morphology observed in this study is consistent with the majority view that clavelinids are closest to the ancestral ascidian but is also consistent with other conclusions, particularly that the Cionidae are closest to the stem ascidian.     

基于宏基因组技术获得的对厌氧氨氧化菌代谢的新理解

厌氧氨氧化菌(Anaerobic ammonium oxidation bacteria,AAOB)是化能自养菌,由于其生理代谢的奇异性、细胞结构的特殊性以及对氮素循环的重要性,已成为环境工程、微生物以及海洋生物学等领域的研究热点.然而,AAOB未能实现纯培养的现状已成为AAOB代谢途径研究的巨大障碍.近年来兴起的宏基因组技术(Metagenomics)为AAOB代谢途径的研究提供了新手段.采用宏基因组技术,可直接研究微生物群体中某特定微生物基因组的结构与功能,摆脱了传统微生物学研究对纯培养的依赖,使未培养微生物的认识和开发成为可能.本文首先简述获取AAOB宏基因组信息的过程,然后通过比较由传统代谢研究方法和宏基因组技术获得的AAOB代谢途径的研究成果,论述基于宏基因组技术获得的对AAOB代谢的新理解,得出以下结果和结论:1)AAOB的碳素固定途径为乙酰辅酶A途径,碳素固定的还原力来自NADH或者QH2;2)AAOB氮素转化的重要中间产物是NO,而非NH2OH,并提出了以NO为核心的AAOB代谢的改进模型;3)AAOB的ATP合成途径为氧化磷酸化,推测的电子传递途径为N2H4—QH2—细胞色素bc1复合体;细胞色素bc1复合体再将电子用于NO2-还原和N2H4合成.AAOB的宏基因组技术使AAOB代谢途径的研究更具方向性.随着分子生物学理论和技术的不断发展,宏基因组学的升级技术(如宏转录组学、宏蛋白质组学)将为AAOB代谢途径的研究提供新的方法与平台.图3表1参51     

Structure of the sperm of some South African archaeogastropods (Mollusca) from the superfamilies Haliotoidea,Fissurelloidea and Trochoidea

The structure of the spermatozoa of 11 species from five families of archaeogastropod (Haliotidae, Fissurellidae, Trochidae, Turbinidae, Phasianellidae) has been examined using transmission electron microscopy. All sperm are of the primitive, or ect-aquasperm type and each species has a sperm head (nucleus and acrosome) with unique morphology. Furthermore, the results from the present study together with those published in the literature reveal that, although there are a few exceptions, the size (length to breadth ratio) and spape of the nucleus and acrosome of sperm of species within each family are similar. It is therefore possible to identify members of a family and differentiate between families using sperm morphology. The intrafamilial similarities and interfamilial differences in sperm structure indicate that a spermiocladistic study of the suborder Vetigastropoda could provide interesting insights into the phylogenetic relationships of this group.     

Spermiogenesis and spermatozoa in the relict bivalve genus Neotrigonia: relevance to trigonioid relationships,particularly Unionoidea

Spermiogenesis and spermatozoa in the relict trigonioid genus Neotrigonia are examined ultrastructurally. Mature Neotrigonia spp. spermatozoa exhibit the following features: (1) a blunt-conical nucleus; (2) an acrosomal complex composed of discoidal vesicles (9 to 15) arranged as a thin layer over the nuclear apex; (3) five (rarely four) spherical mitochondria positioned in depressions at the base of the nucleus; (4) proximal and distal centrioles (surrounded by the mitochondria); (5) a satellite complex anchoring the distal centriole and flagellum to the plasma membrane; and (6) a single flagellum. Spermatozoa of unionoids examined to date (Unionidae and Hyriidae only) exhibit similar features, including a nucleus with a blunt apex capped by a thin acrosomal complex (showing up to three vesicles). Among the Bivalvia, only Neotrigonia spp. and unionoid sperm possess an acrosomal complex composed of multiple vesicles. These data suggest that the Trigonioidea and Unionoidea are phylogenetically linked, but further work particularly on primitive unionoids is required to determine if this relationship is one of common ancestry or whether the Unionoidea have in fact been derived from the Trigonioidea.     

Structural and functional peculiarities of the digestive system of Cucumaria frondosa (Echinodermata: Holothuroidea)

The fine structure of the intestinal epithelium of Cucumaria frondosa has been examined. The digestive cells possessed some specific ultrastructural peculiarities which testify to their participation in the transport of substances and in the processes of intracellular and cavitary digestion. The main units of the intestinal epithelium appear to be multifunctional digestive cells in which the phenomenon of compartmentalization among the organelles' groups, intended to carry out various functions, occurs. The heterogeneity of the structure of populations of the digestive cells is explained. Both primitive and progressive features of the structure of the digestive tract of C. frondosa are indicated.     

In situ mapping of growth lines in the calcitic prismatic layers of mollusc shells using X-ray absorption near-edge structure (XANES) spectroscopy at the sulphur K-edge

The microstructure and composition, including chemical speciation of sulphur (S), of two mollusc shells were investigated using a combination of scanning electron microscopy, X-ray absorption near-edge structure spectroscopy (XANES) and electron probe microanalysis (EPMA). The shell of Pinna is composed of monocrystalline, and Pinctada, of polycrystalline, calcite prisms separated by organic-rich walls. Sulphur speciation information from XANES spectra using a scanning X-ray microscope showed that the protein S content of the interprismatic walls is higher than the SO₄ content, whereas the reverse is true for the intraprismatic structures. High-spatial-resolution XANES maps for the different S species across adjacent calcite prisms confirm their distinctive distributions in the molluscan shells and illustrate the presence of narrow, submicron transverse growth features. On a larger scale, a series of wider growth zones, incorporating the submicron zones, are aligned parallel to each other and cross-cut many calcite prisms. EPMA element maps for magnesium (Mg) and S demonstrate that these growth increments are compositionally zoned, comprising alternating layers of high mineral (Mg-rich) and high organic (S-rich) components. Additionally, these maps confirm that the organic interprismatic walls have lower Mg and higher S than the intraprismatic structures.     

Measurement of Odor-Plume Structure in a Wind Tunnel Using a Photoionization Detector and a Tracer Gas

The patterns of stimulus available to moths flying along pheromone plumes in a 3-m-long wind tunnel were characterized using a high frequency photoionization detector in conjunction with an inert tracer gas. Four contrasting flow regimes and source conditions were produced: odor released in pulses from a vertical and horizontal array of four sources, odor released continuously from a point source, and odor released continuously from a point source into an oscillatory wake. Although the four flow regimes produced plumes of intermittent and fluctuating concentration, there were considerable differences in the structure of the signal presented to the sensor. Pulses of tracer gas released at 10 Hz retained most of their longitudinal and lateral separation. The plume growing in the disturbed flow (`oscillatory'), was broader in its lateral extent than the plume growing in an undisturbed flow (`continuous'), and the concentrations in the former were lower at each downstream position. The signal recorded in the disturbed flow had higher intermittency, but the ratio between the peak concentration and the signal mean was lower than in the continuous plume. Time scales were typically longer in the tunnel than in a field setting, but length scales and the main features of intermittency and fluctuation were similar. Moths flying along plumes of pheromone in this and similar wind tunnels typically slow their velocity and narrow the lateral excursions of their flight track as they approach a pheromone source. Which features of the plumes measured in this study account for these behavioral reactions remains to be determined.     

Molecular population genetics of male and female mitochondrial genomes in subarctic Mytilus trossulus

The doubly uniparental inheritance system allows for the use of two independent mitochondrial genomes for population history investigations. Under this system, two lineages of mitochondrial DNA (mtDNA) exist and males are typically heteroplasmic, having the additional, usually divergent, mitochondrial genome inherited from their male parent. This additional mtDNA typically evolves faster, potentially allowing for insight into more recent events in population history. Few studies did explore this possibility in marine mussels Mytilus showing its usefulness. Recent observations of the Mytilus trossulus mussels who have retained their native mtDNA in European waters posed the question of their origin. Are they part of a population present, but previously undetected, or is this a potentially human mediated, ongoing spread of an invasive species? To tackle this question, we amplified with species-specific primers and sequenced an approximately 1,200-bp-long fragment spanning COIII and ND2 genes from both mitochondrial genomes of mussels sampled at five locations worldwide, covering the whole M. trossulus range. The overall pattern of polymorphisms is compatible with the entirely postglacial history of the whole species, indicating a very deep bottleneck at last glacial maximum, with possible retention of the whole species in a single refugium, and the effective population size of no more than a few thousands. Both analyses of molecular variance and isolation with migration (IM) models point at the West Atlantic as the source of the European M. trossulus mussels, at least the ones who retained their native mtDNA. The hypothesis that this is an ongoing, human-mediated process was considered. To this end, comparison with the well-known case: the introduction of congeneric mussel, Mytilus galloprovincialis, from Mediterranean Sea to Asia was used. This introduction occurred within the last 100 years. The results inferred by the IM model suggest that the timing and structure of transatlantic migration of M. trossulus differs significantly from the M. galloprovincialis case: it is more than 1,000 years old and involves a much larger fraction of the ancestral population. Therefore, most likely, this invasion is not a human-mediated process.