肉足鞭毛类原生动物中宿主—共生体系统的研究

目前已在20多种变形虫和70多种鞭毛虫中发现细菌内共生体。大部分细菌内共生体位于宿主细胞质共生泡中,仅少数鞭毛虫的内共生体位于核质中。变形虫-细菌共生系统形成后,共生体影响宿主细胞基因,对其基因缺陷产生互补作用。灰胞藻类鞭毛虫-蓝绿藻共生体系统的研究表明,叶绿体起源于一种原始的共生蓝细菌。锥体亚目鞭毛虫细胞质内普遍含有双心体,该共生体可能是由来自波豆亚目的锥体类鞭毛虫遗传的。作者推测,继续研究鞭毛虫和原校生物共生关系起源的基本阶段,可阐明原生动物的共生系统起源的基本原则,并为真核细胞起源的理论提供进一步的证据;深入研究变形虫-细菌共生系统,可在遗传精细结构和代谢调节的进化方面为真核细胞内共生起源的理论提供分子水平上的证据。     

纤毛类原生动物中宿主－共生体系统的研究

目前已经在100多种纤毛虫中观察到细菌、藻类和其他微生物等共生体。对纤毛虫中宿主－共生体系统的研究表明,双小核草履虫中卡巴粒的遗传为细胞质遗传理论提供了例证;含细菌共生体的许多厌氧纤毛虫无线粒体,共生体对宿主代谢有重要作用;尾草履虫－钝状全孢螺菌共生作用中,共生菌感染形式的39kDa、15kDa周质蛋白可分别与IF-3-1、IF-3-2两种单抗反应,其共生体早期感染过程中两种抗原的量发生显著变化,并且共生体生殖形式选择性地合成63kDa蛋白质,该蛋白质可能是与共生作用有联系的关键分子;绿草履虫－小球藻共生系统中,共生藻中存在葡糖胺硬性壁是其与草履虫发生共生关系的基本条件,其中,共生藻参与宿主代谢,与宿主形成相互受益的专一性关系,并且藻类共生体的作用可能影响了宿主草履虫基因组有关结构,改变了其基因表达。作者推测,探索共生体对宿主基因结构及其表达产物的影响可能是对纤毛虫中共生作用研究的主要趋势,这对于深入了解真核细胞中宿主－共生体双方的相互作用、物质交流在分子水平上的调控机理、细胞结构与功能的关系等细胞生命活动规律是有意义的。     

纤毛类原生动物中宿主—共生体系统的研究

目前已经在100多种纤毛虫中观察到细菌、藻类和其他微生物等共生体。对纤毛虫中宿主－共生体系统的研究表明,双小核草履虫中卡巴粒的遗传为细胞质遗传理论提供了例证;含细菌共生体的许多厌氧纤毛虫无线粒体,共生体对宿主代谢有重要作用;尾草履虫－钝状全孢螺菌共生作用中,共生菌感染形式的39kDa、15kDa周质蛋白可分别与IF－3－1、IF－3－2两种单抗反应,其共生体早期感染过程中两种抗原的量发生显著变化,并且共生体生殖形式选择性地合成63kDa蛋白质,该蛋白质可能是与共生作有联系的关键分子;绿草履虫－小球藻共生系统中,共生藻中存在葡糖胺硬性壁是其与草履虫发生共生关系的基本条件,其中,共生藻参与宿主代谢,与宿主形成相互受益的专一性关系,并且藻类共生体的作用可能影响了宿主草履虫基因组有关结构,改变了其基因表达。作者推测,探索共生体对宿主基因结构及其表达产物的影响可能是对纤毛虫中共生作用研究的主要趋势,这对于深入了解真核细胞中宿主－共生体双方的相互作用、物质交流在分子水平上的调控机理、细胞结构与功能的关系等细胞生命活动规律是有意义的。     

珊瑚共生体中“细菌-虫黄藻-宿主”三角关系的通讯交流

“细菌-虫黄藻-珊瑚”是生态系统中一对经典的三角关系,其中包含着复杂的物质流、信息流和能量流,三者的平衡与稳定是维护珊瑚礁生态系统健康的重要保障。过去20年里针对共生体交互关系进行了大量研究,并取得了一些重要成果,明确了“细菌-虫黄藻-宿主”三者之间的物质代谢、营养交换以及与环境的交互关系。然而,基于共生系统的复杂性,一些现象背后的机制仍然未被充分揭示,尤其是共生体之间的通讯交流。信号分子介导的相互作用是珊瑚共生体稳态维持和高效运转的内在驱动力。本文以珊瑚共生体系中化学信号为重点,尝试梳理最新的研究进展,包括细菌与细菌、细菌与珊瑚、细菌与虫黄藻以及虫黄藻与珊瑚之间的通讯方式,重点关注了群体感应信号(QS)、二甲基巯基丙酸盐(DMSP)、糖类信号、脂类信号以及非编码RNA。选择性例举了QS信号介导的微生物协作和竞争、DMSP调节下的细菌和宿主的相互作用,以及环境胁迫下珊瑚和虫黄藻对非编码RNA的响应过程,强调了它们在共生体中的作用模式和生态意义。并对今后的研究重点和可能方向进行了提炼,包括研究维度的扩充、新技术-新方法的应用以及生态模型的构建等,旨在提升对三角关系互作方式的认识,增进对珊瑚共生体的理解,探索基于通讯语言的操纵方式为珊瑚礁生态系统的恢复和保护提供新思路。     

昆虫共生菌对宿主功能研究的方法体系

共生菌可参与昆虫的生理生化过程,影响昆虫的营养、生长发育、解毒作用、天敌防御和免疫能力等。在共生菌对宿主功能的研究中,核心问题是如何从组成复杂的共生菌组中筛选出具有特定功能的共生菌。共生菌对宿主功能的研究模式通常包括:通过共生菌多样性分析,提出差异共生菌对宿主功能的假设;分析并验证特定共生菌在宿主体内的功能。本文围绕共生菌对宿主功能的研究模式,系统地总结和比较昆虫共生菌功能研究的方法和技术,构建昆虫对宿主功能研究的方法体系,以期推进共生菌-宿主联系的研究。     

昆虫病原线虫和共生细菌培养系统中噬菌体的检测

本文利用紫外照射、温度、pH、盐度诱导五株溶源性共生菌株Xenorhabdus和Photorhabdus,同时测定斯氏和异小杆属线虫Steinernema carpocapsae A24,S.carpocapsae AIL,S.feltiae English,S.feltiae SN,Heterorbabditis bacterophora H06在体外培养过程中是否存在感染共生细菌的噬菌体.结果未发现噬菌斑,说明实验菌株在实验诱导条件下以及昆虫病原线虫固体培养系统中不可能诱发噬菌体的危害.     

湿地稻-鸭复合系统中水稻纹枯病的变化规律

为了探明稻田养鸭对水稻纹枯病的发生、发展的影响 ,为稻 -鸭复合系统中水稻纹枯病的防治提供依据 ,笔者在中稻、晚稻田进行了稻田养鸭试验。试验结果表明 ,在中稻田每 6 6 6 .7m2 放养体重 15 0 g左右鸭子 15～ 2 0只 ,能使纹枯病病蔸率减少5 6 .0 % ,病株率减少 5 7.74 % ,同时比用井岗霉素防治的小区病蔸率下降 9.0 % ,病株率下降 15 .2 5 % ,病情指数比空白对照下降2 6 .4 6 ,比施用井岗霉素的施药区减少 0 .95 ;防治效果显著 ,基本可控制纹枯病的危害     

稻田土壤－作物－家畜系统中氮的循环研究

本文用Ｎ￣（１５）标记水稻和绿肥研究了稻田土壤－作物－家畜系统中氮的循环。Ｎ￣（１５）标记稻草喂羊,羊体回收饲料稻草Ｎ３１．１６％,羊粪２８．２６％,羊尿５．７２％,总回收６５．１４％,损失３４．８６％。将羊粪尿单施,稻谷回收饲料稻草Ｎ３．１９％,水稻全株回收４．８２％,土壤残留１９．００％,损失１０．１４％。故羊体、水稻及土壤残留共回收饲料稻草Ｎ５４．９８％．将羊粪与尿素配施,则饲料稻草Ｎ的总回收率为５５．８８％。Ｎ￣（１５）标记绿肥喂猪;猪体回收饲料绿肥Ｎ２３．５１％．猪粪２３．８５％,猪尿２８．７６％,总回收率７６．１２％,损失率２３．８８％。将猪粪、尿还田,稻谷回收饲料绿肥Ｎ６．６９％,水稻全株回收１０．０５％,土壤残留１９．１７％,故猪体、水稻和土壤残留共回收饲料绿肥Ｎ５２．７３％,将猪粪与尿素配施,则饲料绿肥Ｎ的总回收率为５２．７５％。     

原核表达系统T7RNA聚合酶/启动子在真核细胞中表达目的基因的实验研究

将目前高表达水平强大的原核表达系统之一T7RNA聚合酶 启动子表达系统通过一系列改进引入真核细胞。通过转染真核细胞实验表明 ,采用真核启动子CMV调控T7RNA聚合酶的表达和在T7启动子下游插入EMCVIRES序列两种解决方案能使该原核表达系统在真核细胞高效表达目的基因 ,且能适应不同的真核细胞环境 ,是一良好的细胞类型非依赖的表达体系。     

原核表达系统T7RNA聚合酶/启动子在真核细胞中表达目的基因的实验研究

将目前高表达水平强大的原核表达系统之一T7 RNA聚合酶/启动子表达系统通过一系列改进引入真核细胞.通过转染真核细胞实验表明,采用真核启动子CMV调控T7 RNA聚合酶的表达和在T7启动子下游插入EMCV IRES序列两种解决方案能使该原核表达系统在真核细胞高效表达目的基因,且能适应不同的真核细胞环境,是一良好的细胞类型非依赖的表达体系.     

Flagellar systems in the euglenoid flagellates

The flagellar apparatus of euglenoids consists of two functional basal bodies, three unequal microtubular roots subtending the reservoir, and a fourth band of microtubules nucleated from one of the flagellar roots and subtending the reservoir membrane. The flagellar apparatus of some euglenoids may contain additional basal bodies, striated roots ("rhizoplasts"), fibrous roots, striated connecting fibers between basal bodies, layered structures, or various electron-dense connective substances. With the possible exception of Petalomonas cantuscygni, nearly all euglenoids are biflagellate although the length of one flagellum may be highly reduced. The flagellar transition zone and number of basal bodies are highly variable among species. In recent years a cytoplasmic pocket that branches off from the reservoir has been discovered. The microtubules of the ventral flagellar root are continuous with the microtubules which line this pocket. Based on positional and structural similarities, this structure is believed to be homologous with the MTR/cytostome of bodonids. Coupled with other ultrastructural and biochemical data, the fine structure of the flagellar apparatus supports the belief that the euglenoid flagellates are descendant from bodonid ancestors.     

Flagellar and cytoskeletal systems in amitochondrial flagellates: Archamoeba,Metamonada and Parabasala

Summary The hypothesis that protists without mitochondria, the so-called Archezoa of Cavalier-Smith, are primitive has received some support from rRNA sequence studies on Microsporidia and Diplomonadida. In spite of the lack of mitochondria the archezoan groups of protists show considerable differences in their organization: mastigont and cytoskeletal system, mitosis, Golgi apparatus, hydrogenosomes. This paper examines the characters of the flagellar apparatus and its associated cytoskeleton to obtain clues used for phylogenetic consideration on the three cited groups of flagellates. Archamoebae of the Pelobiontida order comprising families such as Pelomyxidae and Mastigamoebidae share common features: a rudimentary mastigont system composed of only one basal body giving rise to a poorly motile flagellum and a basal body associated microtubular cone capping the nucleus. No Golgi apparatus has been detected.Metamonada, comprising three orders: Retortamonadida, Diplomonadida, and Oxymonadida, have been tentatively assembled on the basis of the absence of mitochondria, Golgi apparatus, and basal body arrangement. They all have four basal bodies arranged in two pairs with always one recurrent flagellum generally included in a cytostomal depression. The recurrent basal body/flagellum is in relation to recurrent microtubular fibers. However, they display marked differences in their cytoskeletal system and fiber ultrastructure indicating a distant evolutionary relationship. The presence of a corset of microtubules in retortamonads and three microtubular fibers are distinguished in diplomonads, as well as a paracrystalline preaxostyle and axostyle in oxymonads are features that lend support to these groups being highly divergent.Parabasala, comprising the orders Trichomonadida and Hypermastigida, is a monophyletic group with a set of homologous features such as the presence of the same arrangement of four basic basal bodies, the parabasal apparatus (striated fibre supporting Golgi), the microtubular pelta-axostyle complex, the external mitotic apparatus (crypto-pleuro-mitosis), the hydrogenosomes. These three phyla appear distantly related, the Parabasala being a homogeneous group, perhaps also the Pelobiontida, while the Metamonada is heterogeneous and composed of three evolutionary lineages. Additional information such as rRNA and protein sequence data could contribute to a better understanding of the phylogenetic relationships among these groups.Abbreviations EM electron microscopy - MTOC microtubule organizing centre - PF parabasal fibre     

Observations on endonuclear bacteria in euglenoid flagellates

Summary Rod-shaped inclusions in the nuclei and/or cytoplasm of strains ofEuglena spirogyra, Lepocinclis ovum, Strombomonas conspersa andTrachelomonas oblonga var.punctata have been identified as living populations of bacteria by reference to size, shape and ultrastructure. Most strains contain 20–200 bacteria per nucleus. Each bacterial cell is surrounded by a mucilaginous sheath and consists of a double-membraned envelope, dense cytoplasm with small ribosomes, and a filamentous DNA-containing centre. The bacteria multiply to keep pace with euglenoid division but there is no evidence that the euglenoid cells benefit from or are harmed by the association. Attempts to transfer the endonuclear bacteria to uninfected strains and species have been unsuccessful, as have attempts to isolate the bacterium into pure culture. The infected strain ofEuglena spirogyra cannot fix nitrogen. The possible nature of the euglenoid/bacterium association is discussed.Dedicated to Prof. Dr.Lothar Geitler on the occasion of his 70th birthday.     

Ultrastructural characteristics of the host-symbiont interface in nitrogen-fixing peanut nodules

Summary Nitrogen-fixing peanut root nodules are characterized by their unique structural organization, distinct from other legume nodules. The focus of this study has been in and around the hostsymbiont interface, where the bacterioid and the host cell surface (peribacteroid membrane envelope) interact during symbiosis. The infected nodule cells have revealed the presence of lipid bodies (oleosomes) in intimate association with the peribacteroid membrane, which encloses the large spherical bacteroids with a relatively narrow peribacteroid space. Electron dense structures, referred to as dense bodies have been found attached to the bacteroid outer membranes at the host-symbiont interface. The dense bodies are osmiophilic, amorphous and 3,3-diaminobenzidine positive. The isolated intact bacteroids with dense bodies attached to their cell wall showed significant catalase activity. Many microbodies showing DAB-positive reaction have been found in the host cytoplasm, associated closely with the peribacteroid membrane. These ultrastructural and cytochemical characteristics of peanut root nodules suggest that lipids are utilized during symbiosis and the dense bodies and microbodies may be involved in the catabolic process.Abbreviation DAB 3,3-diaminobenzidine     

Strict host-symbiont cospeciation and reductive genome evolution in insect gut bacteria

Host-symbiont cospeciation and reductive genome evolution have been identified in obligate endocellular insect symbionts, but no such example has been identified from extracellular ones. Here we first report such a case in stinkbugs of the family Plataspidae, wherein a specific gut bacterium is vertically transmitted via “symbiont capsule.” In all of the plataspid species, females produced symbiont capsules upon oviposition and their gut exhibited specialized traits for capsule production. Phylogenetic analysis showed that the plataspid symbionts constituted a distinct group in the γ-Proteobacteria, whose sister group was the aphid obligate endocellular symbionts Buchnera. Removal of the symbionts resulted in retarded growth, mortality, and sterility of the insects. The host phylogeny perfectly agreed with the symbiont phylogeny, indicating strict host-symbiont cospeciation despite the extracellular association. The symbionts exhibited AT-biased nucleotide composition, accelerated molecular evolution, and reduced genome size, as has been observed in obligate endocellular insect symbionts. These findings suggest that not the endocellular conditions themselves but the population genetic attributes of the vertically transmitted symbionts are probably responsible for the peculiar genetic traits of these insect symbionts. We proposed the designation “Candidatus Ishikawaella capsulata” for the plataspid symbionts. The plataspid stinkbugs, wherein the host-symbiont associations can be easily manipulated, provide a novel system that enables experimental approaches to previously untouched aspects of the insect-microbe mutualism. Furthermore, comparative analyses of the sister groups, the endocellular Buchnera and the extracellular Ishikawaella, would lead to insights into how the different symbiotic lifestyles have affected their genomic evolution.     

Molecular evidence for host-symbiont specificity in soritid foraminifera

Symbiosis between the dinoflagellate genus Symbiodinium and various invertebrates and protists is an ubiquitous phenomenon in shallow tropical and subtropical waters. Molecular studies undertaken on cnidarian symbionts revealed the presence of several distinctive lineages or subgeneric clades of Symbiodinium whose taxonomic level provides limited information about the specificity between invertebrate hosts and their symbionts. This contrasts with the finding of several Symbiodinium clades being present almost exclusively in foraminifera and belonging to the subfamily Soritinae. To test whether such specificity also exists at a lower taxonomic level within Soritinae, we obtained the SSU rDNA sequences from 159 soritid individuals collected in nine localities worldwide and representing all known morphospecies of this subfamily. For each individual, the symbionts were determined either by sequencing or by RFLP analysis. We distinguished 22 phylotypes of Soritinae in relation with a number of symbiont "groups" corresponding to 3 clades and 5 subclades of Symbiodinium. Among the 22 soritid phylotypes, 14 show strict symbiont specificity and only one was found to be a host for more than two "groups" of Symbiodinium. It is suggested that the strong host-symbiont specificity observed in Soritinae is a combined effect of a selective recognition mechanism, vertical transmission of symbionts, and biogeographical isolation.