银合欢根瘤细胞的光学显微镜和电子显微镜研究

银合欢接种根瘤菌形成根瘤后,应用光镜和电镜技术观察。银合欢根瘤由分生组织细胞、皮层组织细胞、维管束系统和侵染细胞区域四个不同部分组成。根瘤菌借助于侵染线侵染细胞,释放进入宿主细胞质中,转变成固氮类菌体。最初每个包被膜内只含单独的类菌体,随后较老的侵染细胞中,每个包被膜内含有一个以上的类菌体。因此,成熟根瘤的侵染细胞可见有2~5个类菌体群集包被膜里,并且明显地累积PHB物质,显示电子染色透明颗粒。本文还讨论了上述变化的意义与银合欢根瘤细胞结构和功能的关系。     

沙冬青根瘤菌的电子显微镜研究

用透射电镜研究沙冬青根瘤中的根瘤菌。这种根瘤菌的精细结构不仅与生态环境有关,而且还随寄主细胞发育程度不同而异。在早期侵染细胞中,根瘤菌很少,一般呈圆形或椭圆形,电子密度很高,位于细胞壁附近。在成熟侵染细胞中,根瘤菌很多,布满了整个细胞,形态出现多样化。在衰老侵染细胞中,根瘤菌形状不规则,细胞质收缩,电子密度增高,内部结构模糊不清,有的甚至变成一团膜泡状结构     

几种罗汉松属植物根瘤的形态与结构

对分布在广西的几种罗汉松属植物自然结瘤的根瘤形态和显微结构进行了观察。结果表明:几种罗汉松属植物所结的根瘤形态是一致的,均为球形或近球形,直径约1.0～1.5mm。根瘤的组织结构从内向外包括:维管组织、薄壁细胞、侵染细胞和表皮(周皮);侵染细胞与薄壁细胞呈不均匀的相间分布。侵染细胞中所含菌体为细菌。此外还发现细叶罗汉松根瘤的侵染细胞中有变形膨大的细胞核。总体上看,罗汉松属植物根瘤在形态与结构上与豆科植物的根瘤有许多相似之处。     

濒危药用植物降香檀根瘤的解剖学研究

采用显微观察法对国家二级保护植物降香檀根瘤的结构进行细胞学研究。结果表明,降香檀的根瘤生长在侧根上,外表金黄色,呈球形或扁球形;根瘤结构由周皮、皮层、维管系统和侵染组织等4部分组成;根瘤菌在皮层中形成多个侵染区域,增大了根瘤的体积,增强了根瘤菌的固氮能力,更利于降香檀植物的生长发育。     

苜蓿根瘤菌胞外多糖缺损突变体侵染方式的超微结构

应用透射电子显微镜观察到缺损胞外多糖根瘤菌突变体侵染莒蓿根的方式与前人描述的一般根瘤菌的侵染有明显不同。突变菌贴近根毛时,根毛外层壁被降解,突变菌陷入根毛外层壁中。突变菌由外层壁移入内层壁后,在菌体周围形成大量新的壁物质。被新沉积的壁物质形成细胞内生包被的突变菌进一步形成宽的感染线,这种感染线内不舍有细的颗粒基质,突变菌被包埋在壁物质中,以后感染线解体。说明突变菌感染线的发生与一般根瘤菌在巳降解的根毛壁侵染原位直接发生感染线是不一样的。突变菌诱导的根瘤起始于根的维管柱中的薄壁细胞不规则分裂,以及与木质部极相对的皮层细胞。随着根瘤进一步发育在皮层细胞内形成一个宽的分生组织带,根瘤细胞内不含突变菌,说明突变菌诱导的根瘤与野生苜蓿根瘤菌诱导的根瘤的发育途径是十分不同的。     

豌豆根瘤中细菌的扫描电镜观察

用扫描电镜观察了豌豆根瘤的侵染细胞.结果表明,在这些细胞中有大量的细菌,它们主要是杆状细菌,其次是球形、Y形和T形细菌,其它形状的细菌很少.除了细菌形状不同外,还有一些细菌比较特殊,如有的细菌较长,菌体出现部分收缩并形成一个或一个以上的收缩环,其形状类似一条莲根;有的细菌很大,它的体积是普通细菌的2倍或2倍以上;有的细菌粗细不均匀,端部膨大,呈棒槌状.侵染细胞中有许多小泡,它们大小不同,呈球形.它们存于细菌之间,其中一些小泡还位于细菌的表面上,而且附近细菌的表面有时还有各种隆起.     

羊奶果根瘤内生菌的亚显微结构

羊奶果是桤木型根瘤。根瘤的横切面是轴对称的圆形结构,含菌细胞分散于皮层组织中。随着根瘤的发育,内生菌也有不同的形态结构。侵染初期的内生菌是一种分枝、具隔膜的菌丝体。幼龄菌丝中含有电子半透明的颗粒,成熟菌丝没有这种颗粒。成熟菌丝顶端会臌大形成具隔膜的椭圆球状泡囊,直径比菌丝大。不同发育时期的泡囊,其形态结构也有差异。     

2,4-D诱发小麦根瘤及引导根瘤菌进入小麦根瘤细胞的研究

本文通过水培和砂培的冬小麦(丰抗8号),研究用2,4-D处理并接种慢生型大豆根瘤菌61A76(Bradyrhizobium japonicum 61A76)和快生型沙打旺根瘤菌16(Rhitobium sp. astrugllus16),诱发小麦形成含菌根瘤的可能性。结果表明2,4-D可诱发小麦根系产生瘤状结构,此瘤状结构起始于根中柱鞘,2,4-D刺激小麦根瘤细胞的DNA合成,根瘤细胞的核及DNA含量约为正常根部的二倍,2,4-D能引导上述两种根瘤菌进入小麦根瘤细胞并大量繁殖,进入根瘤细咆的根瘤菌其形态有些变化,内含聚β—羟丁酸颗粒,有些菌体被膜囊包围。从接种大豆根瘤菌的小麦根瘤内分离出一株根瘤菌,经镜检、血清学及回接大豆试验证明为原接种的大豆根瘤菌株。     

肯氏相思根瘤亚显微结构观察

肯氏相思根瘤亚显微结构观察结果表明：刚受侵染的寄主细胞的细胞核和质体膨大,线粒体内嵴消失成为圆球状体,内质网膜松散;幼年类菌体细胞外型较小,呈圆形和椭圆形,细胞质浓密,染色深而均匀;成熟类菌体外型较大,形态多样,细胞内聚-β-羟基丁酸（PHB）累积增多;随着根瘤细胞逐渐发育成熟,在类菌体包被内可允1至数个类菌体;在根瘤衰老细胞中,类菌体包被周膜解离破裂,流出电子透明物质。本文还对周膜扩增的问题进行了讨论。     

新疆豆科短命植物弯果胡卢巴根瘤与根瘤菌的特性

[目的]对弯果胡卢巴根瘤增殖特性、根瘤显微结构、根瘤菌遗传聚类以及根瘤菌各种抗性进行观察、分析和鉴定.[方法]分别利用不同基质培养、石蜡切片和树脂半超薄切片以及16S rRNA基因序列扩增和序列分析等技术方法对弯果胡卢巴根瘤和根瘤菌进行研究.[结果]①在混合土(养花土:白杨林下土:沙土=1:1:1)中有明显结瘤且植株结荚最多,多数根瘤呈掌状和姜形;②显微结构显示根瘤由表及里分为表层、皮层、维管束、已侵染细胞与未侵染细胞几部分;③对根瘤菌16S rRNA基因全长序列(1377bp)测序并分析,结果显示其与苜蓿中华根瘤菌16S rRNA基因的同源性达99.9%;④根瘤菌抗逆性鉴定结果显示,在温度为4℃～60℃(20 Min)、pH值为6.0～12.0、NaCl浓度为0%～2%的范围内根瘤菌均可正常生长;低浓度的卡那霉素、链霉素及头孢霉素等抗生素(25μg/mL,)就能完全抑制根瘤菌的生长,但仍能在100μg/mL的氨苄青霉素中正常生长.[结论]弯果胡卢巴结瘤需要较好的土壤及通气条件;根瘤簇生,瘤内含大量被根瘤菌侵染的细胞;弯果胡卢巴根瘤菌与苜蓿中华根瘤菌(sinorhizobium meliloti,)同源性最高,是一类较耐高温和强碱的菌株.     

拟菌体包囊膜的动态变化和宿主细胞与细菌的相互关系

本文初次报道紫云英根瘤的超微结构。用根瘤中段的中心组织作实验材料,以显示受根瘤菌侵染的宿主细胞的一般结构。细菌借助于侵入线进入宿主细胞,发育成拟菌体,为包囊膜所裹。一个包囊膜内一般只有一个拟菌体。包囊膜可以与细胞质内的囊泡和小液泡融合而扩增,导致膜对拟菌体的包裹由紧密到疏松的变化。包囊膜和拟菌体表面都有突起,两者的突起相对接触和融合。对拟菌体包囊膜的动态变化与衰老的关系以及宿主细胞和拟菌体之间物质交换的关系进行了讨论。作者指出包囊膜的扩增和电子透明区域的存在,是拟菌体发育成熟的一个阶段,包囊膜和拟菌体通过互相突起、融合沟通的结构,可能是宿主细胞和细菌之间物质交换功能的一种表现。     

The development and structure of root nodules on bambara groundnut [Voandzeia (Vigna)subterranea]

The infection of Vigna subterranea (formerly Voandzeia subterranea) by Bradyrhizobium strain MAO 113 (isolated from V. subterranea) was examined by light and transmission electron microscopy. Bacteria accumulated on the epidermis close to root hairs, and subsequently entered the latter via infection threads. Most of the steps involved in nodule formation were generally characteristic of determinate nodules, such as those which form on the closely related V. radiata. For example, nodule meristems were induced beneath the root epidermis adjacent to infected root hairs, but prior to infection of the meristem by rhizobia. Moreover, after the infection of some of the meristematic cells by the infection threads, and the release of the rhizobia into membrane-bound vesicles, the infection process ceased and dissemination of the rhizobia was by division of already-infected host cells. However, there were some aspects of this process in V. subterranea which have been more commonly described in indeterminate nodules. These include long infection threads entering a number of cells within the meristems simultaneously and a matrix within infection threads which was strongly labelled with immunogold monoclonal antibodies, MAC236 and MAC265, which recognize epitopes on an intercellular glycoprotein. The MAC236 and MAC265 antibodies also recognized material in the unwalled infection droplets surrounding bacteria which were newly-released from the infection threads. The amount of labelling shown was more characteristic of the long infection threads seen in indeterminate nodules such as pea (Pisum sativum) and Neptunia plena. The structure of mature V. subterranea nodules was similar to that described for other determinate nodules such as Glycine max, Vigna unguiculata and V.radiata, i.e. they were spherical and the infected zone consisted of both infected and uninfected cells. Surrounding the infected tissue was an inner cortex of uninfected cell layers containing the putative components of an oxygen diffusion barrier (including glycoprotein-occluded intercellular spaces), and an outer cortex with cells containing calcium oxalate crystals.     

The development and structure of root nodules on bambara groundnut [Voandzeia (Vigna)subterranea]

The infection of Vigna subterranea (formerly Voandzeia subterranea) by Bradyrhizobium strain MAO 113 (isolated from V. subterranea) was examined by light and transmission electron microscopy. Bacteria accumulated on the epidermis close to root hairs, and subsequently entered the latter via infection threads. Most of the steps involved in nodule formation were generally characteristic of determinate nodules, such as those which form on the closely related V. radiata. For example, nodule meristems were induced beneath the root epidermis adjacent to infected root hairs, but prior to infection of the meristem by rhizobia. Moreover, after the infection of some of the meristematic cells by the infection threads, and the release of the rhizobia into membrane-bound vesicles, the infection process ceased and dissemination of the rhizobia was by division of already-infected host cells. However, there were some aspects of this process in V. subterranea which have been more commonly described in indeterminate nodules. These include long infection threads entering a number of cells within the meristems simultaneously and a matrix within infection threads which was strongly labelled with immunogold monoclonal antibodies, MAC236 and MAC265, which recognize epitopes on an intercellular glycoprotein. The MAC236 and MAC265 antibodies also recognized material in the unwalled infection droplets surrounding bacteria which were newly-released from the infection threads. The amount of labelling shown was more characteristic of the long infection threads seen in indeterminate nodules such as pea (Pisum sativum) and Neptunia plena. The structure of mature V. subterranea nodules was similar to that described for other determinate nodules such as Glycine max, Vigna unguiculata and V.radiata, i.e. they were spherical and the infected zone consisted of both infected and uninfected cells. Surrounding the infected tissue was an inner cortex of uninfected cell layers containing the putative components of an oxygen diffusion barrier (including glycoprotein-occluded intercellular spaces), and an outer cortex with cells containing calcium oxalate crystals.     

Cytoskeletal arrays in the cells of soybean root nodules: The role of actin microfilaments in the organisation of symbiosomes

Summary Within the infected cells of root nodules there is evidence of stratification and organisation of symbiosomes and other organelles. This organisation is likely to be important for the efficient exchange of nutrients and metabolites during functioning of the nodules. Using immunocytochemical labelling and confocal microscopy we have determined the organisation of cytoskeletal elements, micro tubules and actin microfilaments in soybean nodule cells, with a view to assessing their possible role in organelle distribution. Most microtubule arrays occurred in the cell cortex where they formed disorganised arrays in both uninfected and infected cells from mature nodules. In infected cells from developing nodules, parallel arrays of microtubules, transverse to the long axis of the cell, were observed. In incipient nodules, before release of rhizobia into the plant cells, the cells also had an array of microtubules which radiated from the nucleus into the cytoplasm. Three actin arrays were identified in the infected cells of mature nodules: an aster-like array which emanated from the surface of the nucleus, a cortical array which had an arrangement similar to that of the cortical microtubules, and, throughout the cytoplasm, an array of fine filaments which had a honeycomb arrangement consistent with a distribution between adjacent symbiosomes. Uninfected cells from mature nodules had only a random cortical array of actin filaments. In incipient nodules, the density of actin microfilaments associated with the nucleus and radiating through the cytoplasm was much less than that seen in mature infected cells. The cortical array of actin also differed, being composed of swirling configurations of filaments. After invasion of nodule cells by the rhizobia, the number of actin filaments emanating from the nucleus increased markedly and formed a network through the cytoplasm. Conversely, the cytoplasmic array in uninfected cells of developing nodules was identical to that in the cells of incipient nodules. The cytoplasmic network in infected cells of developing nodules is likely to be the precursor of the honeycomb array seen in mature nodule cells. We propose that this actin array plays a role in the spatial organisation of symbiosomes and that the microtubules are involved in the localisation of mitochondria and plastids at the cell periphery in the infected cells of root nodules.     

Early cytological events in the infection of the root hairs ofTrifolium repens byRhizobium leguminosarum biovartrifolii observed by glass slide culture in living seedlings

This report describes the early cytological events in the infection byRhizobium leguminosarum biovartrifolii of the root hairs ofTrifolium repens seedlings kept alive on agar medium in glass slide culture experiment. The infection threads bearing rhizobia were formed as soon as the epidermal cells began to emerge as root hairs. On the top of some of these infected emerging root hairs, there were smoky, cell-debris-like bodies, which appeared to be derived from the cell wall dug by rhizobia. Similar bodies were also observed in longer root hairs. None of the root hair cells along the length of the roots which contained infection threads were curled or distorted. A substantial number of pink-colored nodules were later formed on the roots with non-curled infected root hairs.     

Draft genome sequence of Mesorhizobium alhagi CCNWXJ12-2T, a novel salt-resistant species isolated from the desert of northwestern China

Mesorhizobium alhagi strain CCNWXJ12-2(T) is a novel species of soil-dwelling, nitrogen-fixing bacteria that can form symbiotic root nodules with Alhagi sparsifolia. Moreover, the strain has high resistance to salt and alkali. Here we report the draft genome sequence of Mesorhizobium alhagi strain CCNWXJ12-2(T). A large number of osmotic regulation-related genes have been identified.     

Nodule ultrastructure and initial growth of Anadenanthera peregrina (L.) Speg. var. falcata (Benth.) altschul plants infected with rhizobia

The anatomy and ultrastructure of root nodules of Anadenanthera peregrina var. falcata (Leguminosae-Mimosoideae) were analysed, as was plant growth. To ensure that nodules developed, seedlings were inoculated with a mixture of six strains of rhizobia. Nodules were produced that differed in appearance-and probably also effectiveness-but their structure was similar and they showed characteristics typical of indeterminate nodules, such as persistent meristematic tissue and a gradient of cells at different stages of development. Many starch grains were present in inner cortex cells and interstitial cells of infected tissue. Infected cells were densely packed with bacteroids, which contained many poly-beta-hydroxybutyrate granules. The high incidence of these granules, together with high levels of starch accumulation in interstitial cells, suggested low N2-fixation efficiency of the rhizobia isolates used for inoculation. In the symbiosomes of early-senescent infected cells, reticulum-like structures, small vesicles and a fibrillar material were observed; these may be related to bacteroid degradation. In the cytoplasm of late-senescent infected cells, many vesicles and membrane-like structures were observed, probably associated with membrane degradation of bacteroids and peribacteroids. The total biomass of plants inoculated with rhizobia was low and their xylopodia and shoots had low levels of N compared with noninoculated plants fertilized with ammonium nitrate. However, inoculated plants did not show N-deficiency symptoms and grew better than non-inoculated plants without N fertilization. These growth results, together with ultrastructural observations of nodules, suggest that nitrogen fixation of rhizobia isolates associated with Anadenanthera peregrina var. falcata roots is poor.