Influence of plant polymers on the distribution and cultivation of bacteria in the phylum Acidobacteria

Members of the phylum Acidobacteria are among the most abundant bacteria in soil. Although they have been characterized as versatile heterotrophs, it is unclear if the types and availability of organic resources influence their distribution in soil. The potential for organic resources to select for different acidobacteria was assessed using molecular and cultivation-based approaches with agricultural and managed grassland soils in Michigan. The distribution of acidobacteria varied with the carbon content of soil: the proportion of subdivision 4 sequences was highest in agricultural soils (ca. 41%) that contained less carbon than grassland soils, where the proportions of subdivision 1, 3, 4, and 6 sequences were similar. Either readily oxidizable carbon or plant polymers were used as the sole carbon and energy source to isolate heterotrophic bacteria from these soils. Plant polymers increased the diversity of acidobacteria cultivated but decreased the total number of heterotrophs recovered compared to readily oxidizable carbon. Two phylogenetically novel Acidobacteria strains isolated on the plant polymer medium were characterized. Strains KBS 83 (subdivision 1) and KBS 96 (subdivision 3) are moderate acidophiles with pH optima of 5.0 and 6.0, respectively. Both strains grew slowly (μ = 0.01 h(-1)) and harbored either 1 (strain KBS 83) or 2 (strain KBS 96) copies of the 16S rRNA encoding gene-a genomic characteristic typical of oligotrophs. Strain KBS 83 is a microaerophile, growing optimally at 8% oxygen. These metabolic characteristics help delineate the niches that acidobacteria occupy in soil and are consistent with their widespread distribution and abundance.     

Effect of pH on isolation and distribution of members of subdivision 1 of the phylum Acidobacteria occurring in soil

The pH strongly influenced the development of colonies by members of subdivision 1 of the phylum Acidobacteria on solid laboratory media. Significantly more colonies of this group formed at pH 5.5 than at pH 7.0. At pH 5.5, 7 to 8% of colonies that formed on plates that were incubated for 4 months were formed by subdivision 1 acidobacteria. These colonies were formed by bacteria that spanned almost the entire phylogenetic breadth of the subdivision, and there was considerable congruence between the diversity of this group as determined by the cultivation-based method and by surveying 16S rRNA genes in the same soil. Members of subdivision 1 acidobacteria therefore appear to be readily culturable. An analysis of published libraries of 16S rRNAs or 16S rRNA genes showed a very strong correlation between the abundance of subdivision 1 acidobacteria in soil bacterial communities and the soil pH. Subdivision 1 acidobacteria were most abundant in libraries from soils with pHs of <6, but rare or absent in libraries from soils with pHs of >6.5. This, together with the selective cultivation of members of the group on lower-pH media, indicates that growth of many members of subdivision 1 acidobacteria is favored by slightly to moderately acidic growth conditions.     

Effect of pH on Isolation and Distribution of Members of Subdivision 1 of the Phylum Acidobacteria Occurring in Soil

The pH strongly influenced the development of colonies by members of subdivision 1 of the phylum Acidobacteria on solid laboratory media. Significantly more colonies of this group formed at pH 5.5 than at pH 7.0. At pH 5.5, 7 to 8% of colonies that formed on plates that were incubated for 4 months were formed by subdivision 1 acidobacteria. These colonies were formed by bacteria that spanned almost the entire phylogenetic breadth of the subdivision, and there was considerable congruence between the diversity of this group as determined by the cultivation-based method and by surveying 16S rRNA genes in the same soil. Members of subdivision 1 acidobacteria therefore appear to be readily culturable. An analysis of published libraries of 16S rRNAs or 16S rRNA genes showed a very strong correlation between the abundance of subdivision 1 acidobacteria in soil bacterial communities and the soil pH. Subdivision 1 acidobacteria were most abundant in libraries from soils with pHs of <6, but rare or absent in libraries from soils with pHs of >6.5. This, together with the selective cultivation of members of the group on lower-pH media, indicates that growth of many members of subdivision 1 acidobacteria is favored by slightly to moderately acidic growth conditions.     

Reviewing the DA001-files: a 16S rRNA chase on suspect #X99967, a Bacillus and Dutch underground activist.

A variant of 'the rRNA approach' on uncultured soil bacteria is discussed, which is mainly based on 16S rRNA rather than on genomic 16S rDNA. While the rDNA only reflects the presence of bacteria, the rRNA indicates much more the activity of bacteria. Hence, the presented strategy can indicate the involvement of uncultured bacteria to the metabolic activity of the total microbial community. The potentials and limitations of the applied techniques will be discussed: isolation of ribosomes from soil, temperature gradient gel electrophoresis, cloning and sequencing, and the verification of these data by V6 Southern blot hybridization, dot blot hybridization and in situ hybridization. By this and another novel rRNA quantification approach, the multiple competitive RT-PCR, it could be found that an uncultured Bacillus, recognized as ribotype DA001, contributes approximately 5-10% to all bacterial ribosomes in Dutch Drentse A grassland soils. These bacteria should be major operators of biogeochemical processes in soil.     

Acidobacteria are active and abundant members of diverse atmospheric H2-oxidizing communities detected in temperate soils

Significant rates of atmospheric dihydrogen (H₂) consumption have been observed in temperate soils due to the activity of high-affinity enzymes, such as the group 1h [NiFe]-hydrogenase. We designed broadly inclusive primers targeting the large subunit gene (hhyL) of group 1h [NiFe]-hydrogenases for long-read sequencing to explore its taxonomic distribution across soils. This approach revealed a diverse collection of microorganisms harboring hhyL, including previously unknown groups and taxonomically not assignable sequences. Acidobacterial group 1h [NiFe]-hydrogenase genes were abundant and expressed in temperate soils. To support the participation of acidobacteria in H₂ consumption, we studied two representative mesophilic soil acidobacteria, which expressed group 1h [NiFe]-hydrogenases and consumed atmospheric H₂ during carbon starvation. This is the first time mesophilic acidobacteria, which are abundant in ubiquitous temperate soils, have been shown to oxidize H₂ down to below atmospheric concentrations. As this physiology allows bacteria to survive periods of carbon starvation, it could explain the success of soil acidobacteria. With our long-read sequencing approach of group 1h [NiFe]-hydrogenase genes, we show that the ability to oxidize atmospheric levels of H₂ is more widely distributed among soil bacteria than previously recognized and could represent a common mechanism enabling bacteria to persist during periods of carbon deprivation.Subject terms: Soil microbiology, Biodiversity, Bacterial physiology     

Decreased activity of Blastocladiella emersonii zoospore ribosomes: correlation with developmental changes in ribosome-associated proteins

Ribosomal proteins isolated from dormant zoospores were compared to the ribosomal proteins found in the active growth phase by two-dimensional polyacrylamide gel electrophoresis. Zoospore ribosomes were found to contain a set of five proteins, designated Z1 to Z5, which were not present in growth phase ribosomes. The Z1-Z5 proteins were not removed by high-salt washes using either 1 M KCl or 1 M NH4 Cl. The Z1 protein is found associated with zoospore 60 S subunits while Z2-Z5 are bound to 40 S subunits. Zoospore monoribosomes and polyribosomes contain comparable levels of each of the five proteins. Approximately 60 min. after sporulation is induced, the Z1-Z5 proteins begin to accumulate on the ribosomes with the highest levels of these proteins found associated with ribosomes at the zoospore stage. During germination, the proteins gradually disappear and are not detectable on the ribosomes after 4 hr of germination. The presence of the Z1-Z5 proteins correlates with a decrease in in vitro protein synthetic activity of the fungal ribosomes. The data are consistent with the hypothesis that the proteins regulate translation by completely blocking protein synthesis on a subset of ribosomes while the remainder of the ribosomes function at normal rates.     

藏北高寒草地土壤有机质化学组成对土壤蛋白酶和脲酶活性的影响

土壤酶作为生态系统的生物催化剂, 是土壤有机体的代谢驱动力, 在土壤物质循环和能量转化过程中起着重要作用。该研究以藏北5种不同类型高寒草地(高寒草甸、高寒草原、高寒草甸草原、高寒荒漠草原和高寒荒漠)为研究对象, 利用热裂解气质联用技术(Py-GC/MS)分析不同类型草地土壤有机质化学组成, 并建立其与土壤蛋白酶和脲酶活性之间的相互关系。结果表明, 5种高寒草地土壤(0-15 cm)的酶活性表现出一定差异性, 高寒荒漠草原土壤的脲酶活性显著高于蛋白酶活性, 而其余类型高寒草地的脲酶和蛋白酶活性之间的差异未达到显著水平; 蛋白酶活性在5种高寒草地土壤之间的差异显著, 而脲酶活性在5种草地土壤之间的差异未达到显著水平。相关分析发现, 土壤蛋白酶活性与土壤有机质烷烃、烯烃和芳香烃的相对丰度和糠醛:吡咯的值密切相关, 土壤脲酶活性与土壤有机质化学组成之间相关性未达到显著水平。综上所述, 高寒草地类型和土壤有机质化学组成是影响高寒草地土壤蛋白酶活性的重要因素, 而对土壤脲酶活性的影响均未达到显著水平, 其影响因素有待进一步深入的研究。     

Soil Bacterial and Fungal Community Structure Across a Range of Unimproved and Semi-Improved Upland Grasslands

Changes in soil microbial community structure due to improvement are often attributed to concurrent shifts in floristic community composition. The bacterial and fungal communities of unimproved and semi-improved (as determined by floristic classification) grassland soils were studied at five upland sites on similar geological substrata using both broad-scale (microbial activity and fungal biomass) and molecular [terminal restriction fragment length polymorphism (TRFLP), automated ribosomal intergenic spacer analysis (ARISA)] approaches. It was hypothesized that microbial community structure would be similar in soils from the same grassland type, and that grassland vegetation classifications could thus be used as predictors of microbial community structure. Microbial community measurements varied widely according to both site and grassland type, and trends in the effect of grassland improvement differed between sites. These results were consistent with those from similar studies, and indicated that floristic community composition was not a stable predictor of microbial community structure across sites. This may indicate a lack of correlation between grassland plant composition and soil microbial community structure, or that differences in soil chemistry between sites had larger impacts on soil microbial populations than plant-related effects.     

Plant regulation of soil nutrient distribution in the northern Chihuahuan Desert

Vegetation throughout the southwestern United States has changed from perennial grassland to woody shrubland over the past century. Previous studies on the development of islands of fertility focused primarily on only the most limiting, plant-essential element, soil nitrogen (N). The research presented here addressed the question of whether other plant-essential elements, namely phosphorus (P) and potassium (K), showed similar concentration gradients under the desert shrub Larrea tridentata, creosotebush. It also examined whether the spatial distribution of N, P, and K differed from that of essential, but non-limiting nutrients, namely calcium (Ca), magnesium (Mg), and sulfur (S), and non-essential elements, namely sodium (Na), chloride (Cl), and fluoride (F). Within adjacent grassland and shrubland plots, surface soils were collected under and between vegetation and analyzed for a suite of soil nutrients. Soil nutrient distribution followed a uniform pattern that mirrored the spatial homogeneity of bunchgrasses in the grassland, but followed a patchy distribution that mirrored the spatial heterogeneity of individual shrubs in the shrubland. The main differences were that in the grassland, all elements were uniformly distributed, but in the shrubland the plant-essential elements, nitrogen, phosphorus, and potassium, were concentrated under the shrub canopy, and the non-limiting and non-essential elements were either concentrated in the intershrub spaces or were equally concentrated under shrubs and in the interspaces. Our results show how vegetation shifts from grassland to shrubland contribute to long-term, widespread change in the structure and function of desert ecosystems.     

Population dynamics of annuals in perennial grassland controlled by ants and environmental stochasticity

Questions: In a system of five annual plant species restricted to nest‐mounds of the ant Lasiusflavus in a perennial grassland: 1. Are the population dynamics influenced by ant disturbance? 2. Is the survival of the annuals at the scale of the whole grassland possible under the observed conditions of disturbance dynamics? 3. Which phases in the annuals’ life cycle and patch types contribute most to population growth? Location: Borec hill, northern Czechia, 50°31’ N, 13°59’ E, 446 m a.s.l. Methods: Local population dynamics of the annuals were analysed separately for five patch types that differed in the proportion of bare soil. Vitality rates were assessed directly in the field, but also in a garden experiment, during 2000–2001 and 2001–2002. Population dynamics at the scale of the whole grassland was analysed with a megamatrix approach, combining patch dynamics of the nest‐mounds with patch‐specific population dynamics. Contributions of different phases and patch types to growth rate were estimated by elasticity analysis. Results: Nest‐mounds differed in the percentage of bare soil. Increasing moss cover significantly reduced germination and seed production of all studied annuals and decreased their population growth rates (λ). Although successional processes dominated over ant disturbance, populations of all species could survive well (λ? 1) in the grassland according to the 2000–2001 megamatrix dynamics. Based on the dynamics from the following period, two species would not survive in a long‐term perspective due to random environmental variation. Whereas the A‐A transition (adult plants originating from adults of the previous year) had the highest elasticity under open conditions and ‘good period’ demography, the importance of persistent seeds increased under reverse conditions. This, however, differed among species. Conclusions: Ant‐disturbance was shown to be critical for the population survival of five annual species in the studied grassland. The fate of the annual populations in the grassland system also depends on random environmental variation, which may override the effect of ant activity.     

农药污染对水稻田土壤反硝化细菌种群数量及其活性的影响

在实验室条件下,研究了农药污染对水稻田土壤反硝化细菌(Denitrfying bacteria,DNB)种群数量及其反硝化活性的影响。结果表明,紫色稻田土壤、黄松稻田土壤和红壤稻田土的DNB种群数量范围分别为59．04×10⁴～157．59×10⁴、42．89×10⁴～108．97×10⁴和32．14×10⁴～75．30×10⁴cfu·g^-1,稻田土DNB种群数量和土壤NO3^-的消耗量之间具有正相关性。在1kg干土中加入1mg丁草胺或呋喃丹,能刺激DNB的生长及其反硝化活性,在1kg干土中加入5mg多菌灵、10mg丁草胺或呋喃丹,对DNB的生长及其反硝化活性有明显的抑制作用,丁草胺和呋喃丹施后7d,多菌灵施后14d,对水稻田土壤的DNB种群数量和反硝化活性抑制作用达到最大,然后逐渐减轻,最后呈现一定的促进作用。     

Genomic insights into the Acidobacteria reveal strategies for their success in terrestrial environments

Members of the phylum Acidobacteria are abundant and ubiquitous across soils. We performed a large‐scale comparative genome analysis spanning subdivisions 1, 3, 4, 6, 8 and 23 (n = 24) with the goal to identify features to help explain their prevalence in soils and understand their ecophysiology. Our analysis revealed that bacteriophage integration events along with transposable and mobile elements influenced the structure and plasticity of these genomes. Low‐ and high‐affinity respiratory oxygen reductases were detected in multiple genomes, suggesting the capacity for growing across different oxygen gradients. Among many genomes, the capacity to use a diverse collection of carbohydrates, as well as inorganic and organic nitrogen sources (such as via extracellular peptidases), was detected – both advantageous traits in environments with fluctuating nutrient environments. We also identified multiple soil acidobacteria with the potential to scavenge atmospheric concentrations of H₂, now encompassing mesophilic soil strains within the subdivision 1 and 3, in addition to a previously identified thermophilic strain in subdivision 4. This large‐scale acidobacteria genome analysis reveal traits that provide genomic, physiological and metabolic versatility, presumably allowing flexibility and versatility in the challenging and fluctuating soil environment.     

适度放牧增加内蒙古典型草原甲烷氧化菌丰度和甲烷吸收

微生物氧化是大气甲烷唯一的生物汇.认识草原甲烷(CH₄)通量对不同利用方式的响应是制定低碳高效草原管理体系的基础.本研究通过测定内蒙古中部典型草原在放牧、割草和围封管理下生态系统的CH₄通量和土壤甲烷氧化菌丰度,旨在确定不同利用方式对内蒙古典型草原生态系统CH₄吸收的影响,验证甲烷氧化菌功能基因(pmoA)丰度调控CH₄通量.测定草原是连续5年实施4种不同利用处理的试验草原,4个处理为全植物生长季(5—9月)放牧(T₁)、春夏5月和7月放牧(T₂)、秋季割草(T₃)和围封禁牧(T₀).在测定植物生物量和土壤理化特征的基础上,采用静态箱置法测定草原植物生长季CH₄通量,采用分子技术测定草原表层土壤甲烷氧化菌pmoA功能基因的丰度.结果表明: 放牧显著促进CH₄吸收,增加甲烷氧化菌丰度(即每克干土pmoA功能基因拷贝数),其在生长季的变化范围是6.9×10⁴~3.9×10⁵.T₁处理下植物生长季的CH₄平均吸收量为(68.21±3.01) μg·m^-2·h^-1,显著高于T₂、T₃和T₀处理22.1%、37.5%和30.9%.草原CH₄吸收与甲烷氧化菌丰度呈极显著正相关,与土壤砂粒占比呈显著正相关,而与土壤粉粒占比、土壤水分含量、土壤铵态氮和硝态氮含量,以及植物地上生物量呈显著负相关.表明不同利用方式下内蒙古典型草原都是CH₄的汇,而适度放牧可增加草原土壤砂粒占比,降低土壤无机氮含量和植被生物量,提高土壤甲烷氧化菌丰度和CH₄吸收.本结果对制定低排放草原管理体系具有重要意义.     

青藏高原不同高寒草地类型土壤酶活性及其影响因子

土壤酶活性作为生态系统养分循环的关键因素, 是反映土壤质量和生态系统功能的重要指标, 但是关于高寒草地生态系统中不同草地类型间酶活性的差异研究还很少。因此, 该研究在藏北高寒草地选择高寒草甸、高寒草原、高寒草甸草原、高寒荒漠草原和高寒荒漠5种草地类型进行野外原位调查和采样, 测定了涉及碳(C)、氮(N)和磷(P)循环的14种酶的活性, 并建立了高寒草地酶活性与土壤微生物和土壤理化性质等环境因子的关系。结果表明: C循环酶(蔗糖酶、纤维素酶、β-葡萄糖苷酶、多酚氧化酶和过氧化物酶)和P循环酶(碱性磷酸酶)在不同高寒草地类型间活性差异明显, N循环酶中仅芳香氨基酶和亚硝酸盐还原酶两种酶在不同高寒草地类型间活性差异明显。同时, C、N和P循环酶之间存在一定的相关关系, 其中, 蔗糖酶和碱性磷酸酶、纤维素酶和α-乙酰氨基葡萄糖苷酶活性显著正相关, 多酚氧化酶与亚硝酸还原酶和β-乙酰氨基葡萄糖苷酶活性显著负相关。在测定的19个环境指标中, 土壤有机质(SOM)含量、革兰氏阴性菌数量、土壤N和P含量计量比、革兰氏阳性菌数量、细菌数量、放线菌数量、全氮含量、真菌数量是影响土壤酶活性的关键因子, 且SOM含量的影响最大(解释量为11.9%)。综上所述, 不同高寒草地类型间C循环酶、P循环酶和两种N循环酶(芳香氨基酶和亚硝酸还原酶)活性差异显著, SOM含量、微生物数量和N含量等是影响高寒草地生态系统土壤酶活性的关键因子。     

温带草原退化对土壤剖面微生物学特征的影响

【目的】草地退化已成为我国草原当前面临的最主要问题。土壤微生物量和土壤酶活性是反映土壤养分和土壤环境质量的重要指标。揭示退化程度对温带草原土壤剖面微生物学特征的影响规律。【方法】以内蒙古温带草原为研究对象,选取成熟自然草地以及中、重度退化草地和极度退化草地4种典型不同退化程度的草地,按不同土壤深度分层采样并进行土壤微生物量和土壤微生物酶活性的测定。【结果】表层土壤微生物生物量及其酶活性在不同退化样地中呈现出一致的趋势:成熟自然样地中度退化样地重度退化样地极度退化样地;10-20 cm土层土壤微生物学特征与表层的差异随着退化程度的加深逐渐减少,甚至在极度退化样地中10-20 cm层土壤微生物指标高于表层。【结论】表层土壤微生物生物量及其酶活性随着退化程度的加深而减少。同时,退化程度越严重,表层与10–20 cm土层之间土壤微生物学特征的差异越小。这一结果为评价草地退化程度提供了新思路,为温带草原的恢复和重建提供了重要的理论依据。     

Correlation of structure to antitumor activities of five derivatives of a beta-glucan from Poria cocos sclerotium

A water-insoluble (1-->3)-beta-D-glucan isolated from fresh sclerotium of Poria cocos was, respectively, sulfated, carboxymethylated, methylated, hydroxyethylated, and hydroxypropylated, to afford five water-soluble derivatives. Their weight-average molecular masses (Mw) and intrinsic viscosities ([eta]) were determined by size-exclusion chromatography combined with laser light scattering (SEC-LLS), LLS, and viscometry in phosphate buffer solution (PBS) at 37 degrees C. The antitumor activities, against Sarcoma 180 tumor cell (S-180) and gastric carcinoma cell strain (MKN-45 and SGC-7901) of the native beta-glucan and the five derivatives, were tested in vitro and in vivo. The Mw values of the five derivatives in PBS were determined to be 3.8 x 10(4), 18.9 x 10(4), 16.0 x 10(4), 76.8 x 10(4), and 224.3 x 10(4), respectively. The high Mw values of the hydroxyethylated and hydroxypropylated derivatives in aqueous solution resulted from aggregation, and their true Mw values obtained in dimethyl sulfoxide were 20.1 x 10(4) and 19.1 x 10(4). The sulfated and carboxymethylated derivatives having DS of 1.0-1.3 show good water solubility, and exist as relatively expanded chains in aqueous solution. Interestingly, the native beta-glucan did not show antitumor activity, whereas the sulfated and carboxymethylated derivatives exhibit significant antitumor activities against S-180 and gastric carcinoma tumor cells. This work showed that good water solubility, relatively high chain stiffness, and moderate molecular mass of the derivatives in aqueous solution contribute beneficial to enhancement of antitumor activity.     

内蒙古典型草原土壤固碳潜力及其不确定性的估算

以内蒙古锡林郭勒盟典型草原围封和放牧对照的实测数据为基础,比较了不同植物群落类型的碳储量和固碳量,并估算了重度退化草地完全恢复后的土壤固碳潜力.结果表明: 在局地尺度上,以羊草、大针茅和克氏针茅为优势种的3种植被类型的年均固碳量均随着围封年限的增加而减少.羊草和大针茅草原群落的碳储量显著高于克氏针茅草原群落,而在过牧导致的退化过程中,后者流失了更多的土壤有机碳,其固碳潜力高于前两者.在区域尺度上,内蒙古典型草原土壤表层0～20 cm的固碳潜力在-0.03×10⁴～3.71×10⁴ kg C·a^-1范围内变化,总固碳潜力为12.1×10⁸kg C·a^-1.不确定性分析发现,忽略石砾含量不会造成土壤固碳潜力估算值的较大偏差,而气象数据空间插值造成的误差为±4.7×10⁹ kg C·a^-1.假设未来生长季降水的平均变化值为-3.2 mm·(10 a)^-1时,土壤固碳潜力将减少1.07×10⁸ kg C·(10 a)^-1.     

Characterization of Soil Bacterial Communities in Rhizospheric and Nonrhizospheric Soil of Panax ginseng

A culture-independent approach was used to evaluate the bacterial community in rhizospheric and nonrhizospheric soil in which Panax ginseng had grown for 3?years. For each sample, soil was randomly collected from multiple sampling points and mixed thoroughly before genomic DNA extraction. Universal primers 27f and 1492r were used to amplify 16S rRNA genes. Clone libraries were constructed using the amplified 16S rRNA genes, and 192 white clones were chosen for further sequencing. After digestion with restriction endonuclease, 44 operational taxonomic units (OTUs) were generated for rhizospheric and 21 OTUs for nonrhizospheric soils, and the clones of each OTU were sequenced. Blast analysis showed that bacillus, acidobacteria, and proteobacteria were the dominant populations in rhizospheric soil, and proteobacteria were dominant in nonrhizospheric soil. Phylogenetic results showed that bacillus and acidobacteria were clustered into the group of uncultured bacteria in rhizospheric soil; however, proteobacteria were the unique dominant in nonrhizospheric soil.     

ELECTRON MICROSCOPE STUDY OF MITOCHONDRIAL 60S AND CYTOPLASMIC 80S RIBOSOMES FROM LOCUSTA MIGRATORIA

Purified mitochondrial ribosomes (60S) have been isolated from locust flight muscle. Purification could be achieved after lysis of mitochondria in 0.055 M MgCl₂. Mitochondrial 60S and cytoplasmic 80S ribosomes were investigated by electron microscopy in tissue sections, in sections of pellets of isolated ribosomes, and by negative staining of ribosomal suspensions. In negatively stained preparations, mitochondrial ribosomes show dimensions of ~270 x 210 x 215 Å; cytoplasmic ribosomes measure ~295 x 245 x 255 Å. From these values a volume ratio of mitochondrial to cytoplasmic ribosomes of 1: 1.5 was estimated. Despite their different sedimentation constants, mitochondrial ribosomes after negative staining show a morphology similar to that of cytoplasmic ribosomes. Both types of particles show bipartite profiles which are interpreted as "frontal views" and "lateral views." In contrast to measurements on negatively stained particles, the diameter of mitochondrial ribosomes in tissue sections is ~130 Å, while the diameter of cytoplasmic ribosomes is ~ 180–200 Å. These data suggest a volume ratio of mitochondrial to cytoplasmic ribosomes of 1:3. Subunits of mitochondrial ribosomes (40S and 25S) were obtained by incubation under dissociating conditions before fixation in glutaraldehyde. After negative staining, mitochondrial large (40S) subunits show rounded profiles with a shallow groove on a flattened side of the profile. Mitochondrial small subunits (25S) display elongated, triangular profiles.