Determination of optimal carbon source and pH value for sclerotial formation of Polyporus umbellatus under artificial conditions

Polyporus umbellatus is one of the precious medicinal fungi, with sclerotia used as a diuretic agent and antidote in China for many years. This has led to the present interest in producing sclerotia of P. umbellatus in the laboratory due to a decreased abundance in natural sources. Here, we investigated the determining factors for sclerotial formation in P. umbellatus. Five carbon sources, namely, maltose, fructose, glucose, sucrose and soluble starch with different initial pH values were evaluated for their effects on mycelial growth and sclerotial development of P. umbellatus. Maltose, fructose and glucose could induce sclerotial formation of P. umbellatus. Sucrose and soluble starch could stimulate growth of the fungus but had no effect on sclerotial formation. The most efficient sclerotial production occurred with maltose followed by fructose and a pH of 5. In addition, different macroscopically evident characteristics of sclerotial development of P. umbellatus induced by different carbon sources were also observed. Our findings could provide new insights into further research on sclerotial production in P. umbellatus under artificial cultivation.     

Local adaptation and host race formation of a gall-forming aphid in relation to environmental heterogeneity

Summary The process of host race formation in the aphid Tetraneura yezoensis is examined in relation to its population structure. T. yezoensis induces pouch galls on new leaves of Ulmus davidiana and U. laciniata. Its populations on the two host species are often sympatric. Fundatrices found on one elm species, when reciprocally transplanted to the other, suffered greatly reduced average fitness. This shows that aphid populations associated with the two elm species are genetically differentiated in physiological traits. Individual trees of each elm species showed large differences in susceptibility to gall formation and in bud burst time, and such between-tree variations were consistent over years. Overwintered eggs taken in early spring from four trees (two from each species) were incubated under the same temperature conditions. The average hatching time differed significantly even between populations from conspecific trees, and the sequence of egg hatching paralleled that of the leafing of those four trees. This between-tree difference in hatching time was consistent over years and was found to be genetic, showing that gene flow between aphid populations on separate trees is often restricted. The heterogeneity in host traits may have promoted the evolution of philopatry in this aphid. Of the fundatrices that hatched on a tree of one elm species, a few precent were preadapted to gall formation on the other elm species. This suggests that the formation of a new host race proceeds parapatrically under disruptive selection and at a low level of gene flow. Evidence was actually obtained that a small fraction of Tetraneura alates are passively transported and land on non-host plants.     

海桑次生木质部导管解剖特征与土壤理化因子年内动态变化的关系研究

为揭示海桑次生木质部导管解剖特征随土壤理化因子年内动态变动而变化的适应机制,该研究利用海桑具有生长轮的特点,通过显微技术界定了海桑采样枝条一年内10个不同连续时间段形成的新"生长层"(新形成的次生木质部),观测了10个不同连续时间段新"生长层"的导管解剖特征,并对10个新"生长层"形成阶段所对应的土壤理化因子数量特征进行了测定,用逐步回归法分析了10个不同连续时间段海桑新"生长层"管孔数量解剖特征与对应土壤理化因子数量特征之间的关系。结果显示:(1)10个不同连续时间段海桑形成新"生长层"管孔数量特征指标,除相邻管孔间接触壁长占比无显著差异外,其他8项指标包括管孔径向直径、管孔弦向直径、导管壁厚、导管长度、管孔密度、单孔率、导管聚合度和相邻管孔间接触壁长等均具有显著差异(P0.05);多重比较显示,10个不同连续时间段海桑形成新"生长层"管孔数量特征8项指标具不同程度的变动。(2)海桑新"生长层"形成阶段(10个不同连续时间段)土壤理化因子,包括土壤有机质含量、土壤全氮含量、土壤全磷含量、土壤pH值和土壤全盐量等5项指标均具有显著差异(P0.05);多重比较显示,海桑新"生长层"形成阶段(10个不同连续时间段)土壤理化因子指标均具有不同程度变动。(3)10个不同连续时间段海桑形成新"生长层"的管孔数量解剖特征,与所对应新"生长层"形成阶段土壤理化因子数量特征的逐步回归分析表明,随着土壤全盐含量的升高,海桑导管弦向直径和导管聚合度同时呈显著增大趋势(P0.05)。研究表明,海桑在一年内不同连续时间段,随土壤全盐量增加,土壤渗透势将增大,水分在次生木质部导管中输导的安全性将下降,而海桑导管分子随一年内不同连续时间段土壤全盐量的增加呈增大趋势,根据木材生态解剖学的观点,水分输导的安全性将进一步降低,但导管聚合度随一年内不同连续时间段土壤全盐量的增加而增大,具有增进水分输导安全性的作用,这可能是海桑对土壤盐含量变化的生态适应策略。     

Electron microscopy of spore germination and cell wall formation in Mucor rouxii

Summary The fine structure of ungerminated and aerobically germinated sporangiospores of Mucor rouxii was compared. The germination process may be divided into two stages: I, spherical growth; II, emergence of a germ tube. In both stages, germination is growth in its strictest sense with overall increases in cell organelles; e.g., the increase in mitochondria is commensurate with the overall increase in protoplasmic mass. Noticeable changes occurring during germination are the disappearance of electron-dense lipoid bodies, formation of a large central vacuole and, most strikingly, formation of a new cell wall. Unlike many other fungi, M. rouxii does not germinate by converting the spore wall into a vegetative wall. Instead, as in other Mucorales, a vegetative wall is formed de novo under the spore wall during germination stage I. This new wall grows out, rupturing the spore wall, to become the germ tube wall. Associated with the apical wall of the germ tube is an apical corpuscle previously described. The vegetative wall exhibits a nonlayered, uniformly microfibrillar appearance in marked distinction to the spore wall which is triple-layered, with two thin electron dense outer layers, and a thick transparent inner stratum. The lack of continuity between the spore and vegetative walls is correlated with marked differences in wall chemistry previously reported. A separate new wall is also formed under the spore wall during anaerobic germination leading to yeast cell formation. On the other hand, in the development of one vegetative cell from another, such as in the formation of hyphae from yeast cells, the cell wall is structurally continuous. This continuity is correlated with a similarity in chemical composition of the cell wall reported earlier.     

Multivariate analysis reveals differences in biofilm formation capacity among Listeria monocytogenes lineages

Biofilm formation capacity evaluated under identical conditions differs among Listeria monocytogenes lineages. The approach of using one set of factors or one variable at a time fails to explain why some lineages are more prevalent than others in certain environments. This study proposes the use of multivariate analysis to compare biofilm formation by various strains and describes the ecological niches of L. monocytogenes lineages. Nutrient availability, temperature, pH and water activity (a_w) at three different levels were used to determine biofilm formation by 41 strains. Despite the high degree of similarity (≤ 80%), distinct lineage-associated biofilm formation patterns were identified. A linear regression model for each strain and a principal component analysis of regression coefficients indicated that Lineages I and III have different, but overlapping, ecological niches. This study is the first to report the use of multivariate analyses to compare biofilm formation by various isolates of L. monocytogenes.     

Ethylene‐dependent aerenchyma formation in adventitious roots is regulated differently in rice and maize

In roots of gramineous plants, lysigenous aerenchyma is created by the death and lysis of cortical cells. Rice (Oryza sativa) constitutively forms aerenchyma under aerobic conditions, and its formation is further induced under oxygen‐deficient conditions. However, maize (Zea mays) develops aerenchyma only under oxygen‐deficient conditions. Ethylene is involved in lysigenous aerenchyma formation. Here, we investigated how ethylene‐dependent aerenchyma formation is differently regulated between rice and maize. For this purpose, in rice, we used the reduced culm number1 (rcn1) mutant, in which ethylene biosynthesis is suppressed. Ethylene is converted from 1‐aminocyclopropane‐1‐carboxylic acid (ACC) by the action of ACC oxidase (ACO). We found that OsACO5 was highly expressed in the wild type, but not in rcn1, under aerobic conditions, suggesting that OsACO5 contributes to aerenchyma formation in aerated rice roots. By contrast, the ACO genes in maize roots were weakly expressed under aerobic conditions, and thus ACC treatment did not effectively induce ethylene production or aerenchyma formation, unlike in rice. Aerenchyma formation in rice roots after the initiation of oxygen‐deficient conditions was faster and greater than that in maize. These results suggest that the difference in aerenchyma formation in rice and maize is due to their different mechanisms for regulating ethylene biosynthesis.     

Die Normalisierung Genetisch Bedingter Defekte der Zelltrennung beiParamaecium aurelia Durch Sauerstoffmangel und Kohlenmonoxyd

Summary Of two strains ofParamecium aurelia Syngen4, one differs from the normal form by a gene-induced cytpolasmic mutation (Mo-strain), the other by a simple gene mutation (snaky). Both strains have a very similar character: the cells fail to separate with a variable frequency and monster formation results (Table 1).The phenotype of the two strains can be completely normalised by low oxygen tension (Table 2) or by the presence of carbon monoxide. The CO effect is reversible by light (Table 3, 4). 2,4-Dinitrophenol reduces the frequency of monster formation, sometimes to zero (Table 5). Iodoacetic acid, on the other hand, causes a tenfold increase in monster formation over the controls under certain conditions (Table 6).The facts are discussed in relation to the Pasteur phenomenon.     

Physiological factors affecting yield formation in the canopy of traditional and new morphotypes of triticale plant (Triticosecale Wittmarck)

A canopy of the traditional morphotype of Triticosecale Wittmarck and the new one were analysed in two consecutive years under different climatic conditions. On the basis of dry matter and assimilatory surface of the plant organs during the vegetation period, the following growth indices were determined: RGR, CGR, ULR, ULRc, LAI, LAD, LADc and HI. Radiation use efficiency was also calculated. The two possible strategies of yield formation by analysed triticale plants were discussed. An erratum to this article is available at .     

PYRENOID FORMATION ASSOCIATED WITH THE CELL CYCLE IN THE BROWN ALGA,SCYTOSIPHON LOMENTARIA (SCYTOSIPHONALES,PHAEOPHYCEAE)1

Vegetative cells of the brown alga Scytosiphon lomentaria (Lyngbye) Link characteristically have only one chloroplast with a prominent protruding pyrenoid, whereas zygotes have both paternal and maternal chloroplasts. In zygotes, before cell and chloroplast division, each chloroplast has an old and a new pyrenoid. In this study, we raised a polyclonal antibody to RUBISCO and examined the distribution of RUBISCO by immunofluorescence microscopy, focusing on new pyrenoid formation in vegetative cells of gametophytes and zygotes in Scytosiphon. In interphase, only one old pyrenoid was positively indicated by anti‐RUBISCO antibody in vegetative cells of gametophytes. From mid‐S phase, small fluorescence aggregates reflecting RUBISCO localization started to appear at stroma positions other than adjacent to the old protruding pyrenoid. The fluorescent spots eventually coalesced into a protrusion into the adjacent cytoplasm. We also used inhibitors to clarify the relationship between the cell cycle and new pyrenoid formation, using zygotes after fertilization. When DNA replication was blocked by aphidicolin, new pyrenoid formation was also inhibited. Washing out aphidicolin permitted new pyrenoid formation with the progression of the cell cycle. When mitosis was prolonged by nocodazole, which disrupted the spindle microtubules, the fluorescent masses indicating RUBISCO localization continued to increase when compared with pyrenoid formation in untreated zygotes. During treatment with chloramphenicol, mitosis and cytokinesis were completed. However, there was no occurrence of new RUBISCO localization within the chloroplast stroma beyond the old pyrenoid. From these observations, it seems clear that new pyrenoid formation in the brown alga Scytosiphon depends on the cell cycle.     

Mechanical pressure inhibits vessel development of xylogenic cambial derivatives of beech (Fagus sylvatica L .)

 Segments of standing beech stems (Fagus sylvatica) were put under mechanical pressure from the end of May, in order to investigate the influence of constant external pressure on the development of secondary vascular tissue. After 4 weeks, the new xylem increment was investigated anatomically in cross-sections. The first axial xylem derivatives of the new year’s increment had differentiated into normal vessel elements and fiber-tracheids. Application of the pressure girdle had no effect on fiber-tracheid development, but it inhibited vessel formation. Under pressure, changes in the two dimensional PAGE protein pattern were characterized by the appearance of two new protein species as well as by the absence of one species that occurs under regular growth. Received: 7 June 1996 / Accepted: 14 November 1996     

Processi di Neoformazione di Mitocondri Nella Cellula Vegetale

Abstract

Processes of formation of new mitochondria in plant cells. — While studying with the electron microscope the quiescent centre in the onion (Allium cepa) root tips, the writer was able to observe the endoplasmic reticulum arranged into peculiar concentric ring systems, which are here described. In some of the ring structures, between a ring and the contigous one, mitochondria-like bodies are formed, which are then detached by a kind of budding process. A short review of the current literature on formation of the mitochondria is also reported.     

Characterization of the fixABC region of Azorhizobium caulinodans ORS571 and identification of a new nitrogen fixation gene

Summary The fast growing strain, Azorhizobium caulinodans ORS571, isolated from stem nodules of the tropical legume Sesbania rostrata, can grow in the free-living state at the expense of molecular nitrogen. Five point mutants impaired in nitrogen fixation in the free-living state have been complemented by a plasmid containing the cloned fix-ABC region of strain ORS571. Genetic analysis of the mutants showed that one was impaired in fixC, one in fixA and the three others in a new gene, located upstream from fixA and designated nifO. Site-directed Tn5 mutagenesis was performed to obtain Tn5 insertions in fixB and fixC. The four genes are required for nitrogen fixation both in the free-living state and under symbiotic conditions. The nucleotide sequence of nifO was established. The gene is transcribed independently of fixA and does not correspond to fixX, recently identified in Rhizobium meliloti and R. leguminosarum. Biochemical analysis of the five point mutants showed that they synthesized normal amounts of nitrogenase components. It is unlikely that fixA, fixC and nifO are involved in electron transport to nitrogenase. FixC could be required for the formation of a functional nitrogenase component 2.     

Bacterial Species Diversity in Cigarettes Linked to an Investigation of Severe Pneumonitis in U.S. Military Personnel Deployed in Operation Iraqi Freedom

This report presents results from a study on the bacterial diversity of cigarette brands collected from military personnel during the U.S. Army’s investigation of a series of cases of acute eosinophilic pneumonitis in military personnel deployed in Operation Iraqi Freedom. Eight species of Bacillus, including five new species, and one new species of Kurthia were isolated from the cigarettes. Some of these species have been identified elsewhere as causes of hypersensitivity pneumonitis and other respiratory syndromes. All of the isolates were facultative anaerobes, and many displayed mucoid growth under anaerobic conditions. In addition, many isolates also displayed the ability to form surface biofilms under liquid culture. Although biofilm formation and mucoid growth were not correlated, the former was found to be much more pronounced under anaerobic conditions as opposed to aerobic ones. The implications of these findings are discussed.     

Two New Species of Trypanosomatid Parasites Isolated from Heteroptera in Costa Rica

ABSTRACT. Two new trypanosomatid species (Euglenozoa, Kinetoplastea) isolated from the intestinal tract of heteropteran insect hosts were described based on molecular phylogenetic analyses of Spliced Leader (SL) RNA gene repeats, glycosomal glyceraldehyde phosphate dehydrogenase, and small subunit ribosomal RNA genes, as well as by morphology. Leptomonas barvae n. sp., from a mirid host Collaria oleosa, was found to represent one of the closest monoxenous (one host) relatives of the dixenous (two hosts) parasitic genus Leishmania. This finding further supports the origin of these dixenous parasites from monoxenous progenitors in the Neotropics. Blastocrithidia largi n. sp., from a largid host Largus cinctus, is among a few members of this genus available in culture. The species is a close relative of Blastocrithidia triatomae and is a member of a new monophyletic phylogenetic group characterized by formation of straphanger cysts.     

PERIDINIUM GREGARIUM,A NEW SPECIES OF DINOFLAGELLATE1,2

A dinoflagellate has been found in tidepools on the coast of the Palos Verdes Peninsula in southern California. It is described as a new species, Peridinium gregarium. In the tidepool habitat, great numbers of the organism form cloud-like masses held together by a mucous matrix. The gregarious cloud formation seems to be shared by only one other species of dinoflagellate, Peridinium sociale (Henneguy ex Labbé) Biecheler, 1935.     

Widespread formation of intracellular calcium carbonates by the bloom-forming cyanobacterium Microcystis

The formation of intracellular amorphous calcium carbonates (iACC) has been recently observed in a few cultured strains of Microcystis, a potentially toxic bloom-forming cyanobacterium found worldwide in freshwater ecosystems. If iACC-forming Microcystis are abundant within blooms, they may represent a significant amount of particulate Ca. Here, we investigate the significance of iACC biomineralization by Microcystis. First, the presence of iACC-forming Microcystis cells has been detected in several eutrophic lakes, indicating that this phenomenon occurs under environmental conditions. Second, some genotypic (presence/absence of ccyA, a marker gene of iACC biomineralization) and phenotypic (presence/absence of iACC) diversity have been detected within a collection of strains isolated from one single lake. This illustrates that this trait is frequent but also variable within Microcystis even at a single locality. Finally, one-third of publicly available genomes of Microcystis were shown to contain the ccyA gene, revealing a wide geographic and phylogenetic distribution within the genus. Overall, the present work shows that the formation of iACC by Microcystis is common under environmental conditions. While its biological function remains undetermined, this process should be further considered regarding the biology of Microcystis and implications on the Ca geochemical cycle in freshwater environments.     

Effects of Itraconazole and Micafungin on Aspergillus fumigatus Biofilms

Aspergillus fumigatus (A. fumigatus) is the most common airborne opportunistic fungal pathogen. Biofilm formation is one of the main pathogenic mechanisms of A. fumigatus. During the past decades, A. fumigatus azole resistance has become prevalent due to the medical and agricultural use of antifungal drugs and fungicides. Until now, the role of fungal biofilms in azole resistance of A. fumigatus remains unclear. In the present study, we compared biofilm drug susceptibility and biofilm formation under itraconazole of azole-resistant strains, sensitive strains, and standard strains, separately. The biofilm viability and matrix thickness at the early and the late stage were measured by XTT assay and Calcofluor white. Our results showed that the sessile minimum inhibitory concentration of itraconazole, which describing the inhibition of drugs on fungi sessile with biofilm, was much higher than the traditional minimal inhibitory concentration of itraconazole. Additionally, low concentrations of itraconazole inhibited biofilm formation of A. fumigatus strains. Notably, biofilm formation by azole-resistant strains could not be inhibited by high concentrations of itraconazole but could be effectively restrained by low concentrations of micafungin, revealing the efficacy of a cell-wall inhibitor to disrupt A. fumigatus biofilm formation. However, late-stage biofilms of both azole-resistant strains and standard strains were hard to disrupt using itraconazole. We found that itraconazole was effective to prevent A. fumigatus biofilm formation at the early stage. For the treatment of A. fumigatus biofilm, our findings suggest that an early-stage preventive strategy is preferred and micafungin is effective to control the azole-resistant strain infection.

     

Germinazione «In Vitro» di Carpospore di Petroglossum Nicaeense (Duby) Schotter

Abstract

In vitro development of carpospores of Petroglossum nicaeense (Duby) Schotter. This paper describes the germination of carpospores of P. nicaeense (Phyllophoraceae) and the development on glass of new tetrasporophytes to the size of 3–4 mm. Different patterns of development are observed: 1) formation of one or two rizoidal filaments which give rise to an adhesive disc; 2) formation of a morula, of limited growth, and of rizoidal filaments which then originate adhesive discs; 3) formation of a morula which originates a single frond, and formation of filaments that do not form adhesive discs. Eventually, the discs give rise to one or more outgrowths that develop into erect fronds. These fronds reach a 4 mm length 3 months after germination.     

Enhanced formation of methane in plant cell cultures by inhibition of cytochrome c oxidase

The claim of methane (CH₄) formation in plants has caused much controversy and debate within the scientific community over the past 4 years. Here, using both stable isotope and concentration measurements, we demonstrate that CH₄ formation occurs in plant cell cultures that were grown in the dark under sterile conditions. Under non‐stress conditions the plant cell cultures produced trace amounts [0.3–0.6 ng g^?1 dry weight (DW) h^?1] of CH₄ but these could be increased by one to two orders of magnitude (up to 12 ng g^?1 DW h^?1) when sodium azide, a compound known to disrupt electron transport flow at the cytochrome c oxidase (complex IV) in plant mitochondria, was added to the cell cultures. The addition of other electron transport chain (ETC) inhibitors did not result in significant CH₄ formation indicating that a site‐specific disturbance of the ETC at complex IV causes CH₄ formation in plant cells. Our study is an important first step in providing more information on non‐microbial CH₄ formation from living plants particularly under abiotic stress conditions that might affect the electron transport flow at the cytochrome c oxidase in plant mitochondria.