Cytological Study of Wheat Spike Infection by Bipolaris sorokiniana

The infection of wheat spikelets by Bipolaris sorokiniana , the causal agent of black point on grains and grain shrivelling, was examined by light and electron microscopy. Conidia of the pathogen germinated 6–12 h after inoculation on the surfaces of the different spike tissues. Extracellular sheaths were observed on germ tubes and appressoria attached to the surfaces of lemma, palea and seeds, but were only scarcely detected on the surface of conidia. Appressoria, frequently found over grooves, formed penetration hyphae invading the epidermal cell walls. Infection process was similar on the surface of the lemma, palea and glume. Growth of the fungus in the epidermal and parenchyma cells was found predominantly in the cell walls, and hyphae also extended intercellularly and intracellularly. Infection of seeds appeared to occur via two ways: (i) direct infection of the outer layers of the cell walls of the pericarp and (ii) through entering the stigma into the pericarp cells. Secretion of host cell wall hydrolytic enzymes at the apex of the penetrating hyphae may facilitate the spread of the fungus. In addition, toxins secreted by the fungus might explain the rapid death of host cells in contact with or distant to fungal cells. A host response to fungal infection involved the development of appositions between cell wall and plasma membrane in cells adjacent to fungal cells. Fungal hyphae were sometimes also surrounded by electron dense material.     

Comparative Chemical Characterization of Pigmented and Less Pigmented Cell Walls of Alternaria tenuissima

Alternaria tenuissima, the parasitic fungus, was obtained from the pruned upper-cut surfaces of mulberry stems. This fungus contains dark pigment because of the presence of melanin in the cell wall. To obtain less-pigmented cell walls, this fungus was grown under dark condition. When the pigmented and less-pigmented cell walls were chemically analyzed, no differences were observed in amino-acid composition, hexoses, or pentoses. However, in pigmented cell walls, higher contents of melanin (2.6%) were found than in less-pigmented cell walls (0.3%). Interestingly, a significant difference was observed in the relative fatty-acid compositions between these two types of cell walls. Among the major fatty acids, there were increased concentrations of tetradecanoic acid (C14:0), hexadecanoic acid (C16:0), 9-hexadecenoic acid (C16: 1,Δ9), and 9-octadecanoic acid (C18:1,Δ9) and a concomitant decrease in 9,12-octadecadienoic acid (C18:2,Δ9,12) in less-pigmented compared with pigmented cell walls. This difference in fatty-acid composition may be related to the higher percentage of melanin in the pigmented than the less-pigmented cell walls. Lesser amounts of 9,12-octadecadienoic acid in less-pigmented cell walls may have been caused by the growth of the fungus under environmental stress conditions. An interesting observation was the presence in pigmented cell walls only of methyl-substituted fatty acids with carbon numbers C14 to C17, but their occurrence could not be ascertained in the present study.     

Brow suspension,a minimally invasive technique in facial rejuvenation

People tend to prefer noninvasive or minimally invasive methods of facial rejuvenation, especially when it involves their face, which is the hallmark of a person's identity and impossible to hide. It is widely known that brow ptosis gives the face a "tired look" and also accentuates deformities of the upper eyelid. Most people who are interested in facial rejuvenation may not accept even a minor surgery, such as an endoscopic surgery. The senior author has developed a minimally invasive method of suspending the brow at a higher position. In this technique, there is neither surgical dissection nor a surgical incision except for four stab incisions and suture insertion, which is why we refer to it as a nonsurgical brow suspension. It is done under local anesthesia, and the brows are fixed in the position that they assume when the patient is supine. In the past 6 years, we performed 387 brow suspensions on 324 female and 63 male patients. The youngest patient was 19 years old, and the oldest was 74 years old. A retrospective chart review was done. These 387 cases were reviewed by comparison of preoperative and postoperative photographs. This approach was not only used for patients who were not interested in surgical rejuvenation but was also combined with lipofilling, laser resurfacing, and/or upper blepharoplasty. This technique is useful for correcting postsurgical brow asymmetry. We present this technique as an adjunct to the established techniques of facial rejuvenation. Despite the high patient acceptance and technical ease, it is not a replacement for the established techniques of facial rejuvenation.     

梭绒盘菌(子囊菌门:锤舌菌纲)对寄主植物Medeola virginiana的系统性侵染(英文)

设计了用于扩增梭绒盘菌ITS rDNA片段的特异性引物。研究发现,梭绒盘菌不仅存在于被侵染植物的茎部病斑中,而且存在于寄主Medeola virginiana未被感染的叶片、茎及根部。由于寄主植物通过克隆繁殖,因此病害发生与营养体的系统性侵染有关。文中评述了寄主植物的生长模式以及该菌在其寄主群体中长期生存的状况。     

Production of Pectinase and Cellulase by Cladosporium cucumerinum with Dissolved Carbohydrates and Isolated Cell Walls of Cucumber as Carbon Sources

The scab fungus Cladosporium cucumerinum can use pectins and polygalacturonic acid as sole sources of carbon. Cellulose and Ca-polygalacturonate are not available carbon sources for the fungus. When growing on sucrose or pectin, pectinase is produced. In these cases the production of cellulase is insignificant. On a mixture of pectin and carboxymethylcellulose also cellulase is produced. Both pectinase and cellulase are released into the culture filtrate when the fungus grows on cell walls without ionic proteins, whereas only cellulase is released when cell walls with ionic proteins are the carbon source. Pectinase produced by the pathogen can bind to isolated cell walls. The bound pectinase can be extracted with 1 M NaCl from cell walls without ionic proteins, but not from cell walls with ionic proteins. A water-extract or 1 M NaCl-extract of cucumber hypocotyls with visible disease symptoms contains cellulase but no pectinase activity. Lack of pectinase activity in the 1 M NaCl-extract may be due to inhibition by a component that could be extracted by NaCl from the cucumber cell walls.     

Finished genome of the fungal wheat pathogen Mycosphaerella graminicola reveals dispensome structure, chromosome plasticity, and stealth pathogenesis

The plant-pathogenic fungus Mycosphaerella graminicola (asexual stage: Septoria tritici) causes septoria tritici blotch, a disease that greatly reduces the yield and quality of wheat. This disease is economically important in most wheat-growing areas worldwide and threatens global food production. Control of the disease has been hampered by a limited understanding of the genetic and biochemical bases of pathogenicity, including mechanisms of infection and of resistance in the host. Unlike most other plant pathogens, M. graminicola has a long latent period during which it evades host defenses. Although this type of stealth pathogenicity occurs commonly in Mycosphaerella and other Dothideomycetes, the largest class of plant-pathogenic fungi, its genetic basis is not known. To address this problem, the genome of M. graminicola was sequenced completely. The finished genome contains 21 chromosomes, eight of which could be lost with no visible effect on the fungus and thus are dispensable. This eight-chromosome dispensome is dynamic in field and progeny isolates, is different from the core genome in gene and repeat content, and appears to have originated by ancient horizontal transfer from an unknown donor. Synteny plots of the M. graminicola chromosomes versus those of the only other sequenced Dothideomycete, Stagonospora nodorum, revealed conservation of gene content but not order or orientation, suggesting a high rate of intra-chromosomal rearrangement in one or both species. This observed "mesosynteny" is very different from synteny seen between other organisms. A surprising feature of the M. graminicola genome compared to other sequenced plant pathogens was that it contained very few genes for enzymes that break down plant cell walls, which was more similar to endophytes than to pathogens. The stealth pathogenesis of M. graminicola probably involves degradation of proteins rather than carbohydrates to evade host defenses during the biotrophic stage of infection and may have evolved from endophytic ancestors.     

An infection of cow with Mortierella wolfii

A Japanese black cow from a farm in Gunma prefecture showed pyrexia, anorexia and depression. An autopsy revealed hepatic lesions consisting of numerous approximately spherical pale lesions scattered about on the surface and cut parts. The inflammatory lesions were observed in the right lobe of the lung. A part of the trachea was whitish and thickened. Histologically, necrotic foci were observed throughout most of the liver with fungal proliferation about the blood vessels. The lesions of the lung were seriously exudative and the fungus was also present in some of the bronchiole and on bloodvessel walls. Mortierella wolfii was isolated from the liver. The hyphae in the liver were identical in appearance with those seen in the tissues of rabbits experimentally inoculated with M. wolfii.This report is the first case of M. wolfii infection in Japan     

Direct observation of the structure of mycelial cell walls from the potato blight fungus Phytophthora infestans by solid-state 13C NMR

Mycelial cell walls from the potato blight fungus ( Phytophthora infestans ) (Mont.) de Bary were examined in the solid state by ¹³C nuclear magnetic resonance spectroscopy with cross-polarization and magic-angle spinning. The spectrum was free from interference by spinning sidebands. The main component of the cell walls had the spectral properties of a β -(1,3')-glucan. Protein appeared to be present also. The presence of β -(1,6')-linked glucose residues, cellulose or chitin was not ruled out, but there was no evidence for these as major components of the cell walls.     

A study of pityriasis versicolor in Bangkok (Thailand)

A total of 410 cases of pityriasis versicolor were studied in the Division of Dermatology, Department of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand, during January to December 1987. These comprised 1% of all skin disorders, while infection caused by fungus comprised 17%. Most persons infected with pityriasis versicolor were 12–21 years old (mean age 20 years) and 47% were male and 53% were female. The common sites of infection were back, arms, face and trunk. The colors of lesions found were white (83%), brown (7%), pink (6%) and ashy (4%). Both forms of M. furfur were isolated, the ovale form in 97% and the orbiculare form in 3% of cases. The colors of the lesions did not depend on the form of the organism. Mycological and immunological studies confirmed that the two forms were different stages in the complex developmental cycle of a single species.     

Virulence and the Production of Endo-1,4-β-glucanase by Isolates of Alternaria alternata Involved in the Moldy-core Disease of Apples

Alternaria alternata, is the predominant fungal pathogen responsible for moldy‐core in apple cultivars of the Red Delicious group. Here we report on the association between virulence of natural isolates of A. alternata, their production of endo‐1,4‐β‐glucanase (EG) and moldy‐core development in apple fruits. Based on decay development following wound inoculations of mature fruits, three of 150 isolates, collected in three orchards in northern Israel and representing low, moderate and high virulence, were selected and designated Rm44, Er30 and Sh42, respectively. All three isolates secreted EG when grown on enzyme‐inducing medium (EIM) containing commercial cellulose or apple cell walls and this production was related to their degree of virulence. Polyacrylamide gel electrophoresis (PAGE) revealed quantitative differences between the three isolates, relative to their virulence. When fungal extracts were run in native gels, a single band with a molecular mass of 23 kDa showing EG activity was produced by the high‐ (Sh42) and the medium‐virulence (Er30) isolate but not by the low‐virulence (Rm44) isolate. A commercial cellulase preparation (containing endo‐ and exo‐1,4‐β‐glucanase) placed on pricked fruit led to the formation of symptoms similar to those developing on A. alternata‐inoculated fruits within 2–4 days. Inoculation of bloom clusters at full bloom with the highly virulent isolate (Sh42) of A. alternata resulted in a significantly higher infection in fruits (58%) than in those inoculated with the low‐virulence isolate (Rm44) (30%). Our results suggest that the moldy‐core symptoms caused by A. alternata in apple, could be related to the ability of the fungus to produce EG in developing lesions.     

Differential location of α-expansin proteins during the accommodation of root cells to an arbuscular mycorrhizal fungus

-Expansins are extracellular proteins that increase plant cell-wall extensibility. We analysed their pattern of expression in cucumber roots in the presence and in the absence of the mycorrhizal fungus, Glomus versiforme. The distribution of -expansins was investigated by use of two polyclonal antibodies (anti-EXPA1 and anti-EXPA2, prepared against two different cucumber -expansins) in immunoblotting, immunofluorescence, and immunogold experiments. Immunoblot results indicate the presence of a 30-kDa band specific for mycorrhizal roots. The two antibodies identify antigens with a different distribution in mycorrhizal roots: anti-EXPA1 labels the interface zone, but the plant cell walls only weakly. By contrast, the anti-EXPA2 labels only the plant cell walls. In order to understand the potential role of -expansins during the accommodation of the fungus inside root cells, we prepared semi-thin sections to measure the size of cortical cells and the thickness of cortical cell walls in mycorrhizal and non-mycorrhizal root. Mycorrhizal cortical cells were significantly larger than non-mycorrhizal cells and had thicker cell walls. In double-labelling experiments with cellobiohydrolase–gold complex, we observed that cellulose was co-localized with -expansins. Taken together, the results demonstrate that -expansins are more abundant in the cucumber cell walls upon mycorrhizal infection; we propose that these wall-loosening proteins are directly involved in the accommodation of the fungus by infected cortical cells.     

Development of schematic face preference in macaque monkeys

This study investigated schematic face preferences in infant macaque monkeys. We also examined the roles of whole and partial features in facial recognition and related developmental change. Sixteen infant monkeys, all less than two months old, were presented with two stimulus pairs. Pair A consisted of "face" and "parts," with the components representing facial parts (i.e. eyes, mouth, and nose). Pair B consisted of "configuration" and "linear," each including three black squares. In each pair, one of two stimuli represented a facial configuration, namely "face" and "configuration." Visual following responses toward each stimulus were analyzed. The results revealed an early preference for schematic faces in these nonhuman primates. Infants less than one month of age showed a preference only for a stimulus that contained only whole facial configuration (i.e. "configuration" in Pair B). One-month-old macaque infants showed a preference only for "face" but not for "configuration." This result means that their preference at that age was affected by both the shape of the components and the overall configuration. As the developmental change and the contribution of both facial features are similar to those in human infants, it may suggest that primates share common cognitive processes in early schematic face recognition.     

Host-Pathogen Interactions: I. A Correlation Between alpha-Galactosidase Production and Virulence

Resistance or susceptibility of Red Kidney, Pinto and Small White beans (Phaseolus vulgaris) to the alpha, beta, and gamma strains of Colletotrichum lindemuthianum was either confirmed or established. These fungal strains secrete α-galactosidase, β-galactosidase and β-xylosidase when grown on cell walls isolated from the hypocotyls of any of the above bean varieties. These enzymes effectively degrade cell walls isolated from susceptible 5-day old hypocotyls but degrade only slightly the walls isolated from resistant 18-day old hypocotyls. The amounts of the β-galactosidase and β-xylosidase secreted by the 3 fungal strains are relatively low and are approximately equivalent. The secretion of these 2 enzymes is not dependent upon the bean variety from which the hypocotyl cell walls used as a carbon source were isolated. However, the fungal strains secrete greater amounts of α-galactosidase when grown on hypocotyl cell walls isolated from susceptible plants than when grown on walls from resistant plants. Virulent isolates of the fungus, when grown on hypocotyl cell walls isolated from a susceptible plant, secrete more α-galactosidase than do attenuated (avirulent) isolates of the same fungal strain grown under the same conditions. The α-galactosidase secreted by each of the fungal strains is capable of removing galactose from the hypocotyl cell walls of each bean variety tested. Galactose is removed from the cell walls of each variety at the same rate regardless of whether the cell walls were isolated from a susceptible or resistant plant.     

Cell wall degradation in the autolysis of filamentous fungi

A systematic study on autolysis of the cell walls of fungi has been made on Neurospora crassa, Botrytis cinerea, Polystictus versicolor, Aspergillus nidulans, Schizophyllum commune, Aspergillus niger, and Mucor mucedo. During autolysis each fungus produces the necessary lytic enzymes for its autodegradation. From autolyzed cultures of each fungus enzymatic precipitates were obtained. The degree of lysis of the cell walls, obtained from non-autolyzed mycelia, was studied by incubating these cell walls with and without a supply of their own lytic enzymes. The degree of lysis increased with the incubation time and generally was higher with a supply of lytic enzymes.Cell walls from mycelia of different ages were obtained. A higher degree of lysis was always found, in young cell walls than in older cell walls, when exogenous lytic enzymes were present.In all the fungi studied, there is lysis of the cell walls during autolysis. This is confirmed by the change of the cell wall structure as well as by the degree of lysis reached by the cell wall and the release of substances, principally glucose and N-acetylglucosamine in the medium.     

Isolation and chemical analysis of the cell wall of Morchella sp.

Cell walls from Morchella sp. were isolated after mechanical breakage of the fungus. Analysis of the material revealed that the most prominent compound was protein, phosphate being present in lesser amounts. Non-nitrogenous polysaccharides accounted for 17%. They were mostly soluble in hot HCl. The following monosaccharides were identified by chromatography of hydrolyzed samples: glucose, mannose and galactose. No lipids were detected in the purified cell walls. Chitin was present in a concentration of 16%. It was identified by cytochemical reactions, infrared spectra, and X-ray diffraction analysis. The cell wall of Morchella did not contain chitosan nor cellulose.     

Loss of cell wall alpha(1-3) glucan affects Cryptococcus neoformans from ultrastructure to virulence

Yeast cell walls are critical for maintaining cell integrity, particularly in the face of challenges such as growth in mammalian hosts. The pathogenic fungus Cryptococcus neoformans additionally anchors its polysaccharide capsule to the cell surface via alpha(1-3) glucan in the wall. Cryptococcal cells disrupted in their alpha glucan synthase gene were sensitive to stresses, including temperature, and showed difficulty dividing. These cells lacked surface capsule, although they continued to shed capsule material into the environment. Electron microscopy showed that the alpha glucan that is usually localized to the outer portion of the cell wall was absent, the outer region of the wall was highly disorganized, and the inner region was hypertrophic. Analysis of cell wall composition demonstrated complete loss of alpha glucan accompanied by a compensatory increase in chitin/chitosan and a redistribution of beta glucan between cell wall fractions. The mutants were unable to grow ina mouse model of infection, but caused death in nematodes. These studies integrate morphological and biochemical investigations of the role of alpha glucan in the cryptococcal cell wall.     

The Establishment and Structure of Latent Infections with Monilinia fructicola on Apricots

Latent infections of apricot fruit with Monilinia fructicola were initiated by inoculation at shuck fall and 43 and 63 days after shuck fall. The fungus entered via the stomata and penetrated a guard cell through the thin walled region at the stomatal pore. The fruit tissue responded by death of cells around the point of infection, suberization of walls of surrounding living cells and accumulation o phenolic compounds in cells up to 20 distant. Periderm developed around lesions formed at shuck fall but was absent from those formed 65 days later. When the fruit ripened, approximately 100 days after shuck fall, viable hyphae in latent infections escaped from the lesions by growing out between the cuticle and epidermis or just below the epidermal cells. Outbreak was more efficient from later inoculations and only a small proportion of latents initiated at shuck fall became invasive.