Ultrastructure of spined conidia and hyphae of the rice false smut fungus Ustilaginoidea virens

Spined conidia and hyphae of Ustilaginoidea virens were examined by light and electron microscopy. Bright-field light microscopy showed that conidia were round to elliptical and warty on the surface with diameters approximately ranging from 3 to 5 microm. Scanning electron microscopy revealed the globose to irregularly rounded and ornamented conidia with prominent spines. The spines were pointed at the apex or irregularly curved, and approximately 200-500 nm long. Ultrastructure of spined conidia and hyphae revealed by transmission electron microscopy showed lipid globules and vacuoles in the cytoplasm enclosed by an electron-transparent cell wall. Conspicuous electron-dense spines were evident on the surface of conidia, and had obclavate or irregularly protruding shapes with varying heights along the conidial cell wall. Microfibrillar structures with stretching or branching patterns were evident in the spine matrix. Some conidia were interconnected by spines from the neighboring conidia by their extended outgrowth. Hyphae had concentric bodies that showed an electron-transparent core surrounded by an electron-dense layer. One or more intrahyphal hyphae were found in hyphal cytoplasm. The fungus is thought to form concentric bodies and intrahyphal hyphae as survival mechanisms against the water- and nutrient-deficient environments that may occur in the necrotic regions of host plants.     

Colonization of cashew plants by Lasiodiplodia theobromae: microscopical features

Lasiodiplodia theobromae is a phytopathogenic fungus causing gummosis, a threatening disease for cashew plants in Brazil. In an attempt to investigate the ultrastructural features of the pathogen colonization and its response to immunofluorescence labeling, light, confocal and electron microscope studies were conducted on different severity scale patterns of diseased plants. Lasiodiplodia-antisera was checked for cross reactivity against common cashew plants fungi. Optical microscopy analysis revealed a longitudinally sectioned hyphae located within the xylem vessels, showing an extensive hyphal development in the secondary xylem tissue. SEM images demonstrated that the fungus was found in some asymptomatic samples, particularly within the xylem vessels as confirmed by the optical images. Symptomatic sample images showed an extensive distribution of the fungus along the secondary xylem, within the vessels, infecting xylem parenchyma. A closer look in the secondary xylem parenchyma reveals a heavy and profuse invasion of the cells with a distinguishable cell wall disintegration and fully hyphae dispersal. There was no reactivity of Lasiodiplodia-antisera against mycelial extracts of Colletotrichum gloeosporioides, Phomopsis anardii and Pestalotiopsis guepinii. Following incubation of sections with the polyclonal antisera, the hyphae were intensely and regularly labeled. Rays, vessels and parenchyma cells were the preferred pathway for L. theobromae colonization. Artificial infection provides the information that the vascular cylinder is undoubtedly employed and used by the fungus for hyphae distribution. Immunofluorescence assay employed in situ was applied and the polyclonal antisera produced was able to recognize the fungus and proved to be a sensitive technique to detect it.     

Use of light and scanning electron microscopy to examine colonisation of barley rhizosphere by the nematophagous fungus Verticillium chlamydosporium

Barley roots were readily colonised by the nematophagous fungus Verticillium chlamydosporium. Light microscopy (LM) but also low temperature scanning electron microscopy (LTSEM) revealed details of the colonisation process. Hyphae were found on the rhizoplane often with dictyochlamydospores. Hyphae of V. chlamydosporium penetrated epidermal cells, often by means of appressoria. A hyphal network was formed in epidermal and cortical cells. Likewise, hyphal coils were found within root cells next to transverse cell walls. Cortical cells were the limits of fungal colonisation, since no hyphae were seen in the vascular cylinder. Modifications of root cell contents (phenolic droplets and callose appositions) were common three weeks after inoculation with V. chlamydosporium. These features may indicate induction of plant defence reactions in late stages of root colonisation by the fungus. Both LTSEM and LM have proved extremely useful to describe root colonisation by the fungus. The results found may have implications in the mode action of nematophagous fungi against plant parasitic nematodes.     

Morphological and ultrastructural examination of senescence in Morchella elata

In recent years, the artificial cultivation of Morchella mushrooms that belong to Elata Clade, including Morchella elata, has been developed rapidly in China. However, the prominent problem of spawn aging has been frustrating the morel farming. In this paper, aging in M. elata was achieved from 12 to 17 subcultures and lifespan of 1536–2256 h by successive subculturing. The lifespan can be roughly divided into juvenile phase and senescent phase with respect to the mycelia linear growth rate. After a certain period of rapid growth with almost constant rate at the juvenile phase, the isolate entered the senescent phase characterized by slow down of mycelia growth, producing pigments ahead of time and final death of the apical hyphae. The period of the senescent phase was definitely 240–288 h; while that of the juvenile phase was diverse relying on different isolates. Moreover, microscopic study showed that angles between the leading and primary hyphae increased constantly with aging. In senesced hyphal cells of M. elata, the typical characteristics of autophagy (enlargement of vacuoles and existence of organelles sequestrated lysosomes) and apoptosis (condensation of the cytoplasm and nuclear and plasmolysis) were observed. In addition, in the final stage of senescence, the apical hyphae collapsed with the plasma membrane and all the cellular organelles disrupted, indicated a necrotic mode of cell death. Taken together, these data revealed the involvement of autophagy, apoptosis and necrosis in senescence of M. elata. The characterization and molecular mechanism of autophagy, apoptosis and necrosis need further study and the systematic study of morel aging will be beneficial for the healthy development of morel farming.     

First evidence of an intimate symbiotic association between fungi and larvae in basal attine ants

The most conspicuous ants in all of tropical America are those that belong to the tribe Attini which cultivate fungus. The objective of the present study is to verify the alterations that occur in the cuticle of the worker larvae from Myrmicocrypta, Mycetarotes and Trachymyrmex with the purpose of trying to establish the degree of this association. Attine ants from the Atta genus were used as a control group. The analysis of histological sections showed results about the distribution of the cells and tissues on the epidermis of A. sexdens rubropilosa ant larvae and Mycetarotes parallelus, Trachymyrmex fuscus and Myrmicocrypta sp. A cuticle covering the ants was observed in the larvae of all the species, and this is formed by a simple cubic epithelium, whose cells possibly change its shape to prismatic, depending on their secretory activity. Just under this epithelium large adipose cells with reserve granules in their cytoplasm were found. The presence of a space filled by granulose and acellular material was also observed, indicating that the larvae were in a molt period. The presence of fungal hyphae is observed both on the external side of the basal attine larvae as well as emitting projections to the interior of the cuticle reaching the epithelium and the adipocyte cells. Data obtained in the present study demonstrated that the fungus deposited on the surface of immature ants from attine basal species maintain a close relationship with them, once the fungus hyphae have the ability to disorganize the cuticle lamellas, penetrating the interior of the insect cells through the emission of prolongations transporting the cuticle and epithelium barriers and making substances exchanges between larvae and fungus.     

Thallus structure and isidium development in two Parmeliaceae species (lichenized Ascomycota)

(Thallus structure and isidium development in two Parmeliaceae species (lichenized Ascomycota)). Conventional techniques for structural studies under light microscope and scanning electron microscope were employed to describe and compare the histology of thallus and isidia development in Parmotrema tinctorum and Parmelinopsis minarum. Although presenting layers typical of the heteromerous structure of Parmeliaceae lichens, the studied species have very distinct anatomical details including: the type of upper cortex, medulla thickness, hyphal orientation pattern and distinct strata in this layer, variations of crystal shape, color, location and degree of association with medullary hyphae, rhizines thickness and covering by a cortical layer. Isidium formation starts with cortical cell proliferation, which results in a small protuberance on the thallus surface, followed by the photobiont cell proliferation. Medullary hyphal intrusions occur only after the formation of this protuberance. In this stage medullary hyphae grow, ramifying or not inside the protuberance. In a posterior stage, the isidium base becomes constricted, which facilitates its detachment from the thallus.     

Ultrastructure of the siphonaceous green alga Halimeda cuneata,with emphasis on the cytoskeleton

Cytoplasm streaming is a fundamental process for the transport of molecules and organelles in plant cells. In vegetative filaments of the coenocytic green alga, Halimeda cuneata Hering, the spatial organisation of microtubules in the cytoplasmic layer, was observed under transmission electron microscopy. A cross section of a cortical filament shows a tubular cell wall enclosing a peripheral layer of cytoplasm with numerous chloroplasts, amyloplasts, nuclei, mitochondria and microtubules surrounding a small central vacuole. Towards the thallus medulla the central vacuole enlarges considerably and the cytoplasm becomes gradually reduced to a thin parietal layer, the number of organelles is reduced and the quantity of microtubules increases. Therefore, most of the thallus volume is occupied by the huge central vacuole which extends throughout the coenocytic filaments. Microtubules run longitudinally, being about 0.05 μm from each other. Some microtubules were observed in close association to cell organelles.     

基于共聚焦显微拉曼的真菌菌丝中几丁质的原位检测研究

几丁质是真菌细胞壁中一种重要的结构多糖,本文首次采用共聚焦显微拉曼技术对山茶刺盘孢菌的气生菌丝进行原位检测研究,首先确定了采集菌丝拉曼光谱的最优实验参数,并获得了菌丝,几丁质标准品和背景三种物质的典型拉曼光谱,对其中的特征峰进行归属分析,发现菌丝光谱中有明显的几丁质特征峰。然后对置于载玻片上菌丝的感兴趣区域进行拉曼光谱面扫描,通过主成分分析法发现面扫描区域中,菌丝和背景两种信号可以明显区分开来,结合主成分的载荷因子图得到了菌丝的两个主要的特征差异峰1 622和1 368 cm-1,1 622 cm-1属于菌丝中几丁质的特征峰,而1 368 cm-1是来源于菌丝中的果胶多糖。最后通过对几丁质在1 622 cm-1特征峰波段附近范围积分,绘制了几丁质在菌丝中二维和三维的化学成像图,直观且无损的再现了几丁质在菌丝中的空间分布。     

Biotrophic interaction of Sporisorium scitamineum on a new host—Saccharum spontaneum

Sporisorium scitamineum is a biotrophic smut fungus harbored inside the smut gall on the top internodal region of Saccharum spontaneum, a wild relative of sugarcane (Saccharum officinarum). The interactions of spined conidia of S. scitamineum with S. spontaneum were examined during the different stages of plant growth starting from the bud stage to the decaying stage. The spores in the soil from the polyetic inocula grew into confined epidermal cells of the buds and finally sporulated in the topmost internodal region. Hyphae invasion of the plant tissues were restricted to the point of infection. Culms of infected plants in late October sporulated, notably; hyphal sporulation produced shorter hyphal stolons. Remarkably, the nodal regions of infected plants had no spores and fragmented hyphae. On the basis of microscopic analyses, hyphae and spores were absent in all internodes above the ground till the topmost smut gall region. This result indicated that, S. scitamineum undergoes tissue-confined invasion of S. spontaneum. By associating culture medium method with polymerase chain reaction (PCR) on plant portions void of smut gall, S. scitamineum was not detected, indicating that colonization was not systemic. It was observed that the biotrophic interaction resulted in structural reorganization in the restricted region of infection forming erect cylindrical structure, in which the fungus was sandwiched between the central stalk and sheath, and possibly played a key role in preventing inflorescence. Comparatively, a significant difference in the rate of teliospores germination between reference Ustilago esculenta (26.6%, P < 0.05) and S. scitamineum (62.9%, P < 0.05) at 20° C was observed. This study also provides insights on the effect of different temperature regimes on the germination of S. scitamineum teliospores in vitro.     

Electron microscopic analysis of dairy microbes inactivated by ultrasound

Ultrasonication is a non-thermal method of food preservation that has the advantage of inactivating microbes in food without causing the common side-effects associated with conventional heat treatments, such as nutrient and flavour loss. The aim of this study was to evaluate the use of ultrasound as an alternative to heat pasteurisation and to assess cell damage using transmission electron microscopy (TEM). Three spoilage microbes, previously isolated from pasteurised milk, were used as "test" microbes. Saline solution (SSS) and UHT milk were used as suspension media and were inoculated with exponential growth phase "test" microbes at a microbial concentration of 1 x 10(4) cfu ml(-1). The samples were subjected to power ultrasound (20 kHz, 750 W), at 100%/124 microm wave amplitude for different time intervals. Both Escherichia coli and Saccharomyces cerevisiae were reduced by >99% (for both suspension media) after ultrasonication and Lactobacillus acidophilus was reduced by 72% and 84% in SSS and milk, respectively. Transmission electron microscope micrographs showed that ultrasonication inflicts extensive microbicidal/microbistatic external and internal damage on all three "test" microbes. In E. coli, sonication-induced emulsification caused the formation of unique minute lipopolysaccharide vesicles from the fragmenting cell envelope.     

Spontaneous rotational inversion in Phycomyces

The filamentary fungus Phycomyces blakesleeanus undergoes a series of remarkable transitions during aerial growth. During what is known as the stage IV growth phase, the fungus extends while rotating in a counterclockwise manner when viewed from above (stage IVa) and then, while continuing to grow, spontaneously reverses to a clockwise rotation (stage IVb). This phase lasts for 24-48 h and is sometimes followed by yet another reversal (stage IVc) before the overall growth ends. Here, we propose a continuum mechanical model of this entire process using nonlinear, anisotropic, elasticity and show how helical anisotropy associated with the cell wall structure can induce spontaneous rotation and, under appropriate circumstances, the observed reversal of rotational handedness.     

以最大原子密度定义合金相中的第一近邻团簇

本文提出利用不同壳层所包含的径向原子密度, 即单位体积内的原子个数随着径向的分布, 来方便而精确地定义团簇, 即具有最大径向原子密度的且表面呈现三角密堆结构的完整壳层为第一近邻团簇. 最后以Al-Ni-Zr合金相为例说明了该方法的合理性与适用性, 及此方法所定义的团簇与非晶形成的关系.     

P granules phase transition induced by cytoplasmic streaming in Caenorhabditis elegans embryo

P granules are germ granules contained in Caenorhabditis elegans germ cells.The first germ cell is specified by the one-cell embryo in which P granules localize to the posterior.Previous studies suggested that the mechanism of the localization phenomena is induced by liquid-liquid phase transition(LLPT),in which the polarity proteins control the saturation point of P granules.In the present study,we propose that the P granules phase transition can be triggered by the cytoplasmic streaming.A two-phase flow model is employed to simulate the localization of P granules,i.e.,the cytoplasm is considered as a liquid phase,and the droplet-like P granules are another liquid phase.With the presence of the cytoplasmic streaming,P granules,initially distributing uniformly in the entire one-cell embryo,eventually condense/dissolve in the cytoplasm phase,regulated by difference between the saturation pressure and the hydrodynamic pressure.The numerical results reveal that the cytoplasmic streaming has significant effects on the localization of P granules,as well as the embryo division.     

Occurrence and ultrastructure of Albugo candida on a new host, Arabis alpina in Saudi Arabia

A population of Arabis alpina (Brassicaceae) growing in Saudi Arabia was observed to be infected for the first time by the Oomycete, Albugo candida. Both conventional chemical fixation and high pressure freezing followed by freeze substitution (HPF/FS) were used to prepare zoosporangia, intercellular hyphae, haustoria, invading host cells and host–parasite interface of A. candida for study with both scanning and transmission electron microscopy. Both fixations gave good preservation of ultrastructural details and data from the two sample types were highly complementary. Scanning electron microscopic observation revealed that mature zoosporangia of A. candida are spherical or ellipsoidal in shape and characterized by a smooth surface and faint terminal secession scar at each end. Transmission electron microscopic observation indicated that coenocytic intercellular hyphae are located in intercellular spaces of host leaf tissue forming haustoria in host mesophyll cell. Each haustorium is connected to intercellular hyphae by a narrow, slender neck which enclosed by a collar as a response of host cell to infection. The cytoplasm of the haustorium contains different organelles characteristic of the Oomycetes. No obvious responses are observed in host cell organelles following infection which may be due to the presence of a compatibility between the host and the Oomycete. Modifications of the host plasma membrane around the haustorium are detected. Many tubular elements were found to be continuous with the extrahaustorial membrane. This appears to be the first report of the presence of these tubular elements in case of A. candida haustoria. These tubular elements may increase membrane surface area and consequently increase the efficacy of nutrients uptake by haustoria from host cell.     

超高压处理对枯草芽孢杆菌超微结构的影响

 研究了超高压处理对枯草芽孢杆菌（Bacillus subtilis AS 1.140）超微结构的影响,探讨其营养体以及芽孢的灭活机制。在经过500 MPa、60 ℃下保温保压20 min超高压处理后,采用透射电子显微镜进行观察,比较处理前后超微结构的变化。观察结果表明：超高压处理后,枯草芽孢杆菌的营养体细胞壁皱缩、出现缺口,胞浆泄漏、结构层次感消失、出现大片透电子区;其芽孢外壳被破坏、出现缺口,芽孢内含物结构紊乱、泄漏、出现部分透电子区;甚至内含物质完全泄漏,出现细胞壁或孢子外壳残留。     

Sm(CobalFe0.1Cu0.1Zr0.033)6.9高温永磁合金的矫顽力

对Sm(Co_balFe_0.1Cu_0.1Zr_0.033)_6.9合金, 经810℃等温时效后以0.5℃/min逐渐冷却, 在600℃-400℃温度区间淬火, 研究了不同淬火温度下的磁滞回线、磁畴和矫顽力温度系数β. 发现时效600℃淬火后磁滞回线出现台阶状, 说明畴壁中应存在两处钉扎. 随淬火温度的降低, 合金的室温矫顽力显著增加, 磁滞回线的台阶消失. 通过磁畴形貌发现时效600℃淬火后的磁畴接近条形畴, 1:5相中Cu分布相对均匀, 形成的畴壁钉扎较弱, 从而使磁滞回线出现台阶, 决定矫顽力的畴壁钉扎位于两相界面处; 随时效淬火温度的降低, 磁畴逐渐细化, 畴壁1:5相中的畴壁能降低, 形成了较强的内禀钉扎, 并决定材料的矫顽力, 两相界面处的畴壁钉扎被掩盖. 对不同温度淬火合金的高温矫顽力研究表明, 最强的畴壁钉扎位于两相界面处时, 矫顽力随温度升高逐渐增加, 矫顽力出现温度反常现象; 最强的畴壁钉扎位于1:5相中心时, 矫顽力随温度升高逐渐衰减. 当测试温度达到500℃后不同淬火温度样品的矫顽力几乎相同, 此时最强畴壁钉扎均在两相界面处.     

Scanning electron microscopy of Ancylostoma spp. dog infective larvae captured and destroyed by the nematophagous fungus Duddingtonia flagrans

The interaction between the nematode-trapping fungus Duddingtonia flagrans (isolate CG768) against Ancylostoma spp. dog infective larvae (L₃) was evaluated by means of scanning electron microscopy. Adhesive network trap formation was observed 6 h after the beginning of the interaction, and the capture of Ancylostoma spp. L₃ was observed 8 h after the inoculation these larvae on the cellulose membranes colonized by the fungus. Scanning electron micrographs were taken at 0, 12, 24, 36 and 48 h, where 0 is the time when Ancylostoma spp. L₃ was first captured by the fungus. Details of the capture structure formed by the fungus were described. Nematophagous Fungus Helper Bacteria (NHB) were found at interactions points between the D. flagrans and Ancylostoma spp. L₃. The cuticle penetration by the differentiated fungal hyphae with the exit of nematode internal contents was observed 36 h after the capture. Ancylostoma spp. L₃ were completely destroyed after 48 h of interaction with the fungus. The scanning electron microscopy technique was efficient on the study of this interaction, showing that the nematode-trapping fungus D. flagrans (isolate CG768) is a potential exterminator of Ancylostoma spp. L₃.     

用x射线形貌研究半绝缘砷化镓单晶胞状结构

通过x射线异常透射形貌（XRT），化学腐蚀显微观察和电子显微技术（TEM）研究了液封直拉（LEC）法生长的半绝缘砷化镓（SI-GaAs）单晶中蜂窝状胞状结构，从晶体生长和冷却过程的热应力、弹性形变引起位错的运动和反应考虑，分析了该结构的形成机理与过程.认为这是由高密度位错（EPD）运动和反应所形成的小角度晶界的集合群. 关键词： SI-GaAs 小角度晶界 胞状结构     

Numerical analysis and design of patterned electrode liquid crystal microlens array with dielectric slab

The phase profile of hole patterned electrode liquid crystal (LC) microlens array (MLA) was studied by a numerical method. It indicated that the formation of inverse domain and tilt wall in the liquid crystal cell limits the optical performance of the microlens array. The physical mechanism that determines the formation of inverse domain and tilt wall was elucidated by analyzing the phase profile and the liquid crystal director distribution. According to our numerical results, a new configuration for hole patterned electrode liquid crystal microlens array was proposed by introducing a dielectric slab between the patterned electrode and the liquid crystal. The physical parameters that influence the optical performance of this new configuration LC MLA were studied, which provide fundamental information for the design. The results indicate that the phase profile and focusing power of the new configuration is greatly improved.     

Ultrastructure of Tuta absoluta parasitized eggs and the reproductive potential of females after parasitism by Metarhizium anisopliae

Among the microorganisms used in biological control, the muscadine fungus Metarhizium anisopliae (Metsch.) Sorok. is produced and formulated world wide aiming to control pests from several agricultural crops. This work evaluated effects of M. anisopliae isolate UFRPE-6 on the fecundity and mortality of Tuta absoluta (Lepidoptera: Gelechiidae) females and the mechanism of infection on eggs. The infection of the females by the fungus did not affect their oviposition and fecundity; however it affected the survival with total and confirmed mortality of 54.2% and 37.14%, respectively. The eggs were treated with suspension at concentration of 10(6)conidia/mL. The analysis under scanning electron microscopy showed that conidia germination and penetration processes in the eggs of T. absoluta started within the period of 6h after the inoculation. Several hyphal bodies were observed from 12h and an intense extrusion of the mycelium covering all the external surface of the eggs 72h after inoculation. Despite its moderate activity in adults, the isolate URPE-6 of M. anisopliae showed promising in the control of T. absoluta due to its pathogenicity and virulence to eggs from this pest.