Spore Shape Research Articles

Polygonibacillus indicireducens gen. nov., sp. nov., an indigo-reducing and obligate alkaliphile isolated from indigo fermentation liquor for dyeing

Obligately alkaliphilic and halophilic strains, designated In2-9T and D2-7, were isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample obtained from a craft centre in Date City, Hokkaido, Japan. The 16S rRNA gene sequence phylogeny suggested that strain In2-9T is a member of the genus Bacillus with the closest relatives being the alkaliphilic species of the genus Bacillus, Bacillus hemicellulosilyticusJCM 9152T (96.4 % 16S rRNA gene sequence similarity) and Bacillus alcalophilus DSM 485T (96.5 %). Cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. Strain In2-9T grew between 13 and 45 °C with optimum growth at approximately 35-37 °C. The isolates grew in the pH range of 8-12 with optimum growth at pH 10. The isoprenoid quinone detected was menaquinone-6 (MK-6) and the DNA G+C content was 39.4 mol%. The whole-cell fatty acid profile mainly (>10 %) consisted of iso-C15 : 0, anteiso-C15 : 0 and C16 : 0. Spore shape and location and chemotaxonomic characteristics revealed that the isolates were distinctly different from phylogenetic neighbouring alkaliphilic species of the genus Bacillus. On the basis of phenotypic and chemotaxonomic characteristics and phylogenetic data, the isolates represent a novel species of a new genus, for which the name Polygonibacillusindicireducens gen. nov., sp. nov. is proposed. The type strain of the type species is In2-9T (=JCM 30831T=NCIMB 14982T), and strain D2-7 is an additional strain of the species.

Back-trajectory modelling and DNA-based species-specific detection methods allow tracking of fungal spore transport in air masses

Recent advances in molecular detection of living organisms facilitate the introduction of novel methods to studies of the transport of fungal spores over large distances. Monitoring the migration of airborne fungi using microscope based spore identification is limited when different species produce very similar spores. In our study, DNA-based monitoring with the use of species-specific probes allowed us to track the aerial movements of two important fungal pathogens of oilseed rape (Brassica napus L.), i.e., Leptosphaeria maculans and Leptosphaeria biglobosa, which have identical spore shape and size. The fungi were identified using dual-labelled fluorescent probes that were targeted to a β-tubulin gene fragment of either Leptosphaeria species. Spore identification by Real-Time PCR techniques capable of detecting minute amounts of DNA of selected fungal species was combined with back-trajectory analysis, allowing the tracking of past movements of air masses using the Hybrid Single Particle Lagrangian Integrated Trajectory model. Over a study period spanning the previous decade (2006–2015) we investigated two specific events relating to the long distance transport of Leptosphaeria spp. spores to Szczecin in North-West Poland. Based on the above mentioned methods and the results obtained with the additional spore sampler located in nearby Szczecin, and operating at the ground level in an oilseed rape field, we have demonstrated that on both occasions the L. biglobosa spores originated from the Jutland Peninsula. This is the first successful attempt to combine analysis of back-trajectories of air masses with DNA-based identification of economically important pathogens of oilseed rape in Europe. In our studies, the timing of L. biglobosa ascospore dispersal in the air was unlikely to result in the infection of winter oilseed rape grown as a crop plant. However, the fungus could infect other host plants, such as vegetable brassicas, cruciferous weeds, spring rapeseed and winter rapeseed growing as a volunteer plant.

PRESENCIA DE HONGOS EN EL AIRE DE AULAS DEL DEPARTAMENTO DE INGENIERÍA QUÍMICA Y METALURGIA DE LA UNIVERSIDAD DE SONORA, UNIDAD CENTRO

En este trabajo se evaluó la presencia de hongos en el aire de aulas del Departamento de Ingeniería Química y Metalurgia, recolectándose 90 muestras utilizando el método pasivo en cajas Petri de 90 mm de diámetro, que contenían Agar Sabouraud Dextrosa. Para identificar el tipo de hongo se preparó un frotis, de acuerdo a la técnica de Impronta. Se determinó la forma y ordenamiento característico de las esporas. Los hongos encontrados fueron Alternaria sp., Aspergillus flavus, Aspergillus niger, Cladosporium sp., Drechslera sp., Penicillium sp., y Fusarium sp. De acuerdo al número de unidades formadoras de colonias (UFC) registradas durante los días de monitoreo, los mayores recuentos se encontraron en el turno matutino, antes del inicio de actividades. Además, se observó que el muestreo matutino del día Lunes presentó la mayor cantidad de UFC en las diferentes aulas, lo cual se atribuye principalmente a la inactividad en el fin de semana.

Measuring spore settling velocity for an improved assessment of dispersal rates in mosses.

The settling velocity of diaspores is a key parameter for the measurement of dispersal ability in wind-dispersed plants and one of the most relevant parameters in explicit dispersal models, but remains largely undocumented in bryophytes. The settling velocities of moss spores were measured and it was determined whether settling velocities can be derived from spore diameter using Stokes' Law or if specific traits of spore ornamentation cause departures from theoretical expectations. A fall tower design combined with a high-speed camera was used to document spore settling velocities in nine moss species selected to cover the range of spore diameters within the group. Linear mixed effect models were employed to determine whether settling velocity can be predicted from spore diameter, taking specific variation in shape and surface roughness into account. Average settling velocity of moss spores ranged from 0·49 to 8·52 cm s(-1) There was a significant positive relationship between spore settling velocity and size, but the inclusion of variables of shape and texture of spores in the best-fit models provides evidence for their role in shaping spore settling velocities. Settling velocities in mosses can significantly depart from expectations derived from Stokes' Law. We suggest that variation in spore shape and ornamentation affects the balance between density and drag, and results in different dispersal capacities, which may be correlated with different life-history traits or ecological requirements. Further studies on spore ultrastructure would be necessary to determine the role of complex spore ornamentation patterns in the drag-to-mass ratio and ultimately identify what is the still poorly understood function of the striking and highly variable ornamentation patterns of the perine layer on moss spores.

English

Chroogomphus roseolus is a common mycorrhizal species exclusively found with Pinus wallichiana which has been collected from Himalayan moist forests of Pakistan. Taxonomically, this species is previously not described from Pakistan and also first time reported outside Chinese territory. In this study, Pakistani collection has been characterized on morphological and molecular basis. Macrosopic characters like size, shape and colour of pileus, stipe and spore are compared in Cinese European and North American literature for its identification. Its Internal Transcribed Spacers (ITS) of ribosomal DNA sequence was amplified using universal primer pair (ITS1f and ITS4) and nucleotide sequences queried in GenBank database using BLAST analysis where it matches 99% with sequences of the same species from China. Phylogenetic analysis, description, photographs and line drawings of Pakistani collection confirmed as Chroogomphus roseolus. . The exclusive presence of this species Pinus wallichiana indicates that it can be used in mycorrhization control in afforestation and reforestation of Pinus wallichiana vegetation.

Ultrastructural description of Myxobolus cuneus (Myxosporea) in the skeletal muscle and kidney of tropical farmed fish Piaractus mesopotamicus (Characiformes: Characidae)

This study characterizes by transmission electron microscopy (TEM) and morphometric features the myxozoan Myxobolus cuneus (Myxosporea) in Piaractus mesopotamicus and reports the skeletal muscle and kidney as site of infection. The register was based in 21 young fish from intensive fish farming in Southeast Brazil and the spores were analyzed in fresh-mounted slides of the infected organs stained with Toluidine blue and processed as usual for TEM. It differs from Myxobolus cunhai from the fish host and different polar capsule size, and from Myxobolus serrasalmi on the pyriform spore shape and an oval macrospore, differently to that reported in this study. Morphometric characteristics and TEM study confirmed the present material as M. cuneus.

Phylogenetic study of the genus Kudoa (Myxozoa: Multivalvulida) with a description of Kudoa rayformis sp. nov. from the trunk muscle of Pacific sierra Scomberomorus sierra

Kudoa rayformis n. sp. (Myxozoa; Multivalvulida) was observed in the trunk muscle of Pacific sierra Scomberomorus sierra caught off the coast of Tonosi, Panama. The species formed pseudocysts in myofibers and infection was subclinical. The myxospores possessed four polar capsules and spore valves, one of which had a distinct filamentous extension. This unique morphological characteristic of the myxospore validated this as a new species of Kudoa. Genetically, K. rayformis n. sp. is closest to K. inornata, with 98% and 91% similarity in 18S and 28S rDNA, respectively, but its spore shape was clearly distinct. The 18S rDNA and concatenated sequences from K. rayformis were used in molecular phylogenetic analyses of kudoids to examine the congruence of phylogeny with infection site tropism, spore morphology and cyst/pseudocyst formation. The results demonstrated that the phenotypic traits were correlated with the phylogeny of Kudoidae, and that the biological features of K. rayformis originated from the ancient Kudoidae as exhibited by the non-specific infection site tropism and the ability to infect muscle and form pseudocysts.

Morphological and molecular diversity of endophytic Colletotrichum gloeosporioides from tea plant, Camellia sinensis (L.) O. Kuntze of Assam, India.

Thirty isolates of endophytic fungi were isolated from healthy asymptomatic leaves of tea plant (Camellia sinensis) and identified morphologically based on colony morphology, spore shape and size, growth and sporulation rate. Internal transcribed spacer r-DNA sequence analysis supported for molecular identification of all the isolates. Based on morphological and molecular characteristics the isolates were identified as Colletotrichum gloeosporioides. Variations on colony morphology which included the production of conidial masses, led to divide the isolates into different groups. Variations on spore size, growth rate and sporulation rate were exhibited by all the isolates. With RAPD molecular markers, genetic variations among the thirty isolates were observed. Genetic variations and relatedness among the thirty isolates were analyzed with UPGMA phylogram using NTSYS program. Two major groups were obtained among the thirty isolates. Group I comprised of 16 isolates which included three sub groups (Ia, Ib and Ic) and Group II constituted fourteen isolates and it also had three sub groups (IIa, IIb and IIc). A partial co-relationship among the isolates was established on the basis of morphological and molecular based clustering.

Tuber melosporum smooth spores: an anomalous feature in the genus Tuber

This paper adds new insights on ecology and micromorphology of Tuber melosporum, a rare species with smooth spores. Eight T. melosporum ascomata collected in a 50 y old Pinus halepensis and Quercus ilex plantation in Sardinia, Italy, represent the first recovery of this species outside Spain. In comparison to the T. melosporum holotype, Italian specimens revealed differences in the number of spores in asci and spore shape. We propose an emended description of Tuber to include species without spore ornamentation.

Taxonomic reexamination of two <i>Huperzia</i> species (Lycopodiaceae) in Northeast Asia

A comparison of the external morphology and spores clearly distinguished Huperzia lucidula (Michaux) Trevis. in northeast Asia and North America in terms of leaf shape, leaf margin, distribution of sto- mata on leaves, and spore shape. Therefore, the northeast Asian plants should be treated as a separate species, H. asiatica (Ching) B.-Y. Sun & J. Lim. In addition, we believe that H. javanica (Sw.) C. Yang, regarded as con- specific or infraspecific with H. serrata (Thunb.) Trevis., is a distinct species based on the presence of a leaf pet- iole and serrated leaf margins, and the shape of the gemmiferous branchlets.

Isolation of yeast from soil and different food samples and its characterization based on fermentation

Yeasts are eukaryotic microorganisms which can also be used for bioethanol production. In this modern era of increasing demand for energy and fuel, the microbial biosynthesis of ethanol has gained importance. In this study, the potential yeasts for ethanol production from pentose and hexose sugars were identified. Yeasts were isolated from soil and different food samples. They were identified and characterized based on cell morphology (e.g., mode of cell division and spore shape) and physiology (e.g., sugar fermentation tests). Furthermore, quantification of ethanol and cell concentration was performed throughout the fermentation. Spot plate count method was followed to determine the viable yeast count whereas Potassium dichromate oxidation method was used for determining the ethanol concentration. Six different species of yeasts were cultured in three sets of broth for 24, 48, 72, and 96 hours for bioethanol production. The yeasts isolated from black and green grapes relatively synthesized higher concentration of ethanol.Nepal Journal of Biotechnology. Dec. 2015 Vol. 3, No. 1: 29-34

DIVERSITY OF EXINE’S ORNAMENTAL CHARACTER STUDIES IN FERN AND FLOWERING PLANT

<p>Pollen or pollen wall consists of two layers, namely exine and intine. Exine (outer layer) composed of sporopolenin, and intine (inner layer) composed of cellulose. Pollen wall structure, especially the Exine, is one of the characters used for identification. The shape and type of pollen or spores of each specific plant species. Exine’s ornamentation have variations in form and structure. That like are thorns, scratches, basins and other types of ornamentation. Pollen and spore sampling is done randomly in some plants that are supposed to represent members berspora ferns (Asplenium nidus, Platycerium bifurcatum, and Microsorum diversifolium) and pollen of flowering plants (Ipomea sp., Tridax procumben. And flowers Crinum asiaticum). Spores of Asplenium nidus to have a corrugate apertura monocalpalae with ornaments. Platycerium bifurcatum spores aperture shape gemenete dicolpale with ornaments, while the spores Microsorum diversifolium has the form apertura psilatum dicolpalae with ornaments. Polen has Tridax procumben apertura echinatus peripole with ornaments, Ipomea sp. has the form apertura saculate peripole with ornaments, while the Crinum asiaticum has the form apertura clevate peripole with ornaments. The diversity of spore of plant species in nature have affect the diversity of forms of ornamentation owned. </p><p><strong>Keywords</strong>: Diversity, exine, ornament’s type, spores and pollen.</p>

Synopsis of the species of the genus Sphaeromyxa Thélohan, 1892 (Myxosporea: Bivalvulida: Variisporina: Sphaeromyxidae).

Sphaeromyxa spp. are parasites of marine fishes, infecting the gall-bladders or bile ducts. The spores of these species possess characteristic ribbon-like polar filaments, a unique character among myxozoans. This unique character is also a synapomorphy consistent with estimates of phylogeny for this group which forms a lineage distinct from other myxozoans. There are 49 nominal species of Sphaeromyxa Thélohan, 1892 for which a synopsis is provided, reporting spore dimensions, spore shape, locality, and host species. A line drawing is also provided for each species.

Pathogenicity of Microencapsulated Insecticide fromBeauveria bassianaandMetarhizium anisopliaeagainst Tobacco Budworm,Heliothis virescens(Fabricius)

Abstract. Pathogenicity of microencapsulated spores of Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metsch) Sorokin at a dose of 1 × 108 spores per milliliter were used against tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae). Toxicity of non-microencapsulated spores at the same dose was also assessed. Bovine gelatin was used as a matrix to produce microcapsules by the spray-drying technique with an inlet temperature of 85°C and an outlet temperature of 33°C in the spray dryer process. Scanning electron microcopy was used to assist with measuring the microcapsule size and determining spore shape. Spore viability and moisture were also evaluated. Both fungal formulations were applied by immersing first-instar tobacco budworm larvae for 30 seconds. The number of larvae that died was recorded 48 hours post inoculation. Microcapsules were <20 μm. Spore viability was 16.0 ± 0.7% for B. bassiana and 13.2 ± 0.8% for M. anisopliae. Moisture contents of the fungi were...

Spelling out Jaapia species

Jaapia is a wood-saprobic genus of corticioid fungi for which two species have been recognized: J. argillacea Bres. and J. ochroleuca (Bres.) Nannf. & J. Erikss. Whereas the first one is easily recognized by its characteristic spores, the descriptions of the second indicated variable spores, which once led us to believe that J. ochroleuca could be a species complex rather than a single species. Eleven new ITS nrDNA sequences of J. ochroleuca were aligned with two obtained from GenBank and four of J. argillacea. The molecular results, parsimony analysis and KP2 distances clearly delimitate one highly supported Jaapia clade, with two subclades that correspond to the two described species. Morphological studies, including the holotype and isotype of J. ochroleuca, show significant differences between the clades concerning the size and shape of spores. The present study corroborates two species in this genus and also confirms that J. ochroleuca is a well-defined species in which spores show great morphological variability. Based on the Jaapia “species hypothesis”, the J. ochroleuca reference sequence has been selected. A comprehensive key to two Jaapia species is also provided.

Genetic characterization of microsporidians infecting Indian non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) by using PCR based ISSR and RAPD markers assay

This study established the genetic characterisation of 10 microsporidian isolates infecting non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) collected from biogeographical forest locations in the State of Assam, India, using PCR-based markers assays: inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD). A Nosema type species (NIK-1s_mys) was used as control for comparison. The shape of mature microsporidian spores were observed oval to elongated, measuring 3.80 to <TEX>$4.90{\mu}m$</TEX> in length and 2.60 to <TEX>$3.05{\mu}m$</TEX> in width. Fourteen ISSR primers generated reproducible profiles and yielded 178 fragments, of which 175 were polymorphic (98%), while 16 RAPD primers generated reproducible profiles with 198 amplified fragments displaying 95% of polymorphism. Estimation of genetic distance coefficients based on dice coefficients method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was done to unravel the genetic diversity of microsporidians infecting Indian muga and eri silkworm. The similarity coefficients varied from 0.385 to 0.941 in ISSR and 0.083 to 0.938 in RAPD data. UPGMA analysis generated dendrograms with two microsporidian groups, which appear to be different from each other. Based on Euclidean distance matrix method, 2-dimensional distribution also revealed considerable variability among different identified microsporidians. Clustering of these microsporidian isolates was in accordance with their host and biogeographic origin. Both techniques represent a useful and efficient tool for taxonomical grouping as well as for phylogenetic classification of different microsporidians in general and genotyping of these pathogens in particular.

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One hundred and five (105) corneal scraping samples were taken from patients suffering of microbial keratitis referred to Ibn Al-Haitham Eye Teaching Hospital in Baghdad. After culturing on the related selective media, results showed that 73 of the samples were positive for occurrence of microorganisms; of these, 68 (64.76%) were belonged to bacterial and 5 (6.85%) to fungi. After the bacterial isolates were identified by cultural, microscopic and biochemical examinations, the following species and percentages were recorded: Pseudomonas aeruginosa 29 (39.73%), Staphylococcus aureus 22 (30.14%) and Staphylococcus epidermidis 17 (23.28%). While the fungal isolates were belonged to Aspergillus niger 3 (4.12 %) and Alternaria spp. 2 (2.73 %) according to their identification through the cultural and microscopic examinations, in addition to the types and shapes of fungal spores. Upon such findings, P.aeruginosa was the predominant bacteria and A.niger the prevalent fungi. Regarding to the patients gender, it was found that cases of microbial keratitis in the samples were more abundant in males 41 (56.16%) than in females 32 (42.85%). Patients group of age (41-59) years was the most microbial keratitis infected group with a percentage of (30.20 %), while the lowest infected group was (≥ 60) year with a percentage of (17.80 %).

Molecular phylogeny of Colletotrichum (Sordariomycetes: Glomerellaceae) inferred from multiple gene sequences.

The genus Colletotrichum contains many important plant pathogens mainly distributed in tropical areas. Previous studies that utilized morphology or single genes have failed to resolve the phylogenetic relationships among the species. In this study, sequences of β-tubulin, 28S ribosomal DNA, and ITS region from nine species were analyzed separately and combined to establish a fast method to infer the phylogeny of Colletotrichum using maximum parsimony, maximum likelihood, and Bayesian inference methods. The tree topologies inferred from the combined data set received higher bootstrap and posterior probability support than those inferred from the individual data sets. Obtained phylogenies highly supported C. capsici as the earliest diverging lineage followed by C. nymphaeae. The remaining seven species clustered into two distinct clades. Clade 1 consists of two monophyletic subclades: C. circinans, C. trichellum, and C. caudatum form one subclade and three accessions of C. dematium form another subclade. In Clade 2, C. incarnatum is in the basal-most clade. Three accessions of C. musae and C. caricae form a strongly supported clade indicating their close relationship. Spore shape analysis reveals an interesting evolutionary trend in the spore shape from acute- to obtuse-ended conidia and from curved to straight conidia in the sampled group of species. A quick and reliable way to infer the phylogeny of Colletotrichum based on combined DNA sequence data is presented in this paper.

Nosema sp. PM-1, a new isolate of microsporidian from infected Papilio machaon Linnaeus, based on ultrastructure and molecular identification.

A new microsporidium, Nosema sp. PM-1, was first isolated from Papilio machaon Linnaeus. The spore shape of the PM-1 isolate was a long oval with an average size of 3.22 μm × 1.96 μm. Ultrastructure observation showed that PM-1 had a typical Nosema common diplokaryotic nuclei structure with 10-13 polar filament coils, spore wall, plasma membrane, and anchoring disk. The complete rRNA gene sequences were obtained by polymerase chain reaction amplification and each rRNA unit was arrayed as follows: 5'-LSU (2497 bp)-ITS (179 bp)-SSU (1232 bp)-IGS (278 bp)-5S (115 bp)-3', which was the same as typical Nosema. The phylogenetic trees of rRNA, DNA-directed RNA polymerase II subunit, and tubulin genes all show that PM-1 was a sister to the clade comprising Nosema bombycis, Nosema spodopterae, and Nosema sp. PX1. The spore morphology, ultrastructure, and complete rRNA structure indicate that this isolate assigned to the ˝true˝ Nosema group, can parasitized in Papilio machaon Linnaeus, which provides a wider host range for Nosema.

Aeromycology: studies of fungi in aeroplankton

Air is a natural environment for spores of many genera and species of fungi. Despite its small size and a significant dispersion they have a great impact on human health and different areas of our activities, such as agricultural production. The study on spores of fungi that belong to aeroplankton or bioaerosole is called aeromycology. The most frequent fungi present in the air are Cladosporium and Alternaria species. Their numbers are abundant regardless of latitude and height above the sea level and above the ground. They mostly originate from agricultural environment. Other frequently listed species of fungi, whose spores are present in the air include of Aspergillus, Penicillium, Fusarium, Sclerotinia and Ganoderma. The concentration of spores in the air strongly depends on the abundance of their formation during the studied period. This in turn relates to geobotanical region, vegetation, degree of urbanization, climatic conditions, season, current weather, wind force and direction, local microclimate, and many other factors. Changes in humidity affect the concentration of different types of fungal spores. In general they are divided to ‘dry’ (Alternaria, Cladosporium, Puccinia, Ustilago, Melampsora, Epicoccum, Drechslera) and ‘wet’ (Didymella, Fusarium, Ganoderma, Gliocladium, Leptosphaeria, Verticillium). Study of the composition of species and genera are being done using different types of spore samplers, mostly volumetric instruments. Visual identification is based on colony morphology of the fungus and the shape and size of spores. The identification at the species level is possible with molecular tools. Methods based on DNA/RNA amplification are very sensitive and accurate. They allow the identification below the species level, e.g. chemotypes, mating types or isolates with genes or alleles of interest. Aerobiological monitoring is widely used in the epidemiology of human diseases (inhalant allergies) and infections of arable crops (decision support systems for the protection of cultivated plants). Aeromycology is interconnected with such diverse areas as industrial aerobiology, bioterrorism, ecology, climatology or even speleology and cultural heritage.