Karyology and the possible function of the dual conidia ofDissoconium (Hyphomycetes)

The morphological development and nuclear behaviour of two species ofDissoconium are described. The fungi are monokaryotic in all stages of their life cycle. Liberated pairs of conidia may form anastomoses, probably serving survival under hostile environmental conditions. The function of the one-celled conidia inD. dekkeri remains unexplained.     

Nuclear distribution in Alternaria tenuis

Nuclear distribution and behaviour during vegetative growth and spore formation inAlternaria tenuis was studied utilising the HCl — Giemsa staining technique. The vegetative mycelium and conidia are predominantly monokaryotic. Anastomoses, followed by nuclear migrations, have been recorded. Intercellular nuclear migrations have only been observed in germinating conidia. Nuclear behaviour during conidial formation indicates that the conidia are homokaryotic. Cytological differences have been found in different monocoidal isolates of A. tenuis.     

Pathogenesis of anAspergillus flavus infection ofGalleria mellonella eggs

Egg masses of the greater wax moth,Galleria mellonella L. experimentally infected with conidia ofAspergillus flavus Link became rapidly covered with the mycelium and were eventually destroyed. Germ tubes from conidia penetrated not only infertile, but also damaged eggs. Healthy eggs were often penetrated both by conidial germ tubes and mycelia elements. Once penetrated, eggs become filled with mycelium in as little as two days, after which conidiophores and mycelia protrude outwardly through the chorion.

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Résumé Des pontes de la Teigne des ruches,Galleria mellonella, infectées expérimentalement avec des conidies deAspergillus flavus ont été rapidement recouvertes de mycélium et furent éventuellement détruites. Les tubes germinatifs issus des conidies ont pénétré non seulement les œufs infertiles mais aussi les œufs blessés. Les œufs sains furent souvent pénétrés à la fois par des tubes germinatifs conidiens et des éléments mycéliens. Une fois pénétrés, les œufs se remplissent de mycélium en moins de 2 jours, après lesquels le mycélium et les conidiophores ressortent à l'extérieur à travers le chorion.

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Authorized for publication on November 7, 1968 as paper No. 3503 in the Journal Series of the Pennsylvania Agricultural Experiment Station.     

Carbohydrate and lipid components of hyphae and conidia of human pathogenFonsecaea pedrosoi

The carbohydrate and lipid components of mycelium and conidia ofFonsecaea pedrosoi (Brumpt) were analysed by paper, thin-layer and gas-chromatography, mass spectrometry and ultraviolet spectroscopy. Glucose, mannose, galactofuranose, rhamnose and glucosamine were polysaccharide components identified inF. pedrosoi. Significant changes in the carbohydrate pattern occurred during the conversion of mycelium into conidia. Rhamnose was predominant in conidia whereas galactose was prominent in mycelium. Palmitic, stearic, oleic, linoleic, and arachidonic acids were the fatty acids identified in the total lipid fraction. Palmitic and oleic acids were major fatty acids. Marked alterations in the fatty acid constituents were observed between the cell types ofF. pedrosoi. Arachidonic acid was detected only in conidia and linoleic acid was preferentially identified in mycelium. Differences in the sterol composition was also associated with morphogenesis inF. pedrosoi. Two main sterols, ergosterol and another less polar sterol, not fully characterized, were found in mycelium whereas in conidia only the latter sterol was present.     

Physiological effects of nanosilver on vegetative mycelium,conidia and the development of the entomopathogenic fungus,Isaria fumosorosea

An assessment of the influence of water nanosilver suspension was made at a concentration of 10 mg·L^?1 on biological material (i.e., vegetative mycelium and conidia of Isaria fumosorosea entomopathogen) on a background of the silver nitrate ionic form used. Conidia of I. fumosorosea treated with silver nitrate for more than 168 h were completely deactivated. The application of nanosilver on Isaria hyphae resulted in a quantitative limitation of mycelium growth and its weaker sporulation after culturing compared to the control. The pathogenicity of a conidial suspension obtained from such culturing towards test insects did not diverge from that observed in the standard culture. No obvious toxic effects of nanosilver were observed on I. fumosorosea conidia. Isaria conidia, after exposure to nanosilver over a period between 1 and 800 h, initially demonstrated weaker vegetative mycelium formation in culture on solid medium and, as a consequence, this mycelium often sporulated in a poorer manner. In one case, there was a significant stimulation of the sporulation process for nanosilver treatment before culture for 168 h. Concurrently, conidial suspensions obtained from the culture after exposure of over 168 h to nanosilver exhibited enhanced pathogenicity towards test insects, which may be considered a beneficial phenomenon in terms of the function played by Isaria in the whole environment. The reaction of conidia with nanosilver indicates the deactivation of conidia cells in suspensions and a possibility of selection in increased pathogenicity.     

双孢蘑菇疣孢霉病的发病过程及病原菌的核相研究

【目的】确定有害疣孢霉的传播途径,明确双孢蘑菇受有害疣孢霉侵染后发病症状和微观形态变化,以及有害疣孢霉的核相。【方法】将有害疣孢霉喷施于培养料及覆土材料的不同深度,观察记录双孢蘑菇的发病情况;将有害疣孢霉接种于不同生长阶段的双孢蘑菇子实体,观察记录其发病情况;使用光学显微镜及扫描电镜观察双孢蘑菇子实体受有害疣孢霉侵染前后的形态变化;通过DAPI(4′,6-二脒基-2-苯基吲哚)染色的方法对有害疣孢霉核相进行观察。【结果】将有害疣孢霉接种于培养料及覆土层的不同深度得到双孢蘑菇发病率如下:覆土层表面覆土层中间覆土与培养料交界处培养料中间层;有害疣孢霉可以侵染双孢蘑菇的任意阶段,将其接种于原基直径小于3 mm子实体表面时,得到不能正常分化的"马勃状"组织;对有害疣孢霉的侵染过程进行观察得到:其孢子可粘附于双孢蘑菇表面,并萌发长出芽管,接种处双孢蘑菇表面产生褐色病斑,双孢蘑菇菌丝体发生质壁分离,最后菌丝体膨大,细胞壁变薄甚至溢裂,菌丝体内部中空;有害疣孢霉产生两种类型的分生孢子,Ⅰ类无隔膜含1个细胞核;Ⅱ类具1隔膜含2个细胞核,2个细胞核被隔膜分开;细胞核的第1次有丝分裂发生于分生孢子母细胞中;厚垣孢子由上下2个细胞构成,上胞中含有2个细胞核。下胞含1–2个细胞核。有害疣孢霉的厚垣孢子萌发可产生1–2个芽管,芽管中细胞核的数目不断变化,一般0–2个细胞核。【结论】双孢蘑菇受其侵染后发生显著的细胞学变化;我们对有害疣孢霉做遗传分析时,进行单孢分离需挑取无隔膜的分生孢子为实验材料进行遗传分析。     

Fusarium nisikadoi, a new species from Japan

A new species ofFusarium, F. nisikadoi, isolated fromPhyllostachys nigra var.henonis (bamboo) andTriticum aestivum (wheat) in Japan, is described, illustrated and discussed. This species is differentiated from other known species of the genus by the following characteristics: whitish colony color, long zigzag-like chains of 0–3(-5)-septate clavate conidia, intermixed with pyriform conidia, produced mostly from monophialides and rarely from polyphialides in the aerial mycelium, very long and slender sporodochial conidia, and no chlamydospores. The long chains of septate conidia are known only in this species of the genusFusarium. The conidiophores on the aerial mycelium sometimes proliferate sympodially. The species is tentatively placed in the form-sectionLiseola.     

Efficacy of Exserohilum longirostratum on barnyard grass (Echinochloa crus-galli spp. crusgalli) under field conditions

An isolate of Exserohilum longirostratum obtained from Rottboelia cochinchinensis in Malaysia was highly pathogenic to barnyardgrass (Echinocloa crus-galli,) a serious weed in rice fields in Malaysia. In glasshouse trials, high levels of barnyardgrass control were achieved when E. longirostratum was applied as a conidial concentration at 10⁵ conidia/mL. This conidial concentration may not be sufficient for field use; therefore, a mini plot trial was carried out to assess the bioherbicidal potential of the fungus and the efficacy of different inoculum types under field conditions. Excellent barnyard grass disease as indicated by Area Under Disease Progress Curve (AUDPC) was achieved with mycelium (AUDPC = 583.8 unit²); mycelium + pretichlaclor (AUDPC = 610.4 unit²) and conidia alone (AUDPC = 468.3 unit²) compared to conidia + pretichlaclor (AUDPC = 395.8 unit²). Although conidia caused lower disease severity on barnyard grass compared to mycelium, this inoculum equally reduced the competition from barnyard grass as indicated by lower mean dry weight and fewer barnyard grass tiller numbers. Positive correlations between AUDPC of rice and its tiller number and AUDPC of rice and its dry weight were recorded while negative correlations were observed for the AUDPC of barnyard grass and its tiller number and AUDPC of barnyard grass and its dry weight. These findings confirmed that E. longirostratum applied either in the form of conidia or mycelium was effective against barnyard grass under field conditions.     

Involvement of the glutamine synthetase/glutamate synthase pathway in ammonia assimilation by the wood-rotting fungusPleurotus ostreatus

The mycelium of the wood-rotting fungus,P. ostreatus, contains NAD-dependent glutamate synthase inhibited by azaserine.l-Glutamine andl-glutamate are the most important free amino acids in the mycelium. Feeding of the mycelium with nitrogenous substrates showed thatl-glutamate,l-aspartate andl-alanine are interconnected by way of transaminases. After the inhibition of glutamine synthetase by methionine-S-sulfoximine the synthesis ofl-glutamate was inhibited and the level of all free amino acids decreased. The¹⁵N-NMR spectra of mycelia after the addition of¹⁵NH₄Cl confirmed that the GS/GOGAT is the only pathway of ammonia assimilation inP. ostreatus and NAD-glutamate dehydrogenase should be the deaminating enzyme.     

AN ODOROUS BASIDIOBOLUS OFTEN PRODUCING CONIDIA PLURALLY AND FORMING SOME DICLINOUS SEXUAL APPARATUS

A coarse fungus isolated from decaying plant detritus originating in northern Wisconsin and western Maryland is newly described as Basidiobolus magnus. Like B. ranarum, it gives off an odor recalling benzene hexachloride. Its mycelium, especially while young, includes many long segments markedly wider in the middle than at the ends. Its distally phototropic conidiophores sometimes are branched in their procumbent or submerged portions, and each propulsive distention may bear 1–3 globose conidia, which may be shot off forcibly or may fall through collapse of the supporting hypha. The primary globose conidia of B. magnus are generally larger than those of B. ranarum, from which they differ further in sometimes giving rise to plural secondary conidia of either the globose or the elongated adhesive type. The zygospores, like those of B. ranarum, show abundantly undulated sculpture but differ in their generally larger size, their occasional origin through conjugation between cells of separate hyphae, their rather frequent binary development, and their germination by production, often, of 2 globose uninucleated conidia. The elongated conidia, which seem generally no bigger than those of B. ranarum and are often only meagerly tipped with adhesive material, sometimes undergo conversion into sporangia through internal segmentation.     

Effect of relative humidity,temperature and fungicide on germination of conidia of Cercospora canescens caused the Cercospora leaf spot disease in mungbean

The aim of the present investigation was to determine the impact of relative humidity (RH) and temperature on conidial germination, nuclear position and effect of important fungicides on growth and conidial germination of Cercospora canescens. Germination of conidia was observed at RH range 92–100% at 5–35°C. Significant interaction between temperature and RH indicated that higher humidity and high temperature promoted quick germination both in the presence and absence of free moisture. Although in absence of free moisture at 92–95% RH higher temperatures 25–35°C promoted quick evaporation of moisture and no conidial germination. Number of germtube was increased significantly at the optimum temperature 25–30°C and higher humidity (98–100%). But higher temperature 25–35°C with lower RH did not support the conidial germination. This finding is very important for disease forecasting using meteorological data. The spray of Carbendazim as contact fungicide may not be useful since it is not effective against the conidia of C. canescens. Triadimefon did not inhibit the conidia germination but completely inhibited mycelium development at 50 μg/ml. Propriconazole inhibited both conidia germination and mycelial development. Therefore, Propiconazole may be taken as protective as well as curative spray. In non-systemic fungicide, Copper oxychloride gave anticipated result by inhibiting both conidial germination and mycelium development. Therefore, copper oxychloride can be used as protectant fungicides for Cercospora leaf spot caused by C. canescens.     

Segregation of heterokaryons in the asexual cycle of Phycomyces

Summary Artificial heterokaryons between carotene mutants ofPhycomyces blakesleeanus Bgff. have been prepared by squeezing cytoplasm out of two different mutant sporangiophores and allowing the fused droplets to regenerate.These heterokaryons are used to study the distribution of nuclei at different stages of the asexual life cycle. It is proposed that the nuclear ratio is constant in all parts of the mycelium, sporangiophores and sporangia, and that random samples of nuclei are packaged into spores. This model permits quantitative predictions regarding the proportions of phenotypes in the asexual progeny and these predictions are corroborated by experiments. The nuclear ratio remains constant during repeated mycelial transfers.     

Peculiarities of Exogenous Dormancy of Aspergillus nigerConidia

Aspergillus nigerconidia are characterized by exogenous dormancy: the first stage of their germination is accomplished in twice-distilled water. However, germ tube formation requires the availability of carbon and nitrogen sources. Exogenous dormancy in A. nigerconidia exhibits the following peculiar features: (i) nitrogen-containing substances are active stimulators of germination; (ii) temperature-dependent changes in the lipid bilayer and in the neutral lipid composition of conidia are virtually identical to those occurring in growing mycelium under temperature stress; and (iii) the spore viability threshold does not exceed 45°C; i.e., the spores are more heat-resistant than the mycelium, but they are less heat-resistant than the spores that are in the state of endogenous dormancy. According to the current classification of the types of cell metabolism arrest, the exogenous dormancy of A. nigerconidia resembles the pattern of metabolism characteristic of vegetative cells during the idiophase.     

Effect ofAspergillus parasiticus soil inoculum on invasion of peanut seeds

Environmental control plots adjusted to late season drought and elevated soil temperatures where inoculated at peanut planting with low and high levels of conidia, sclerotia, and mycelium from a brown conidial mutant ofAspergillus parasiticus. Percentage infection of peanut seeds from undamaged pods was greatest for the subplot containing the high sclerotial inoculum (15/cm² soil surface). Sclerotia did not germinate sporogenically and may have invaded seeds through mycelium. In contrast, the mycelial inoculum (colonized peanut seed particles) released large numbers of conidia into soil. Soil conidial populations of brownA. parasiticus from treatments with conidia and mycelium were positively correlated with the incidence of seed infection in undamaged pods. The ratio ofA. flavus to wild-typeA. parasiticus in soil shifted from 7:3 to 1:1 in the uninoculated subplot after instigation of drought, whereas in all subplots treated with brownA. parasiticus, the ratio of the two species became approximately 8:2. Despite high levels of brownA. parasiticus populations in soil, nativeA. flavus often dominated peanut seeds, suggesting that it is a more aggressive species. Sclerotia of wild-typeA. parasiticus formed infrequently on preharvest peanut seeds from insect-damaged pods.     

Morphology,development and cytology of Taphrina maculans Butler

Morphology, development and nuclear behavior of the ascogenous stroma and asci in the infection spots have been described inTaphrina maculans Butler. The fungus forms subcuticular and intercellular mycelium in the leaf tissues and the ascogenous layers originate through division of the subcuticular hyphal cells in the infection sites. Germination of ascogenous cells starts with their elongation in the uppermost layer forming asci and ascospores without formation of stalk cells. Meiosis of the fusion (diploid) nucleus occurs in the young ascus as in otherTaphrina species devoid of stalk cells. The haploid chromosome complement in this species consists of 3 chromosomes (n=3). All the cells in the stromatic layer are potential ascogenous cells and ascus formation continues, until all of them are exhausted in the infection spot. Eight ascospores are normally formed in each ascus, but multi-plication of ascospores may occurin situ later. Three morphologically distinct types of ascus opening are encountered, which are apparently not correlated with prevalent environment. Multiplication of ascospores after their discharge from mature asci occurs by budding proceded by a mitotic division of the spore nucleus. Blastospores (budded cells) germinate into short hyphae and binucleate condition of cells originates by mitotic division of the nucleus. Occurrence of giant cells containing 2 nuclei is often observed. Possible origin of Uredinales fromTaphrina-like ancestors has been indicated due to their close resemblance.     

Production ofBeauveria bassiana [Deuteromycotina: Hyphomycetes] in different liquid media and subsequent conidiation of dry mycelium

Mycelium ofBeauveria bassiana can be grown in liquid culture, filtered, and the mycelium dried. After rehydration the mycelium sporulates. Two carbohydrate sources (sucrose and maltose), and one nitrogen/vitamin source (yeast extract) were tested for mycelium growth and subsequent conidial production. Maximum mycelium growth (12.31 mg/ml), in liquid culture, was in the sucrose (3.5%)/yeast extract (3.5%) medium, but mycelium from a maltose (2%)/yeast extract (0.75%) medium produced the maximum of 4.62×10⁶ conidia/mg dry mycelium after incubation in moist Petri dishes. Using the data on mycelium yield (in liquid culture) and conidial production (by dry mycelium) it is calculated that the sucrose (3.5%)/yeast extract (3.5%) and the maltose (2%)/yeast extract (0.75%) media produce most conidia per media volume (an equivalent of 3.52–3.72×10⁷ conidia/ml).      

Beauveria bassiana submerged conidia production in a defined medium containing chitin,two hexosamines or glucose

Summary Beauveria bassiana in liquid culture can produce blastospores and occasionally submerged conidia. For use as a bioinsecticide, conidia have definite advantages. Numerous studies have investigated conidia production in liquid cultures using synthetic and industrial grade media supplemented with glucose. We have studied growth, development and sporulation in microcultures using growth media containing chitin monomers. For the production of submerged conidia growth media containing N-acetyl-d-glucosamine (GlcNAc) proved to be better than yeast extract-peptone-glucose (YPG), glucose plus ammonium salts (Glc+NH₄Cl) or N-acetyl-d-galactosamine (GalNAc). Sixty-one percent of the spores in the GlcNAc medium were submerged conidia with the remainder being blastospores. The concentration of submerged conidia reached 8.0 × 10⁵/ ml after two days in GlcNAc medium as compared to 8.9 × 10⁵/ml in YPG medium. Therefore, in terms of percentage of submerged conidia produced, GlcNAc medium generated more submerged conidia in spite of its lower cell yields. Growth in a medium containing chitin, a polymer of GlcNAc, resulted in 86.3% of the spores as submerged conidia exceeding 10⁶/ml after 48 h. Growth under phosphate limitation resulted in an increased percentage of submerged conidia for all media tested. Electron microscopy and spore protein analysis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed that structural and compositional differences exist between the spore types.     

The effects of culture media on Saprolegnia antigens

In a previous paper (Hodkinson &Hunter, 1970a) we reported that salmon, suffering from a disease called ulcerative dermal necrosis, appeared to have precipitating antibodies to the mycelium of a fungus associated with their diseased skin lesions. This was confirmed when antigens were prepared from pure cultures of the fungus, a species ofSaprolegnia (Hodkinson &Hunter, 1970b). In this paper we describe the effects of culture media on the antigenicity ofSaprolegnia mycelium, as detected by double gel diffusion with salmon sera.     

Five additional genera of conidial lichen-forming fungi from Europe

Three remarkable new genera of conidial lichen-forming fungi, with pycnidial or acervular conidiomata are described and illustrated.Hastifera tenuispora gen. et spec. nov. from the South-Tyrol, Italy, is characterized by very long and narrow hyaline conidia produced in pycnidial conidiomata immersed in thalline warts.Lichingoldia gyalectiformis gen. et spec. nov. from rocks subject to inundation by freshwater in Norway has long sigmoidly curved conidia which appear to be adapted to dispersal in water.Woessia fusarioides gen. et spec. nov. from aQuercus stump in the Burgenland, Austria, has a finely granular thallus and disc-like conidiomata producing falcate conidia; this species is also of interest in that it hasChlorella as the photobiont. The recently described hyphomyceteCheiromycina flabelliformis B. Sutton is reported from Austria for the first time, and is also lichenized. Recent collections ofNigropuncta rugulosa D. Hawksw. from Austria and Italy also show that this species is lichenized rather than parasymbiotic. In all five cases the fungal hyphae of the conidiogenous structures are continuous with those in intimate contact with cells of the algal partners and the biological relationship appears to be mutualistic giving rise to stable crustose lichen thalli.     

Changes in enzyme activity of germinating conidia of Neurospora crassa

The conidia of wild-type Neurospora crassa are shown to have a drastically lower activity for three enzymes of the isoleucine-valine pathway—acetohydroxy acid synthetase, dihydroxy acid dehydratase, and aminotransferase—than the actively growing mycelium. Lower activity was also found in the conidia for ornithine transcarbamylase and aspartate transcarbamylase. Lower activity (10- to 100-fold) was found for the overall synthesis of valine from pyruvate in the conidia as compared to the mycelium as expected.In addition it is also apparent that the distribution of the isoleucine-valine enzymes is different in conidia from the mycelium as regards activity in the mitochondria as compared to the cytosol. In conidia their activity in the mitochondria is lower than in the cytosol, but the opposite holds in the mycelium. These differences are also reflected in the overall activity.Cycloheximide inhibits the increase in total activity of the acetohydroxy acid synthetase and the dehydratase during germination of the conidia.