White rust of chrysanthemums

Teleutospores of Puccinia horiana Henn. germinate and discharge sporidia between 4 and 23 °C. At the optimum temperature of 17 °C sporidia discharge starts within 3 h. Maximum germination of the sporidia takes place within 2·5 h between o and 30 °C, there being no clear optimum. High humidity and a film of moisture appear to be necessary for germination of both teleutospores and sporidia. Sporidia can penetrate either leaf surface of chrysanthemum to cause infection between 4 and 24 1°C and within the optimum temperature range, 17–24 °C, effectively penetrate within 2 h. The sporidia are very sensitive to desiccation at below 90 % relative humidity. Methods are described, using leaf discs and whole plants, for screening chrysanthemum cultivars for susceptibility to white rust. Cultivars were placed in five classes ranging from susceptible to immune. Leaf discs of immune cultivars can be distinguished within 30 h by a brown discolouration at the point of inoculation. The early stages of development of the fungus in susceptible, resistant and immune hosts are described. The incubation period in susceptible plants is normally 7–10 days, teleutospores being formed a few days later. Leaves become less susceptible with age but the oldest leaves on 5-month-old plants could still be infected. The maximum survival time of teleutospores in the sori on detached leaves was 8 weeks but was considerably less under moist conditions or buried in soil. Low doses of a mancozeb with zineb fungicide controlled infection by preventing penetration rather than by inhibiting sporidial germination.     

Ustilago maydisMating Hyphae Orient Their Growth toward Pheromone Sources

Snetselaar, K. M., Bölker, M., and Kahmann, R. 1996.Ustilago maydismating hyphae orient their growth toward pheromone sources.Fungal Genetics and Biology20,299–312. When small drops ofUstilago maydissporidia were placed 100–200 μm apart on agar surfaces and covered with paraffin oil, sporidia from one drop formed thin hyphae that grew in a zig-zag fashion toward the other drop if it contained sporidia making the appropriate pheromone. For example,a2b2mating hyphae grew towarda1b1anda1b2mating hyphae, and the filaments eventually fused tip to tip. Time-lapse photography indicated that the mating hyphae can rapidly change orientation in response to nearby compatible sporidia. When exposed to pheromone produced by cells in an adjacent drop, haploid sporidia with thea2allele began elongating before sporidia with thea1allele. Sporidia without functional pheromone genes responded to pheromone although they did not induce a response, and sporidia without pheromone receptors induced formation of mating hyphae although they did not form mating hyphae. Diploid sporidia heterozygous atbbut not ataformed straight, rigid, aerial filaments when exposed to pheromone produced by the appropriate haploid sporidia. Again, thea2a2b1b2strain formed filaments more quickly than thea1a1b1b2strain. Taken together, these results suggest that thea2pheromone diffuses less readily or is degraded more quickly than thea1pheromone.     

Ultrastructure of primary sporidia of a wheat-bunt fungus,Tilletia caries,during ontogeny and mating

Summary Primary sporidia ofTilletia caries (DC.) Tul. are borne on denticles at the tips of promycelia. The promycelia contain many small vacuoles and mitochondria and numerous lipid bodies. As the primary sporidia develop, the promycelial cytoplasm passes into the nascent cells. Septa develop between the bases of mature sporidia and the tips of the denticles. Sporidia that abscise from the denticles commonly have prominent birth scars at their bases. The sporidia have very thin walls, few vacuoles, attenuated mitochondria, and numerous lipid bodies. Conjugation pegs are generally produced by both members of a conjugating pair of sporidia and there are bud scars where they emerge from the sporidia. The sporidial walls are apparently hydrolyzed during emergence of the pegs. Vesicles are sometimes present at the tips of the conjugation pegs and, before fusion, electron-dense accumulations are sometimes observed between the tips of adjacent pegs. The approaching conjugation pegs are precisely aligned prior to fusion, suggesting polar communication. The walls of the conjugation pegs fuse and then are hydrolyzed. Fused sporidia are relatively homogeneous in content. The nucleus in a sporidum is often close to the conjugation tube and occasionally is partly within the fusion tube.Cooperative investigations of the Oregon Agricultural Experiment Station, Brigham Young University, and Science and Education Administration-Agricultural Research, U.S. Department of Agriculture. Technical Paper No. 4,934 of the Oregon Agricultural Experiment Station. Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable.     

Successful inoculation of mature pine with Tricholoma matsutake

Our finding demonstrates, for the first time, that the roots of mature pine trees can be successfully inoculated with a symbiotic ectomycorrhizal fungus, the valuable matsutake mushroom. Long root segments (ca. 5–10 mm in diameter, ca. 50 cm in length) of 50-year-old Pinus densiflora trees were excavated, washed, auxin-treated (2–5 mg indole butyric acid, IBA, per root) and incubated in moist Spagnum moss. Twelve months later, short roots were regenerated, of which approximately 90% were free of mycorrhizae. Mycorrhiza-free short roots were inoculated with mycelial pieces of Tricholoma matsutake and incubated further in a sterilized substrate. Four-and-a-half months later, roots putatively colonized by Matsutake were sampled near the inoculation points. A T. matsutake-specific ITS-rDNA fragment was amplified by nested PCR from approximately 80% of the root samples analyzed, whereas approximately 66% of the root samples processed for staining with Chlorazol black E displayed characteristic T. matsutake Hartig net structures. These results confirm the symbiotic infection of mature P. densiflora roots by matsutake.     

Dibutyryl c-AMP as an inducer of sporidia formation: Biochemical and antigenic changes during morphological differentiation of Karnal bunt (<Emphasis Type="Italic">Tilletia indica</Emphasis>) pathogen in axenic culture

Effect of dibutyryl adenosine 3′,5′-cyclic monophosphate (dbc-AMP), an analogue of c-AMP, was investigated on growth and morphological differentiation ofTilletia indica. Exponential growth was observed up to 21 days in both presence and absence of dbc-AMP; however, increasing concentration of dbc-AMP was deleterious to mycelial growth in liquid culture. A slow increase of mycelial biomass up to 21 days and decline at 30 days in the presence of 2.5 mM dbc-AMP was observed, therefore, this concentration was chosen in subsequent investigations. The inhibitory influence of dbc-AMP was further substantiated by decrease in soluble protein. The fungus on exposure to dbc-AMP experienced morphological differentiation from vegetative mycelial phase to sporogenous mycelial phase, and was induced to produce filiform sporidia. Use of quantitative ELISA further suggested that sporidia formation took more than 21 days in the presence of dbc-AMP. Variations of proteins during different stages ofT. indica grown in the presence and absence of dbc-AMP suggested the expression of stage-specific proteins or differential expression of proteins induced by dbc-AMP. The changes in expression of cell surface antigens as evidenced from decrease and increase binding of anti-mycelial and anti-sporidial antibodies in dbc-AMP treated culture by ELISA was further interpreted on the basis of morphological differentiation from mycelial to sporidial phase     

Influence of experimental conditions on nitrogenous excretion by Lake MichiganMysis relicta (Lovén): laboratory studies with animals acclimated inFragilaria

In laboratory experiments, rates of excretion of ammonia and urea by Lake MichiganMysis relicta were compared for animals incubated in the presence and the absence of algal food (Fragilaria crotonensis chemostat outflow). Prior to experiments, all animals were acclimated to laboratory conditions and the experimental food for 2–4 weeks. Algae used in experiments were enriched in the dark with nutrients (N and P) prior to experimental incubation.There were no significant differences in the ammonia and urea excretion rates of mysids incubated in filtered water compared to those inFragilaria cultures, or in ammonia excretion rates either as a function of individual size or sex of the mysid or of its having been held at 5 °C, 10 °C, or 15 °C. Ammonia excretion rates measured between 4 and 8 h of incubation were significantly higher than those between 1–3 h and 8–15 h, for mysids held both in filtered water and inFragilaria cultures. The results are compared to those from shipboard incubations in a previous study and are discussed with respect to physiological regulation of nitrogenous waste production.Contribution No. 234, Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin.Contribution No. 234, Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin.     

Genetic analysis of Karnal bunt (Neovossia indica) resistance in wheat

Embryos excised from seeds of six generations (P₁, P₂, F₁, BC₁, BC₂ and F₂) of a cross WH 283 x WH 533 were cultured on modified MS medium already inoculated with secondary sporidia ofNeovossia indica. Significant variations for callusing response (CR) (54.55–75.55%) were observed among generations but the presence or absence ofN. indicia did not affect callusing response. A clear inhibition zone (IZ) was formed around each embryo showing callusing. The diameter of IZ varied significantly among generations and was maximum in the resistant genotype, WH 283 (3.60 cm). Fresh weight and dry weight of calli, initiated from embryo cultured and inoculated withN. indica, varied significantly among generations. Coefficient of infection as well as percentage of infection reflected the overdominance of susceptibility. Generation mean analysis showed that the three parameter model was adequate for diameter of IZ only. Six-parameter model showed that additive (in presence ofN. indica), additive and additive x dominance (in absence ofN. indica) effects were also significant. Complementary type of epistasis for fresh weight of calli and dominance, and dominance x dominance effects for dry weight of calli were observed in the presence ofN. indica. Magnitude of additive effects was higher for diameter of IZ in three parameter model. Therefore, selection might assist in improving this trait and thus indirectly help in attaining the resistance towardsN. indica.     

Chitinase production by Beauveria felinaRD 101: optimization of parameters under solid substrate fermentation conditions

Major parameters affecting the production of chitinase by Beauveria felinaRD 101 under solid substrate fermentation conditions have been optimized. Wheat bran moistened with 100 MS-HCl medium adjusted to pH 5.0, inoculated with 1 × 10¹⁰ conidia g^–1 initial dry bran and incubated at 28 °C for 6 days produced maximum chitinase activity of 6.34 U g^–1 initial dry substrate.     

Storage and formulation of the entomopathogenicnematodes Heterorhabditis indica and H. bacteriophora

Successful control of insect pests through theapplication of entomopathogenic nematode dauerjuveniles of H. bacteriophora and H.indica can only be achieved when the nematodematerial reaches the end user in good condition.Storage and formulation techniques must provideoptimum conditions to guarantee a maximum survival andinfectivity of the nematodes. Nematode survival wastested at temperatures ranging between 5–25 °C.A maximum survival of H. indica was achieved at15 °C and the highest mortality at 5 °C.H. bacteriophora survived best at 7.5 °Cand least at 25 °C. An increase of the saltconcentration had positive effects on dauer juvenilesurvival in aqueous suspensions. Low pH between 6 and4 reduced the bacterial growth and prolonged survivalof stored dauer juveniles. Of the organic acidsascorbic, benzoic, citric and sorbic acid, onlyascorbic acid had a positive effect on H. indicasurvival. Extracts of the dried spice plants cinnamon,cloves, rosemary and oregano were tested. Enhancementof H. indica survival was recorded for cinnamonand cloves. Survival and infectivity of nematodesstored in attapulgite and bentonite clays and spongewere recorded over several weeks at different storagetemperatures. Infectivity was not influenced by thedifferent formulation materials. When stored insponge at 25 °C nematodes survived less than 1week and the formulation in clay could only prolongthis period for another week. At 5 °C thesurvival of H. bacteriophora in sponge wassuperior to that in clay, whereas H. indicasurvived less well in sponge than in clay at15 °C. Storage in aerated water at 5 °Cfor H. bacteriophora and at 15 °C for H. indica resulted in the lowest mortality. Forstorage at controlled conditions (temperature, pH andosmolarity), aerated water is superior to all othermethods tested and the addition of preservatives willincrease survival.     

Production of neutral and alkaline extracellular proteases by the thermophilic fungus, Scytalidium thermophilum, grown on microcrystalline cellulose

Extracellular proteases produced by Scytalidium thermophilum, grown on microcrystalline cellulose, were most active at pH 6.5–8 and 37–45 °C when incubated for 60 min. Highest protease activity was at day 3 where endoglucanase activity was low. Protease activity measurements with and without the protease inhibitors, p-chloromercuribenzoate, PMSF, antipain, E-64, EDTA and pepstatin A, suggest production of thiol-containing serine protease and serine proteases. Endoglucanase and Avicel-adsorbable endoglucanase activity in culture medium was not significantly affected by protease inhibitors.     

Stomatal responses in isolated epidermis of the crassulacean acid metabolism plant Kalanchoe daigremontiana Hamet et Perr.

The optimal conditions for opening of stomata in detached epidermis of the Crassulacean Acid Metabolism (CAM) plant Kalanchoe daigremontiana were determined. Stomatal opening in CO₂–free air was unaffected by light so subsequently all epidermal strips were incubated in the dark and in CO₂–free air. Apertures were maximal after 3 h incubation and were significantly greater at 15° C than 25° C. Thus stomata in isolated epidermis of this species can respond directly to temperature. Stomatal opening was greatest when the incubating buffer contained 17.6 mol m^–3 K⁺, but decreased linearly with increasing K⁺ concentrations between 17.6 and 300 mol m^–3; the decrease in aperture was shown to be associated with increasing osmotic potentials of the solutions. Reasons for this behaviour, which differs from that of many C₃ and C₄ species, are discussed. Stomatal apertures declined linearly upon incubation of epidermis on buffer solutions containing between 10^–11 and 10^–5 mol m^–3 abscisic acid (ABA). Hence stomata on isolated epidermis of K. daigremontiana respond to lower concentrations of ABA than those of any species reported previously.     

<Emphasis Type="Italic">In vitro</Emphasis> screening of sugarcane to evaluate smut susceptibility

Tissue-cultured plantlets of three sugarcane (Saccharum spp.) cultivars having a known field smut reaction were screened for susceptibility to Ustilago scitaminea H&P Sydow. Plantlets were inoculated with 0.5 l of a suspension of equally mixed quantities of plus and minus mating type sporidia of U. scitaminea at concentrations ranging from 1×10¹ to 1×10⁶ cells. Fungal sori (whips) were produced in cultivar N12 (intermediate) 6weeks following inoculation with 1×10⁵ mixed sporidia and thereafter in cultivar NCo310 (susceptible) but not in cultivar N19 (resistant). Sori bearing teliospores were produced up to 3months following inoculation and incubation at 26°C. No sori were produced at mixed sporidial concentrations lower than 1×10⁵cells. The in vitro soral production in cultivars N19, N12 and NCo310 was 0, 27.5 and 47.5% respectively. Plantlets inoculated with 1×105sporidia of only one mating-type did not produce sori in any of the three cultivars tested. Blind scoring of an unknown sugarcane cultivar by this method corresponded exactly with its field smut rating.     

CaCl2 extractable N fractions and K2SO4 extractable N released on fumigation as affected by green manure mineralization and soil texture

Repeated mild wet-dry cycles were imposed on a sandy loam to accelerate the mineralization of organic C involved in stabilising macro-aggregates. Soil maintained continually moist (control soil) was compared to that subjected to a series of 6 wet-dry cycles. Two patterns of rewetting and drying were investigated: (1) incubated dry at 25°C for six days between each wet-dry cycle (dry-incubated), or (ii) incubated moist for six days at 25°C between each cycle (moist-incubated). Changes in the proportion of >2 mm, 1–2 mm, 0.5–1 mm and 0.25–0.5 mm aggregates, and carbohydrate C extracted by hot-water or hot-1.5 M H₂SO₄, were measured after each wet-dry cycle, or weekly in the continuously moist control soil. Respiration rates (CO₂ efflux) were measured during the incubation of the moist soil between the wet-dry cycles and compared with the continually-moist control soil.The wet-dry treatments did not increase soil respiration in soil after re-wetting compared to soil kept continually moist and incubated for the same period of time. Despite this, the treatments caused changes in the amounts of acid- and water-extractable carbohydrate C fractions and substantial changes in aggregation. Macro-aggregation and the proportion of soil in each fraction did not change in the soil maintained continuously-moist for 6 weeks (control). However, effects of the two wet-dry treatments on total macro-aggregation were similar to those in the >2 mm, 1–2 mm and 0.25–0.5 mm aggregate fractions: there was a rapid decline in aggregation by 48–65% over the first two cycles, a sharp recovery to 78–100% of the initial aggregation after three cycles, and a further decline after 4–6 cycles.The resistance of organic C mineralization to mild wet-dry cycles confirmed that the organic C in this soil is very stable and resistant to decomposition. Despite aggregates being disrupted, the organic C stabilising these aggregates was resistant to decomposition as determined by CO₂ efflux. When soil was re-moistened and incubated to allow microbial re-colonization, aggregation was similar to that in the soil where microbial re-colonization was limited by rapid drying treatments. Short term changes in the aggregation of this soil appear to be dominated by chemical and/or physical processes.     

Use of agarose-entrapped Aspergillus niger cells for the production of citric acid from soy whey

Aspergillus niger cells immobilized in agarose beads were utilized for the productin of citric acid from soy whey, a by-product generated during the tofu-making process. Soy when samples supplemented with 10% sucrose were inoculated with 10% (w/v) free and immobilized cells and incubated at 30°C in a shaker water bath at a speed of 200–220 rpm. Maximal citric acid yields of 21 g/l and 27 g/l with free and immobilized cells, respectively, were recorded on the 10th day under repeated batch conditions.     

Effects of temperature and relative humidity on development times and mortality of eggs from laboratory and wild populations of the European house-dust miteDermatophagoides pteronyssinus (Acari: Pyroglyphidae)

As part of a study on passive physical control of house-dust mites, a total of 6000 eggs from a population ofDermatophagoides pteronyssinus (Trouessart, 1897) from 17-year-old laboratory cultures were incubated at 60 temperature and relative humidity combinations between 10–35°C and 55–100% RH. Eggs hatched at every combination, although mortality and development time increased between 10–20°C and 30–35°C and below 65% RH. Optimum conditions were 35°C and 80–85% RH. In temperate dry conditions, eggs from a wild population were found to be more resistant to mortality: they developed faster, with 7 times lower mortality than eggs from the laboratory population. This may have been because the laboratory population had become acclimated to the constant near-optimum conditions at which it was kept. Therefore it has been suggested that where laboratory cultures have been used in studies relating to passive physical control, caution should be taken in applying the conclusions to wild populations in the natural house-dust environment.     

Tapesia yallundae collected from wheat stubble in South Africa

Tapesia yallundae var.yallundae is newly recorded from wheat stubble collected near Moorresburg in the western Cape Province of South Africa. Apothecia were observed on wheat stubble incubated for 8 months at 10°C under near-ultraviolet light. Single ascospore isolates produced colonies typical of the Wheat (W)-type in culture. Apothecia were also induced after single-conidial isolates were mated on wheat stubble and incubated at 10–15°C for 6 months. All single-conidial isolates were of the W-pathotype (Ramulispora herpotrichoides var.herpotrichoides).     

Microhabitat segregation and physiological differences in co-occurring tiger beetle species,Cicindela oregona and Cicindela tranquebarica

Summary The daily movements of two co-occurring tiger beetle species were monitored in conjunction with changes in microclimate along streams in Northeast Arizona. Cicindela oregona and C. tranquebarica temporarily segregated across areas of beach exhibiting different microclimates. C. oregona progressively moved from the dry upper beach to the wet stream edge as beach temperatures increased and humidity decreased. The actively foraged throughout the day in this moist habitat at air temperatures between 25 and 38°C. C. tranquebarica remained on the dry, upper portions of the beach and shuttled between sun and shade at air temperatures above 35°C. Only when stream edge temperatures exceeded 30°C was tranquebarica found in this subhabitat. Both species exhibited physiological tolerances in the laboratory that were consistent with their microhabitat preferences in the field. Although both species had similar high lethal temperatures (47–48°C) in saturated air, oregona died at lower temperatures (39–43°C) than tranquebarica (46–47°C) under dry (0% RH) conditions. C. oregona was considerably more active than tranquebarica at body temperatures below 30°C and exhibited higher levels of active metabolism between 25 and 40°C. In addition, C. tranquebarica exhibited significantly lower water loss rates than oregona at 30, 35 and 40°C.     

Teliospore germination and compatibility groups in Sphacelotheca tanglinensis

Teliospores ofS. tanglinensis germinate readily in tap water, each producing a 4-celled promycelium bearing 4 sporidia. Sporidia become 2 or more celled after detaching from the promycelium. Fusion between compatible sporidia occurs in a number of ways resulting in hyphae formation. Teliospores form in 3-month old compatibly mated sporidial cultures. Compatibility inS. tanglinensis is a bipolar type.

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Zusammenfassung Sphacelotheca tanglinensis (Tracy &Earle)Zundel (syn.S. ischaemicola Ling) infiziert die Infloreszenzen vonIschaemum indicum (Houtt.)Merr. undI. timorense Kunth. in Singapore und Malaya. Die Keimung von Teliosporen, die nachfolgenden Wachstumphasen und die Kompatibilitätsgruppen sind bisher nicht untersucht worden, außer einigen Beobachtungen der Keimung der Teliosporen. Diese Phasen sind deshalb untersucht worden.

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Adapted from thesis submitted by the junior author to the University of Singapore for the M. Sc. degree.     

Species specific chlorophyll a fluorescence-temperature profile at high temperatures in the leaves

Chlorophyll a (Chl a) fluorescence-temperature profile in the region of 20–80°C was recorded for fourteen different plant species. In all the species studied, there was a rise in the fluorescence intensity in the region of 45–50°C and around 55°C the fluorescence intensity started to decline. In four of the species (Acacia melanoxylon, Ervatamia montana, Eucalyptus tertecornius and Azardicta indica) tested, there was a secondary rise in the fluorescence intensity around 65–70°C whereas in all other species a sharp decline in the fluorescence intensity was observed at this point. These changes in the fluorescence intensity at high temperatures (65–70°C) appear to be species specific and cannot be explained either in terms of changes in the stoichiometry between the two photosystems or in terms of Chl a fluorescence emission from photosystem I (PS I) at higher temperatures. This conclusion is supported by following observations: (1) there was no definite correlation between the Chl a/Chl b ratio and the pattern of fluorescence-temperature profile at high temperatures; (2) the sun and shade plants of the same species had a similar pattern of fluorescence-temperature profile; and (3) preferential excitation of PS I did not alter the fluorescence-temperature profile.Abbreviations Chl chlorophyll - PS photosystem     

Effects of ambient Lake Mohave temperatures on development,oxygen consumption,and hatching success of the razorback sucker

Synopsis Spawning of razorback suckers,Xyrauchen texanus, in Lake Mohave occurred from 10–22°C and larvae were collected at water temperatures from 10–15°C in 1982 and 1983. In the laboratory, hatching success was similar from 12–20°C, but reduced hatching success was found at 10°C while none hatched a 8°C. Development rate and oxygen consumption were positively related to incubation temperature. Direct effects of ambient Lake Mohave water temperatures on hatching success of razorback sucker embryos are considered minimal. Historical spawning temperatures for the species are hypothesized based upon successful incubation temperatures and comparison to the white sucker,Catostomus commersoni.