A comparison of methods for inoculating bean plants with Elsinoë phaseoli and some factors affecting infection

Bean (Phaseolus vulgaris) leaves were inoculated with a conidial suspension of Elsinoe phaseoli by spraying, by applying a 10 μl drop, by a cotton wool technique or by using a camel hair brush. Spray application resulted in a uniform distribution of lesions over the leaf surface and gave consistent levels of infection between repetitions. Drop application allowed good control of the amount of inoculum applied. Although brush or cotton wool techniques gave high levels of infection it was not possible to control the amount of inoculum. Only immature tissues were susceptible to infection. Five cultivars were inoculated with two isolates and disease was scored on the basis of numbers of lesions and lesion type (scab severity). The small white cvs PAN 181 and Teebus were the most resistant; lesions were small and generally fewer than on the large-seeded red speckled sugar bean cvs Umlazi and Umvoti. The value of the different inoculation techniques in studies on bean scab is discussed.     

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.     

STEM INFECTION OF COTTON BY XANTHOMONAS MALVACEARUM (E. F. SM.) DOWSON

Varying the position of stem inoculation, the concentration of inoculum and the age of plant affected the reaction of cotton, Gossypium sp., to infection with Xanthomonas malvacearum (E. F. Sm.) Dowson.
The extent of stem discoloration, internal and external, and the probability of disease ocurring in leaves by bacteria moving within the plant increased ( a ) the nearer the point of stem inoculation was to the apex, and ( b ) the higher the concentration of inoculum. The leaf symptoms were not the angular spots typical of primary leaf infection. Instead, bacteria seemed to lodge in, discolour and blacken sections of leaf veins. Then tissue next to the affected veins became water-soaked and leaf sectors dependent upon these veins died and dried. These symptoms usually developed 14 to 55 days after inoculation in the expanding leaves.
The amounts of stem discoloration and the probabilities of leaf symptoms developing were less when hypocotyls of old plants were inoculated than when hypocotyls of young plants were inoculated. The probabilities of leaf symptoms developing were similar, however, when young tissues in young and old plants were inoculated.
American cotton, Gossypium hirsutum , was less affected by stem inoculation than Egyptian cotton, G. barbadense. Of the resistance factors against primary leaf infection only B_6m gave appreciable stem resistance.     

Foliar application of selenium increases fertility and grain yield in bread wheat under contrasting water availability regimes

The study was conducted to assess whether selenium (Se) application modulates fertility to alter grain yield in bread wheat grown under different moisture regimes. Seeds of wheat cultivar Millat-2011 were sown in the plots using a randomized complete block design with three replicates per treatment. After germination, the plants were exposed to six moisture regimes, viz. no irrigation after germination, irrigation at boot stage, irrigations at boot and grain-filling stages, irrigations at crown root, boot and grain-filling stages, irrigations at crown root, boot, heading and grain-filling stages and irrigations at crown root, stem elongation, boot, heading and grain-filling stages. At the heading stage, foliar spray of sodium selenate (0, 2 and 4 mg Se L^?1) was done. Withholding water at early growth stages significantly increased oxidative stress and decreased growth and grain yield. Irrespective of moisture regimes, foliar application of Se (2 mg L^?1) decreased oxidative stress, modulated photosynthetic pigments and fertility and increased grain yield in wheat. The Se-mediated increase in grain yield was attributed to the increase in chlorophyll and ascorbic acid contents and fertility coupled with decrease of oxidative stress under different moisture regimes. The results could be helpful to manage wheat production in the semi-arid environments.     

Effect of inoculum density, stage of plant growth and dew period on the incidence of black point caused by Alternaria alternata in durum wheat

Glasshouse studies showed that the incidence of black point caused by Alternaria alternata in durum wheat was positively correlated with both the density of the inoculum and the growth stage of the wheat plants at the time of inoculation. A curvilinear relationship of the form Y=a + log X was found between inoculum density and disease incidence. The incidence of black point was linearly related to the stage of plant growth, between anthesis and the late milk stages of development, at the time of inoculation. A better relationship between growth stage and disease incidence was found when plant growth was expressed as days after anthesis than when the Romig scale was used (R²= 0.30 and 0.24 respectively). A threshold dew period of 3–6 h was required for black point symptoms to develop. The incidence of black point increased with increasing duration of the dew period until maximum disease incidence occurred after exposure to a dew period of 48 h.     

Optimum conditions for initiation of filamentation in Candida albicans.

A medium that initially produced filaments from almost all of its Candida albicans blastospore inoculum contained 1% mycological peptone and 0.2% glucose, final pH 7.4-7.5. The medium was inoculated to 10-6 cells/ml and incubated at 40 degrees C. Reversion to secondary blastospores began at a mean of 2.4 h after inoculation. The patterns of utilization of growth nutrients during optimal mycelial growth showed no correlation with the events of filamentation.     

Factors Affecting the Persistence of Staphylococcus aureus on Fabrics

The persistence of Staphylococcus aureus (Smith) on wool blanket, wool gabardine, cotton sheeting, cotton knit jersey, cotton terry cloth, and cotton wash-and-wear fabrics was studied. The fabrics were exposed to bacterial populations by three methods: direct contact, aerosol, and a lyophilized mixture of bacteria and dust having a high content of textile fibers. The contaminated fabrics were held in 35 or 78% relative humidities at 25 C. In general, the persistence time of S. aureus populations on fabrics held in 35% relative humidity was substantially longer when the fabrics were contaminated by exposure to aerosolized cultures or to dust containing bacteria than when contaminated by direct contact. In a 78% relative humidity, bacterial populations on the fabrics persisted for substantially shorter periods of time regardless of the mode of contamination or fabric type. Cotton wash-and-wear fabric (treated with a modified triazone resin) was the material on which populations of S. aureus persisted for the shortest time. This organism retained its virulence for Swiss mice after being recovered from wool gabardine swatches held 4 weeks in 35% relative humidity and 6 weeks in 78% relative humidity.     

Cell size, macromolecular composition, and O2 consumption during agitated cultivation of Naegleria gruberi.

Cell size, macromolecular composition, carbohydrate utilization patterns, and O2 concentrations were measured throughout the growth stages of Naegleria gruberi in agitated culture in a complex medium. Biphasic logarithmic growth occurred during the intial 83 hr of growth and the mean generation time was 7.0 hr and 19 hr during initial and secondary log growth stages, respectively. The maximum yield was 5 X 10(6) amebae/ml. The pH rose rapidly (1 pH unit) during the secondary log growth phase (52-83 hr) and continued into the stationary growth phase (83-120 hr). Dry weight, total protein, carbohydrate, and RNA per ameba increased just before the secondary log growth phase. RNA increase 31% to 35% per ameba at the end of each phase of log growth. DNA increased approximately 2-fold throughout the different growth phases. Average cell size increased 90% during biphasic log growth then decreased during stationary phase. O2 tension decreased from 100% to 18% of saturation during the biphasic growth phase, then increased during stationary growth to near 100% saturation. Glucose and total carbohydrate assays showed little utilization of those substrates throughout the growth stages. Naegleria gruberi presumably has a predominantly aerobic metabolism, also its metabolism may change during the different growth phases.     

BACTERIAL CANKER OF STONE-FRUITS III. INOCULUM CONCENTRATION AND TIME OF INOCULATION IN RELATION TO LEAF-SCAR INFECTION OF CHERRY

Leaf scars on the fruiting spurs of the cherry varieties Roundel (high resistance) and Napoleon (low resistance) were inoculated with Pseudomonas mors-prunorum on four separate occasions in the autumn, using, on each occasion, the same range of five different inoculum concentrations. The results, recorded the following year, showed that the percentage diseased spurs (disease incidence) and the severity of the disease symptoms (disease severity) both increased with inoculum concentration.
The median threshold concentration of inoculum (T. C. 50), defined as the concentration necessary to give 50% diseased spurs, varied with time of inoculation, but on all occasions was considerably higher for Roundel than for Napoleon.
In another experiment the leaf scars at the nodes of the current year's growth, inoculated at weekly intervals throughout the autumn, were found to be susceptible from the beginning of September to the latter part of October. During this period disease incidence varied considerably with time of inoculation. There was evidence that this variation was related to two factors which influenced the numbers of bacteria penetrating into leaf scars, namely, (1) the rate of evaporation of the infection drop, and (2) the rate of suction of inoculum into the vessels of the leaf traces.
The experiments provided evidence of a long infection period beginning early in the autumn. It is suggested, therefore, that the timing of bactericidal sprays in the autumn be advanced and that the present concept of 'protective' sprays in disease control be replaced by one based on the eradication of external sources of inoculum.     

Bacterial canker of poplar

Bacterial canker is a serious disease of poplar and it is important for commercial planting to select those clones of poplar which show resistance to the disease. In inoculation tests, natural bacterial slime and a pure bacterial suspension gave similar extent of invasion. A study of the life-cycle of the disease indicated the presence of inoculum and of potential infection courts throughout the growing season. It is suggested that the susceptibility of a poplar clone should be determined by assessment of both artificial inoculations and natural infection.     

The life cycle of the smut fungus Moesziomyces penicillariae is adapted to the short-cycle of the host, Pennisetum glaucum

Moesziomyces penicillariae (Brefield) Vànky is a basidiomycete fungus responsible for smut disease on pearl millet, an important staple food in the sub-Sahelian zone. We revisited the life cycle of this fungus. Unlike other Ustilaginales, mating of sporidia was never observed and monoclonal cultures of monokaryotic sporidia were infectious in the absence of mating with compatible partner. These data argued for an atypical monokaryotic diploid cell cycle of M. penicillariae, where teliospores only form solopathogenic sporidia. After inoculation of monoclonal solopathogenic strains on spikelets, the fungus infects the ovaries and induces the folding of the micropilar lips, as observed during early pollination steps. The infected embryo then becomes disorganized and the fungus invades peripheral ovary tissues before sporulating. We evaluated the systemic growth abilities of the fungus. After root inoculation, mycelium was observed around and inside the roots. As argued by transmission electron microscopy (TEM) observations and polymerase chain reaction (PCR) detection using specific primers for M. penicillariae, the fungus can grow from roots to the caulinar meristems. In spite of this systemic growth, no sori were formed on the varieties of pearl millet tested after root inoculation. All together, these data suggest that the reduced life cycle of M. penicillariae – i.e. dispersal of ‘ready to infect’ solopathogenic sporidia, floral infection – is an adaptation to the aetiology of this disease to short-cycle pearl millet varieties from the sub-Sahel.     

Infection of potato stems by the gangrene pathogen, Phoma exigua var. foveata

Limited transport of pycnidiospores of Phoma exigua var. foveata within transpiring stems was demonstrated by isolating from cut mature stems placed for 24 h in a spore suspension. Pieces of shoot dipped in spore suspension and subsequently rooted contained the pathogen which was redistributed as the plants grew but without causing symptoms until after desiccation. The fungus was only rarely detected within stems growing from rotted seed tubers suggesting that spore transport in the vascular system was relatively unimportant in these experiments. Stems were readily infected from spore suspensions applied to unwounded epidermis especially when the inoculum was held in cotton wool and secured with wax film. The progressive colonisation of growing stems was demonstrated by isolations made at different positions on the stem and at different times after inoculation. In field experiments in two years stems were inoculated with four spore concentrations on four dates from late June to late August. After desiccation the extent of pycnidial lesions was greatest from the earliest inoculation date and the largest spore concentration and these differences were reflected subsequently in the incidence of gangrene on damaged progeny tubers. Younger stems may be more susceptible and infection in early summer probably results in extensive growth within the stem before desiccation and thus in massive pycnidial production.     

Influence of larval stage and virus inoculum on virus yield in insect host Neodiprion abietis (Hymenoptera: Diprionidae)

Virus yield produced by dead larvae of balsam fir sawfly, Neodiprion abietis (Harris) (Hymenoptera: Diprionidae), that had been infected at four different larval stages (second, third, fourth, or fifth instar) with two virus concentrations (10(5) polyhedral inclusion bodies (PIB) /ml or 10(7) PIB/ml), were analyzed and compared to determine the effects of instar and amount of virus inoculum on virus production. The results indicate that both larval stage and inoculation dosage significantly affect virus yield. On average, each dead larva produced 1.36-12.21 x 10(7) PIB, depending upon larval age and virus concentration of inoculation. Although each dead larva produced more PIB when it was inoculated in the fourth or fifth stage, inoculation of these larvae did not result in the highest virus yield because of low larval mortality. In terms of net virus return, third instars would maximize virus yield when they are inoculated with a virus concentration that can cause 95-100% larval mortality.     

Quantitative Studies on Fabrics as Disseminators of Viruses: II. Persistence of Poliomyelitis Virus on Cotton and Wool Fabrics

The length of time that poliovirus could be recovered from wool gabardine and blanket, and from cotton sheeting, terry cloth, and knit jersey fabrics was determined under conditions of controlled temperature and humidity (25 C in 35 and 78% relative humidities). Three types of exposure of the fabrics to viruses were used: direct contact, aerosol, and virus-containing household dust having a high content of textile fibers. When held in 35% relative humidity, virus persisted for 20 weeks on wool fabrics, but only 1 to 4 weeks on cotton fabrics. At this relative humidity, virus titers on wool fabrics decreased rapidly to low but detectable levels which persisted for long periods of time, whereas in 78% relative humidity the decrease in virus titer was less rapid, but the period of viral persistence was shorter. Generally, virus titers on cotton fabrics held in both relative humidities decreased exponentially to an undetectable level. The method of exposure to virus had a definite effect on the duration of viral persistence on a given fabric. Virus contained in household dust was least stable.     

Enhancement of Infectivity of Murine Cytomegalovirus in Vitro by Centrifugal Inoculation

Centrifugation of murine cytomegalovirus inocula from a variety of sources onto secondary mouse embryo cell monolayers at 1,900 x g for 30 min regularly revealed 10- to 100-fold more infectious virus than could be found in the same materials using standard inoculation methods. Virus demonstrable only by centrifugation was present throughout the entire growth cycle in a constant proportion to virus measured without centrifugation. Extracellular growth curves of both populations revealed an 18- to 21-hr latent period, followed by a long-linear increase over the next 12 hr; final yield was 30 plaque-forming units (PFU) per cell. Centrifugation of cells prior to inoculation or after standard adsorption and removal of inoculum failed to result in any significant change in measured virus titer. However, even after 4-hr adsorption, the supernatant inoculum could be transferred and centrifuged onto a fresh monolayer resulting in the same increment of measurable virus. Neutralizing antibody and interferon were equally efficacious against 100 PFU of virus as defined by either method. Thus, this newly identified population of cytomegalo-virus represents the vast majority of potentially infectious units and appears to differ solely in ease of adsorption onto cell monolayers.     

Relationship of inoculum ofVerticillium dahliae to disease in tomato plants

Summary Tomato (Lycopersicon esculentum Mill.) seedlings were inoculated by dipping roots in suspensions of conidia ofVerticillium dahliae Kleb. There was a linear relationship between inoculum concentration and infection at 21 days after inoculation in steam-disinfested soil. Thereafter the number of infections per plant increased. At higher concentrations data indicated a synergistic interaction between conidia. Synergism was more pronounced and was detected earlier in soil not disinfested before inoculation. When conidia from 3-, 7-, and 14-day-old cultures were used, the greatest response was from 3-day-old cultures. A lower total response and lower rate of response to older inoculum indicated a decrease in aggressiveness of conidia with age. Older plants were less affected by the pathogen when plants inoculated at the fourth and sixth leaf stages with minimum root disturbances were compared. There was also an increase in infection with an increase in the volume of root zone infested. In field experiments infection reduced stand when roots were disturbed at inoculation. With minimum root disturbance yields were reduced without a reduction in stand. Part of thesis to be submitted by the senior author to the University of Stellenbosch in partial fulfilment of the Ph.D. (Agric.) degree.     

Histological and cytological characterization of adult plant resistance to wheat stripe rust

Wheat cultivar Xingzi 9104 (XZ) possesses adult plant resistance (APR) to stripe rust caused by Puccinia striiformis f. sp. tritici (Pst). In this study, histological and cytological experiments were conducted to elucidate the mechanisms of APR in XZ. The results of leaf inoculation experiments indicated that APR was initiated at the tillering stage, gradually increased as the plant aged and highly expressed after boot stage. The histology and oxidative burst in infected leaves of plants at seedling, tillering and boot stages were examined using light microscopic and histochemical methods. Subcellular changes in the host–pathogen interactions during the seedling and boot stages were analyzed by transmission electron microscopy. The results showed that haustorium formation was retarded in the adult plants and that the differentiation of secondary intercellular hyphae was significantly inhibited, which decreased the development of microcolonies in the adult plants, especially in plants of boot stage. The expression of APR to stipe rust during wheat development was clearly associated with extensive hypersensitive cell death of host cells and localized production of reactive oxygen species, which coincided with the restriction of fungal growth in infection sites in adult plants. At the same time, cell wall-related resistance in adult plants prevented ingression of haustorial mother cells into plant cells. Haustorium encasement was coincident with malformation or death of haustoria. The results provide useful information for further determination of mechanisms of wheat APR to stripe rust. Key message The expression of APR to stipe rust in wheat cultivar Xingzi 9104 (XZ) was clearly associated with extensive hypersensitive cell death of host cells and the localized production of reactive oxygen species.     

Internal water balance of barley under soil moisture stress

Leaf water potential, leaf relative water content, and relative transpiration of barley were determined daily under greenhouse conditions at 3 growth stages: tillering to boot, boot to heading, and heading to maturity. The leaf moisture characteristic curve (relative water content versus leaf water potential) was the same for leaves of the same age growing in the same environment for the first 2 stages of growth, but shifted at the heading to maturity stage to higher leaf relative water content for a given leaf water potential. Growth chamber experiments showed that the leaf moisture characteristic curve was not the same for plants growing in different environments.

Relative transpiration data indicated that barley stomates closed at a water potential of about −22 bars at the 3 stages studied.

The water potential was measured for all the leaves on barley to determine the variation of water potential with leaf position. Leaf water potential increased basipetally with plant leaf position. In soil with a moisture content near field capacity a difference of about 16.5 bars was observed between the top and bottom leaves on the same plant, while in soil with a moisture content near the permanent wilting point the difference was only 5.6 bars between the same leaf positions.

     

Effects on plant growth of inoculation of stored stripmining topsoil in North Dakota with mycorrhizal fungi contained in native soils

Stored topsoil from stripmining operations in western North Dakota was inoculated with mycorrhizal fungi contained in native prairie soil. The effects on plant mycorrhizal infection percentage, growth as shoot dry weight, and phosphorus uptake were determined. The studied topsoil piles were found to contain little or no vesicular arbuscular mycorrhizal (VAM) fungal inoculum at a depth of 120 cm. The inoculum soil was mixed into the stored soil at rates of 10% and 1%, or surface-applied at 1%. In control pots, sterilized inoculum soil was used. Corn plant (Zea mays) bioassays were used. After 30 days growth the percent VAM fungal infection of the test plants increased with both the 10% and 1% soil inocula. Phosphorus concentrations were generally increased by inoculation with 10% soil mixtures but not 1%. Shoot dry weights of the plants were not measurably different between 10% and 1% inoculation. However, when the plant growth period was increased to 60 days, all three parameters were increased over the check plants. When the inoculum was not mixed into the soil, but layered on the surface, there were no differences in any of the parameters.     

Crop Rotation and Races of Meloidogyne incognita in Cotton Root-knot Management

The influence o f various crop rotations and nematode inoculum levels on subsequent population densities of Meloidogyne incognita races 1 and 3 were studied in microplots. Ten different 3-year sequences o f cotton, corn, peanut, or soybean, all with cotton as the 3rd-year crop, were grown in microplots infested with each race. Cotton monoculture, two seasons o f corn, or cotton followed by corn resulted in high race 3 population densities and severe root galling on cotton the 3rd year. Peanut for 2 years preceding cotton most effectively decreased the race 3 population and root galls on cotton the 3rd year. Race 1 did not significantly influence cotton growth or yield at initial populations of up to 5,000 eggs/500 cm³ soil. At 5,000 eggs/500 cm³, cotton growth was suppressed by race 3 but yield was not affected.