A comparison of inoculation techniques for assessment of germplasm susceptibility to Karnal bunt (Tilletia indica) disease of wheat

Three inoculation techniques for Karnal bunt (Tilletia indica) disease of wheat were compared: 1) boot inoculation - injection of inoculum with a hypodermic syringe into the boot; 2) spray inoculation - inoculum sprayed at growth stages between heading and anthesis, and 3) cotton wool inoculation - small pieces of cotton wool saturated in inoculum placed either inside the floret or between the spikelet and rachis. Each inoculation technique was assessed using susceptible cultivars to determine the optimum inoculum concentration, the ideal plant growth stage and the humidity requirements for successful infection.
Boot inoculation did not require high humidity and gave reliable infection with low secondary sporidia concentrations (1000–10 000/ml). The ideal plant growth stages for inoculation were early-boot and mid-boot. Spray inoculation required high secondary sporidia concentrations (50 000/ml) and 48 h of high humidity, but infection was initiated over a range of growth stages throughout heading and anthesis. Cotton wool inoculation gave low levels of infection at growth stages throughout heading and anthesis, even with high secondary sporidia concentrations (100 000/ml).     

Responses of potato cultivars to the common scab pathogens, Streptomyces scabies and S. turgidiscabies

A glasshouse experiment was conducted to study the responses to Streptomyces scabies and S. turgidiscabies in potato cultivars Bellona, Matilda and Sabina (Solanum tuberosum). Potatoes were grown in a peat‐sand mixture inoculated with one of the two strains of either S. scabies or S. turgidiscabies. Logit models were used to analyse the data on disease incidence and severity, whereas the data on emergence and yield were tested by analysis of variance. S. turgidiscabies, a recently described potato pathogen in Finland, possessed a high ability to cause superficial, raised and pitted lesions on all three cultivars tested. Symptoms induced by S. turgidiscabies were similar to those of S. scabies, regardless of the cultivar, which suggests that the two causal organisms of common scab cannot be distinguished based on symptoms. Infection by S. turgidiscabies and S. scabies delayed emergence, had the tendency to decrease the yield, and increased the proportion of small tubers in the yield, regardless of the potato cultivar. Differences in the levels of resistance to common scab were evident between potato cultivars, since cvs. Matilda and Bellona showed higher disease incidence and more severe scab symptoms than cv. Sabina.     

Characterisation of the distribution of individual response times in bacterial infection of plants

The distribution of time of visible response to infection was followed after inoculation of Pseudomonas syringae pv. syringae onto the flowers of pear cv. Williams, Pseudomonas syringae pv. phaseolicola into the leaves of bean cvs Borlotto di Vigevano and Saluggia, Corynebacterium michiganense subsp. michiganense into the stem of tomato cvs Fiorentina and C 1402 and Agrobacterium sp. biovar 3 into the stem of sunflower cv. Egnazia at a dosage of either 0.5–32 times the median effective dose (ED₅₀) or 1–64 ED₅₀. The distribution contracted and the median response time decreased as the dose of inoculum was increased for each host-pathogen combination. Modified Weibull regressions with shape parameter 1< c < 2.5 gave a highly significant fit to the observed distributions of individual response times in all dose groups and host-pathogen combinations.     

Transmission of Alternaria macrospora in Cotton Seeds

Alternaria macrospora was isolated from seeds only after the natural opening of the bolls and exposure of seeds to an environment in which the fungus was present. The fungus lacks the ability to penetrate the boll wall and reach the seed site. Attempts to isolate the pathogen from seeds of immature bolls at different developmental stages failed. Internal infection by slow injection resulted in seed infection and partial shedding of the injected plant parts which was high in buds and decreased with the ripening to mature bolls. Severity of seed infection was not dependent on either inoculum level, boll physiological age or even if the boll itself was not diseased. Infection of flowers under field conditions caused flower shedding. Naturally infected seeds or inoculated seeds with inoculum levels of 100 spores/ml and above resulted in diseased cotyledons, the incidence of which was, for inoculated seeds, positively correlated with inoculum level. A small difference was observed between cultivars in susceptibility to artificial inoculation at the cotyledon stage. A. macrospora survived on commercial cotton seeds and on post-season plants left growing at the field edges. Survival in plant debris under field conditions was minimal and may only have a minor effect on field reinfestation.     

Experimental Histomoniasis in the Guinea Fowl,Numida meleagris*

Guinea fowl were inoculated rectally with Histomonas meleagridis to produce histomoniasis. The birds were infected readily by this unnatural route. Severe cecal involvement was frequent and long-lasting but liver lesions and death were rare. Turkeys given the same inoculum had high levels of liver involvement and mortality. Guinea fowl responded much less severely to infection with H. meleagridis when infected by a natural route (ingestion of a vector) rather than by rectal inoculation. Thus, naturally acquired infections with H. meleagridis appear to be of small consequence in the guinea fowl.     

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.     

Scab disease of cowpea (Vigna unguiculata) caused by Sphaceloma a species of the fungus

Scab disease of cowpea (Vigna unguiculuta) was shown to be caused by Sphaceloma sp. It affects all above ground parts of the plant. The first symptom of the disease, appearing within 3 to 6 days of inoculation, is puckering of the lamina. Spots on mature leaves are white with or without brown margins. Typical scab lesions on petiole, stem, peduncle and pod are white turning dark brown when chlamydospores are formed and are oblong-elongate except for pod lesions that are ovoid. The most destructive phase is scab of the flowering axis which causes flower and, or, pod abortion or completely prevents flower formation. Inoculation of asparagus pea (V. sesquipedalis) with a cowpea isolate of Sphaceloma sp. produced symptoms similar to those on cowpea. Inoculated hyacinth bean (Lablab niger) produced atypical mild lesions. The following legumes were not affected when artificially inoculated with the fungus: black gram (Phaseolus mungo), green gram (P, aureus), French bean (P. vulgaris), Lima bean (P. lunatus), groundnut (Arachis hypogaea), and soyabean (Glycine max). The characteristics of the fungus on potato dextrose agar are described.     

The effects of post-inoculation wet and dry periods, and inoculum concentration, on lesion numbers of Septoria nodorum in spring wheat seedlings

Lesion numbers in seedlings of spring wheat cvs Kolibri and Maris Butler, inoculated with Septoria nodorum in a growth cabinet, varied with the length of wet or dry period following inoculation. Increasing the length of dry period led to a curvilinear decrease in lesion numbers at 15° C (60% r.h.) and 20° C (40% r.h.), but some infection still occurred with a dry period of 72 h. Increasing the length of wet period gave a linear increase in lesion numbers in cv. Maris Butler, but lesion numbers levelled off at 12 h at 15° C (60% r.h.) in cv. Kolibri. Increasing the inoculum concentration gave an increase in lesion numbers in both cultivars irrespective of whether wet or dry periods followed inoculation. On occasion, Kolibri and Maris Butler differed significantly in lesion numbers, but differences were not consistent between experiments, and there were interactions with the length of wet period.     

Occurrence of Didymella fabae,the Teleomorph of Ascochyta fabae,on Faba Bean Straw in Spain

Pseudothecia of Didymella fabae, the teleomorph of Ascochyta fabae, were first observed on faba bean (Vicia faba) debris in Spain during autumn 1995. Most pseudothecia were mature by December–February. The ascospores gave rise to typical cultures of A. fabae, and conidia from these cultures caused ascochyta blight symptoms on inoculated faba bean plants. Placing straw‐bearing pseudothecia over the plants to allow ascospore discharge also resulted in typical ascochyta blight symptoms. Pseudothecia maturity and discharge of ascospores from the infested faba bean straw overlapped with the vegetative stage of the faba bean crop, which occurs in southern Spain during winter as the crop is sown in autumn and harvested in spring. These observations indicate that ascospores may serve as primary inoculum for the disease.     

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.     

Modified techniques for the isolation of Dermatophilus spp. from infected material.

Two improved techniques are described for the isolation of Dermatophilus spp. The first is suitable for samples of scab from lesions of the disease in animals, except for sheep affected with lumpy wool. The second is for the isolation of the organism from samples of lumpy wool in sheep. Both techniques are simple, rapid and reliable and are based on releasing the organism by pulverizing the scab and then by suppressing contaminants by including polymixin B sulphate in the medium. Excellent results are obtained with or without pre-incubation exposure to CO2.     

Healthier seed potatoes. I. Effects of inoculating stem cutting stocks with Polyscytalum pustulans and Rhizoctonia solani on growth, yield and disease

Potato seed tubers of seven cultivars derived from stem cuttings in 1965 (healthier seed) were grown in 1969–72 at two sites, one clay with flints soil (Rothamsted) and the other sandy loam soil (Woburn). Inoculating sprouted tubers at planting with Polyscytalum pustulans did not affect the number of stems/plant or total yield but increased stem base and tuber infection. The yields of large tubers (57–83 cm) were increased and small tubers (>57 cm) decreased, indicating a decrease in tuber numbers. Rhizoctonia solani inoculated at planting decreased numbers of stems/plant and yield by up to 14% at Woburn but not at Rothamsted. At both sites, yields of large tubers were increased with cvs Majestic and Record and decreased with Pentland Crown. Stem canker and tuber infection were increased but infection was also prevalent on tubers from non-inoculated seed at Woburn. When both pathogens were inoculated together yields of large tubers were increased in cvs King Edward, Majestic and Record and decreased in Pentland Crown. Infection of stem bases and tubers was sometimes less than when either pathogen was inoculated singly.     

BACTERIAL BOLL ROT OF COTTON (XANTHOMONAS MALVACEARUM (E. F. SMITH) DOWSON)

Two inoculation techniques were compared for their value in assessing resistance of cotton to bacterial boll rot. In the first method, bacterial suspensions were applied with a fine brush to the outsides of young bolls immediately after corolla drop. In the second, suspensions were inoculated into the wall tissue of bolls 3–4 weeks old with a fine needle. The second method is the simpler and produces infections in all weather conditions; other advantages are that bolls are inoculated after the period of natural shedding and the circular lesions are easily measured, but a disadvantage compared with the brush method is that the puncture method does not satisfactorily indicate damage to the lint.
Results from the two methods of inoculation suggest that resistance lies in some component of the boll-wall tissue that affects bacterial growth and not in a physical factor that prevents the bacteria entering the boll. Variability in resistance occurs in local strains of cotton, and this could probably be exploited to give high resistance without the need of a large crossing programme to introduce new genes.     

Control of pearl millet downy mildew by seed treatment with metalaxyl*

Three formulations of the systemic fungicide metalaxyl were tested in various seed treatments for the control of pearl millet downy mildew in three field experiments with downy mildew-susceptible pearl millet hybrid NHB-3. Uniform, high levels of sporangial inoculum of the causal fungus, Sclerospora graminicola, were provided throughout the growth of the test crops from inoculated infector rows of NHB-3, planted earlier between the test plots. Significant reductions in downy mildew were obtained with all fungicide treatments. Best control was obtained when seed was soaked in a 0.5% aqueous solution of a liquid formulation (mean infection index of 9.8% compared with 94.8% in the untreated check). The degree of control with the wettable powder formulations was directly related to fungicide dosage, and there were no significant effects of application method. Simple dusting of seed at 2 g a.i./kg, a rapid and simple operation requiring small quantities of fungicide and no special application equipment, gave a high level of control (infection index of 12.6% compared with 78.9% in the untreated check). In two experiments grain yields from all the treated plots were significantly greater than from the untreated plots (means of 1234 and 1534 kg/ha for treated plots compared with 485 and 743 kg/ha, respectively), and in the third, the treatment with the least downy mildew gave significantly more grain than the untreated check (1228 compared with 727 kg/ha).     

Control of coffee berry disease*

Field experiments comparing six spraying programmes and two fungicides for control of coffee berry disease were carried out at three sites during 1968. These established that protection of the crop by sprays applied throughout the long rains (February–July) gave the best control and increased yields threefold. Early season spray programmes, which reduced the sporulating capacity of the pathogen in coffee twigs, but which ceased before the end of the rainy season, gave no disease control in the late crop (harvested October-December) and yields were less than in unsprayed controls. All programmes which gave protection at the onset of the rains gave some disease control in the early crop (harvested June-August). Captafol (‘Ortho-Difolatan’ 80 WP) was better than a 50% copper formulation (‘Perenox’). Conditions suitable for infection, as determined from meteorological records, occurred frequently during both rainy seasons. Estimates of sporulating capacity and data from spore trappings showed that although inoculum from the bark might initiate an epidemic, subsequent disease progress was more dependent upon spores derived from diseased berries; these accounted for most of the inoculum dispersed during the greater part of the season. Removal of the early crop, which was very small, had no detectable effect on disease in the late crop. Although losses were greatest with early season spray programmes, these did nevertheless delay the onset of the epidemic. However, the subsequent rate of disease increase was greatest and ‘scab’ lesion formation least in these treatments. ‘Scab’ lesions were most abundant in those treatments which caused least delay; in unsprayed coffee, it appeared that early exposure to infection resulted in ‘scab’ lesion formation which subsequently retarded the rate of disease progress. Tree height was positively correlated with disease incidence, but there was little effect of tree aspect. Crop density had a small effect on disease incidence within branches.     

Inoculum Density and Population Dynamics of Suppressive and PathogenicStreptomycesStrains and Their Relationship to Biological Control of Potato Scab

SeveralStreptomycesstrains are capable of suppressing potato scab caused byStreptomyces scabies.Although these strains have been successful in the biocontrol of potato scab in the field, little is known about how populations of pathogenicStreptomycesin the potato rhizosphere are influenced by inoculation of the suppressive strains. The effects of inoculum densities of pathogenic and suppressiveStreptomycesstrains on their respective populations on roots and in rhizosphere soil were examined during the growing season. The relationships between inoculum density or rhizosphere population densities and disease severity were also investigated. Populations of suppressiveStreptomycesstrain 93 increased significantly on roots with increasing inoculum dose. At its highest inoculum dose, the suppressive strain reached a population density greater than 10⁶CFU/g root 14 weeks after planting. The ability of the suppressive strain to increase its populations with increasing inoculum density was hindered at high inoculum doses of the pathogen, suggesting that density-dependent competitive interactions may be occurring between the two antagonists. Strain 93 was most effective at preventing scab early in the growing season (8 weeks after planting), when tubers were most susceptible to the scab disease. Population densities of the suppressive strain in soil were more highly negatively correlated with scab severity than were populations on roots, suggesting that rhizosphere soil rather than potato roots may be the primary source of inoculum of the suppressive strain for tubers.     

NaCl胁迫下AM真菌对棉花生长和叶片保护酶系统的影响

利用盆栽实验研究了 Na Cl胁迫条件下 AM真菌对棉花生长和叶片保护酶系统的影响。结果表明 :在土壤中加入 0、0 .1%、0 .2 %、0 .3%浓度 Na Cl条件下 ,Na Cl胁迫对 AM真菌的接种效果有显著影响。接种 AM真菌提高了棉花根系菌根侵染率 ,增加了棉株的生物产量 ,以 0～ 0 .2 % Na Cl浓度时 AM真菌接种效果最好。 AM真菌对棉株生理参数和保护酶活性的影响因生育期和 Na Cl浓度不同而异 ,现蕾期和低盐浓度 (0～ 0 .1% )下叶片叶绿素含量明显增加 ;中高盐水平 (0 .2 %～ 0 .3% )和生育后期叶片可溶性蛋白质含量和 SOD、POD、CAT等保护酶活性显著提高 ,MDA含量明显降低 ;棉株 K、Ca、Mg含量因植株部位和盐浓度不同而变化。 AM真菌增强宿主植物的耐盐性可能源于促进宿主根系对土壤矿质元素吸收的直接作用和改善植物体内离子平衡和生理代谢活动、提高保护酶活性的间接作用     

Formation and Development of Pseudothecia of Venturia nashicola

Conidia are believed to be the main source of primary inoculum for pear scab, caused by Venturia nashicola, in northern China. Experiments were conducted to investigate the development and potential role of V. nashicola ascospores in northern China. Leaves with pear scab lesions were collected from commercial orchards in November 2003 and 2004 to monitor pseudothecia formation under various environments. Pseudothecium production was shown to occur readily in northern China. The key requirement for pseudothecium production is the occurrence of rain during the winter and early spring, although the exact timing of these rain events appeared not to affect their development. Excess water may lead to the accelerated leaf decay and hence lead to production of fewer pseudothecia. More than 80% scabbed leaves, placed in a pear orchard, produced pseudothecia. Leaves with only non‐sporulating scab lesions in autumn were also able to produce a large number of pseudothecia. Both airborne ascospores and conidia of V. nashicola were caught in a pear orchard. Most ascospores were released by late‐May, a month after pear blossom. These results suggest that ascospores may play an important role in the early stage of pear scab epidemics in spring in northern China.     

Effect of leaf age and inoculum concentration on the symptoms produced by Gymnosporangiumjuniperi-virginianae on apple

Gymnosporangium juniperi-virginianae, the incitant of cedar apple rust, induced the greatest number of lesions (flecks, pycnial lesions, aecial lesions) on 6- and 8-day-old leaves of single-shoot potted apple plants inoculated and incubated under controlled conditions. The largest pycnial lesions developed on 2- and 4-day-old leaves and the largest aecial lesions on 6-day-old leaves. The number of lesions was positively correlated with the concentration of basidiospores in the inoculum, but lesion size was constant at different inoculum levels. Elevated inoculum concentrations failed to induce pycnia on cv. McIntosh which normally bears only flecks; very low concentrations induced the development of pycnia but not aecia on cv. Rome Beauty, which normally bears aecia. An ‘infection rating’, log₁₀ (10 nd²), where n is the highest number of pycnial lesions per leaf and d is the largest mean diameter of pycnial lesions on any leaf, calculated for four cultivars in greenhouse tests showed a positive correlation with a ‘susceptibility rating’ developed from observations of natural infection of these cultivars in the field.