High Inoculum Density Research Articles

Evaluation of Onion Cultivars for Resistance to Aspergillus niger, the Causal Agent of Black Mold

Black mold (BM), caused by Aspergillus niger Tiegh., is a common onion (Allium cepa L.) postharvest disease under hot and humid storage conditions. This study aimed to develop a BM screening protocol, to evaluate onion cultivars for resistance. The effects of pathogen isolate, inoculum density, incubation temperature, and inoculation method on severity of BM were investigated in designing the screening protocol. Variation in virulence was observed and two highly virulent subcultures, An4-2 and An39-3, were selected for study. Disease severity, both in lesion size and sporulation degree, increased with higher inoculum densities and incubation temperatures. In contrast, small lesions and no sporulation were observed when onion scales were inoculated with 1 × 104 spores/mL or incubated at temperatures lower than 20 °C. Inoculation of onion scales or entire bulbs were compared and scale inoculation correlated better with BM incidence in storage. The procedure for BM screening protocol included: wound inoculation of onion slices with 40 μL inoculum suspension of 1 × 105 spores/mL, incubation at 25 °C with 12 hour photoperiod, and rating lesion diameter and sporulation 4 days after inoculation. Using this screening protocol, 42 onion cultivars were evaluated and cultivars with better storability such as `Red Pinoy', `Serrana', `Dehydrator No.3', and `Moonlight' were tolerant to A. niger. These cultivars showed small lesions and no sporulation during screening. On the other hand, cultivars with poor storability like `Texas Early Grano 502', `Granex 429', and `Explorer' exhibited larger lesions with profuse sporulation. Lesion diameter caused by the scale inoculation correlated significantly with bulb storage loss (r = 0.51, P < 0.01) as well as with bulb dry matter (r = -0.48, P < 0.01) and total soluble solids (r = - 0.51, P < 0.01). The screening protocol and tolerant materials are now successfully used in the Asian Vegetable Research and Development Center onion breeding program for resistance to black mold.

The influence of the symbiotic plasmid pRL1JI on the distribution of GM rhizobia in soil and crop rhizospheres, and implications for gene flow.

The distribution of two genetically modified Rhizobium leguminosarum strains was investigated in the field. One, RSM2004, released in 1987, carries a TnS marker on its conjugative symbiotic plasmid (pSym). The second, CT0370, released at the same site in 1994, has a gusA gene integrated into its chromosome but no pSym. Plate counts indicated that the CT0370 population became established at a higher level than RSM2004. However, when peas, alfalfa and barley were grown, RSM2004 was found to outnumber CT0370 on all roots and by 100-fold on pea. Although the transfer of pSym from RSM2004 to CT0370 could be detected on plates and in microcosm studies with high inoculum densities, no transfer was detected in the field. Subsequent transfer of pSym from RSM2004 to CT0370 demonstrated that it conferred an advantage in the rhizosphere. In addition to increasing host fitness, plasmids may transfer, or mobilise other genetic elements, to other bacteria. This is more likely in sites such as the rhizosphere, where cells are active and numbers are high. The distribution of pSym and other genetic elements associated with rhizobia, in bacterial sub-populations from the soil and roots of the different plants, was investigated using PCR. The genetic elements studied were: ISRm3, an insertion element from Sinorhizobium meliloti; pSB 102, a broad host range mer plasmid; the Rhizobium nodC gene (carried on pSym) and plasmid replication origins repCI and repCII. As expected, ISRm3 was detected in rhizoflora cultured from alfalfa but not the other plants. The mer gene was ubiquitous but the transfer region of pSB 102 was not detected. The nodC and both repC primers amplified products from all the plants, giving further evidence for the occurrence of plasmids originating from Rhizobium in the rhizoflora of non-host plants. Despite the abundance of elements associated with transferable plasmids in rhizobia, none was detected in either inoculant strain.

γ-Asarone: the fungitoxic principle of the essential oil of Caesulia axillaris

The essential oil of Caesulia axillaris has exhibited its fungitoxicity against Aspergillus flavus at its minimum inhibitory concentration of 1300 mg/l. It showed the potentiality of an ideal fungitoxicant because of its long shelf life, thermostable nature, broad fungitoxic spectrum and persistence of fungitoxicity even on introduction of high inoculum density of the test fungus. The fungitoxic principle of the oil was standardized as γ-asarone which showed fungitoxicity against the test fungus at 500 mg/l.

Effect of a Single or Double Soil Solarization to Control Verticillium Wilt in Established Olive Orchards in Spain.

Four soil solarization experiments were completed in three commercial olive orchards infested with Verticillium dahliae in southern Spain. Three of the experiments used lines of trees and one used individual plants. Plantations had different initial inoculum densities of the pathogen. Initial studies indicated that highly virulent (cotton-defoliating) isolates of the pathogen were present in Marinaleda (experiment I), which represents the first record of such isolates affecting olive trees in Europe. Solarization treatments were applied to lines of trees for either one (single) or two consecutive (double) years. Solarization significantly reduced pathogen populations in the top 20 cm of soil for at least 3 years in relation to control plots. Pathogen reduction after the single solarization obscured effects of the second solarization treatment. Decrease of inoculum density in soil by solarization did not correspond to a similar reduction in disease severity. Disease severity was reduced only in orchards with medium or high initial inoculum densities. A second soil solarization treatment did not improve the effect of single solarization on Verticillium wilt control. In orchards with low inoculum densities, soil solarization did not result in significant differences in disease incidence and severity, but improved recovery of trees from the disease. Soil-solarized plots remained free of weeds, but tress in solarized plots did not show significant growth increase measured by trunk perimeter.

The Effect of Wounding, Temperature, and Inoculum on the Development of Pink Rot of Potatoes Caused by Phytophthora erythroseptica

The effect of wounding, temperature, and inoculum on the development of pink rot caused by Phytophthora erythroseptica, was studied for its potential impact on postharvest infection. Tissue plugs cut from pink rot infected tubers and plugs of similar size from laboratory cultures of the pathogen were highly effective inoculum sources on wounded tubers. Severe wounding, temperatures of 15 to 25°C, and high inoculum density affected the infection risk. Regardless of source or amount of inoculum, any degree of wounding greatly increased incidence of infection of tubers by P. erythroseptica. Infections in unwounded tubers started at 15°C, whereas in wounded tubers infection started at 10°C. Incidence of pink rot was high when two or three of the factors (severe wounding, high temperature, high inoculum level) were favorable. Incidence of pink rot was intermediate when only one factor was favorable. Incidence of pink rot was low or absent without a favorable factor (no wounding, low temperature, and low inoculum), Since infected tuber tissue may serve as potential inoculum source for postharvest infection of tubers by P. erythroseptica, the removal of pink rot infected tubers at harvest is desirable. Avoidance of wounding and rapid cooling of storage bins to 10°C may also help control pink rot.

Isolation and characterisation of ethoprophos-degrading bacteria.

An enrichment culture technique was used to isolate bacteria responsible for the enhanced biodegradation of ethoprophos in a soil from Northern Greece. Restriction fragment length polymorphism patterns of the 16S rRNA gene, partial 16S rRNA sequence analysis, and sodium dodecylsulfate-polyacrylamide gel electrophoresis total protein profile analysis were used to characterise the isolated bacteria. Two of the three ethoprophos-degrading cultures were pure and both isolates were classified as strains of Pseudomonas putida (epI and epII). The third culture comprised three distinct components, a strain identical to P. putida epI and two strains with 16S rRNA sequence similarity to Enterobacter strains. Isolate epI effectively removed a fresh ethoprophos addition from both fumigated and non-fumigated soil when introduced at high inoculum density, but removed it only from fumigated soil at low inoculum density. Isolates epI and epII degraded cadusafos, isazofos, isofenphos and fenamiphos, but only at a slow rate. This high substrate specificity was attributed to minor (cadusafos), or major (isazofos, isofenphos, fenamiphos) structural differences from ethoprophos. Studies with (14)C-labelled ethoprophos indicated that isolates epI and epII degraded the nematicide by removing the S-propyl moiety.

Isolation and characterisation of ethoprophos-degrading bacteria

An enrichment culture technique was used to isolate bacteria responsible for the enhanced biodegradation of ethoprophos in a soil from Northern Greece. Restriction fragment length polymorphism patterns of the 16S rRNA gene, partial 16S rRNA sequence analysis, and sodium dodecylsulfate–polyacrylamide gel electrophoresis total protein profile analysis were used to characterise the isolated bacteria. Two of the three ethoprophos-degrading cultures were pure and both isolates were classified as strains of Pseudomonas putida (epI and epII). The third culture comprised three distinct components, a strain identical to P. putida epI and two strains with 16S rRNA sequence similarity to Enterobacter strains. Isolate epI effectively removed a fresh ethoprophos addition from both fumigated and non-fumigated soil when introduced at high inoculum density, but removed it only from fumigated soil at low inoculum density. Isolates epI and epII degraded cadusafos, isazofos, isofenphos and fenamiphos, but only at a slow rate. This high substrate specificity was attributed to minor (cadusafos), or major (isazofos, isofenphos, fenamiphos) structural differences from ethoprophos. Studies with 14C-labelled ethoprophos indicated that isolates epI and epII degraded the nematicide by removing the S-propyl moiety.

The inconsistent effect of soil disturbance on colonization of roots by arbuscular mycorrhizal fungi: a test of the inoculum density hypothesis

Reducing the tillage of agricultural soils can increase early-season crop-P uptake. Consistent increases in plant-P have been found in both field- and laboratory-systems with undisturbed (U) compared to disturbed (D) soil. A concomitant stimulatory effect on colonization of roots in U soil by arbuscular mycorrhizal (AM) fungi has been found in some cases, but in others the colonization has been similar in U and D treatments. Disruption of the extraradical mycelium that remains from the previous crop is the mechanism by which soil disturbance restricts mycorrhizally mediated P uptake for the subsequent crop, with a tandem change in colonization not necessary, but sometimes seen. Nonetheless, a complete account of these processes will need an understanding of the conditions under which the extent of colonization is affected. Soil-P does not explain when a difference in colonization will appear. Among ecosystems in Western Australia, high inoculum density in a pasture was reported previously to preclude the appearance of a difference in colonization in response to soil disturbance, whereas for other ecosystems with lower inoculum densities a difference in colonization was seen. Here, we determined if a similar mechanism operates for an agricultural soil collected mid-season during the growth of a maize ( Zea mays L.) crop in Ontario, Canada. Blending various proportions of pasteurized and non-pasteurized soil gave a range of inoculum densities. Maize was taken through two 3-week growth cycles in pots, and for the D treatment the soil was passed through a 5 mm sieve between cycles. All plants became colonized with AM fungi. Reducing the inoculum density served to limit colonization to similar low levels in both U and D soils. Stimulation of colonization and of shoot-P uptake in the U-compared with the D-treatment was greater for plants under the higher inoculum conditions tested. We conclude that the inoculum density during crop growth of the soil studied here is moderate, and that this density makes it possible, if other conditions are met, for a reduction of colonization of roots in response to soil disturbance. Whether or not a difference in colonization will appear following disturbance of a soil such as the one studied here probably depends on the interaction between the environment and the plant. Possible interactions are discussed. The high inoculum density of ecosystems such as the pasture studied in Australia likely overrides any effect of soil disturbance and ensures roots of all plants become well-colonized by AM fungi.

Different measures of human hematopoietic cell culture performance are optimized under vastly different conditions

Hematopoiesis, the formation of mature blood cells from stem (LTC-IC) and progenitor (CFU-GM) cells in the bone marrow, is a complex tissue-forming process that leads to many important physiological functionalities. Consequently, a functioning ex vivo hematopoietic system has a variety of basic scientific and clinical uses. The design and operation of such a system presents the tissue engineer with challenges and choices. In this study, three culture variables were used to control ex vivo human hematopoiesis. Systematic variation of inoculum density (ID), medium exchange interval (MEI), and the use of preformed stroma (PFS) showed that (1) all three variables significantly influenced culture performance, (2) the three variables interacted strongly, and (3) the variables could be manipulated to achieve the optimization of different performance criteria. Donor-to-donor variability in culture performance was great at low ID but was minimized at higher ID. PFS had a large positive effect on cell and CFU-GM output at low ID, but had minimal effect at higher ID. In fact, PFS caused a decrease in LTC-IC output at high ID. The effects of PFS indicated that stromal cell elements became more limiting than proliferative cell elements as ID was reduced.In cultures without PFS, maximum cell output was obtained with high ID using a short MEI, whereas the greatest cell expansion ratio was obtained at low ID with an intermediate MEI. Maximum CFU-GM output was obtained from cultures with high ID using a short to intermediate MEI, whereas the greatest CFU-GM expansion ratio was obtained at intermediate ID with an intermediate MEI. The addition of PFS altered the locations of these maxima. In general, PFS moved the maxima to lower ID, and culture output became more sensitive to MEI. Therefore, the optimization of one performance criterion always resulted in a decline of the others. This study demonstrates that ex vivo tissue function is sensitive to many culture variables in an interactive fashion and that systematic multivariable studies are required to characterize tissue function. Once the effects of individual variables and their interactions are known, this knowledge can be used to optimize tissue performance with respect to desired criteria.

Influence of Inoculum Density of Verticillium dahliae on Root Discoloration of Horseradish

The relationship between inoculum density (number of microsclerotia per gram of air-dried soil) of Verticillium dahliae at the time of planting and the severity and incidence of root discoloration of horseradish at harvest was investigated in a 2-year study conducted in the greenhouse, microplots, and commercial production fields. The objective of the study was to develop a disease-forecast system that would assist growers in assessing the risk of the disease before planting horseradish in a particular field. Significant correlations were observed between inoculum density and severity and incidence of root discoloration in the greenhouse and microplots, although the form of the relationship varied with trials from linear to quadratic and negative exponential. No correlation was found between inoculum density of V. dahliae and severity and incidence of root discoloration in commercial production fields. In some fields with low inoculum densities, high ratings of severity and incidence of root discoloration were observed even with the partially resistant cultivar 769A. Conversely, in other fields with high inoculum densities, low ratings of severity and incidence of discolored roots were observed even with the susceptible cultivar 647A. These results suggest that a disease-forecast system based solely on inoculum densities of V. dahliae would be unreliable under field conditions when the other factors affecting the inoculum density-disease relationships cannot be controlled. Knowing the amount of initial inoculum may, however, save growers from planting horseradish in highly infested fields, but it would not guarantee a disease-free crop in fields with low levels of infestation.

Effects of the Plant-pathogenic Fungus Mycocentrospora acerina (Hartig) Deighton on Growth and Competition of Viola arvensis (Murr.) in Spring Wheat

Mycocentrospora acerina (Hartig) Deighton can kill or suppress the growth of Viola arvensis (Murr.) in spring wheat in pot-grown plants in both glasshouse and outdoor studies. The level of damage to V. arvensis and the effect on its competition with wheat were affected by inoculum density and environmental conditions. High inoculum density (105 macroconidia/ml) caused high mortality in V. arvensis and eliminated its competitive effect on wheat. At a lower density (104 macroconidia/ml), the V. arvensis fresh weight and its competition with the wheat were suppressed by up to 38 and 28% respectively, depending on environmental conditions.

Concentric leaf spot of yam (Dioscorea spp) in South-western Nigeria

The etiology and epidemiology of concentric leaf spot and seedling blight diseases of yam (Dioscorea spp) was investigated at Jbadan, south western Nigeria, in the low land humid tropics. Sclerotium rolfsii was associated with the concentric leaf spot and seedling blight of yam. The pathogen was also harbored by weeds namely Mucuna pruriensis, Commelina erecta, ipomea triloba, I. involucrataand Ipomoea sp found growing within the yam plots. A high mean inoculum densities of 3.0 ± 0.5 × 106 colony forming unit/g of soil was estimated in the soil samples obtained from the yam plots

Dynamics of Primary and Secondary Infection in Take-All Epidemics

ABSTRACT Using a combination of experimentation and mathematical modeling, the effects of initial (particulate) inoculum density on the dynamics of disease resulting from primary and secondary infection of wheat by the take-all fungus, Gaeumannomyces graminis var. tritici, were tested. A relatively high inoculum density generated a disease progress curve that rose monotonically toward an asymptote. Reducing the initial inoculum density resulted in a curve that initially was monotonic, rising to a plateau, but which increased sigmoidally to an asymptotic level of disease thereafter. Changes in the infectivity of particulate inoculum over time were examined in a separate experiment. Using a model that incorporated terms for primary and secondary infection, inoculum decay, and host growth, we showed that both disease progress curves were consistent with consecutive phases dominated, respectively, by primary and secondary infection. We examined the spread of disease from a low particulate inoculum density on seminal and adventitious root systems separately. Although seminal roots were affected by consecutive phases of primary and secondary infection, adventitious roots were affected only by secondary infection. We showed that the characteristic features of disease progress in controlled experiments were consistent with field data from crops of winter wheat. We concluded that there is an initial phase of primary infection by G. graminis var. tritici on winter wheat as seminal roots grow through the soil and encounter inoculum, but the rate of primary infection slows progressively as inoculum decays. After the initial phase, there is an acceleration in the rate of secondary infection on both seminal and adventitious roots that is stimulated by the increase in the availability of infected tissue as a source of inoculum and the availability of susceptible tissue for infection.

Relationships Between Verticillium dahliae Inoculum Density and Wilt Incidence, Severity, and Growth of Cauliflower

ABSTRACT Microplot and field experiments were conducted to evaluate the effects of inoculum density on Verticillium wilt and cauliflower growth. Soil containing Verticillium dahliae microsclerotia was mixed with various proportions of fumigated soil to establish different inoculum densities (fumigated soil was used as the noninfested control). Seven inoculum density treatments replicated four times were established, and the treatments were arranged in a randomized complete block design. Soil was collected from each microplot immediately after soil infestation for V. dahliae assay by plating onto sodium polypectate agar (NP-10) selective medium using the Anderson sampler technique. Five-week-old cauliflower was transplanted into two beds within each 1.2- by 1.2-m microplot. At the same time, several extra plants were also transplanted at the edge of each bed for destructive sampling to examine the disease onset (vascular discoloration) after planting. Cauliflower plants were monitored for Verticillium wilt development. Stomatal resistance in two visually healthy upper and two lower, diseased leaves in each microplot was measured three times at weekly intervals after initial wilt symptoms occurred. At maturity, all plants were uprooted, washed free of soil, and wilt incidence and severity, plant height, number of leaves, and dry weights of leaves and roots were determined. The higher the inoculum density, the earlier was disease onset. A density of 4 microsclerotia per g of dry soil caused 16% wilt incidence, but about 10 microsclerotia per g of soil caused 50% wilt incidence. Both wilt incidence and severity increased with increasing inoculum density up to about 20 microsclerotia per g of soil, and additional inoculum did not result in significantly higher disease incidence and severity. A negative exponential model described the disease relationships to inoculum levels under both microplot and field conditions. Stomatal resistance of diseased leaves was significantly higher at higher inoculum densities; in healthy leaves, however, no treatment differences occurred. The height, number of leaves, and dry weights of leaves and roots of plants in the fumigated control were significantly higher than in infested treatments, but the effects of inoculum density treatments were variable between years. Timing of cauliflower infection, crop physiological processes related to hydraulic conductance, and wilt intensity (incidence and severity) were thus affected by the inoculum density. Verticillium wilt management methods used in cauliflower should reduce inoculum density to less than four micro-sclerotia per g of soil to produce crops with the fewest number of infected plants.

Influence of Inoculum Density, Wetness Duration, Plant Age, Inoculation Method, and Cultivar Resistance on Infection of Pepper Plants by Colletotrichum coccodes.

Effects of inoculum density, wetness duration, plant age, inoculation method, and cultivar resistance on the development of pepper anthracnose caused by Colletotrichum coccodes were evaluated under controlled environmental conditions. As inoculum density of C. coccodes increased from 103 to 106 conidia/ml, symptoms of anthracnose developed. Disease severity also increased with increasing time of wetness duration from 0 to 60 h. Wetness duration above 48 h and a high inoculum density at 106 conidia/ml caused severe defoliation and blight symptoms in pepper seedlings. Inoculum at 104 conidia/ml and 24 to 36 h of continuous wetness duration was enough to form irregular or circular brown lesions on pepper leaves. Age-related resistance was expressed as pepper plants matured. To evaluate resistance, the foliar-inoculation method was more reliable than the soil-drench method, which did not induce symptoms on older plants. The responses of 35 pepper cultivars or accessions tested to C. coccodes were quantitative rather than qualitative. No hypersensitive symptoms were observed in any of the cultivars inoculated with any of the two isolates. No significant differences in susceptibility to anthracnose were found among cultivars from Korea, the United States, India, and Thailand or accessions tested, irrespective of genetic or country origin of cultivars. The Korean cultivar Kwangbok was relatively less susceptible to anthracnose, compared with the other cultivars tested.

Effects of cover crops, weeds and plant debris on development ofMycocentrospora acerinain caraway

SummaryThis paper reports on the search for inoculum sources ofMycocentrospora acerinaon caraway (Carum carviL.). Obvious suspects are cover crops of biennial caraway and preceding crops of annual caraway. Other suspects are weeds in or alongside the field. Finally, survival structures of the fungus, chlamydospore chains, packed in plant debris or naked, are suspected.M. acerinais able to infect many plant species, including cover crops of caraway such as spinach for seed production and peas. However, the agronomical suitability of a crop to serve as a cover crop of biennial caraway proved to be a more important factor in determining caraway yield than the susceptibility of the cover crop toM. acerina.This finding was corroborated by the fact that spinach and peas as preceding crops had no significant effects onM. acerinadevelopment in spring caraway sown the next year. Dill, barley and four weed species were found as new hosts ofM. acerina.The role of weed hosts, susceptible crops and plant debris in the survival of the fungus in years without caraway is discussed. Caraway sown on soil containing infested caraway straw, infested debris of other plant species or chlamydospores grown in pure culture, became infected byM. acerina.Only high inoculum densities of chlamydospores in the soil caused severe damping‐off of caraway seedlings. The opportunity for disease management by agronomical means is quite limited.

Inoculum Density and Expression of Major Gene Resistance to Fusiform Rust Disease in Loblolly Pine.

Inoculum densities of 25 × 103 to 200 × 103 per ml of basidiospores from single aeciospore isolates avirulent or virulent to the Fr1 (fusiform resistance-1) gene were used to inoculate a control-pollinated loblolly pine family heterozygous for this gene. With two avirulent isolates, the regression curve of gall frequency 9 months after inoculation went from 26 to 50% as inoculum density increased to 100 × 103 spores. The regression curve flattened at higher inoculum densities. With two virulent isolates, gall frequency increased from 47% to a plateau at 97% as spore density increased. A double-blind element of the study correlated the occurrence of the genetic marker (RAPD marker J7-485A) for Fr1 resistance in haploid megagametophyte tissuend the presence or absence of galls on seedlings after artificial inoculations. With avirulent isolates at the two higher densities of 100 × 103 and 200 × 103, marker presence-absence and phenotypic assessments of gall presence-absence agreed for 95% of the seedlings. At the 50 ×103 level, marker-phenotype agreed for 86% of the seedlings. The increased marker-phenotype association resulted from a reduction or elimination of disease escapes as Fr1 resistance remained stable even at higher spore densities. The double-blind study indicates that resistant individuals can be identified from the megagametophyte tissue of germinating seedlings. With virulent isolates, marker and disease phenotype did not correlate, even at the lowest inoculum density. The virulent isolates appear to be homozygous for virulence because infection of marker-positive resistant seedlings equaled or exceeded that of marker-negative susceptible seedlings at the lowest inoculum density.

Spatial Patterns of Microsclerotia ofVerticillium dahliaein Soil and Verticillium Wilt of Cauliflower

ABSTRACT The spatial patterns of microsclerotia of Verticillium dahliae in soil and wilt symptoms on cauliflower were determined at three sites in each of two fields in 1994 and 1995. Each site was an 8 x 8 grid divided into 64 contiguous quadrats (2 by 2 m each). Soil samples were collected to a depth of 15 cm with a probe (2.5 cm in diameter), and samples from four sites in each quadrat were bulked. Plants in each quadrat were cut transversely, and the number of plants with vascular discoloration and the number without discoloration were recorded. The soil was assayed for microsclerotia by the modified Anderson sampler technique. Lloyd's index of patchiness (LIP) was used as an indicator to evaluate the aggregation of microsclerotia in the field. Spatial autocorrelation and geostatistical analyses were also used to assess the autocorrelation of microsclerotia among quadrats. The LIP for microsclerotia was greater than 1, indicating aggregation of propagules; however, the degree of aggregation at most sites was not high. Significant autocorrelation within or across rows was detected in some spatial autocorrelograms of propagules, and anisotropic patterns were also detected in some oriented semivariograms from geostatistical analyses for microsclerotia, indicating the influence of bed preparation in the fields on pathogen distribution. The parameter estimates p and theta in the beta-binomial distribution and the index of dispersion (D) associated with the distribution were used to assess the aggregation of diseased plants at each site. A random pattern of wilt incidence was detected at 7 of 12 sites, and an aggregated pattern was detected at 5 of 12 sites. The degree of aggregation was not high. A regular pattern of wilt severity was detected at all sites. The high disease incidence (77 to 98%) observed at 11 of the 12 sites could be explained by high inoculum density.

Comparison of NAA and 2,4-D induced somatic embryogenesis in Cassava

NAA and 2,4-D were compared for their ability to induce somatic embryogenesis in cassava (Manihot esculenta Crantz). In all seven cultivars tested, only 2,4-D had the capacity to induce primary somatic embryos from leaf explants, however, both NAA and 2,4-D were capable of inducing secondary somatic embryos. More secondary somatic embryos were formed in NAA than in 2,4-D medium. Furthermore, the maturation period for secondary somatic embryos was shorter in NAA medium than in 2,4-D medium. In some cultivars, repeated subculture of secondary somatic embryos in NAA medium resulted in a gradual shift from somatic embryogenesis to adventitious root formation. This shift could be stopped and reversed by subculture of the material in 2,4-D medium. In NAA medium the most secondary somatic embryos were formed when they were subcultured every 15 days whereas in 2,4-D a 20 day subculture interval was optimal. Subculture of secondary somatic embryos at a high inoculum density (>1.5 g jar−1) in NAA medium did not result in the formation of secondary somatic embryos, whereas in 2,4-D it lead to the formation of globular secondary somatic embryos. With 2,4-D the newly induced secondary somatic embryos were connected vertically to the explant and with NAA medium horizontally. For all cultivars tested, desiccation stimulated normal germination of NAA-induced somatic embryos. However, the desiccated, secondary somatic embryos required a medium supplemented with BA for high frequency germination. The concentration of BA needed for high frequency germination was higher when the desiccated secondary somatic embryos were cultured in light instead of dark. In only one cultivar desiccation enhanced germination of 2,4-D induced secondary somatic embryos and in three other cultivars it stimulated only root formation.

Modified Behavior in Baculovirus-Infected Lepidopteran Larvae and Its Impact on the Spatial Distribution of Inoculum

Abstract Quantifying the rate of dispersal of target insects when infected with a disease agent will aid the development of biorational pest control programs. The effect of nucleopolyhedrovirus (NPV) infection on the mobility of second and fourth instar Mamestra brassicae larvae was investigated in the laboratory and field. NPV infection altered larval mobility, with the changes in behavior varying with the timecourse of infection. Diseased larvae moved three to five times further than healthy ones during the middle stages of infection. By the 7th day postinfection diseased larvae were less mobile than healthy counterparts. The same pattern of modified behavior was observed in both instars. Fourth instar larvae moved further than second instars under laboratory and field conditions. In the field, infected larvae tended to die on the apex of the cabbage leaves. Bioassay of the leaves showed a linear decrease in inoculum from central to peripheral plants within the plots, which occurred to the same extent for second and fourth instars. Leaves from plots where infected fourth instar larvae had been introduced had higher inoculum density than those from plots with second instars.