Infectivity,inoculum density and germination of Spongospora subterranea resting spores: a solution-culture test system1

Spongospora subterranea, causal agent of powdery scab of potatoes and vector of potato mop-top furovirus, survives in the soil as balls of resting spores (cystosori). So far, the factors affecting longevity, germination and infectivity of cystosori have not been investigated. A rapid and versatile bioassay with tomatoes as bait plants has been developed to quantify the infectivity of cystosorus inoculum or infested soil. The intensity of root infection, as a measure of infectivity, was determined by evaluating the quantity of zoosporangia present in epidermal cells and root hairs of the whole, stained root system. A correlation was obtained between the intensity of root infection and the cystosorus inoculum density in nutrient solution. Sterile soil suppressed the inoculum potential of pure cystosori. Infectivity of untreated soil decreased with increasing time of storage. Root infection was not influenced by the pH level of the nutrient solution.     

Effect of <Emphasis Type="Italic">Meloidogyne ethiopica</Emphasis> parasitism on water management and physiological stress in tomato

Root-knot nematodes (RKN) are obligate endoparasites that severely damage the host root system. Nutrient and water uptake are substantially reduced in infested plants, resulting into altered physiological processes and reduced plant growth. The effect of nematode infestation on the morphological changes of roots and subsequent physiological plant responses of infested tomatoes with the RKN Meloidogyne ethiopica was studied in a pot experiment. Plants were infested with two inoculum densities (10 or 50 eggs per cm³ substrate) and its effect was evaluated 74 and 102 days post inoculation (DPI). Morphological changes and root growth was determined by analysing scanned images of the whole root system. Nematode infestation reduced the portion of fine roots and increased that of coarse roots due to gall formation. Fine roots of non-infested control plants represented around 51% of the area of the whole root system at 74 and 102 DPI. In comparison to controls, plants inoculated with low and high nematode density had 2.1 and 3.2-times lower surface area of fine roots at 102 DPI. Root analyses revealed that plants had a very limited ability to mitigate the effects of the root-knot nematodes infestation by altering root growth. Root galls had a major influence on the hydraulic conductivity of the root system, which was significantly reduced. The low leaf water potential of infested plants coincided with decreased stomatal conductivity, transpiration and photosynthesis. The latter two were reduced by 60–70% when compared to non-infested control plants.     

Inoculum potential and host range of Polymyxa betae and beet necrotic yellow vein furovirus1

The inoculum potential of Polymyxa betae and BNYVV was studied from 52 random samples of Belgian soils and 10 samples from other European countries, by culture of bait plants in tubes under controlled conditions on serial dilutions of the soils in sterile sand. P. betae was detected in all samples within the range of 0.01 to 27.1 infection units per g of soil. BNYVV was detected by ELISA on root extracts of bait plants grown on three Belgian soil samples. All the tested samples from rhizomania-infested areas in France, FRG, the Netherlands, Switzerland and Austria, were found to be infested by BNYVV by this technique. For BNYVV survey, the plant bait technique appears more reliable than the analysis of rootlets collected in the field and observation of external symptoms in case of low BNYVV infestations or non-expression because of unfavourable environmental conditions. P. betae isolates from various origins heavily infected Beta spp. but only moderately spinach. Chenopodium album was slightly infected by 2 of the 7 isolates, C. murale by 4 of them.     

Infection of sugar beet by Polymyxa betae in relation to soil temperature

The effects of soil temperature on infection of sugar-beet roots by the soil-borne fungus Polymyxa betae were investigated in controlled environments. Pre-germinated seeds were sown in pots of naturally infested soil and seedlings sampled at frequent intervals over a period of several weeks. Within the range 10-30°C, the optimum soil temperature for infection was c. 25°C; the time between sowing and the first detectable infection was shortest and the subsequent rate of infection most rapid at this temperature. No infection was observed over 80 days at 10°C.
Both root and shoot dry weight were reduced on plants growing in infested soil at 15, 20 and 25 C compared with those growing in uninfested soil. In general, root growth was more severely affected than shoot growth and the effects were most pronounced at 20°C. These results were confirmed in a subsequent experiment in which P. betae -infected root material was used as the inoculum. In addition to its role as the vector of beet necrotic yellow vein virus (the cause of Rhizomania disease), the significance of P. betae as a plant pathogen in its own right is discussed.     

Spread of Phytophthora Root and Crown Rot in Saintpaulia, Gerbera and Spathiphyllum Pot Plants in ebb-and-flow-systems

Spread of Phytophthora root and crown rot in three pot plant species was studied on ebb-and-flow benches where the nutrient solution was recirculated. The plant species and their respective pathogens were: Saintpaulia ionantha – P. nicotianae, Gerbera jamesonii – P. cryptogea, and Spathiphyllum wallissii – Phytophthora spp. Ebb-and-flow benches were infested with the pathogen using different methods: 18–25% of the plants on a bench were inoculated or potted in soil infested with the pathogen or the nutrient solution was infested by either zoospores or mycelium fragments. More than 80% of the inoculated Saintpaulia plants and 22% of plants potted in infested soil developed disease but no spread of the disease was observed. Infestation of the nutrient solution did not result in any diseased Saintpaulia plant. More than 70% of the Gerbera plants developed disease as a result of spread of the pathogen irrespective of the infestation method used. No significant spread of the disease was observed with inoculated Spathiphyllum plants nor from plants potted in infested soil. A few Spathiphyllum plants developed disease symptoms after infestation of the nutrient solution with zoospores. In one experiment, nearly all Spathiphyllum plants were diseased after infestation of the nutrient solution with mycelium fragments. The presence of an irrigation mat significantly reduced the spread of the Phytophthora disease in Gerbera and Spathiphyllum. The possibility of an irrigation mat acting as a filter for zoospores is discussed.     

Effect of Temperature on and Histopathology of the Interaction Between Meloidogyne incognita and Thielaviopsis basicola on Cotton

ABSTRACT Controlled environments were used to study the relationship between the root-knot nematode (Meloidogyne incognita) and Thielaviopsis basicola on cotton. Temperature treatments were continuous 20, 24, and 28 degrees C or two cyclic linear regimes with ranges of 14 to 32 or 18 to 28 degrees C over 24 h. Cotton seeds were planted in fumigated soil infested with T. basicola, M. incognita, or both. After 42 days, pathogen effects on plant growth and pathogen development were evaluated. Histology was conducted on roots collected 14, 28, and 42 days after planting in the continuous 24 degrees C treatment. Reductions in plant height-to-node ratio and total fresh weight were observed for soils infested with both pathogens compared with the control or with soils infested with either pathogen, except for M. incognita-infested soil at 28 degrees C. T. basicola reduced root galling and reproduction of the nematode at all temperatures. Vascular discoloration caused by T. basicola was greater in the presence of M. incognita compared with that by T. basicola alone. At 2 and 4 weeks, histological studies showed that plants grown in all T. basicola-infested soils contained chlamydospore chains on the root surface and in cortical cells. The fungus was not observed inside the vascular cylinder. Roots from 4-week-old plants from soils infested with T. basicola and M. incognita showed fungal sporulation in vascular tissue and localized necrosis of vascular tissue adjacent to the nematodes. At 6 weeks, plants grown in soil infested with T. basicola alone exhibited no remaining cortical tissue and no evidence of vascular colonization by the fungus. Six-week-old plants grown in T. basicola + M. incognita-infested soils exhibited extensive vascular necrosis and sporulation within vascular tissue. These studies suggest that coinfection expands the temperature ranges at which the pathogens are able to cause plant damage. Further, M. incognita greatly increases the access of T. basicola to vascular tissue.     

Detection of Colletotrichum coccodes and Helminthosporium solani in soils by bioassay

The sensitivity of a bioassay in detecting soil inoculum of Colletotrichum coccodes and Helminthosporium solani was examined using potato minitubers and microplants. Tests were conducted on soils which were collected from fields in which the interval after a previous potato crop differed, and which were also artificially infested with conidia or microsclerotia. For C. coccodes , determining plant infection based on the occurrence of infected roots after 9–12 weeks was a sensitive method for detecting and quantifying the amount of inoculum in soil. Infestations of less than 0·4 microsclerotia per g soil were detected in artificially infested soils. A semiselective medium, developed for isolating C. gloeosporioides from pepper, detected soil infestations by C. coccodes as low as nine conidia or one microsclerotium per g soil in artificially infested soil. For H. solani , infection on minitubers was a sensitive measure, with soil inoculum of fewer than 10 conidia per g soil being detected. Soil infestation could be quantified by assessing the percentage surface area of minitubers covered by sporulating lesions, which was strongly related to the amount of soil infestation. The results of these bioassay tests were compared with published results for real-time quantitative PCR assays on the same soils. The two methods were in good agreement in artificially infested soils, but the bioassay appeared to be more sensitive with naturally infested soils.     

Host status of selected cultivated fruit crops to <Emphasis Type="Italic">Meloidogyne enterolobii</Emphasis>

Meloidogyne enterolobii (syn. M. mayaguensis) has been reported to cause severe damage in commercial guava orchards and other plants in Central and South American countries. Considering the risk of introduction and dissemination of this pest in the European region, M. enterolobii was placed on the EPPO A2 list in 2010. The use of non-host fruit species is a recommended strategy to manage root-knot nematodes in infested guava orchards. This study screened 89 plant genotypes from 25 fruit plants of economic importance, plus two susceptible controls (guava and tomato) for its host status to M. enterolobii. Three to eight months after inoculation, nematode reproduction factor (RF) was used to characterize host suitability of fruit crops to this nematode. Ten banana genotypes, six Barbados cherries, one fig, two grape rootstocks and six melons were rated as good hosts for this nematode. Sixteen fruit plants behaved either as non-hosts or poor hosts to M. enterolobii, including assaí, atemoya, avocado, cashew nut, citrus, coconut, grape, jabuticaba, mango, mulberry, papaya, passion fruit, sapodilla, soursop, starfruit and strawberry. For the future, field experiments in areas infested by this nematode are essential to confirm the greenhouse results. These non-host fruit species can replace in the future eradicated guava trees in fields severely infested by this nematode and become an economic option for growers where M. enterolobii is considered a serious problem.     

Root infection of sugar beet by <Emphasis Type="Italic">Cercospora beticola</Emphasis> in a climate chamber and in the field

Sugar beet root infection by Cercospora beticola, the causal agent of Cercospora leaf spot (CLS), was studied in a climate chamber and in the field. In the climate chamber, root incubation of susceptible seedlings with a conidial suspension resulted in disease incidences that were significantly different for two sugar beet cultivars (Auris: 0.8 ± 0.14 and A00170: 0.5 ± 0.18; P < 0.05) with regard to the control treatment 35 days after root incubation in a standard potting soil-fine river sand mixture. In a field trial with susceptible cv. Savannah with soil-incorporated CLS-infested leaf material, disease developed four weeks earlier in the infested plots than in the control plots. The probability that disease develops in the field was significantly higher for the infested than for the control plots (P < 0.05). Symptomless plants from infested field plots transferred to the glasshouse to induce leaf spot symptoms showed a significantly higher probability to induce symptom development (0.4 ± 0.08), than plants from control plots (0.02 ± 0.02) (P <0.05) 14 days after transfer. This probability was significantly higher than for plants that remained in three of the infested field plots (0.2 ± 0.04; 0.2 ± 0.05 and 0.2 ± 0.04 respectively), except for one infested field plot (0.4 ± 0.05) on July 5. We conclude that C. beticola is able to infect sugar beet seedlings through their roots and that latent CLS infections in sugar beet lead to symptom development at high temperatures (> 20 °C) and high relative humidity (> 95) in our climate chamber or after canopy closure in the field. Quantification of root infection and long term survival in soil is necessary to assess its contribution to the epidemiology and life cycle of Cercospora beticola. Cultural methods such as a wider crop rotation, management of crop debris and ploughing systems may provide control strategies alternative to or reducing fungicide input.     

Interaction of Fusarium solani f. sp. glycines and Heterodera glycines in Sudden Death Syndrome of Soybean

ABSTRACT Sudden death syndrome (SDS) of soybean is caused by the soilborne Fusarium solani f. sp. glycines (synonym F. virguliforme). In a sequential approach, two multifactor factorial-design microplot experiments were conducted to investigate the effects of fungal infestation levels and soil moisture on both root necrosis and foliar SDS severity, and the interaction between F. solani f. sp. glycines and Heterodera glycines in fumigated versus nonfumigated soil. In 2003, soybean cv. Spencer was grown in nonfumigated or methyl bromide-fumigated soil and infested with increasing levels of F. solani f. sp. glycines, either under rainfall or irrigated after growth stage V6/R1. In 2004, interactions between F. solani f. sp. glycines and H. glycines were explored in a factorial inoculation design in fumigated or nonfumigated soil, planted to Williams 82 or Cyst-X20-18. In both years, higher levels of foliar SDS severity and root necrosis were found in F. solani f. sp. glycines-infested soils with H. glycines than in soils without the nematode on the soybean cultivars susceptible to both pathogens. Both natural infestations of H. glycines in 2003 and artificially amended populations of H. glycines in 2004 contributed to higher foliar SDS severity. More severe foliar SDS symptoms always were associated with more root necrosis, but elevated levels of root necrosis did not predict severe leaf symptoms. In contrast to the critical role of H. glycines, increasing fungal infestation levels had no significant effects on increasing either foliar SDS symptoms or root necrosis. Effects of moisture regime and fungal infestation levels also were examined in factorial greenhouse and growth chamber experiments. High soil moisture resulted in higher levels of SDS root necrosis. In the greenhouse, root necrosis increased at a higher rate in low soil moisture than the rate in high soil moisture. The two pathogens acted as a complex and the disease development was strongly dependent on high soil moisture.     

列当杂草及其防除措施展望

根寄生杂草列当Orobanche spp.每株可产生大量细小的种子,易于传播,且在出苗前已经对寄主作物造成危害,使列当的防除成为一个世界性难题。本文详述了列当的生活史、为害现状以及防除措施,并突出了生物防除的生态效益与经济效益。基于＂诱捕＂作物及＂捕获＂作物诱导列当种子＂自杀发芽＂理论的防除机制,本文提出了新型可持续生态防除思想,为列当科杂草防除提供参考。     

土壤处理对西洋参根际线虫数量动态的影响及对防治茎线虫根腐病的效果

 Root rot of American ginseng (Panax quinquefolium) caused by Ditylenchus destructor is a novel disease found recently in Beijing area. The effects of three soil treatments (fumigating with chloropicrin, chloropicrin + lvfeng organic manure and nematicide fosthiazate) on the number of rhizospheric nematodes, survived plants, root yield and root rot of 3-year ginseng plant were compared. The effects were also investigated at the second year after treatment. The results indicated that the number of parasitic nematodes in rhizosphere of treated soil reached the peak value in late June to early July as the soil temperature raised in the growing season. Compared with the regular treatment, the number of plant parasitic nematodes was reduced while non-plant parasitic nematodes increased. The number of non-plant parasitic nematodes in the soil treated with chloropicrin + lvfeng organic manure was 2 times than that treated with chloropicrin only. The ratio of non-parasitic to parasitic nematodes of three treatments was higher than the control. Percent of survived plants was 94.8%-96.4% and diseased root was decreased obviously. The control efficacy was more than 89% at the first year after treatment. The survived plants and plot yield of ginseng increased significantly and the control efficacy was around 40% at the second year. The best of the three treatments was by chloropicrin + organic manure.     

Rhizosphere Competence of Pythium oligandrum

ABSTRACT The associations of Pythium oligandrum with the root cortex, rhizoplane, and rhizosphere were measured with 11 crop species. This work was expedited by the use of a semiselective technique for isolation of P. oligandrum from soil and plant material. Cortical colonization of roots by P. oligandrum was not detected, and the rhizoplanes of the roots of most crops were free of the fungus. However, P. oligandrum was detected in large quantities with every crop tested when roots with adhering soil (rhizosphere soil) were assayed. Different crop species and cultivars of cantaloupe, cauliflower, and tomato varied in rhizosphere densities of P. oligandrum, but rhizosphere population densities of the fungus were consistently higher than in nonrhizosphere soils with plants grown in P. oligandrum-infested sterilized potting mix or an unsterilized mineral soil. After transplanting tomatoes into potting mix infested with P. oligandrum, increases in CFU occurred over time in the rhizosphere but not in the nonrhizosphere soil. In trials on delivery methods of inoculum of P. oligandrum, the rhizosphere populations of tomato plants grown in potting mix were about sixfold higher compared to seed-coat treatments when ground, alginate pelleted oospores were applied to seedlings growing in plug containers prior to transplanting or to pots containing potting mix before direct seeding.     

Suppression of Rhizoctonia root-rot of tomato by Glomus mossae BEG12 and Pseudomonas fluorescens A6RI is associated with their effect on the pathogen growth and on the root morphogenesis

Rhizoctonia solani root-rot is a major soilborne disease causing growth and yield depression. The ability of Glomus mosseae BEG12 and Pseudomonas fluorescens A6RI to suppress this soilborne disease in tomato was assessed by comparing the shoot and root growth of plants infested with R. solani 1556 when protected or not by these beneficial strains. The epiphytic and parasitic growth of the pathogenic R. solani 1556 was compared in the presence and absence of the biocontrol agents by microscopical observations allowing the quantification of roots with hyphae appressed to epidermal cells (epiphytic growth) and of roots with intraradical infection (parasitic growth). The root architecture of the tomato plants under the different experimental conditions was further characterized by measuring total root length, mean root diameter, number of root tips and by calculating degree of root branching. G. mosseae BEG12 and P. fluorescens A6RI fully overcame the growth depression caused by R. solani 1556. This disease suppression was associated with a significant decrease of the epiphytic and parasitic growth of the pathogen together with an increase of root length and of the number of root tips of inoculated tomato plants. The combined effects of G. mosseae BEG12 and P. fluorescens A6RI on pathogen growth and on root morphogenesis are suggested to be involved in the efficient disease suppression.     

Evaluation of eradication measures against Anoplophora chinensis in early stage infestations in Europe

Eradication efforts are described at 5 sites where infestations of Anoplophora chinensis have been detected in Europe since 2003. The infestations were in France (declared eradicated in 2006), in Croatia, in Italy (Rome) and in the Netherlands (Westland and Boskoop). Each of these infestations was on or nearby a nursery or location where plants originating from Eastern Asia had (probably) been stocked or sold. Infested plants were all found within a distance of 30 m from the presumed source of infestation except in the Rome‐infestation where infested plants, in total 40, were found within 500 m of the presumed source of infestation. The history of plant imports and local observations at 3 sites (France, Croatia and Westland in the Netherlands) suggests that larvae arrived in imported plants and adults emerged and laid eggs on plants in the surroundings between 1 and 5 years before the infestation was detected. It was not possible to make such an estimate for the Boskoop infestation in the Netherlands and the Rome infestation in Italy.     

Phytophthora root and collar rot of alders in Bavaria: distribution, modes of spread and possible management strategies

A survey of symptoms of phytophthora root and collar rot of common ( Alnus glutinosa ) and grey alder ( A. incana ) in riparian and forest stands in Bavaria was conducted by the Bavarian State Forestry and river authorities. Symptoms were seen in 1041 out of 3247 forest alder stands. The majority of the affected stands (80·9%) were less than 21 years old; 46% of these young stands were growing on nonflooded sites and 92% had been planted. The riparian survey showed that symptoms were widespread along more than 50% of the river systems. Along some rivers the disease incidence exceeded 50%. The 'alder Phytophthora ' was recovered from 166 of 185 riparian and forest alder stands with symptoms. In 58 of the 60 rivers and streams investigated in detail, the source of inoculum was traced back to infested young alder plantations growing on the river banks or on forest sites that drain into the rivers. Once introduced to a river system, the 'alder Phytophthora ' infects alders downstream. Baiting tests showed that the 'alder Phytophthora ' was present in rootstocks of alders from three out of four nurseries which regularly bought in alder plants for re-sale, but not in rootstocks from four nurseries that grew their own alders from seed. In addition, the infected nurseries used water from infested water courses for irrigation. The Bavarian State Ministry for Agriculture and Forestry has developed a code of practice for producing healthy alder plants in forest nurseries. This includes a 3-year fallow period between bare-rooted alder crops because of poor survival of the 'alder Phytophthora ' in soil.     

Previous infestation affects recolonization of cotton byAphis gossypii: induced resistance or plant damage?

The effect of previous infestation by the cotton aphid (Aphis gossypii Glover) on resistance of cotton seedlings to further infestation by the same insect, was examined in the laboratory. Seedlings which survived a heavy infestation and produced new leaves, were recolonized artificially and the fate of the colonizers was monitored. The two cotton varieties used in the experiments differed in resistance to aphid colonization, and a few seedlings were initially unsuitable for aphid colonization. Aphids transferred to new leaves on previously infested plants were not so successful as on uninfested seedlings and repeated attempts over a few weeks were sometimes required until a colony became established. Reinfested plants often showed no visible signs of stress, but the aphid colonies were frequently of the ‘yellow dwarf’ morph, indicating that the plants had been stressed after the first infestation. We suggest that damage caused by the first infestation — and not the production of some excess metabolites — is the reason for the lower success on previously infested plants.     

Guava decline: effect of root exudates from Meloidogyne enterolobii-parasitized plants on Fusarium solani in vitro and on growth and development of guava seedlings under controlled conditions

In guava decline, Fusarium solani-immune guava trees become susceptible to extensive root rot caused by this fungus after parasitism by Meloidogyne enterolobii. To understand the mechanisms involved in this disease, root exudates were collected from nematode-inoculated (NI) or uninoculated (UN) guava plants cultivated in sand. After filtration through a Millipore® membrane, NI and UN exudates were used: i) to prepare media to assess their effect on mycelial growth and production of propagules of F. solani isolate UENF/CF 163, and ii) to incubate macro- and microconidia to assess their effect on germination. NI exudates promoted (P?<?0.05) more mycelial growth and production of propagules than UN exudates or water. NI and UN exudates were used to water guava seedlings laid over seed germination paper inside plastic boxes. Half of the seedlings had an agar plug colonized by the fungus positioned in the collar region. Upon watering with NI exudates the fungus caused (P?<?0.05) extensive rotting of the seedlings’ rootlets. NI and UN exudates, either unlyophilized or lyophilized and re-suspended to the original concentration, were used to water guava seedlings grown in sterile sand before being inoculated (or left uninoculated) as described before. Solely upon watering with NI exudates, in its unlyophilized form or after lyophilization, the fungus caused a reduction (P?<?0.05) of shoot and root biomass associated with rotting of roots. These results suggest that M. enterolobii induces chemical changes in the root exudates of guava trees, which are necessary for root invasion causing root rotting by F. solani.     

Influence of feeding-damaged plants on the oviposition responses within a community of female moths

Competitive or facilitative interactions characterise phytophagous insect communities that utilise the same resources. These interactions are often mediated by the host plant. Plant mediation influences the oviposition choices that a community of insects, sharing the same host plant make. In this context, the oviposition choices of females within a community of lepidopteran cereal stemborers namely Busseola fusca, Sesamia calamistis and Chilo partellus were studied in plant choice-experiments under laboratory and field conditions. Gravid females of each species were presented with a choice between maize plants infested by conspecific or heterospecific larvae and uninfested maize plants. The number of eggs and egg batches laid on plants were used to quantify oviposition. Results showed that none of the three stemborer species avoided oviposition on infested maize plants. In some cases a significant preference for infested maize plants were observed. Similarly, data from field trials under natural stemborer infestation, with B. fusca as the only species, showed that the wild ovipositing moths were not avoiding infested plants. Host plant mediation may influence the incidence of multi-species infestations by stemborer species often found in the field. The potential roles of herbivore-induced and egg-deposited-induced plant volatiles in this mechanism are discussed.     

The relationship between soil inoculum density and plant infection as a basis for a quantitative bioassay of Verticillium dahliae

Using potato, eggplant and thorn apple as test plants, the relationship between soil inoculum density and plant infection was studied as a basis for the development of a quantitative bioassay of Verticillium dahliae. A linear relationship was demonstrated (P < 0.05) between soil inoculum density and population density on roots for all three test plants and for soil inoculum density and population density in sap extracted from stems for eggplant. Correlation coefficients were higher with densities on or in roots (R² varying from 0.45 to 0.99) than with densities in stems (R² varying from 0.04 to 0.26). With eggplant, population densities on/in root and in sap extracted from stems were significantly correlated at 20 and 25°C with Pearson's correlation coefficients of 0.41 and 0.53, respectively. For potato, root colonization was higher at 15 than at 20°C, whereas the reverse applied to eggplant. Stems of potato were less colonized than stems of eggplant. The pathozone sensu Gilligan (1985) was calculated to be <300 µm, indicating that infection was caused by microsclerotia which were located close to the roots. To assess the density of V. dahliae in plant tissue pipetting infested plant sap on solidified ethanol agar medium without salts yielded higher densities than using pectate medium or mixing sap with molten agar. A bioassay for determining effects of (a)biotic factors on development of V. dahliae in the plant is recommended with eggplants as a test plant, grown in soil infested with 300 single, viable microsclerotia g^-1 soil at a matric potential of –6.2 kPa, and incubated at 20°C for 8 weeks.