Effect of indole-acetic acid (IAA) on the development of symptoms caused by <Emphasis Type="Italic">Pythium ultimum</Emphasis> on tomato plants

The effect of indole-acetic acid (IAA) on the development of symptoms caused by Pythium ultimum on tomato plants was investigated using different bioassays. Application of IAA (5 μg ml⁻¹) on tomato seedlings inoculated with P. ultimum did not affect their emergence suggesting that IAA did not affect the severity of Pythium damping-off. However, IAA was shown to influence the development of P. ultimum symptoms on tomato plantlets. Low concentrations of IAA (0–0.1 μg ml⁻¹) within the rhizosphere of plantlets increased the severity of the symptoms caused by P. ultimum, while higher concentrations (10 μg ml⁻¹), applied either by drenching to the growing medium or by spraying on the shoot, reduced the symptoms caused by this pathogen. In addition, the study demonstrated that P. ultimum produces IAA in liquid culture amended with L-tryptophan, tryptamine or tryptophol (200 μg ml⁻¹) and in unamended culture.     

Vascular blackening of wasabi rhizomes caused by <Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> subsp. <Emphasis Type="Italic">carotovorum</Emphasis>

Wasabi (Wasabia japonica) is grown for its highly-valued rhizome which is used as a condiment in Japanese food. Symptoms of vascular blackening in the rhizome were first observed in 2005 in plants grown in British Columbia, Canada. Microscopic observations and microbial isolation from infected tissues revealed that most of the xylem tracheid cells were blackened and bacteria were consistently associated with symptomatic plants. The bacterium most frequently recovered was identified as Pectobacterium carotovorum subsp. carotovorum (Pcc) using BioLog™ and sequencing of a specific ~510 bp IGS region. Pathogen-free plants obtained using meristem-tip micropropagation were inoculated with a wasabi isolate of Pcc. Vascular blackening symptoms developed in the rhizome after 8 weeks when the rhizome was first wounded by stabbing or cutting, or if the roots were pre-inoculated with Pythium species isolated from rhizome epidermal tissues, followed by inoculation with Pcc at 1 × 10⁸ cells ml⁻¹. Xylem tracheid cells were blackened and Pcc was reisolated from all diseased tissues. The highest frequency of rhizome vascular blackening occurred at 22°C and 27°C and these tissues occasionally succumbed to soft rot at higher temperatures, but not when inoculated tissues were incubated at 10°C. The rooting medium used by growers for vegetative propagation of wasabi was shown to contain Pcc but the pathogen was not recovered from the irrigation water. Entry of Pcc through wounds on wasabi rhizomes and the host tissue response result in symptoms of vascular blackening.     

Biological Control of Sclerotinia Diseases of Rapeseed by Aerial Applications of the Mycoparasite Coniothyrium minitans

Indoor and field experiments were conducted to evaluate the efficacy of applying the mycoparasite Coniothyrium minitans to the aerial parts of rapeseed plants at the flowering stage to control sclerotinia diseases caused by Sclerotinia sclerotiorum. Under controlled conditions, a petal inoculation technique was used to determine the effect of conidial suspensions of C. minitans on suppression of sclerotinia leaf blight. Results showed that C. minitans was effective in inhibiting infection initiated by ascospores of S. sclerotiorum on flower petals by restricting mycelial growth of the pathogen. Suppression of lesion development was related to the conidial concentration of C. minitans, with larger lesions at low concentration (5×10³conidia ml⁻¹), but smaller lesions at high concentration (5×10⁴ conidia ml⁻¹ or higher). When C. minitans-treated rapeseed leaves were inoculated with mycelia of S. sclerotiorum, C. minitans failed to prevent infection of leaves, but caused a significant reduction in number of sclerotia produced on the diseased leaves. No significant difference in efficacy was detected between the two isolates of C. minitans, LRC 2137 and Chy-1, on the two rapeseed cultivars, Westar (spring type) and Zhongyou 821 (winter type). Results of field trials showed a significant reduction of stem rot of rapeseed in four (1997, 1999, 2003 and 2004) out of five years by aerial application of C. minitans, compared with controls. No significant difference in suppressive efficacy was observed between the treatments of C. minitans (10⁶ conidia ml⁻¹), C. minitans (10⁶ conidia ml⁻¹) + benomyl (50 μg ml⁻¹) and benomyl (100 μg ml⁻¹) in 2003, and between the treatments of C. minitans (10⁶ conidia ml⁻¹), C. minitans (10⁶ conidia ml⁻¹) + vinclozolin (100 μg ml⁻¹) and vinclozolin (500 μg ml⁻¹) in 2004. Sclerotia of S. sclerotiorum collected from diseased plants in plots treated with C. minitans in 1999, 2000 and 2003, or with C. minitans + benomyl in 2003 were infected by C. minitans at frequencies ranging from 21.3 to 54.5%. This study concludes that aerial spraying of C. minitans is an effective method for controlling sclerotinia diseases of rapeseed.     

Use of sclerotia of<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> for efficient production of conidia of<Emphasis Type="Italic">Coniothyrium minitans</Emphasis> in liquid culture

A study was conducted to determine the feasibility of using sclerotia ofSclerotinia sclerotiorum for producing conidia ofConiothyrium minitans in liquid culture. The medium (SST) was made of water containing 2.0, 1.5, 1.0 or 0.5% (w/v) ground sclerotia ofS. sclerotiorum and 100 μgl ⁻¹ thiamine hydrochloride (HCl). One ml of conidial suspension (2 × 10⁷ conidia ml⁻¹) ofC. minitans LRC 2534 was inoculated into 100 ml of SST medium or control (thiamine HCl in water) and incubated at 20 ± 2°C on a shaker at 200 rpm. Subsamples were removed periodically and examined under a compound microscope. Conidia in the SST media germinated within 24 h, developed into branched hyphae within 48 h, produced pycnidia after 3–4 days, and the pycnidia released mature conidia after 7 days. Production of conidia varied with the concentration of sclerotia in the SST medium. It was lower (3.6 × 10⁶ conidia ml⁻¹) at 0.5% but higher (1.2 × 10⁸ conidia ml⁻¹) at 2%. The new conidia were viable and the colonies developing from them showed the original morphological characteristics. It was concluded that using SST liquid medium as a substrate for mass production of conidia ofC. minitans has potential for use in commercial development of this mycoparasite as a biocontrol product. http:www.phytoparasitica.org posting Jan. 23, 2007.     

Evaluation of nematicidal properties of saponins from <Emphasis Type="Italic">Medicago</Emphasis> spp.

The nematicidal activity of saponins from Medicago arborea (tops), M. arabica (tops and roots) and M. sativa (tops and roots) against the plant-parasitic nematode Xiphinema index was investigated. Nematicidal activity of related prosapogenins and sapogenins on X. index is also described. Saponins from Medicago spp. at different concentrations were all nematicidal, those from M. arborea tops being the less effective. In general, saponins induced 100% mortality at 500 μg ml⁻¹ between 8 and 48 h, while prosapogenins resulted in toxicity starting at 125 μg ml⁻¹. Differences in the effects on X. index induced by prosapogenins and sapogenins were less pronounced, although prosapogenins displayed a larger range of activity. Assays with purified sapogenins demonstrated the relationship of the observed nematicidal activity of M. sativa and M. arborea to the content of the main aglycones (medicagenic acid and hederagenin, respectively) in the saponin extracts. Hederagenin displayed the highest bioactivity, giving 38% mortality after 1 h at 125 μg ml⁻¹.     

Note: Evaluation of antibacterial activity of essential oil of<Emphasis Type="Italic">Rosa damascena</Emphasis> on<Emphasis Type="Italic">Erwinia amylovora</Emphasis>

Significant antibacterial activity was observed in the essential oil (E.O.) ofRosa damascena Mill. and the Minimum Bactericidal Concentration (MBC) of the E.O. was determined as 1386.5 μg ml⁻¹ forErwinia amylovora, the causal agent of fire blight disease. Ooze formation on immature pears and lesion formation in artificially inoculated shoots were completely (100%) prevented by the essential oil ofR. damascena (1500 μg ml⁻¹), essential oil ofThymbra spicata var.spicata (500 μg ml⁻¹) and streptomycin (100 μg ml⁻¹). Copper oxychloride plus maneb significantly reduced ooze formation and lesion formation, but the control was less than that obtained with the essential oils or streptomycin. The essential oil ofR. damascena may be a useful natural bactericide for the control of the fire blight pathogen,E. amylovora. http://www.phytoparasitica.org posting July 14, 2004.     

<Emphasis Type="Italic">Ulocladium atrum</Emphasis> as a biological control agent for white mold of bean caused by<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>

Field studies were conducted near Lethbridge, Alberta, Canada, in 2001, 2004 and 2005 to determine the efficacy of the antagonistic fungusUlocladium atrum for control of white mold of bean caused bySclerotinia sclerotiorum. Results of the 3 years of field trials showed that, compared with the untreated control, foliar application of a spore suspension ofU. atrum (300 ml m⁻² of 10⁶ spores ml⁻¹ suspension) significantly reduced incidence and severity of white mold, increased seed yield and reduced contamination of bean seed by sclerotia ofS. sclerotiorum. The level of control of white mold observed in the treatment ofU. atrum was similar to that of the mycoparasitic fungusConiothyrium minitans, but lower than the fungicide treatments of Ronilan (vinclozolin) at the rate of 1200 g ha⁻¹ per application in 2001, or Lance (boscalid) at the rate of 750 g ha⁻¹ per application in 2004 and 2005. The potential for use ofU. atrum as a biological control agent for sclerotinia diseases is discussed. http://www.phytoparasitica.org posting Nov. 12, 2006.     

The potential of combining <Emphasis Type="Italic">Pasteuria penetrans</Emphasis> and neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) formulations as a management system for root-knot nematodes on tomato

Initial applications of 10⁴ spores g⁻¹ of Pasteuria penetrans, and dried neem cake and leaves at 3 and 2% w:w, respectively, were applied to soil in pots. Juveniles of Meloidogyne javanica were added immediately to the pots (500, 5,000 or 10,000) before planting 6-week-old tomato seedlings. The tomatoes were sampled after 64 days; subsequently a second crop was grown for 59 days and a third crop for 67 days without further applications of P. penetrans and neem. There was significantly less root-galling in the P. penetrans combined with neem cake treatment at the end of the third crop and this treatment also had the greatest effect on the growth of the tomato plants. At the end of the third crop, 30% of the females were infected with P. penetrans in those treatments where spores had been applied at the start of the experiment. The effects of neem leaves and neem cake on the nematode population did not persist through the crop sequences but the potential for combining the amendments with a biological control agent such as P. penetrans is worthy of further evaluation.     

Induced resistance to<Emphasis Type="Italic">Cladosporium cucumerinum</Emphasis> in cucumber by pectinases extracted from<Emphasis Type="Italic">Penicillium oxalicum</Emphasis>

Pectinases extract (PE) from the fermentation product ofPenicillium oxalicum BZH-2002 was tested for its ability to induce protection against scab caused byCladosporium cucumerinum on cucumber (Cucumis sativus L.) plants. Seedlings with one true leaf were sprayed with various concentrations of PE (10–200 units ml⁻¹) 3 days before inoculation withC. cucumerinum. Results showed that the induced local protection against the pathogen was dose-dependent when the concentrations of PE were between 20 and 120 units ml⁻¹; systemically induced resistance against the pathogen was not observed. Boiled PE had a slight effect on disease reduction. Commercial pectinases prepared fromAspergillus niger showed lower protection against scab compared with PE when they were used at the same concentration of enzyme activity. No inhibitory activity was observed on conidial germination or germ-tube growth ofC. cucumerinum. PE was further evaluated for its enhancement of defense-related enzymes. Peroxidase (PO), polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) increased in cucumber seedlings after treatment with PE. PO and PPO remained at a higher level in PE-pretreated seedlings throughout the experiment period whether pathogen-inoculated or non-inoculated, whereas PAL activity began to decrease 2 days after PE treatment. http://www.phytoparasitica.org posting July 14, 2004.     

<Emphasis Type="Italic">Pseudomonas</Emphasis> sp. LSW25R,antagonistic to plant pathogens,promoted plant growth,and reduced blossom-end rot of tomato fruits in a hydroponic system

A Gram-negative rhizobacterial isolate (LSW25) antagonistic to Pseudomonas corrugata, a vein necrosis pathogen of tomato, and promotes the growth of tomato seedlings was isolated from surface-sterilised tomato roots. A spontaneous rifampicin-resistant mutant (LSW25R) was selected to facilitate its tracking, and identified as Pseudomonas sp. and named as Pseudomonas sp. LSW25R (LSW25R), based on its sequences of the internal transcribed spacer (ITS) region and 16 S rRNA gene. LSW25R inhibited mycelial growth of 12 other plant fungal pathogens such as Botrytis cinerea on V8 agar plates. By using a scanning electron microscope, LSW25R colonised not only the root surface around the natural aperture of tomato radicles but also under epidermal cells like endophytic bacteria. LSW25R successfully colonised the roots of tomato, eggplant and pepper seedlings, significantly promoted the fresh weight, height and dry matter of tomato plants at 10⁸ cfu·ml⁻¹, and increased the plant growth of eggplants and peppers at 10⁴ cfu·ml⁻¹, suggesting that the optimal population density of LSW25R for growth promotion varies from species to species. Moreover, densities of LSW25R inside roots and the lowest leaf of tomato plants were > 9.3 × 10³ cfu·g⁻¹. Although the growth promotion of tomato by LSW25R was observed under N- or Ca-deficient conditions as well as a standard nutrient condition, the uptake of calcium was increased only under the standard nutrient condition. In a hydroponic system, LSW25R not only successfully colonised the rhizosphere during cultivation due to its broad spectrum of antifungal activity and endophytic colonisation, but also reduced blossom-end rot of tomato fruits presumably through increasing calcium uptake.     

Efficacy and dose–response relationship in biocontrol of Fusarium disease in maize by Streptomyces spp.

Two isolates of Streptomyces spp. DAUFPE 11470 and DAUFPE 14632 were evaluated to determine the antagonist–pathogen inoculum concentration relationship under greenhouse conditions. Pathogen and antagonist concentration, significantly (P < 0.05) affected development of Fusarium disease in maize with a significant interaction between pathogen and antagonist concentration. Dose–response relationship also differed significantly (P < 0.05) between the two isolates, but both isolates demonstrated effective control of Fusarium disease, regardless of pathogen concentration. The isolate DAUFPE 11470 provided the most effective control. The lowest value for disease incidence occurred at low pathogen (10³ chlamydospore g⁻¹ soil) and high antagonist concentration (10⁶ cfu ml⁻¹) for both isolates. The disease incidence for control plants ranged from 19% to 76%. However, in relation to control the lowest disease reduction occurred at low pathogen (10³ chlamydospore g⁻¹ soil) and high antagonist concentrations (10⁶ cfu ml⁻¹). These reductions were 10.6% and 13% for DAUFPE 14632 and DAUFPE 11470, respectively. The highest disease reductions, in relation to control plants, occurred at high pathogen (10⁶ chlamydospore g⁻¹ soil) and antagonist (10⁶ cfu ml⁻¹) concentrations for both isolates. These values were 55% and 62.2% for DAUFPE 14632 and DAUFPE 11470, respectively. The chlamydospore germination of Fusarium moniliforme was affected by glucose addition, antagonist isolates and type of inoculum. The lowest chlamydospore germination was observed with bacterial suspensions of the isolates, for all glucose additions. The results suggested that both Streptomyces spp. isolates were effective at different doses as biocontrol agents against F. moniliforme. Also, there was evidence for at least two mechanisms of biocontrol and that apparently, both isolates showed the same mechanisms of biocontrol action related to production of bioactive compounds and competition for carbon. Further studies will be developed to improve the level and effectiveness of control by these isolates.     

Efficacy of bacterial antagonists and different commercial products against Fusarium wilt on rocket

Seven experimental trials were carried out to evaluate the efficacy of the bacterial strains Achromobacter xylosoxydans AM1 and Serratia sp. DM1 obtained from suppressive soils and from soilless used rockwool substrates (Pseudomonas putida FC6B, Pseudomonas sp. FC7B, Pseudomonas putida FC8B, Pseudomonas sp. FC9B and Pseudomonas sp. FC24B) against Fusarium wilt on rocket caused by Fusarium oxysporum ff. spp. raphani and conglutinans. Along with these strains, two commercial bioproducts (RootShield—Trichoderma harzianum T22; Cedomon—Pseudomonas chlororaphis MA342) were also tested. Different application strategies such as soil treatment (trials I to VI; 10⁷ and 10⁸ CFU ml⁻¹) and root dipping (trial VII; 10⁸ and 10⁹ CFU ml⁻¹) were performed in a glasshouse in order to test the efficacy of the bacterial strains against Fusarium oxysporum ff. spp. raphani and conglutinans. The lowest disease incidence (16.7%) was observed with the application of Achromobacter sp. AM1, Serratia sp. DM1 at 10⁸ CFU ml⁻¹ and Pseudomonas sp. FC9B at 10⁷ CFU ml⁻¹ against F. oxysporum f. sp. conglutinans (experiment I). Maximum plant biomass (5.0 g/plant) was registered in Serratia sp. DM1 at 10⁸ CFU ml⁻¹ treated plants in trial IV. The trials against F. oxysporum f. sp. raphani (experiment II) showed that the application of Pseudomonas sp. FC7B, P. putida FC8B at 10⁸ CFU ml⁻¹ and P. chlororaphis MA342 at 7.5 × 10⁶ CFU ml⁻¹ significantly reduced disease incidence to values ranging between 87% and 92%. The highest plant biomass was recorded with the application of Achromobacter sp. AM1 and P. putida FC6B at 10⁷ CFU ml⁻¹ (3.9 to 4.2 g) carried out 7 days before the artificial inoculation of the pathogens (trial IV). The present study showed the potential biocontrol activity of the bacterial strains Achromobacter sp. AM1, Serratia sp. DM1 and Pseudomonas sp. FC9B against F. oxysporum f. sp. conglutinans and of Pseudomonas sp. FC7B, P. putida FC8B, P. chlororaphis MA342, Achromobacter sp. AM1 and P. putida FC6B against F. oxysporum f. sp. raphani. Growth-promoting activity of biocontrol bacteria used during the trials was observed.     

Weed hosts of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in cotton fields in Turkey and characterization of <Emphasis Type="Italic">V. dahliae</Emphasis> isolates from weeds

A weed survey conducted in 2004 and 2005 in Aydin province of Turkey showed that Solanum nigrum, Xanthium strumarium, Amaranthus retroflexus, Portulaca oleracea, Sonchus oleraceus and Datura stramonium were the most prevalent weeds in the cotton fields exhibiting Verticillium wilt. Verticillium dahliae Kleb. was recovered from A. retroflexus and X. strumarium in those cotton fields. This is the first report of V. dahliae occurring naturally in A. retroflexus in Turkey. Pathogenicity tests on cotton and weeds showed that the virulence of V. dahliae isolates from weeds was higher on cotton plants than on weeds, with the disease severity ranging from 31.7% to 98.0%. Disease severity of V. dahliae isolates was 54.7–93.9% on eggplant, 23.7–51.6% on cucumber and 11.0–16.4% on tomato, whereas it did not cause any disease symptoms, or only low levels, on pepper and bell pepper. Two vegetative compatibility groups (VCGs) were identified among seven tested weed isolates: VCG2A (two isolates) and VCG2B (three isolates) using international reference strains.     

Pathogenicity and mycotoxin production by <Emphasis Type="Italic">Fusarium proliferatum</Emphasis> isolated from onion and garlic in Serbia

Fusarium proliferatum can occur on a wide range of economically important vegetable plants but its role in disease is not always well established. In 2000 and 2001, from forty-one field samples of wilting onion and garlic plants in Serbia, F. proliferatum as the predominant fungal species was isolated from root and bulbs. Seventy isolates were firstly characterized for their sexual fertility and were shown to be mostly members of Gibberella intermedia (sixty-seven of seventy isolates, the remaining three isolates were unfertile), the sexual stage of F. proliferatum (syn. mating population D of G. fujikuroi complex). A selected set of eleven F. proliferatum isolates from both hosts were also tested for their pathogenicity and toxigenicity. Although onion and garlic plants were susceptible to all isolates, onion plants showed a significantly higher disease severity index. Six of the eleven isolates of F. proliferatum produced fumonisin B₁ from 25 to 3000 μg g⁻¹, and beauvericin from 400 to 550 μg g⁻¹; ten isolates produced fusaric acid from 80 to 950 μg g⁻¹ and moniliformin from 50 to 520 μg g⁻¹. Finally, all isolates produced fusaproliferin up to 400 μg g⁻¹. These results confirm F. proliferatum as an important pathogen of garlic and onion in Europe and that there is a potential mycotoxin accumulation risk in contaminated plants of both garlic and onion.     

Seedborne fungi detected on stored solanaceous berry seeds and their biological activities

We isolated 629 fungi from 1296 berry seeds of solanaceous plants, including tomato (Lycopersicon esculentum), eggplant (Solanum melongena), bell pepper (Capsicum annuum), and red pepper (Capsicum annuum var. annuum) preserved for long and short terms. The isolates were classified into 22 genera excluding unidentified fungi, and the fungal floras were divided into two types: the tomato–eggplant and pepper groups. The results of cluster analysis with unweighted pair-group method with arithmetic average also supported these groups. Most tomato seeds infested with Geotrichum candidum germinated and grew the same as uninfested seeds. Cladosporium sphaerospermum and Arthrinium sp. isolated from eggplant seeds strongly suppressed germination, and Penicillium variabile suppressed seminal root elongation on eggplant. Alternaria alternata, Botrytis cinerea, and Myrothecium verrucaria detected from red pepper or bell pepper seeds were pathogenic to the fruits and the seedlings after artificial inoculation.     

Improved control of moldy-core decay (<Emphasis Type="Italic">Alternaria alternata</Emphasis>) in Red Delicious apple fruit by mixtures of DMI fungicides and captan

The sterol biosynthesis inhibitors bromuconazole and difenoconazole and tank mixes of each fungicide with captan were applied to apples and evaluated as controls for moldy-core and fruit decay caused by Alternaria alternata. Effectiveness of a mixture of bromuconazole and captan in controlling colonization by the fungus was also evaluated. Decay formation by A. alternata on mature detached fruits was partially inhibited by bromuconazole at 0.5 μg ml⁻¹ and was completely inhibited at 50 μg ml⁻¹; it was significantly affected by either bromoconazole at 5 μg ml⁻¹ or captan at 1,250 μg ml⁻¹, and was completely inhibited by their mixture. In general, three foliar applications of bromuconazole or difenoconazole in the field, during the bloom period, reduced the numbers of infected fruits by 40–60% compared with untreated control trees. However, tank mixes of either fungicide with captan improved control of moldy-core in fruits at harvest. Tank mixtures of bromuconazole and captan also significantly reduced the percentage of fruits colonized by A. alternata when sampled at various days after full bloom. Artificial inoculations in the orchard at full bloom did not change the inhibitory effects of the tank mixtures. Large-scale demonstration trials in commercial orchards supported these findings. The inhibitory effects of tank mixes on decay development in detached fruits, and on moldy-core in the field indicate that a control programme based on mixtures of either bromuconazole or difenoconazole with captan during the bloom period can effectively reduce moldy-core on Red Delicious apples.     

Relationship between the incidence of latent infections caused by <Emphasis Type="Italic">Monilinia</Emphasis> spp<Emphasis Type="Italic">.</Emphasis> and the incidence of brown rot of peach fruit: factors affecting latent infection

Five field experiments were performed in commercial orchards located in Lleida (Spain) over three growing seasons, 2000–2002, in order to estimate the relationship between the incidence of latent infection caused by Monilinia spp. in peaches and the incidence of post-harvest brown rot. No latent infection was recorded at popcorn and the maximum incidence occurred pre-harvest; in some orchards a second peak was detected during the pit hardening period. Monilinia laxa is the most prevalent species isolated from peaches with brown rot. There was a positive correlation between the incidence of latent infection and that of post-harvest brown rot. The average incidence of latent infection during the crop season explained 55% of the total variation in the incidence of post-harvest brown rot. The effect of temperature (T) and duration of wetness (W) on the incidence of latent infection in peach and nectarine orchards was analysed using multiple regression. The regression analysis indicated that T and W jointly explained 83% of the total variation in the incidence of latent infection. The model predicts no latent infections when T < 8°C, and >22 h wetness are required when T = 8°C but only 5 h at 25°C are necessary for latent infection to occur. The incidence of brown rot and latent infection of peaches caused by M. laxa under controlled experimental conditions were also affected by T and W, as well as by fruit maturity and inoculum concentration. Latent infections were produced in fruit when T was not suitable for the development of brown rot symptoms. In these experiments more than 4–5 h of daily wetness were required after embryo growth in fruit sprayed to run-off with an inoculum concentration higher than 10⁴ conidia ml⁻¹ of M. laxa for brown rot and latent infections to develop. The fitted model obtained from the field data was able to predict the observed data obtained under controlled environmental conditions.     

Host-pathogen interaction of root-knot nematode <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> on pepper in the southeast of Spain

The feeder roots of pepper plants (cv. California Wonder) in Campo de Cartagena (southeast Spain) were found to be severely infected by Meloidogyne incognita. Morphometric traits, differential host test and DNA analysis based on PCR were used to characterize the nematode. Naturally and artificially infected pepper plants showed severe yellowing and stunting, with heavily deformed and damaged root systems. Root galls were spherical and commonly contained more than one female and egg masses with eggs. Typical giant cells with a granular cytoplasm and many hypertrophied nuclei were observed in histological preparations. The relationship between initial nematode population density (Pi) and pepper plant growth was tested in greenhouse experiments with inoculum levels that varied from 0 to 64 eggs and second-stage juveniles (J₂) ml⁻¹ soil. A Seinhorst model was fitted to plant height and top fresh weight data of inoculated and non-inoculated plants. The tolerance limit with respect to plant height and fresh top weight of pepper to M. incognita was estimated as 0.85 eggs and J₂ ml⁻¹ soil. The minimum relative values (m) for plant height and top fresh weight were 0.15 and 0.16, respectively, at Pi ≥ 64 eggs and J₂ ml⁻¹ soil. The maximum nematode reproduction rate (Pf/Pi) was 315.4 at an initial population density (Pi) of 4 eggs and J₂ ml⁻¹ soil. The obtained results could be used as a base to establish field experiments that allow strategies to prevent surpassing the threshold of nematodes in fields that are infested.     

Host-parasite relationships in tobacco plants infected with a root-knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) population from the Azores

During a nematode survey, severe infections of tobacco feeder roots and heavy soil infestations byMeloidogyne incognita race 1 were found in S. Miguel (Azores islands, Portugal). This is the first record ofM. incognita infection of tobacco in Azores. Morphology of various life stages, analysis of the esterase electrophoretic pattern and differential host tests were used for nematode characterization and identification. Nematode-induced mature galls were spherical and/or ellipsoidal and usually contained more than one female, males and egg masses with eggs. Feeding sites were characterized by the development of giant cells that contained granular cytoplasm and many hypertrophied nuclei. Giant cell cytoplasm was aggregated along a thickened cell wall. Vascular tissues within galls appeared disorganized. The relationship between the initial nematode population density and growth of tobacco plants was tested in a glasshouse experiment in which inoculum levels varied from 0 to 512 eggs and juveniles (J₂) cm⁻³ of soil. Seinhorst’s model was fitted to height and top fresh weight data of the inoculated and control plants. Tolerance limits with respect to plant height and fresh top weight of tobacco cv. ‘Erzegovina’ plants toM. incognita race 1 were estimated as 1.25 eggs and J₂ cm⁻³ of soil. The maximum nematode reproduction rate was 404.7 at an initial population density of 4 eggs and J₂ cm⁻³ of soil. http://www.phytoparasitica.org posting March 2, 2004.     

Occurrence and distribution of resistance to QoI fungicides in populations of Podosphaera fusca in south central Spain

Cucurbit powdery mildew caused by Podosphaera fusca limits crop production in Spain. Since its management is strongly dependent on chemicals, the rational design of control programmes requires a good understanding of the fungicide resistance phenomenon in field populations. Fifty single-spore isolates of P. fusca were tested for sensitivity to three quinone-outside inhibiting (QoI) fungicides: azoxystrobin, kresoxim-methyl and trifloxystrobin. Minimum inhibitory concentration (MIC) values for QoI-sensitive isolates were found to range from 0.25 to 10 μg ml⁻¹ for azoxystrobin to 5–25 μg ml⁻¹ for kresoxim-methyl, using a leaf disc-based bioassay. High levels of cross-resistance to QoI fungicides were found. Eleven isolates showed resistance to the three QoI fungicides tested with MIC and EC₅₀ values >500 μg ml⁻¹ resulting in RF values as high as >715 and >1000 for trifloxystrobin and azoxystrobin, respectively. A survey of P. fusca QoI resistance was carried out in different provinces located in the south central area of Spain during the cucurbit growing seasons in 2002, 2003 and 2004. Examination of a collection of 250 isolates for QoI resistance revealed that 32% were resistant to the three fungicides tested; the provinces of Ciudad Real, Córdoba and Murcia being the locations with the highest frequencies of resistance (44–74%). By contrast, no resistance was found in Badajoz, and relatively low frequencies were observed in Almería and Valencia (10–13%). Nearly 50% of resistant isolates were collected from melon plants. Based on these data, recommendations about the use of QoI fungicides for cucurbit powdery mildew management in the sampled areas are made.