Nematicidal efficacy of isothiocyanates against root-knot nematode Meloidogyne javanica in cucumber

Isothiocyanates (ITCs), a series of new nematicides of the -NCS group, were evaluated for their efficacy against root-knot nematode Meloidogyne javanica. Of the compounds tested, AllylITC, AcITC, EtITC, BzTC, BzITC, 1-PEITC and 2-PEITC showed in vitro irreversible nematicidal activity against second-stage juveniles of M. javanica, following exposure for 72 h at concentrations as low as 5 μg mL⁻¹. When exposed to AllylITC, AcITC and EtITC at lower concentrations, motile juveniles also became irreversibly immobile in 3 days, with a LC₅₀ value at 2.76, 2.53 and 3.05 μg mL⁻¹, respectively. In the pot experiments, 1.0 ml AllylITC and 1.1 ml AcITC per kg of soil controlled M. javanica, similarly to or better than metam sodium at its recommended dose. Similar results were obtained in the field experiments using 1.0 kg AllylITC or 1.0 kg AcITC ha⁻¹. Based on the results of this study, AllylITC and AcITC have potential to be used as new bio-fumigant nematicides.     

Wilt disease management and enhancement of growth and yield of Cajanus cajan (L) var. Manak by bacterial combinations amended with chemical fertilizer

Root nodulating Sinorhizobium fredii KCC5 and Pseudomonas fluorescens LPK2 were isolated from nodules of Cajanus cajan and disease suppressive soil of tomato rhizosphere, respectively. Both strains produced IAA, siderophore, solubilized insoluble phosphate, showed chitinase and β-1,3-glucanase activities, and strongly inhibited the growth of Fusarium udum. It also caused degradation and digestion of cell wall components, resulting in hyphal perforations, empty cell (halo) formation, shrinking and lysis of fungal mycelia along with significant degeneration of conidia. LPK2 produced volatile cyanogen (HCN). Combinations of S. fredii KCC5 and P. fluorescens LPK2 with half dose of chemical fertilizers showed a significant increase in seed germination (94%) while seed germination with co-inoculated strains (KCC5 + LPK2), KCC5 and LPK2 alone was 90, 84 and 82% respectively as compared to control 77%. After 120 days of sowing, per plant number of pods, nodules, shoot length, root length, shoot weight and root weight were greater for the combination with half dose of chemical fertilizers compared to the control. Combinations of S. fredii KCC5 and P. fluorescens LPK2 with half dose of chemical fertilizers resulted in an 82% increase in grain yield per hectare compared to the control. Both strains KCC5 and LPK2 led to proto-cooperation as evidenced by synergism, aggressive colonization of the roots, and enhanced growth, suggesting potential biocontrol efficacy against Fusarium wilt in C. cajan.     

Mycoparasitic Trichoderma viride as a biocontrol agent against Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes and as a growth promoter of soybean

Trichoderma viride was proved as an effective biocontrol agent against two fungal pathogens, Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes, infecting soybean. During an in vitro biocontrol test, Trichoderma showed mycoparasitism and destructive control against the tested fungal pathogens. Both the pathogens significantly influence the germination and P. arrhenomanes had a severe effect (only 5% germination). The root system of the soybean plant was poorly developed due to the infection and it exerted a negative influence on the nodulation and further growth phases of the plant. During pot assay along with biocontrol activity, Trichoderma showed growth promoting action on the soybean plant. Trichoderma enhanced growth of shoot and root systems and fruit yield after 12 weeks of growth. Pythium and Fusarium infected plants treated with Trichoderma had ∼194% and 141% more height than pathogens alone. The fruit yield treated with Trichoderma was ∼66 per plant whereas the yield was only 41 for a control plant. The plants infected with Pythium and Fusarium and treated with Trichoderma had fruit yields of 43 and 53 respectively and those were 5 and 1.6 times higher than plants infected with pathogens.     

Actinomycetes mediated biochemical responses in tomato (Solanum lycopersicum) enhances bioprotection against Rhizoctonia solani

Six actinomycetes isolates, namely Streptomyces toxytricini vh6, Streptomyces flavotricini vh8, S. toxytricini vh22, Streptomyces avidinii vh32, Streptomyces tricolor vh85 and vh41, an isolate of an unknown species of Actinomycetales, were tested for their efficacy in protecting tomato (Solanum lycopersicum) against Rhizoctonia solani under green house conditions. Actinomycetes treated plants showed better growth in terms of high chlorophyll content, higher phenylalanine ammonia lyase (PAL) activity and high total phenolic content. Qualitative and quantitative estimation of phenolic compounds from tomato leaves showed significant accumulation of six phenolic acids, gallic (29.02 μg g⁻¹ fresh leaf wt), ferulic (11.44 μg g⁻¹ fresh wt), cinnamic (56.84 μg g⁻¹ fresh wt), gentisic (24.19 μg g⁻¹ fresh wt), chlorogenic acid (1.72 μg g⁻¹ fresh wt) and salicylic (0.39 μg g⁻¹ fresh wt) acid, in actinomycetes treated plants. Biochemical profiling, when correlated with plant mortality in actinomycetes treated and untreated plants, indicated that isolates vh6 and vh8 offered 44.55% and 40.14% disease reductions, respectively compared to the control. These results established that these organisms have the potential to act as biocontrol agents.     

Biocontrol of Rhizoctonia solani and Sclerotium rolfsii on tomato by delivering antagonistic bacteria through a drip irrigation system

In a 2-year assessment carried out on tomato crops of Central and Southern Italy, a high incidence of Rhizoctonia solani and Sclerotium rolfsii was found. These fungal pathogens attack horticultural crops and are responsible for severe crown and stem rot. Because of technical, economical and environmental issues their chemical control is an arduous task. To find alternative and eco-compatible control methods, the effectiveness of two new antagonistic bacterial isolates (Burkholderia cepacia, T1A-2B, and Pseudomonas sp., T4B-2A), previously selected from suppressive organic amendments, were tested on tomato plants grown under both growth chamber and field conditions. The potential antagonists were compared with two commercial biofungicides, based on Bacillus subtilis (BSF4) and Trichoderma asperellum (TV1), and four synthetic fungicides (tolclofos-methyl, azoxystrobin, fosetyl-Al and fosetyl-Al + propamocarb). In 2-year field experiments carried out on tomato plants, the biocontrol bacteria as well as the other treatments were applied to the soil, proximal to the plant crowns and main roots, by means of an effective and specific system of drip irrigation. In all the experiments the novel selected biocontrol bacteria significantly reduced both incidence and severity of the diseases caused by S. rolfsii or R. solani, with results demonstrating effectiveness equal to TV1, better than BSF4 and comparable with the synthetic fungicides, except for tolclofos-methyl which was the most effective treatment. In field experiments, carried out for two consecutive years, isolate T1A-2B reduced up to 58.33% and up to 63.8% the severity of the diseases caused by S. rolfsii and R. solani respectively; whereas isolate T4B-2A gave reduction of S. rolfsii and R. solani diseases severity up to 73.2% and up to 62.7%, respectively.     

Efficacy of plant extracts and biocontrol agents against Pythium aphanidermatum inciting chilli damping-off

The fungitoxic effects of 66 medicinal plants belonging to different families were evaluated in vitro on Pythium aphanidermatum, the causal agent of chilli damping-off. Of these, Zimmu leaf extract (Allium sativum L. × Allium cepa L.) showed the highest inhibition of mycelial growth of P. aphanidermatum (13.7 mm). The antimicrobial compounds were isolated from Zimmu leaf extract and 22 compounds were identified through gas chromatography mass spectroscopy (GC–MS). Biocontrol agents Trichoderma viride and Pseudomonas fluorescens and Zimmu extract were also tested alone and together in vitro and in vivo experiments for control of P. aphanidermatum. The in vitro studies revealed that combination of T. viride + P. fluorescens + Zimmu leaf extract showed the highest mycelial growth inhibition over the control. Both antagonists were compatible with each other and with Zimmu leaf extract. The pot culture studies revealed that seed treatment with combined application of T. viride + P. fluorescens + Zimmu leaf extract was superior in reducing the pre and post-emergence damping-off incidence (8.3 and 17.0%, respectively), and increased the plant growth and yield (shoot length and root length of 13.7 and 6.3 cm, 146 g/plant, respectively) of chilli when compared to control.     

Rapid detection of Meloidogyne spp. by LAMP assay in soil and roots

Loop-mediated isothermal amplification (LAMP), a novel DNA amplification technique, has been used to detect a variety of pathogens including viruses, fungi, bacteria and parasites. However, diagnosis of sedentary plant-parasitic nematode (PPN) species has not yet been attempted. In this study, we developed a universal LAMP set (RKN-LAMP) for the diagnosis of four common Meloidogyne species (Meloidogyne incognita, M. arenaria, M. javanica and M. hapla), and M. incognita-specific LAMP set (Mi-LAMP). In both assays, a typical ladder-like pattern on gel electrophoresis was observed in all positive samples but not in the negative controls. Amplification products were further confirmed using restriction analysis of the Hpa II enzyme, detection by visual inspection using SYBR Green I and the lateral flow dipstick (LFD) assay. The two LAMP sets were specifically able to detect four common Meloidogyne species and M. incognita populations having several different geographical origins and pathotypes. No cross reaction with DNA of other PPNs was observed. Sensitivity of the RKN- and Mi-LAMP was 10 and 100 fg of pure genome DNA respectively. Both LAMP sets could also amplified crude DNA isolated from the galled root tissue and from soil containing juveniles of M. incognita. The RKN- and Mi-LAMP sets offer the advantages of simplicity, rapidity and cost effectiveness. Both LAMP sets will be instrumental for the diagnosis of Meloidogyne spp. by local extension and regulatory personnel.     

Use of two PGPR strains in the integrated management of blast disease in rice (Oryza sativa) in Southern Spain

Biocontrol capacity of two plant growth-promoting rhizobacteria (PGPR) strains, against blast disease in rice paddy fields in Southern Spain was studied in three cropping seasons. Both strains (Pseudomonas fluorescens Aur 6 and Chryseobacterium balustinum Aur 9) had already shown biocontrol capacity against pathogens, ability to induce systemic resistance against leaf pathogens and against salt stress in different plant species. Bacterial treatments were carried out on seeds and/or on leaves. Strains were inoculated individually and in combination. Protection against natural disease incidence was evaluated, and rice production and quality measured in 2005 and 2006 trials. In 2004, natural disease incidence was low (between 0.1% and 0.35% of damaged leaf surface) due to environmental conditions; under these conditions, both strains significantly protected plants against rice blast. In 2005, disease incidence was higher than in 2004, reaching higher values of affected leaf surface in controls. In these conditions, each strain individually protected rice against rice blast, although the combination of both strains was the most effective treatment. All three treatments (Aur 6, Aur 9 and Aur 6 + Aur 9) reached 50% protection in panicles, with Aur 9 being the most effective. In 2006, the most effective treatment was the combination of both strains on leaves in three physiological stages, suggesting a biocontrol mediated protection. On the other hand, when bacteria were applied to seeds, disease incidence decreased up to 50%, suggesting induction of systemic resistance. Finally, a direct relation between protection mediated by the PGPR and the increase in rice productivity (mT/ha) and quality (weight of 1000 seeds and number of intact grains after milling) was found.     

Effects of leaf scales of different pineapple cultivars on the epiphytic stage of Fusarium guttiforme

The fungus Fusarium guttiforme (Syn. F. subglutinans f. sp. ananas) is responsible for fusariosis, one of the main phytosanitary threats to pineapple (Ananas comosus var. comosus). A structural study comparing epidermal differences in pineapple cultivars resistant and susceptible to fusariosis was performed, relating properties of the epidermis to known susceptibility to the disease. The basal, non-chlorophylled, portions of mature leaves of pineapple plants were analyzed by light and electron microscopy. All cultivars showed common morpho-anatomic aspects characteristic of Bromeliaceae, such as scutiform scales and unstratified epidermis. However, cultivar Vitoria (resistant) had less scales than cultivars Smooth Cayenne (susceptible, intermediate severity) and Perola (susceptible, with extreme severity of fusariosis symptoms). Inoculation of conidia suspension (10⁵ conidia ml⁻¹) of the fungus F. guttiforme to leaves and harvesting 24 h later yielded numbers of viable colonies related to the density of leaf scales. This suggests that scales can act as havens for fungal conidia and favour the epiphytic stage of the fungus on pineapple plants, and are involved in the interaction of plant and pathogen. A reduction in scale numbers was related to lower infection levels and is relevant to the future breeding programme for development of new pineapple cultivars resistant to fusariosis and their involvement in integrated control strategies.     

Screening of three sweet potato (Ipomoea batatas L.) cultivars for resistance to different virulence groups of root-knot nematodes (Meloidogyne spp.) under controlled conditions

The susceptibility of three sweet potato cultivars (Ipomoea batatas L.) C4, TIS 3290 and TIS 9162 was evaluated against 156 isolates of Meloidogyne spp. with the aim to include resistant/tolerant sweet potato cultivars in a crop rotation scheme for the management of root-knot nematodes. The nematode isolates corresponded to races 1, 2 and 3 of Meloidogyne arenaria (n = 7), races 1, 2, 3 and 4 of M. incognita (n = 131) and Meloidogyne javanica (n = 18). Also, the isolates of M. incognita were differentiated in virulence groups: Pepper (n = 35), Pepper-Mi (n = 25), Tomato (n = 41) and Tomato-Mi (n = 30), depending on their ability to parasitize resistant pepper and tomato cultivars. The tested isolates of M. javanica parasitized C4 and TIS 3290, but not TIS 9162, whereas M. arenaria parasitized C4 and TIS 9162, but not TIS 3290, and M. incognita was able to parasitize the three sweet potato cultivars tested. C4 was the most susceptible cultivar to all nematode species tested, especially M. incognita, TIS 3290 was the most resistant and TIS 9162 was in between (7.2, 62.9 and 26.9% of resistant plants, respectively). Susceptibility of the sweet potato cultivars showed slight variations depending on the race or virulence group of M. incognita. The results suggest that sweet potato cultivars TIS 3290 and TIS 9162 may be used as rotation crops in fields where root-knot nematodes are present, their selection depending on the Meloidogyne isolates present. The use of resistant sweet potato cultivars would be preferably combined with other management practices to avoid virulence selection in nematode isolates.     

Endophytic Lecanicillium lecanii and Beauveria bassiana reduce the survival and fecundity of Aphis gossypii following contact with conidia and secondary metabolites

Lecanicillium lecanii and Beauveria bassiana are important entomopathogens of Aphis gossypii. Their capacity to colonize crop plants is also becoming widely recognized. Their presence in crop plants indicates the possibility of a much greater potential for contact between insect and fungus than previously recognized. The present experiment aimed to study the effects of endophytic strains of these fungi on the survival, and reproduction of A. gossypii. Contact with conidia of both fungi significantly reduced the rate and period of reproduction of A. gossypii. The culture filtrates of L. lecanii and B. bassiana significantly increased mortality and feeding-choice experiments indicate that insects may be able to detect metabolites of the fungi. The culture filtrate of L. lecanii also significantly reduced the reproduction of the aphid. The ethyl acetate and methanolic fractions of the culture filtrate and of mycelia of L. lecanii also caused significant mortality and reduced fecundity of A. gossypii. The methanolic fractions of mycelia of B. bassiana caused significant mortality of A. gossypii. The present investigations indicated that A. gossypii is affected by contact with both conidia and fungal metabolites. This broad influence indicates that these fungi may have a role in regulating insect pest populations.     

Population dynamics of Xanthomonas campestris pv. vitians on different plant species and management of bacterial leaf spot of lettuce under greenhouse conditions

The population dynamics of Xanthomonas campestris pv. vitians (Xcv) was studied both externally and internally in lettuce, tomato and pepper plants. In addition, the application of bactericides during transplant production period was carried out for the management of bacterial leaf spot of lettuce under greenhouse conditions. Epiphytic populations of Xcv were recovered on leaves of lettuce plants (10⁵ CFU/g) 5 weeks after sprayed than the other plant species when inoculated with 10⁸ CFU/ml of Xcv. When plants of each crop species infiltrated with the bacterium at 10⁵ CFU/ml, the highest populations were developed in lettuce (10⁸ CFU/cm²) followed by pepper with 10⁶ CFU/cm² and tomato with 10⁵ CFU/cm² 10-days after infiltration. Application of a mixture of Maneb and Kocide or Kocide alone as a foliar spray on lettuce significantly reduced the incidence and disease severity of bacterial leaf spot by 29 and 27% respectively. Spread of the bacterium and development of the disease may partly be managed by avoiding intercropping of these plants commonly grown in close proximity to lettuce. For the management of leaf-associated populations of Xcv in lettuce, use of a mixture of Maneb and Kocide is advocated to minimize the effect of attacks.     

Disinfection of Tomato chlorotic dwarf viroid by chemical and biological agents

We investigated the effectiveness of various chemical or biological agents in disinfecting Tomato chlorotic dwarf viroid (TCDVd), which causes extensive damage to tomato crops. Among seven tested chemicals, sodium hypochlorite (NaOCl) was the most effective in disinfecting TCDVd-contaminated scalpels with a 15-s dipping. TCDVd-contaminated scalpels were effectively disinfected by sodium hypochlorite solution at a concentration of 0.5% or more. Sodium hypochlorite at 0.25% and 0.125% could also reduce rates of infection via contaminated scalpels, suggesting that solutions of more than about 0.2% could also be used to suppress infection via TCDVd-contaminated tools. Low-pH sodium hypochlorite solution, which has powerful oxidizing activity, was found to degrade TCDVd dramatically even at low concentrations (below 0.1%). However, the effectiveness of such low-pH solutions in disinfecting TCDVd decreased to a level similar to that of high-pH solutions in the presence of plant tissue residues on tool surfaces, probably because of a reduction in cleaning activity. Although trisodium phosphate (Na₃PO₄) at a concentration of 5% was effective against TCDVd, a 2.5% solution of this chemical was not sufficiently effective against the viroid. Among the biological agents tested, crude sap of pepper plants significantly suppressed TCDVd infection.     

Control of tomato powdery mildew (Leveillula taurica) in the Comarca Lagunera, Coahuila State, Mexico, supported by the spray forecast model Tomato.PM

Tomato powdery mildew [Leveillula taurica (Lev.) Arm.] is a common disease of tomato crops in the Comarca Lagunera region of the state of Coahuila, Mexico. To control the disease, fungicides are commonly sprayed up to 18 times on a weekly basis in this region. Effectiveness of the spray forecast model Tomato.PM, developed for California conditions, to reduce fungicide spraying, was evaluated. Disease dynamics on plants treated with three different fungicides (azoxystrobin, myclobutanil and wettable sulfur), with and without model recommendations, was analyzed in three experiments at three locations. When spraying followed model recommendations, it was possible to eliminate 11-16 fungicide applications with no significant effect on fruit yield. All three fungicides reduced the severity of disease but their effects varied with the cropping cycle. A significant increase in total yield due to fungicide spraying was observed in one out of three experiments.     

Development of a formulation of Trichoderma asperellum to control black rot disease on pineapple caused by (Thielaviopsis paradoxa)

A local isolate of Trichoderma asperellum was tested for its antagonistic activity against Thielaviopsis paradoxa (telemorph = Ceratocyctis paradoxa). The highest antagonistic activity was achieved when the concentration of T. asperellum conidia was 1 × 10⁷ conidia/mL. The highest biomass and number of colony forming unit/mL of the T. asperellum peaked at 144 h after incubation in yeast waste residue medium. The minimum inhibition concentration value of the formulation was observed as 1% on growth of Th. paradoxa incubated at 28 ± 2 °C for 10 d. In the soil fungicide-screening test, the effect of concentrations 100-1600 μg/mL on mycelia growth was not significant (P < 0.05). Complete mycelial growth inhibition occurred at concentration above 52,600 μg/mL. Results of the fruit application tests clearly showed that all treated fruits were free of disease at the end of the incubation period. No significant differences (P > 0.05) in pH, total soluble solids and titratable acidity were observed between fruits treated with formulation of T. asperellum and the control formulation treated pineapples.     

The effects of nitrogen rate and the ratio of NO3:NH4 on Bemisia tabaci populations in hydroponic tomato crops

Nitrogen fertilization is one of the factors that influences Bemisia tabaci (Gennadius) population density. The aim of this study was to determine the effects of three N application rates (75, 205 and 335 mg/l) and three ratios of NO₃⁻:NH₄⁺ ions (92:8, 75:25 and 55:45) in standard nutrient solution (205 mg/l N) on the population density of B. tabaci. The experiments were conducted on spring-summer hydroponic crops of tomato. The effect of plant stratum on the whitefly population was also determined. The aggregation of B. tabaci adults as well as their oviposition rate was higher at 205 and 335 mg/l N than on plants grown at 75 mg/l N. By the end of the experiment (60 d after infestation), the number of nymphs on plants at 205 mg/l N was higher than on plants at 75 mg/l N. The number of pupae was lowest on plants supplied with 75 mg/l N. An increase in NH₄⁺ percentage in standard nutrient solution (from 25% to 45% of the total N) reduced adult population density and oviposition rate. The density of nymphs and pupae, at 60 d after infestation, was lower on the tomato plants grown at 75:25 and 55:45 NO₃⁻:NH₄⁺ ratios compared to the 92:8 ratio. The 75:25 and 55:45 NO₃⁻:NH₄⁺ ratios resulted in a higher incidence of blossom-end rot of tomato fruit, with a lower incidence of disorder at 75:25 than at the 55:45 ratio. Plant stratum influenced adult whitefly distribution in two years of the study. Middle stratum leaves were more attractive to adults in both years. The results demonstrate the effects of N fertilization (N rate and the ratio of NO₃⁻:NH₄⁺) and plant stratum on B. tabaci population density.     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Effect of plant growth promoting rhizobia on seed germination, growth promotion and suppression of Fusarium wilt of fenugreek (Trigonella foenum-graecum L.)

Five bacterial strains (TR1 to TR5) isolated from root nodules of fenugreek (Trigonella foenum-graecum) were tested for their plant growth promotory traits and biocontrol potential against Fusarium oxysporum. On the basis of morphological, physiological, biochemical and molecular characteristics, strains TR1 and TR3 - TR5 were identified as Ensifer meliloti, and TR2 as Rhizobium leguminosarum. All bacterial isolates utilized phosphate in vitro. Except TR5, all isolates produced IAA and none of them showed volatile cyanogens production. Except TR3, all isolates produced in vitro siderophore. Isolate TR1 and TR4 showed chitinase production while only TR2 showed β-1,3-glucanase activity. Isolates TR1, TR2 and TR5 exhibited ACC deaminase activity. Isolates TR1, TR2 and TR4 inhibited the growth of F. oxysporum, causing loss of structural integrity of the mycelium, hyphal perforation, lysis, fragmentation and degradation. The potential for nodulation and nitrogen fixation of the strains were confirmed by amplification of 500 bp nodC and 781 bp nifH fragments. The application of the TR1 + TR2 combination resulted in increased grain yield by 35% and 36% of fenugreek in two consecutive field trials, respectively as compared to control. Maximum increments in vigour index, nodule number and root and shoot biomass were recorded with seed inoculated with consortium (TR1 + TR2) followed by single inoculation as compared to control. The antibiotic resistant marker strain of E. meliloti TR1^strep+ and R. leguminosarum TR2^tet+ confirmed the efficient colonization of fenugreek roots. This study showed that these rhizobial isolates have properties of biocontrol agents and may be applied to promote the growth of fenugreek.     

Effects of middle-viscosity chitosan on Ramularia cercosporelloides

The inhibitory properties of middle-viscosity Chitosan on the growth of fungus Ramularia cercosporelloides were studied in vitro. The inhibitory concentration that delayed 50% of the radial growth (IC₅₀) was 3.4 g l⁻¹ for middle-viscosity Chitosans with molecular weights of about 133 and 187 kDa (middle molecular weight) dissolved in lactic and acetic acids, respectively. At 96 h of incubation and under the same growing conditions, inhibitions of 91.79% and 73.13% of the radial growth of the fungus were observed when 3.4 g l⁻¹ of Chitosan was dissolved in 0.05 M acetic and 0.05 M lactic acid, respectively. The biomass production was significantly lower than that observed in the controls at 72 h. Based on these in vitro results; Chitosan could be a good alternative to control the disease caused by R. cercosporelloides on safflower.     

Response of two tephritid species, Bactrocera oleae and Ceratitis capitata, to different emission levels of pheromone and parapheromone

Attractants and pheromones are commonly used in integrated pest management programs in crop systems. However, pheromone dispensers employed in monitoring traps and lure and kill devices are not usually well studied and attractants are released at uncontrolled rates leading to low treatment efficacies and misleading monitoring estimations. Fruit flies are pests of economic importance and monitoring is essential in order to program insecticidal treatments. Moreover, lure and kill techniques are being increasingly used, but the cost of these techniques depends on the number of required traps and, therefore, on the efficacy of the attractants. Ceratitis capitata and Bactrocera oleae are the two main fruit flies in Mediterranean countries, and the effect of different doses of trimedlure and spiroacetal on fly attraction has been studied. Results showed that a release rate over 1.28 mg/day of spiroacetal reduces B. oleae attraction and emission values over 2.4 mg of trimedlure per day did not increase C. capitata catches. Under the environmental conditions of our study, an optimum release rate for pheromone attraction in B. oleae was determined. Emission values over this optimum level reduced B. oleae attraction. However, when a parapheromone was used with C. capitata, a fruit fly of the same family, the optimum emission value was not found and higher quantities of parapheromone attracted the same number of flies. The saturation effect of high concentrations of pheromone and parapheromone is discussed.