Integrated pest management in faba bean

Successful production of faba bean crops in the presence of a wide range of disease-causing fungi, parasitic weeds, nematodes, insects, mites and other pests depends on the integration of genetic resistance, hygienic management, monitoring of the target organisms and timely application of appropriate chemical and biological treatments. This paper reviews the progress in this area.     

Multiple-disease resistance in Vicia faba: Multi-environment field testing for identification of combined resistance to rust and chocolate spot

The aim of this work was to identify Vicia faba germplasm resistant both to rust and chocolate spot. A collection of 43 accessions of V. faba previously identified as chocolate-spot resistant was evaluated for rust and chocolate spot resistance in Egypt and in Spain. The genotype and genotype × environment (GGE) biplot analyses allowed the selection of 11 accessions resistant to both diseases in those locations. These 11 accessions were evaluated for rust in an additional field trial in Spain, all performing better than the susceptible check. The joint analysis of the 11 accessions in the four field trials where they had been evaluated revealed no significant effects either for genotype, environment or the genotype × environment interaction. They are hence promising sources of resistance, both for their low severities and their stable responses across the studied environments. This collection was also tested under controlled conditions against the Egyptian and the Spanish isolates of rust present in the field trials further underlining the stable performance of these genotypes. Finally, the effect of previous infection with chocolate spot on rust resistance was assessed under controlled conditions and no influence of it was found.     

Broomrape management in faba bean

Broomrapes (Orobanche spp. and Phelipanche spp.) are weedy root parasites that represent a major constraint for faba bean (Vicia faba) cultivation in the Mediterranean area. Control methods are being developed that comprise techniques ranging from agronomical practices to precision farming, including chemical and biological control, genetic and induced resistance, modelling, and probably nanotechnology in a nearby future. However, the main concern is that to date, no single method of control provides complete protection against these parasites. For that reason, an integrated approach is needed in which a variety of such techniques are combined, in order to maintain parasite populations below threshold levels of damage.     

Effects of fungicide seed treatments on germination, population, and yield of maize grown from seed infected with fungal pathogens

Seedborne fungi can reduce survival, growth, and yield of maize (Zea mays L.). Laboratory, field, and growth chamber experiments were conducted to determine the effects of the seed treatment fungicides fludioxonil, mefenoxam, and azoxystrobin on germination, plant population, and grain yield of maize grown from low-quality hybrid seed infected with seedborne fungal pathogens. Study I used seed of four hybrids infected at 0-54% incidence with Fusarium spp., Stenocarpella maydis, Penicillium spp., Rhizopus spp., and/or Aspergillus spp. Study II used three seed lots for each of two hybrids infected at 7-37% incidence with S. maydis. Warm and cold germination for untreated seed varied among hybrids in both studies. Warm germination of the seed lot with the highest incidence of S. maydis in study II treated with azoxystrobin and fludioxonil was significantly greater (+7%) than the nontreated control. Plant population in study I was significantly affected by seed treatment, hybrid, and their interactions. Populations were greater (≥9%) for fludioxonil, fludioxonil + mefenoxam, and fludioxonil + mefenoxam + azoxystrobin treatments compared to controls. In growth chamber experiments with pasteurized soil, emergence (≥5%) and plant dry weight (≥14%) were both greater than controls only with the triple seed treatment. Plant populations in study II for all seed treatments except mefenoxam and azoxystrobin alone were greater (≥4%) than controls. Yield in study I was significantly affected by hybrid and seed treatment. Yield for one hybrid was higher (≥20%) than the control with all seed treatments except fludioxonil, whereas yield with another hybrid was consistently greater (≥26%) only with the triple seed treatment. Yield in study II was significantly affected by hybrid, seed treatment, and their interactions. Yield was greater (≥8%) than the controls for all seed treatments with one hybrid and with all (≥5%) except azoxystrobin for the other hybrid. Highest yields occurred with the triple seed treatment. Results indicate that fludioxonil and azoxystrobin can increase germination, population, and yield of maize grown from seed infected by S. maydis and other fungi.     

Yield stability of faba bean lines under diverse broomrape prone production environments

Faba bean (Vicia faba L.) is one of the important pulse crops grown in many parts of the world. Its production and productivity are affected by different biotic and abiotic stresses. Broomrape (Orobanche and Phelipanche spp.) causes yield losses of 7- 80% in major faba bean growing countries mainly in the Mediterranean Basin and Nile Valley countries. Host plant resistance is found to be the most effective and economic method of broomrape control. Efforts made at the International Center for Agriculture Research in the Dry Areas over many years to develop advanced breeding lines with acceptable resistance and tolerance levels to broomrape have resulted in the identification of several useful breeding lines. These lines were shared with National Agricultural Research Systems (Egypt, Sudan, Tunisia and Portugal) as part of the Faba Bean Orobanche International Nursery, in order to test their stability across three seasons (2005, 2006 and 2007). The results showed that Orobanche plant number, dry weight, Orobanche index, flowering date, maturing date and grain yield were significantly different among lines and locations. AMMI analysis showed that some lines derived from hybrid bulk populations “HBP/DS0/2000” and “HBP/ES0/2000” showed acceptable levels of yield stability in the presence of Orobanche infections across all environments compared with susceptible check. In highly infested soils, the tolerant and resistant lines gave acceptable levels of yield compared with the susceptible check. However, the yield potential of resistant and tolerant lines in non-infested soils was lower than the highly susceptible check.     

Physiology of flowering and grain filling in faba bean

The development of flowers and then that of seeds are key processes in the formation of yield in faba bean (Vicia faba L.), as in other grain legumes. Winter faba bean generally has a quantitative vernalization requirement, allowing flowering to occur at a lower node than in unvernalized plants. Some germplasm is day-neutral, other germplasm is long-day with a critical daylength between 9.5 and 12 h. Progress toward flowering follows a conventional thermal-time model, with 830–1000 °C-d above 0 °C required for the onset of flowering and an optimum temperature of 22–23 °C. Flowers may abscise from the crop because of lack of pollination, because proximal flowers on the same raceme are fertilized, because of vegetative–reproductive competition for assimilate, or because of stresses such as drought.     

The effect of hail protection nets on Glomerella leaf spot in ‘royal Gala’ apple

In commercial apple orchards in southern Brazil, it is common to avoid hail damage by covering the trees with nets. This study was carried out to assess the effects of both black and white hail netting on the temporal dynamics of Glomerella Leaf Spot disease (GLS) in a commercial ‘Royal Gala’ apple orchard in a high-altitude region of southern Brazil during two growing seasons (2003/2004 and 2004/2005). Apple trees on M9 rootstock, trained to a slender spindle and with a spacing of 5.5 m × 3 m, were left uncovered (control) or covered with black or white hail netting. The incidence and severity of GLS were quantified biweekly from October to February by evaluating 100 leaves distributed randomly on four medium-height branches per plant on each 12 replications per treatment. Disease progress curves were constructed from the data and the epidemics were compared by using four epidemiological measures: the beginning of symptom appearance (BSA), the time to reach the maximum disease intensity (TRMDI), the maximum value of disease severity (Y_max), and the area under the disease progress curve (AUDPC). The highest intensity of GLS was observed in the plants under the black hail netting. Significant reductions in BSA and TRMDI and significant increases in Y_max and AUDPC were found among treatments in both growing seasons. However, no significant differences of GLS were found between the white hail netting and the control in either growing season.     

Study of faba bean (Vicia faba L.) winter-hardiness and development of screening methods

In cool-temperate regions, faba bean (Vicia faba L.) is mainly grown as a spring crop despite the higher yield potential of the winter type, because of the insufficient winter-hardiness of the present winter genotypes. The objective of this study was to assess winter-hardiness and frost tolerance, to quantify the hardening effect on physiological traits, and to identify auxiliary traits for winter-hardiness. To do so, 31 representative entries were tested in controlled frost tests for frost tolerance and in 12 European environments for winter-hardiness. Total fatty acid composition, proline content, and electrolyte leakage of leaves were analysed. Across all environments, five European winter genotypes were identified with superior winter-hardiness. Controlled frost tests indicated that frost tolerance is a significant, but not an exhaustive component of winter-hardiness (0.021 < |r| < 0.737**). These tests revealed the high frost tolerance of several poorly winter-hardy experimental lines and the limited frost tolerance of well-known winter types. Fatty acid changes due to hardening, proline content, and electrolyte leakage were more strongly correlated with frost tolerance than with field-based winter-hardiness. Although frost tolerance, fatty acid composition, proline content, and electrolyte leakage were significantly correlated with winter-hardiness, the rather low correlation values do not allow a general use of one of them alone to indirectly select for winter-hardiness.     

Effects of crop rotation of soybean with corn on severity of sudden death syndrome and population densities of Heterodera glycines in naturally infested soil

Sudden death syndrome, caused by Fusarium virguliforme, and the soybean cyst nematode, Heterodera glycines, combined cause the highest yield losses in soybean. The objective of this study was to determine the effectiveness of corn rotated annually with soybean on reducing severity of sudden death syndrome (SDS) and if such crop rotation is beneficial to soybean root health and thus improves disease management strategies. Experiments were conducted from 2003 to 2006 through two cycles of a corn–soybean rotation on two commercial fields in Indiana. With one exception, the rotation of soybean with corn did not provide yield benefits compared to monoculture of soybean. Severity of foliar and root symptoms of SDS in rotation plots were never less than in soybean monoculture plots. At one location, soybean monoculture resulted in suppression of SDS compared to the corn–soybean rotation, while H. glycines reproduced freely. At the other location, monoculture of soybean resulted in suppressiveness against H. glycines, while SDS was limited in all treatments. The data suggest that soil suppressiveness can independently impact the pathogens that are important in SDS development. Because H. glycines can increase SDS symptoms, its suppression may also reduce severity of SDS. Current production systems consisting of yearly rotation of soybean with corn are highly vulnerable to the development of severe soil-borne disease complexes. The simple year-to-year rotation of corn and soybean is not considered sustainable. While monoculture of soybean resulted in some disease suppression in these trials, reliance on monoculture may be detrimental due to other environmental considerations beyond the scope of these trials. Including other crops may be beneficial in improving the sustainability of soybean and corn production systems.     

Use of plant extracts in the control of common bean anthracnose

Three hundred (300) methanol extracts of barks, leaves, flowers and stems of 200 plant species from Alto Rio Grande and Vale do São Francisco regions, State of Minas Gerais, Brazil, were prepared and submitted to an in vitro growth test with the fungus Colletotrichum lindemuthianum, causal agent of anthracnose in common bean. Extracts from 13 plant species (Astronium fraxinifolium, Inga marginata, Malva sylvestris, Matayba elaeagnoides, Miconia argyrophylla, Myrcia fallax, Ocimum gratissimum, Origanum vulgare, Rollinia emarginata, Siparuna arianeae, Styrax pohlii, Tabebuia serratifolia and Trichilia pallida) presented antifungal activity and were used in other in vitro assays. Extracts from M. argyrophylla, M. fallax, O. vulgare, S. arianeae and S. pohlii were the most promising for the inhibition of the mycelial growth while the extracts of M. argyrophylla, M. elaeagnoides and O. gratissimum presented the best results for the inhibition of conidial germination. Under greenhouse conditions, the extracts of M. argyrophylla and O. vulgare caused the greatest reductions (41.82% and 37.65%, respectively) in disease severity when a local effect assay was carried out. In the systemic effect assay, also in a greenhouse, the most promising extracts were those from I. marginata, M. argyrophylla, M. fallax, M. sylvestris, O. gratissimum, O. vulgare and S. arianeae, which reduced the severity of the anthracnose to values below 35% of the observed for the control. Therefore, future studies with these plant species should be carried out to develop new products to control the common bean anthracnose.     

Genetic analysis of field resistance to tan spot in spring wheat

Tan spot, caused by Pyrenophora tritici-repentis (Died.) Drechs., is an important constraint to wheat (Triticum aestivum L.) yield in many countries. Since the inheritance of field resistance to tan spot is poorly understood, this study was conducted to determine the genetic control of resistance in the field. Resistance was measured as disease severity caused by P. tritici-repentis race 1 in four crosses involving five wheat parents: parent 1 (P₁) = catbird; parent 2 (P₂) = Milan/Shanghai-7; parent 3 (P₃) = Alondra/Coc//Ures; parent 4 (P₄) = Bcb//Dundee/Gul/3/Gul); parent 5 (P₅) = ND/VG9144//Kal/BB/3/Yaco/4/Chil. P₁, P₂ and P₃ were resistant and P₄ was moderately resistant, whereas P₅ was susceptible to tan spot. The F₂-derived F₃ families and the parents were field evaluated at El Batán, Mexico, in 1996. When all the plants within a F₃ family expressed low levels of disease severity similar to that of the resistant parent it was classified resistant (R), otherwise the progeny was classified as susceptible (S). The progeny of the three crosses of the susceptible parent with the resistant and moderately resistant parents P₂, P₃, and P₄ segregated as 3R:13S whereas the progeny in the cross with P₁ showed a segregation ratio of 1R:15S. This suggests that each resistant parent possessed two genes conditioning resistance to tan spot severity caused by P. tritici-repentis race 1. Information on the inheritance of resistance measured as disease severity on adult plants under field conditions is of practical importance for wheat breeding programs seeking improvement in tan spot resistance.     

Impact of intercropping of medicinal and aromatic plants with organic farming approach on resource use efficiency in arecanut (Areca catechu L.) plantation in India

The present investigation was conducted at Vittal, Karnataka, India during 2004-2007 to study the feasibility of intercropping of medicinal and aromatic plants (MAPs) in arecanut plantation. The results revealed that MAPs can be successfully grown as intercrops in arecanut plantation with increased productivity and net income per unit area. Kernel equivalent yield of MAPs varied between 272 kg ha⁻¹ in case of Piper longum to 1218 kg ha⁻¹ in Cymbopogon flexuosus. Pooled data indicated that Asparagus racemosus produced fresh root yield of 10,666 kg ha⁻¹ of arecanut plantation and contributed to maximum kernel equivalent yield of 1524 kg ha⁻¹ among all medicinal and aromatic plants. Intercropping of MAPs in arecanut was found economical. The net return per rupee investment was highest in C. flexuosus (4.25) followed by Bacopa monnieri (3.64), Ocimum basilicum (3.46) and Artemisia pallens (3.12). The total system productivity of arecanut + MAPs intercropping system varied from 2990 to 4144 kg ha⁻¹. Arecanut + O. basilicum intercropping system registered significantly higher production efficiency 8.2 kg ha⁻¹ day⁻¹ than other systems. Intercropping of MAPs had more positive effect on soil pH in arecanut based cropping system. The soil pH was 5.6 in 2004 and it was 0.3-0.9 units higher in 2007. Soil organic carbon (SOC) content varied significantly due to intercropping of MAPs at the end of experiment. The SOC content increased in Aloe vera, A. pallens, P. longum and B. monnieri, while it depleted in grasses and rhizomatic MAPs. Based on demand and marketing opportunities for MAPs, farmers are advised to grow aromatic plants in large areas on a community basis to meet huge industrial demand and variety of medicinal crops in small areas to meet the requirement of traditional systems of medicine.     

Effect of herbicide and cover crop on weed control in no-tillage jack-o-lantern pumpkin (Cucurbita pepo L.) production

Field experiments were conducted to evaluate cover crop (none, winter wheat, or winter rye) and pre-emergence (PRE) applications of clomazone plus ethalfluralin alone or with halosulfuron [PRE or post-emergence (POST)] for smooth crabgrass [Digitaria ischaemum (Schreb. ex Schweig) Schreb. ex Muhl.] and redroot pigweed (Amaranthus retroflexus L.) control in no-tillage ‘Aspen’ jack-o-lantern pumpkin (Cucurbita pepo L.). At pumpkin harvest, cover crops had reduced smooth crabgrass density, but not redroot pigweed. Although PRE or POST applications of halosulfuron alone were more effective at reducing redroot pigweed density than clomazone plus ethalfluralin PRE, the combination of halosulfuron plus clomazone and ethalfluralin PRE reduced redroot pigweed density to the greatest extent. Pumpkin yields were not affected by cover crop, although average pumpkin sizes were greater with the inclusion of cover crop residues. Pumpkin fruit sizes and yields were the greatest with clomazone and ethalfluralin PRE in combination with halosulfuron applied PRE or POST. Overall, cover crop had relatively little influence on pumpkin yields compared with herbicide treatments. The addition of halosulfuron to clomazone and ethalfluralin provided greater broadleaf weed control resulting in greater jack-o-lantern pumpkin yields.     

Evaluation of electrolyzed oxidizing water for phytotoxic effects and pre-harvest management of gray mold disease on strawberry plants

Near neutral (pH = 6.3-6.5) electrolyzed oxidizing water (EO water) has been demonstrated to inactivate fungi in pure culture and to mitigate infection on fruit surfaces. One possible alternative or supplement to traditional pre-harvest crop management practices that currently rely on the use of large quantities of fungicides is near neutral EO water. In the present work, treatment of Botrytis cinerea or Monilinia fructicola with near neutral EO water (50 or 100 ppm total residual chlorine (TRC)) in pure culture resulted in a 10⁶ reduction and 100% inactivation as evidenced by negative broth enrichment. When applied in concert with 50 or 100 ppm EO water, treatments of Captan 50WP (captan), Rovral (iprodione), Iprodione 4LAG (iprodione), or Switch 62.5 WDG (cyprodinil and fludioxonil) effectively inhibited fungal growth of B. cinerea as evidenced by a 10⁶ reduction on the direct plate and negative broth enrichment. Treatments of Captan 50WG (captan), Rovral (iprodione), Iprodione 4LAG (iprodione), Switch 62.5 WDG (cyprodinil and fludioxonil), Captan 80 WDG (captan), or Captevate (captan and fenhexamide) when applied in concert with 50 or 100 ppm EO resulted in a 10⁶ reduction of M. Fructicola and 100% inactivation as evidenced by negative broth enrichment. Strawberry plants sprayed with EO water (pH = 6.3-6.5) at concentrations of 50 and 100 ppm TRC once per week, did not result in significant (P > 0.05) phytotoxicity relative to a water (0 ppm TRC) treatment. In this study, the application of 100 ppm EO water (pH = 6.3-6.5) twice per week to strawberry plants infected with B. cinerea was more effective (P ≤ 0.05) than a once per week Captan application and as effective as a once per week captan/once per week EO treatment. The once per week captan/once per week EO treatment was significantly more effective (P ≤ 0.05) than the captan once per week treatment. Dip treatments of strawberries in near neutral EO solutions (50 and 100 ppm TRC; pH = 6.3-6.5) did not leave a chlorine residue on the fruit relative to a water dip. The results from this study suggest that near neutral EO solutions could be used to manage infection of B. cinerea on strawberry plants in the field and also as a disinfection solution for harvesting equipment, greenhouses, packing houses and in commercial facilities to prevent or manage infections of B. cinerea and M. fructicola.     

Effect of intercropping pepper with sugarcane on populations of Liriomyza huidobrensis (Diptera: Agromyzidae) and its parasitoids

The effect of intercropping pepper with sugarcane on populations of the leafminer Liriomyza huidobrensis Becker was investigated over two seasons (2007, 2008) in Yunnan, China. Populations of adults, the female:male sex ratios and the densities of larval mines were significantly higher in pepper monoculture than in intercropping in a 2:2 pattern (2 rows of pepper to 2 rows of sugarcane), which in turn were significantly higher than intercropping in a 1:2 pattern (1 row of pepper to 2 rows of sugarcane). The densities of the parasitoids of the leafminer were significantly higher in intercropping than in the pepper monoculture. The total parasitism rates were 71.6%, 22.5% and 5.9% in 2007, and 69.5%, 20.6% and 5.9% in 2008, in the 1:2, 2:2 and pepper monoculture, respectively. The lower densities of leafminer adults, the lower female:male sex ratio, and smaller numbers of larval mines in the intercropping systems were attributed to the intercropping pattern and the growth of pepper. These results indicate that intercropping is suitable for the control of L. huidobrensis on pepper plants. These results demonstrate the utility of intercropping of pepper with sugarcane in the management of L. huidobrensis.     

Effects of soil amendment with poultry manure on carnation Fusarium wilt in greenhouses in southwest Spain

In a search for alternatives to methyl bromide for controlling carnation vascular wilt caused by Fusarium oxysporum f. sp. dianthi (Fod), poultry manure plus soil solarization was studied in soil under greenhouse conditions in four 2-year experiments. These were conducted in naturally infested soil to compare the effects of this treatment with soil solarization alone and methyl bromide. Soil treatments were performed during June 2000 for Experiment 1, from July to mid-August 2002 for Experiment 2, from late July to late August 2006 for Experiment 3 and from late May to late June 2008 for Experiment 4. Additionally, a treatment with commercial poultry manure pellet plus soil solarization was included in the two latter experiments. Poultry manure caused reductions of Fod viability in soil samples at depths of 15 and 30 cm, ranging respectively from 93 to 100% and 89 to 100% for Experiments 1, 2 and 3. Carnations planted in plots treated either with poultry manure, methyl bromide or soil solarization had lower final disease incidences, smaller areas under their disease progress curves and higher yields in comparison with untreated plots in Experiments 2, 3 and 4. In Experiment 1, soil solarization was performed under suboptimal conditions, and it provided disease levels and yields similar to those of the untreated control plots. Nevertheless, under the same conditions, previous amendment of Fod-infested soil with poultry manure increased disease control over soil solarization alone, improved carnation yield and quality and also increased plant vigor, thus providing a satisfactory alternative to methyl bromide. The application of organic amendment to the same plot before every crop cycle is recommended to ensure continuous disease control, but the rates of application could be reduced to half for the third and fourth crop cycles, thereby reducing undesirable environmental effects.     

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.     

Predicting rodent impact in crop fields by near-infrared reflectance spectroscopy analysis of their diet preferences

Prediction of impact by near-infrared reflectance spectroscopy (NIRS) analysis of rodent diet preferences was evaluated in four dominant field rodents in Moravia farmland (Czech Republic). These include the common vole (Microtus arvalis), pygmy field mouse (Apodemus uralensis), wood mouse (Apodemus sylvaticus), and black-striped field mouse (Apodemus agrarius). Using fecal samples, this method predicts the proportions of food components consumed, based on a calibration equation developed through feeding tests in the laboratory. We used the wood mouse as a model species for NIRS device calibration. The two types of monofood feces collected from mice that were fed with either barley or wheat grain were mixed to form a series of increasing proportions of wheat feces subsequently measured by NIRS. Because of a good fit, the calibration equation was then applied to other mice, whereas a separate calibration equation had to be developed for the common vole. The percentage diet preferences were determined by comparing feces from preference choice tests with the calibration equation. Whereas all mice preferred wheat to barley, the common vole showed no preference for either crop. Thus proportion of wheat preference by all Apodemus sp. predict stronger impact of these in wheat fields in ripe stage than in barley one.     

Seed and juice characterization of pomegranate fruits grown in Tunisia: Comparison between sour and sweet cultivars revealed interesting properties for prospective industrial applications

Tunisian pomegranate genetic resources consist of sweet and sour cultivars, showing large morphometric variability. In the present work we characterized seeds and juice contents of sugars and organic acids of 5 sour and 7 sweet pomegranate cultivars. Results showed that citric acid was predominant in sour pomegranates, while malic acid was the most prevalent in sweet ones. Paradoxically, sour cultivars have higher sugar content than the sweet ones. A strong correlation was found between sourness and citric acid content, which is assumed to be the major factor that determines sour taste in pomegranate fruits. Besides, some of the seed parameters showed a significant positive correlation with acidity. Sweet cultivars were appropriate for fresh consumption and juice production due to several attributes in addition to their sweetness. Equally, sour pomegranate showed several characteristics that could be of great interest for food and nutraceutical industries.     

Isolation of entomopathogenic Bacillus from a biodynamic olive farm and their pathogenicity to lepidopteran and coleopteran insect pests

The occurrence of Bacillus entomopathogenic bacteria on a Tunisian biodynamic farm was determined by examining 75 samples from olive tree (Olea europaea L.) habitats. A total of 40 Bacillus isolates were characterized according to their phenotypic, physiological and biochemical parameters. Isolates of the species Bacillus subtilis, Bacillus mycoides, Brevibacillus brevis, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus sp. (1), Bacillus sp. (2) and a standard strain Btk HD-1 were used separately in feeding bioassays on fresh artificial diet against larvae of lepidopterans Prays oleae (Bernard) and Palpita unionalis (Hübner) and coleopterans Hylesinus oleiperda (F.) and Phloeotribus scarabaeoides (Bernard), which are olive tree pests. Larvae were successfully reared on an artificial diet with 25 g powdered olive tree leaves. Compared to the control data, only Btk and the isolates of B. licheniformis, P. polymyxa and B. brevis were entomopathogenic. Larval mortality assessed 7 days post-treatment showed high mortality rates with Btk to lepidopteran larvae (86.6% for P. oleae and 80.9% for P. unionalis) and low mortality against coleopteran pests. B. brevis isolates showed high mortality rates against P. oleae (up to 67.9%). B. licheniformis isolates caused up to 59.2% larval mortality for P. oleae and 43.6% for P. unionalis. Highest coleopteran mortality was achieved by P. polymyxa isolates (up to 55%). According to the 16S rDNA results, isolates of each of the three entomopathogenic strains were similar. Proteins in the strain supernatants were toxic to P. oleae larvae with LC50 values of 10.0 (B. brevis), 12.5 (B. licheniformis) and 37.6 μg/ml (P. polymyxa). Also, P. polymyxa showed an LC50 of 12.4 mg/l against P. scarabaeoides. Our results suggest that entomopathogenic Bacillus present locally in the biodynamic farm could be used in biological control programmes of olive tree pests.