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.     

Evaluation of the Ugandan sorghum accessions for grain mold and anthracnose resistance

Sorghum accessions from Uganda were evaluated for grain mold and anthracnose resistance during the 2005 and 2006 growing seasons at the Texas A&M University Research Farm, near College Station, TX. Accession PI534117 and SC719-11E exhibited the lowest grain mold severities of 2.4, whereas, accessions PI534117, PI534144, PI576337, PI297199, PI533833, and PI297210, with SC748-5 were highly resistant to anthracnose in both years. Accessions PI534117, PI297134, PI297156 exhibited low grain mold severities in 2006. Significant negative correlation was recorded between grain mold and percent germination and high temperature in both years. In 2006, daily precipitation recorded significant positive correlation with grain mold. The seed mycoflora was analyzed across sorghum lines and treatments. In 2005, Curvularia lunata and Fusarium thapsinum were the most frequently recovered fungal species with 31 and 21% incidence, respectively, followed by Alternaria spp. (19%) and F. semitectum (13%). In 2006, predominant colonizers were F. thapsinum (58%), followed by Alternaria and F. semitectum with 15 and 10, percent respectively, while C. lunata had a 6% incidence. In this study, PI534117 holds promise for multiple disease resistance, as it had the lowest disease severity of grain mold and was highly resistant to anthracnose in both years. It also has a high germination rate, a high seed weight, and its short stature is more advantageous for the new A-line conversion program.     

In vitro activity of eighteen essential oils and some major components against common postharvest fungal pathogens of fruit

The development of natural crop protective products as alternatives to synthetic fungicides is currently in the spotlight. In vitro experiments are valuable precursors to more costly in vivo trials, allowing the identification of effective essential oils and establishing the concentrations required for inhibition of a specific, or spectrum of decay pathogens. In this study, the antifungal properties of eighteen essential oils were evaluated in vitro by addition to the fungal growth medium of five pathogens (Lasiodiplodia theobromae, Colletotrichum gloeosporioides, Alternaria citrii, Botrytis cinerea and Penicillium digitatum) isolated from mango, avocado, citrus, grapes and cactus pear. The inhibitory properties of some of the major compounds of the oils, identified and quantified by gas chromatography-mass spectroscopy and gas chromatography-flame ionization detection were also determined. Most of the oils were selected on the basis of commercial availability and for containing a predominant compound. Visual inspection of fungal growth was done and the lowest concentration where fungal growth was completely inhibited on all replicates was recorded. Thyme oil proved to be the most effective inhibitor, totally inhibiting all of the pathogens tested at concentrations of 1000 μl/l and lower, with the exception of a resistant Penicillium strain. Cinnamon oil, rich in eugenol (81.2%), demonstrated good fungicide potential, while the carvone-rich oils displayed promising activity against the citrus pathogens. Oils characterized by high concentrations of S-carvone were less effective than those containing the R-enantiomer. Essential oil of Lippia citriodora was active against all of the pathogens, excluding L. theobromae from avocado. These essential oils, applied alone or in combination, are good candidates for further in vivo testing and for investigations concerning their modes of action.     

Pre- and post-inoculation activity of a protectant and a systemic fungicide for control of anthracnose fruit rot of strawberry under different wetness durations

A protectant fungicide (Captan, a.i. captan) and a systemic fungicide (Switch, a.i. fludioxonil + cyprodinil) were evaluated as pre- and post-inoculation applications for control of anthracnose fruit rot (AFR), caused by Colletotrichum acutatum, under a short (6 or 8 h) or long (18 or 24 h) wetting period. Evaluations were conducted for two seasons in Maryland and for two seasons in Florida. Both Captan and Switch were very effective for control of AFR when applied prior to inoculation, but control was more effective under the shorter wetting period. Switch was as effective when applied 4, 8, or 24 h post-inoculation as when applied before inoculation, but control was better under the short wetting period. Captan was effective when applied 4 or 8 h after inoculation under the short wetting period, but was ineffective at 24 h post-inoculation. Post-inoculation sprays of Captan were ineffective at any time under the long wetting period. The post-infection activity of Switch allows greater flexibility for managing AFR when fungicide applications are scheduled based on weather-based decision-support systems.     

Feeding behavior of a potential insect pest, Lygus hesperus, on four new industrial crops for the arid southwestern USA

Camelina (Camelina sativa), guayule (Parthenium argentatum), lesquerella (Physaria fendleri), and vernonia (Centrapalus pauciflorus [formerly Vernonia galamensis]) are either under limited commercial production or being developed for production in the southwestern USA. Insect pests are a potential economic threat to all these new crops, with Lygus hesperus, the western tarnished plant bug, among the most prominent due to its regional abundance and propensity to feed on reproductive plant tissue. The objectives of this study were to establish baseline data on the feeding behavior and potential impact of L. hesperus on camelina, guayule, lesquerella and vernonia. Behavioral observations of adult females and males, and nymphs of this insect were made in the laboratory. Insects spent ≈35% of their time either probing (=tasting) or feeding on various reproductive and vegetative tissues of guayule, lesquerella or vernonia, but only 20% on camelina. When insects did probe and feed they preferred reproductive tissues, primarily flowers and siliques/achenes, and there were differences in these behaviors relative to crop but not generally to insect stage or sex. Insects probed and fed more on flower tissue of guayule and vernonia compared with camelina and lesquerella, and more on siliques of lesquerella compared with achenes of vernonia. When probing and feeding on vegetative tissue, there was generally a preference for stems compared with leaves in all crops except guayule. Results show that L. hesperus will readily feed on the economically important tissues of all crops, and although research has shown that this feeding did not consistently affect lesquerella yield, further work will be needed to determine if such feeding poses a risk to commercial production of camelina, guayule or vernonia.     

Effect of Trichoderma asperellum and arbuscular mycorrhizal fungi on cacao growth and resistance against black pod disease

Arbuscular mycorrhizal fungi (AMF) and members of the genus Trichoderma have emerged as promising groups of microbial inoculants that can induce plant growth and resistance to disease. This study aimed at investigating the potential of AMF and a strain (PR11) of Trichoderma asperellum to promote cacao growth and induce resistance against Phytophthora megakarya. Cacao seedlings were either non-inoculated, or inoculated with the saprophytic fungus T. asperellum and/or a mixture of two different mycorrhizal fungi, Gigaspora margarita and Acaulospora tuberculata. Eighteen weeks after planting, a series of morphological as well as biochemical changes, which are considered to be part of the plant defense response, were measured after a challenge inoculation of the leaves with zoospores of P. megakarya. Inoculation with AMF and T. asperellum alone was essential for the promotion of plant growth. Significant increase in plant height, root and shoot fresh weights, as well as phosphorous uptake was recorded in comparison to non-inoculated control plants. However, dual inoculation of cacao seedlings with T. asperellum and AMF did not always positively benefit the plants. Leaf inoculation showed variation among the treatments, with the lowest disease index (highest level of resistance) recorded in plants inoculated with either AMF or T. asperellum only. This came along with a high synthesis of amino acids and phenolic compounds in both healthy and infected leaves, suggesting that these metabolites are implicated in disease resistance.     

A new tray-type arena to mass rear Neoseiulus baraki, a predatory mite of coconut mite, Aceria guerreronis in the laboratory

Neoseiulus baraki Athias-Henriot (Acari: Phytoseiidae) is an important predatory mite of the coconut mite, Aceria guerreronis Keifer (Acari: Eriophyidae) and attempts are now being made to evaluate the effectiveness of augmenting N. baraki to control coconut mite in the field. These studies require a steady supply of N. baraki in large numbers. One major constraint of the existing method to mass rear N. baraki on Tyrophagus putrescentiae Shrank (Acari: Acaridae) in a closed arena without a water barrier was frequent contamination of cultures by other mites. This paper describes the development of an efficient method to mass rear N. baraki in the laboratory with less monitoring and relatively minimal contaminants. Three box-type arenas and a tray-type arena were tested to mass rear T. putrescentiae. Mites were successfully developed on two box-type arenas and the tray-type arena, but the production (2197 mites per tray in 4 weeks) of T. putrescentiae was significantly higher on the tray-type arena. A 110-fold increase of T. putrescentiae was achieved in tray-type arenas in 4 weeks. None of the box-type arenas could be maintained more than 5 weeks without acarine and fungal contaminations. N. baraki could be successfully reared on the tray-type arena at least for 6 weeks. More than 4800 mites per tray, a 240-fold increase was achieved in 5 weeks when T. putrescentiae was added at 3-week intervals. Advantages and disadvantages of the new method are described.     

Improvements in the qualities of gluten-free bread after using a protease obtained from Aspergillus oryzae

From a nutritional perspective, rice flour is one of the most valuable flours and it is suitable for preparing food for people suffering from wheat allergy. However, bread made from rice flour is very difficult to bake because it lacks gluten. We found that pre-fermenting of rice flour using Aspergillus oryzae facilitated a better formulation of gluten-free rice bread. Bread swelling was remarkably improved with a longer pre-fermenting period at 55 °C. The specific loaf volume (SLV) without polymeric thickeners after a 12 h fermentation was approximately 2.2-fold (2.98 ml/g) higher than that after 0 h (1.36 ml/g). An enzymatic assay of the batter indicated that protease activity during the pre-fermentation period increased from 0.38 to 1.44 U/ml and this activity correlated with bread swelling. Furthermore, a commercial protease from A. oryzae also produced similar results with an adequate SLV of 3.03 ml/g. Rheological analysis showed that batter treated with protease had an increased batter viscosity and decreased flour settling behavior because of the aggregation of flour particle after partial cleavage of storage proteins. These results indicated that the improved SLV was mainly because of an A. oryzae protease, which affected the batter rheology thereby improving gas retention before baking.     

Improved control of lettuce drop caused by Sclerotinia sclerotiorum using Contans combined with lignin or a reduced fungicide application

The use of Contans (Coniothyrium minitans) in Belgian commercial lettuce greenhouses has so far not always fulfilled the expectations of growers. With the aim of improving the efficacy of control of lettuce drop (Sclerotinia sclerotiorum), Contans applied alone and Contans combined with Radix (Trichoderma asperellum and Trichoderma gamsii), Kraft pine lignin and Rovral was compared in three successive lettuce crops in a naturally infested greenhouse. At harvest of each crop, the effect on disease incidence, and the effect on viability and fungal infection of sclerotia buried prior to each crop was evaluated. Contans applied alone negatively affected the sclerotial viability, but did not reduce lettuce drop symptoms. The combination of Contans with Kraft pine lignin, on the other hand, reduced sclerotial viability and lettuce drop compared with the untreated control in the last crop. Furthermore, when Contans was combined with Kraft pine lignin, a higher number of infected sclerotia was observed compared with Contans applied alone. When Radix was added to the combination, there was no extra benefit for the control of lettuce drop. The combination of one Rovral application and Contans resulted in a significant control of lettuce drop together with a negative effect on sclerotial survival. Two Rovral applications, by contrast, reduced the ability of C. minitans to parasitize and kill the sclerotia. To conclude, integration of Contans with a reduced Rovral application and with Kraft pine lignin has potential to improve the control of lettuce drop in commercial greenhouses.     

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.     

Effects of free air carbon dioxide enrichment and nitrogen supply on growth and yield of winter barley cultivated in a crop rotation

The increase in atmospheric CO₂ concentration [CO₂] has been demonstrated to stimulate growth of C₃ crops. Although barley is one of the important cereals of the world, little information exists about the effect of elevated [CO₂] on grain yield of this crop, and realistic data from field experiments are lacking. Therefore, winter barley was grown within a crop rotation over two rotation cycles (2000 and 2003) at present and elevated [CO₂](375 ppm and 550 ppm) and at two levels of nitrogen supply (adequate (N2): 262 kg ha⁻¹ in 1st year and 179 kg ha⁻¹ in 2nd year) and 50% of adequate (N1)). The experiments were carried out in a free air CO₂ enrichment (FACE) system in Braunschweig, Germany. The reduction in nitrogen supply decreased seasonal radiation absorption of the green canopy under ambient [CO₂] by 23%, while CO₂ enrichment had a positive effect under low nitrogen (+8%). Radiation use efficiency was increased by CO₂ elevation under both N levels (+12%). The CO₂ effect on final above ground biomass was similar for both nitrogen treatments (N1: +16%; N2: +13%). CO₂ enrichment did not affect leaf biomass, but increased ear and stem biomass. In addition, final stem dry weight was higher under low (+27%) than under high nitrogen (+13%). Similar findings were obtained for the amount of stem reserves available during grain filling. Relative CO₂ response of grain yield was independent of nitrogen supply (N1: +13%; N2: +12%). The positive CO₂ effect on grain yield was primarily due to a higher grain number, while changes of individual grain weight were small. This corresponds to the findings that under low nitrogen grain growth was unaffected by CO₂ and that under adequate nitrogen the positive effect on grain filling rate was counterbalanced by shortening of grain filling duration.     

Composition and biological activity of the essential oil from Thapsia minor, a new source of geranyl acetate

The essential oils from aerial parts of Thapsia minor Hoffmanns. & Link (Apiaceae), obtained by hydrodistillation, were analyzed by gas-chromatography and gas-chromatography-mass spectrometry. Oxygenated monoterpenes represented the main fraction with geranyl acetate (82.3-83.0%) as the major constituent, making this plant a good source of this compound.Biological activity of the essential oil was evaluated against several fungal strains. The highest activity was found against dermatophyte strains, Crytococcus neoformans and Candida guillermondii with minimal inhibitory concentrations values from 0.32 μl/ml to 1.25 μl/ml. The antifungal activity could be associated with the significant contribution of the oxygenated monoterpene geranyl acetate.Furthermore, this essential oil demonstrated to have potent anti-inflammatory in vitro activity, at concentrations that did not affect macrophages and keratinocytes viability, being the anti-inflammatory properties of the oil mainly due to geranyl acetate.These favorable results emphasize the antifungal and anti-inflammatory abilities of the molecule geranyl acetate and further support the exploitation of T. minor oil for pharmaceutical industry, in order to find new phytopharmaceuticals.