Influence of dosage,storage time and temperature on efficacy of metalaxyl-treated seed for the control of pearl millet downy mildew

Metalaxyl (Apron 35WS) as a seed treatment has been used extensively to control downy mildew (caused by Sclerospora graminicola) in pearl millet in India. However, the extent of disease control has varied across cultivars, years and locations. We investigated the effects of fungicide dosage, storage time and storage temperature of metalaxyl-treated seed on disease incidence in four pearl millet lines having varying levels of resistance. A linear relationship was found between fungicide dosage (0.5, 1.5 and 2 g a.i. kg⁻¹ seed) and reduction in disease incidence up to 40 days after emergence in all the lines. The normal fungicide dose (2 g a.i. kg⁻¹ seed) protected the crop for up to 20, 40 and 50 days after emergence in highly susceptible (7042S), moderately susceptible (4042R), and moderately resistant (ICMP 451) lines, respectively. However, the quarter and half the normal dosage of fungicide provided protection only up to 20 days after emergence in 7042R and 40 days after emergence in ICMP 451. Storage duration of metalaxyl-treated seed (2 g a.i. kg⁻¹) up to 9 months at 25 ± 2°C did not affect fungicide efficacy. Storage temperatures (5, 25 and 40°C) and duration (30, 60 and 90 days) of metalaxyl-treated seed (2 g a.i. kg⁻¹) showed differential effects in two pearl millet lines 7042S and 843B with downy mildew incidence being significantly lower in 7042S than in 843B. Metalaxyl-treated seed of 7042S and 843B stored at 40°C for different durations showed phytotoxic effects and it was more pronounced in 843B stored for 60 and 90 days where seed germination was inhibited in pot soil.     

Chitosan enhances disease resistance in pearl millet against downy mildew caused by Sclerospora graminicola and defence-related enzyme activation

BACKGROUND: The present study investigated the effect of chitosan seed priming on the induction of disease resistance in pearl millet against downy mildew disease caused by Sclerospora graminicola (Sacc.) Schroet. RESULTS: Pearl millet seeds were primed with chitosan at different concentrations: 0.5, 1.5, 2.5 and 3 g kg^?1 seed. Of the different concentrations, 2.5 g kg^?1 was found to be optimum, with enhanced seed germination of 99% and seedling vigour of 1782, whereas the untreated control recorded values of 87% and 1465 respectively. At optimum concentration, chitosan did not inhibit sporulation and release of zoospores from sporangia. Furthermore, pearl millet seedlings raised after seed treatment with chitosan showed an increased level of the defence‐related enzymes chitosanase and peroxidase as compared with the untreated pearl millet seedlings on downy mildew pathogen inoculation. The effect of chitosan in reducing downy mildew incidence was evaluated in both greenhouse and field conditions, in which respectively 79.08 and 75.8% disease protection was obtained. CONCLUSION: Chitosan was effective in protecting pearl millet plants against downy mildew under both greenhouse and field conditions by inducing resistance against the pathogen. Thus, chitosan formulation can be recommended for seed treatment in the management of downy mildew disease. Copyright © 2008 Society of Chemical Industry     

Passalora blight of anise (Pimpinella anisum) and its control in Turkey

Passalora blight of anise, caused byPassalora malkoffii (Bubák) U. Braun, is an important disease of anise in Turkey. The disease affects all the aboveground parts of plants including flower clusters. Infected seeds have dark, linear stromata. Detection of the pathogen on seeds was studied by the blotter method, agar method, washing test and sowing infected seeds in disease-free soils. The pathogen was recovered only by the washing test and to a limited extent by water agar + seed decoction agar. Sixteen of 24 seed samples from diseased regions were found to be infected. The pathogen was not detected by any other methods. However, several indigenous fungi,e.g. Alternaria alternata, were isolated, which may have prevented the growth of the pathogen. Seed washings of infected seed samples had typical spores of the pathogen up to 10⁶ conidia per gram of seed. Transmission of the pathogen was shown by sowing infected seeds in disease-free field soils in two locations where anise had not been grown previously. Azoxystrobin, chlorothalonil + carbendazim and flutriafol seed treatments at 0.04 g a.i. kg⁻¹ seed, 1.0 g + 4.5 g a.i. kg⁻¹ seed and 0.015 g a.i. kg⁻¹ seed reduced the disease by 92.5%, 89.6% and 36.2% in 2002 and by 78.9%, 75.8% and 41.2% in 2003, respectively. Three foliar applications of axosystrobin, chlorothalonil + carbendazim and flutriafol at the rates of 187.5 g a.i. ha⁻¹, 1500 g + 6750 g a.i. ha⁻¹ and 31.3 g a.i ha⁻¹ reduced disease incidence by 92.5%, 86.0% and 96.8% in 2002 and by 97.5%, 90.8% and 97.0% in 2003, respectively. http://www.phytoparasitica.org posting May 17, 2005.     

Spore cell wall components ofAspergillus niger elicit downy mildew disease resistance in pearl millet

Elicitors derived from the cell wall of fungi are shown to be active in eliciting resistance in plants against a wide range of pathogens. In the present study carbohydrate components from the autoclaved spore cell wall ofAspergillus niger were prepared as aqueous suspensions and tested for defense response in pearl millet (Pennisetum glaucum (L.) R.Br.) against the oomycetous downy mildew pathogenSclerospora graminicola (Sacc.) Schroet. The aqueous suspension derived from the spore cell wall ofA. niger was used as a seed soak treatment at concentrations of 0.25, 0.5, 1.0, 1.5 and 2.0 mg ml⁻¹ for time intervals of 3, 6, 9 and 12 h. The concentration of 0.5 mg ml⁻¹ for a 6 h soaking period offered 94% seed germination and seedling vigor index increased to 1526. The seed germination and the seedling vigor were significantly higher than the untreated check. Spore cell wall suspension as seed treatment at a concentration of 0.5 mg ml⁻¹ required a 3-day time interval to provide 67% protection against downy mildew. Histological and biochemical studies were conducted to elucidate the mechanism of defense response in treated seedlings uponS. graminicola infection. Resistance host response was detected in the form of lignin and callose deposition in the epidermal cell wall of pearl millet seedlings, which is the site ofS. graminicola infection. A time course study showed rapid and localized deposition of lignin and callose in epidermal cell wall of carbohydrate components-treated pearl millet seedling coleoptiles. Increased levels of the defense-related enzyme peroxidase were detected in the treated seedlings. Peroxidase activity in elicitor-treated samples reached a peak at 8 h post-infection, which was 45% more than in their respective uninoculated control. Characterization of peroxidase isoforms by isoelectric focusing revealed 16 different isoforms, of which pI 6.8, 7.2 and 8.7 increased in elicitor-treated samples uponS. graminicola infection. http://www.phytoparasitica.org posting Nov. 14, 2005.     

Selection,characterization and genetic analysis of laboratory mutants of <Emphasis Type="Italic">Botryotinia fuckeliana</Emphasis> (<Emphasis Type="Italic">Botrytis cinerea</Emphasis>) resistant to the fungicide boscalid

Resistance to the fungicide boscalid in laboratory mutants of Botryotinia fuckeliana (Botrytis cinerea) was investigated. The baseline sensitivity to boscalid was evaluated in terms of colony growth (EC₅₀ = 0.3–3 μg ml⁻¹; MIC = 10–30 μg ml⁻¹) and conidial germination (EC₅₀ = 0.03–0.1 μg ml⁻¹; MIC = 1–3 μg ml⁻¹) tests. Mutants were selected in vitro from wild-type strains of the fungus on a fungicide-amended medium containing acetate as a carbon source. Mutants showed two different levels of resistance to boscalid, distinguishable through the conidial germination tests: low (EC₅₀ ∼ 0.3 μg ml⁻¹, ranging from 0.03 to 1 μg ml⁻¹; MIC > 100 μg ml⁻¹) and high (EC₅₀ > 100 μg ml⁻¹) resistance. Analysis of meiotic progeny from crosses between resistant mutants and sensitive reference strains showed that resistant phenotypes were due to mutations in single major gene(s) inherited in a Mendelian fashion, and linked with both the Daf1 and Mbc1 genes, responsible for resistance to dicarboximide and benzimidazole fungicides, respectively. Gene sequence analysis of the four sub-units of the boscalid-target protein, the succinate dehydrogenase enzyme, revealed that single or double point mutations in the highly conserved regions of the iron-sulphur protein (Ip) gene were associated with resistance. Mutations resulted in proline to leucine or phenylalanine replacements at position 225 (P225L or P225F) in high resistant mutants, and in a histidine to tyrosine replacement at position 272 (H272Y) in low resistant mutants. Sequences of the flavoprotein and the two transmembrane sub-units of succinate dehydrogenase were never affected.     

Aggressiveness and mycotoxin production of eight isolates each of <Emphasis Type="Italic">Fusarium graminearum</Emphasis> and <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> for ear rot on susceptible and resistant early maize inbred lines

Fusarium graminearum and F. verticillioides are among the most important pathogens causing ear rot of maize in Central Europe. Our objectives were to (1) compare eight isolates of each species on two susceptible inbred lines for their variation in ear rot rating and mycotoxin production across 3 years, and (2) analyse two susceptible and three resistant inbred lines for potential isolate x line interactions across 2 years by silk-channel inoculation. Ear rot rating, zearalenone (ZEA) and deoxynivalenol (DON) concentrations were evaluated for all F. graminearum isolates. In addition, nivalenol (NIV) concentrations were analysed for two NIV producers. Fumonisin (FUM) concentrations were measured for all F. verticillioides isolates. Mean ear rot severity was highest for DON producers of F. graminearum (62.9% of the ear covered by mycelium), followed by NIV producers of the same species (24.2%) and lowest for F. verticillioides isolates (9.8%). For the latter species, ear rot severities differed highly among years (2006: 24%, 2007: 3%, 2008: 7%). Mycotoxin concentrations among isolates showed a broad range (DON: 100–284 mg kg⁻¹, NIV: 15–38 mg kg⁻¹, ZEA: 1.1–49.5 mg kg⁻¹, FUM: 14.5–57.5 mg kg⁻¹). Genotypic variances were significant for isolates and inbred lines in all traits and for both species. Isolate x line interactions were significant only for ear rot rating (P < 0.01) and DON concentration (P < 0.05) of the F. graminearum isolates, but no rank reversals occurred. Most isolates were capable of differentiating the susceptible from the resistant lines for ear rot severity. For resistance screening, a sufficiently aggressive isolate should be used to warrant maximal differentiation among inbred lines. With respect to F. verticillioides infections, high FUM concentrations were found in grains from ears with minimal disease symptoms.     

Phytotoxic effects of fumonisin B1 on cowpea seed

The cultivation of cowpea (Vigna unguiculata) plays a vital role in the livelihood of many subsistence farmers and rural communities in tropical and subtropical countries. The seeds are prone to fungal infestation and mycotoxin contamination during sub-optimal storage conditions. Fumonisin B₁ (FB₁), produced byFusarium proliferatum, has been detected in cowpea seeds. Surface-disinfected seeds were imbibed in sterile distilled water amended with FB₁ at various concentrations. Percentage germination was determined according to the International Seed Testing Association rules. All the toxin concentrations significantly decreased seed germination and the two highest concentrations — 50 and 100μg ml⁻¹ FB₁ — inhibited root and shoot elongation. FB₁-treated embryonic tissues evinced compaction of the protoplasm and separation of the plasmalemma from the cell wall. Lipid bodies accumulated, which seemed to be lining the cell wall. This is the first study to demonstrate the phytotoxic effects of FB₁ on cowpea seeds.     

Sensitivity of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> to propamidine: in vitro determination of baseline sensitivity and the risk of resistance

The baseline sensitivity of Botrytis cinerea to propamidine and assessment of the risk of propamidine resistance in vitro are presented in this article. The baseline sensitivities of 41 wild-type strains were distributed as a unimodal curve with EC₅₀ values of mycelial growth ranging from 0.182 to 1.460 μg ml⁻¹, with a mean of 0.79 ± 0.27 μg ml⁻¹. A total of 10 resistant mutants, obtained from one parental strain, were induced by UV irradiation and selected for resistance to propamidine with an average frequency of 1.98 × 10⁻⁹ and 0.025 respectively. These mutants were divided into three classes of resistant phenotypes with low (LR), moderate (MR) and high (HR) levels of resistance, determined by the EC₅₀ values of 5.0–15.0 μg ml⁻¹, 15.1–75.0 μg ml⁻¹ and more than 75.0 μg ml⁻¹ respectively. Neither positive cross-resistance nor negative cross-resistance was detected between propamidine and the fungicides, benzimidazole carbendazim, anilino-pyrimidine pyrimethanil, dicarboximide iprodione or procymidone. All 10 propamidine-resistant mutants showed reduced mycelial growth in vitro, sporulation, spore germination and pathogenicity when compared with the parental strain. These studies demonstrated that propamidine possesses a low risk of resistance developing. However, as B. cinerea is a high-risk pathogen, appropriate precautions against resistance development should be taken.     

Effects of some fungicides on <Emphasis Type="Italic">Isaria farinosa</Emphasis>, and <Emphasis Type="Italic">in vitro</Emphasis> growth and infection rate on <Emphasis Type="Italic">Planococcus citri</Emphasis>

The effects of some fungicides used against citrus diseases, on mycelial growth and conidial germination of Isaria farinosa (Holmsk.) Fries [Sordariomycetes: Hypocreales] and also on the pathogenicity of the fungus on citrus mealybug, Planococcus citri (Risso), were determined. Systemic fungicides such as tebuconazole, penconazole and nuarimol were the most effective as regards both conidial germination and mycelial growth. Protective fungicides such as captan, chlorothalonil, mancozeb and propineb inhibited conidial germination at between 1 and 5 μg ml⁻¹ concentration, but captan, chlorothalonil and propineb did not inhibit the mycelial growth at 5,000 μg ml⁻¹. Mancozeb inhibited mycelial growth between 2,500 and 5,000 μg ml⁻¹. Sulphur and copper oxychloride did not inhibit the fungus even at very high concentrations. Sulphur, copper oxychloride, fosetyl-al, chlorothalonil and carbendazim did not decrease the mortality percentage caused by I. farinosa. Tebuconazole, penconazole and mancozeb were the most effective and respectively reduced the mortality from 83% to 33%, 28% and 30% in the ovisacs, from 81% to 29%, 27% and 29% in the 1st instar larvae, and from 84% to 34% in the adult females.     

<Emphasis Type="Italic">Paecilomyces lilacinus</Emphasis>, a potential biocontrol agent on apple rust mite <Emphasis Type="Italic">Aculus,schlechtendali</Emphasis> and interactions with some fungicides <Emphasis Type="Italic">in vitro</Emphasis>

The apple rust mite Aculus schlechtendali (Nal.) (Acari: Eriophyidae), is a main pest in apple-growing areas in Ankara, Turkey, and chemical control applications have some limitations. Entomopathogenic fungi have a potential for biological control of mites. In this study, an entomopathogenic fungus, Paecilomyces lilacinus (Thom) Samson (Deuteromycota: Hyphomycetes), was first isolated from the mite cadavers on Japanese crab apple leaves and pathogenicity of the fungus was observed in different inoculum densities and relative humidities. The pathogen caused up to 98.22% mortality of the mite population. The effects of some fungicides on the entomopathogenic fungus were determined in in vitro studies. Carbendazim, penconazole and tebuconazole were the most effective fungicides on mycelial growth of P. lilacinus, with EC₅₀ values under 3 μg ml⁻¹. In spore germination tests, captan, mancozeb, propineb were the most effective fungicides, followed by tebuconazole, penconazole, nuarimol and chlorothalonil. Sulphur could not inhibit the conidia germination totally at 5,000 μg ml⁻¹. Copper oxychloride and fosetyl-al prevented conidia formation at concentrations above 1,000 μg ml⁻¹.     

Seed dormancy and soil seedbank of the invasive weed Chenopodium hybridum in north‐western China

Seed dormancy and persistence in the soil seedbank play a key role in timing of germination and seedling emergence of weeds; thus, knowledge of these traits is required for effective weed management. We investigated seed dormancy and seed persistence on/in soil of Chenopodium hybridum, an annual invasive weed in north‐western China. Fresh seeds are physiologically dormant. Sulphuric acid scarification, mechanical scarification and cold stratification significantly increased germination percentages, whereas dry storage and treatments with plant growth regulators or nitrate had no effect. Dormancy was alleviated by piercing the seed coat but not the pericarp. Pre‐treatment of seeds collected in 2012 and 2013 with sulphuric acid for 30 min increased germination from 0% to 66% and 62% respectively. Effect of cold stratification on seed germination varied with soil moisture content (MC) and duration of treatment; seeds stratified in soil with 12% MC for 2 months germinated to 39%. Burial duration, burial depth and their interaction had significant effects on seed dormancy and seed viability. Dormancy in fresh seeds was released from October to February, and seeds re‐entered dormancy in April. Seed viability decreased with time for seeds on the soil surface and for those buried at a depth of 5 cm, and 39% and 10%, respectively, were viable after 22 months. Thus, C. hybridum can form at least a short‐lived persistent soil seedbank.     

Dormancy,germination and emergence of Urochloa panicoides regulated by temperature

Urochloa panicoides is an annual weed of summer crops. In Argentina, in subhumid areas with monsoon rainfall, it germinates and establishes in a single flush. To (i) identify the environmental factors that modify its seed dormancy level and germination and (ii) quantify the parameters describing the thermal behaviour of the germination and emergence dynamics of this weed under non‐limiting water conditions, we established a set of germination experiments performed (i) under controlled conditions using seeds after ripened for 3 or 6 months in different thermal and hydric conditions and (ii) under field conditions, where the soil temperature was modified by applying different shading levels. Seed dormancy level remained high with 3 months after ripening in all treatments. After 6 months, seeds stored at 4°C in dry conditions did not germinate at any temperature, while seeds stored at 25°C in dry conditions and in situ germinated c. 20% and 60% respectively. Germination percentage was higher in seeds harvested before their natural dispersal. The base, optimum and maximum temperatures for seed germination were 6, 35 and 45°C respectively. Shading reduced the number of emerged seedlings, possibly by reducing the soil thermal amplitude. The results explained the dormancy‐breaking mechanism of U. panicoides that allows a high germination rate in the field when rainfall occurs.     

Effect of phosphate‐based seed priming on strigolactone production and Striga hermonthica infection in cereals

Strigolactones, plant‐secreted underground signalling molecules, play an important role in agricultural ecosystems, because they mediate the interaction of crops with symbiotic AM fungi and parasitic weeds like Striga hermonthica. Cereal host plants secret these signalling molecules particularly under nutrient‐deficient conditions and especially when phosphate (P) is limiting. The objective of the present study was to see the potential of P seed priming for S. hermonthica management in cereals in relation to strigolactone production. It has been demonstrated that P fertiliser application down‐regulates the production of these signalling molecules in the rhizosphere, which results in lower S. hermonthica infection of cereals. The laboratory study showed maximum production of strigolactones from dry and water‐soaked seeds, while seed soaking in P solution reduced their production. Similarly, maximum S. hermonthica infection was observed under control treatments with dry sowing or water soaking, while P seed soaking decreased S. hermonthica germination, emergence and dry biomass in all cereal crops. Our study shows that P seed priming resulted in lower exudation of strigolactones, which induced less S. hermonthica seeds germination and hence may lead to lower S. hermonthica infection. P‐based seed priming could prove to be an effective and affordable strategy to reduce S. hermonthica infection in cereals. Further research for practical field application is needed.     

Black nightshade (<Emphasis Type="Italic">Solanum nigrum</Emphasis>) emergence and development is affected by seed density and burial depth

Black nightshade (Solanum nigrum L.) is a troublesome weed worldwide, affecting a large number of crops. As weed development is affected by a variety of factors, advanced knowledge on the plant’s biological and ecological qualities can support favorable management strategies. Seed density is an ecological factor affecting emergence and early growth of many weed species. For that reason, the current study characterizes and quantifies the seed density impact on black nightshade seedling-emergence and early growth, and the interaction between seed density and burial depth on these growth parameters. Greenhouse studies conducted in 2005, 2006 and 2010 examined black nightshade seed densities of 1, 5, 10, 25 and 50 seeds per pot, buried at, 1, 2, 3, 4 and 5 cm. Impact of seed density on weed emergence and growth was characterized by number of emerged plants and leaves, followed by biomass measurements. Seed density had no impact on emergence at the tested burial depths. There was a strong logistic relationship (P < 0.001, and R ²  > 0.97) between time and emergence, and a strong linear relationship (P < 0.002 and R ²  > 0.86) between time and number of leaves produced in all depths and seed densities. A negative relationship was observed between seed density and plant development and growth rate. Results of this study extend the knowledge on black nightshade biology and support decision-making concerning rational weed management.     

Effect of botanical extracts on the biological activity of granulosis virus against <Emphasis Type="Italic">Pieris brassicae</Emphasis>

A granulosis virus strain infecting Pieris brassicae (PbGV) was isolated from the dry temperate region of northwestern Himalayas as a potential microbial agent for its management. The effect of different botanicals (having insecticidal action against P. brassicae) on the bioefficacy of PbGV was evaluated under laboratory conditions using leaf disc bioassays on cabbage for improving the insecticidal performance of the PbGV. The synergistic action of different botanical extracts was evident in terms of reduction in LC₅₀ values against different botanical extracts. Among different extracts, petroleum-ether extract of neem seed kernel (NSK) when combined with PbGV resulted in maximum reduction of LC₅₀ value (4.39 × 10² occlusion bodies [OBs] ml⁻¹) followed by methanolic extract (7.38 × 10² OBs ml⁻¹) and aqueous extract (9.36 × 10³ OBs ml⁻¹) as compared with PbGV alone (1.85 × 10⁴ OBs ml⁻¹) for 2nd instar larvae of the test insect. These trends were found analogous in cases of 3rd and 4th instars of P. brassicae with different solvent extracts of NSK. The other botanicals evaluated, viz., Eupatorium and Artemesia, also resulted in reduction of LC₅₀ values for 2nd, 3rd and 4th instars as compared with PbGV alone when different extracts were combined with virus for bioassays. The studies suggest that the PbGV in combination with botanical pesticides could be more useful as a bio-pesticide against cabbage butterfly (P. brassicae) in IPM programs.     

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.     

Effect of cropping systems and crop successions on fumonisin levels in corn from Northern Paraná State,Brazil

In this study the effect of different cropping systems and crop successions was evaluated on natural Fusarium sp. contamination and fumonisin levels in corn. The cropping systems consisted of a conventional and no-tillage area cultivated with corn in the summer following either oats or fallow in the winter (2006 and 2007 growing seasons). In addition, the effect of applying nitrogen fertilizer (0, 22.5, 45.0, 90.0 and 90.0 kg ha⁻¹ nitrogen supplemented with potassium oxide) on fumonisin contamination was evaluated in the 2006 growing season. Fusarium sp. was detected in 90% samples in 2006 and in 100% samples in 2007. In both growing seasons, no-till corn following oats showed the highest mean fumonisin levels and differed significantly (P < 0.05) from all the others (2006) and from conventional till corn following either oats or fallow in the winter (2007). Fumonisin levels ranged from 0.13 to 19.52 μg g⁻¹ (mean 6.97 μg g⁻¹) and from 3.70 to 7.75 μg g⁻¹ (mean 6.29 μg g⁻¹) in no-till corn following oats from the 2006 and 2007 growing seasons, respectively. Plots treated with 0 kg ha⁻¹ and 22.5 kg ha⁻¹ nitrogen showed the highest mean fumonisin levels and differed significantly from those with 45.0 and 90 kg ha⁻¹ nitrogen. Fumonisin levels correlated negatively (P < 0.05) with the nitrogen fertilization rates. Although no-till is advantageous from a soil conservation standpoint, it may enhance the potential for fumonisin contamination in corn.     

Grass weed control with herbicide-treated crop seeds

Fluazifop-butyl applied in lung oil at rates of 4.4 to 0.5 g a.i. kg^?1 soybean seeds was evaluated in the glasshouse for control of Eleuisine indica. Soybean seeds pretreated with herbicide at 4.4 to 2.1 g a.i. kg^?1 gave 100% control of E. indica at the highest sowing rate of four seeds per pot and 90 to 80% control when sowed at one seed per pot. Soybeans were not injured by the seed treatment. Cotton seeds pre-treated with fluazifop at 2.2 g a.i. kg^?1 seeds and sown 4 cm apart in a row across a 20 ± 20cm tray of soil containing seeds of Echinochloa crus-galli produced a weed-free band 12 cm wide centred on the row of cotton, without injury to cotton. CGA-82725 (2-propynyl 2-(4-((3, 5-dichloro-2-pyridinyl)oxy)phenoxy) propanoate) at 2–3 g a.i. kg^?1 seeds was as effective as 4–4 g fluazifop-butyl in controlling E. indica. but growth of soybean was retarded. Sethoxydim gave less control than fluazifop butyl at comparable rates and did not injure soybeans.     

Effects of garlic (<Emphasis Type="Italic">Allium sativum</Emphasis>) juice containing allicin on <Emphasis Type="Italic">Phytophthora infestans</Emphasis> and downy mildew of cucumber caused by <Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis>

The volatile antimicrobial substance allicin (diallylthiosulphinate) is produced in garlic when the tissues are damaged and the substrate allicin (S-allyl-l-cysteine sulphoxide) mixes with the enzyme alliin-lyase (E.C.4.4.1.4). Allicin undergoes thiol-disulphide exchange reactions with free thiol groups in proteins and it is thought that this is the basis of its antimicrobial action. At 50 μg ml^-1, allicin in garlic juice inhibited the germination of sporangia and cysts and subsequent germ tube growth by Phytophthora infestans both in vitro and in vivo on the leaf surface. Disease severity in P. infestans-infected tomato seedlings was also reduced by spraying leaves with garlic juice containing allicin over the range tested (55–110 μg ml⁻¹) with an effectiveness ranging from approximately 45–100%. Similarly, in growth room experiments at concentrations from 50–1,000 μg ml⁻¹, allicin in garlic juice reduced the severity of cucumber downy mildew caused by Pseudoperonospora cubensis by approximately 50–100%. These results suggest a potential for developing preparations from garlic for use in specialised aspects of organic farming, e.g. for reducing pathogen inoculum potential and perhaps for use under glass in horticulture.     

Detection of viridiofungin A and other antifungal metabolites excreted by <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> active against different plant pathogens

Antibiosis is assumed to be an essential mechanism exerted by potential biocontrol agents (BCAs) of Trichoderma spp. Therefore, in the present study, we report for the first time on the elucidation and production of viridiofungin A (VFA) from T. harzianum isolate T23 cultures and investigate the antifungal potential of VFA and some other secondary metabolites purified from T. harzianum cultures against Fusarium moniliforme. The bioautography assay revealed that T. harzianum isolates T16 and T23 excreted several secondary metabolites with antifungal activity. Following isolation and purification of the antifungal zones, three fractions (F223, F323 and F423) from extracts of isolate T23 and two fractions (F416 and F516) from extracts of isolate T16 exhibited pronounced fungitoxic activity in the bioautography and antibiotic disk assays against Cladosporium spp. and F. moniliforme, respectively. The structure of the antifungal metabolite in fraction F323 was identified as viridiofungin A (VFA), the first report of production of VFA by isolate T23 of T. harzianum. Following cultivation of isolate T23 in PDB medium for 9 days, 94.6 mg l⁻¹ of VFA were determined. VFA and fraction F516 retarded the mycelial growth of F. moniliforme in the non-volatile phase assay by >90% for each 250 μg ml⁻¹ 7 days post-inoculation (dpi). While VFA and fraction F416 showed both volatile and non-volatile effects, fraction F516 seemed to exhibit mainly non-volatile activity. Microscopic examination revealed that hyphae of F. moniliforme grown on VFA-amended medium were less branched and appeared thicker than untreated hyphae. Furthermore, in the presence of VFA, formation of chlamydospores by F. moniliforme was increased. Finally, the antifungal spectrum of VFA towards various important plant pathogens was evaluated. Germination of propagules of a variety of fungal pathogens in vitro was differentially inhibited by VFA. While in the presence of 100 μg ml⁻¹ VFA conidial germination of V. dahliae was completely inhibited, a slightly higher concentration (150 μg ml⁻¹) of the inhibitor was required to suppress germination of Phytophthora infestans sporangia or sclerotia of Sclerotinia sclerotiorum. Contrary to several reports in the literature, VFA proved to be fungistatic rather than fungicidal. However, neither VFA nor the other Trichoderma metabolites, such as 6PAP, F416 and F516, exhibited any antibacterial activity against Gram-positive and Gram-negative bacteria.