The mechanism of fungistatic action of sec-butylamine: I. Effects of sec-butylamine on the metabolism of hyphae of Penicillium digitatum

Growth of Penicillium digitatum was inhibited after a 40-min incubation in a culture medium containing 0.5 mM sec-butylamine, and the dry weight of the hyphae was 50% of the control value after 180 min. Respiration of the hyphae was reduced 13% after a 20-min contact with 0.5 mM sec-butylamine but this treatment did not influence the uptake of amino acids, glucose, or phosphate nor intensify the efflux of ³³P- or ¹⁴C-labeled metabolites from the cells. The syntheses of cell walls and total lipids were inhibited 20–30% after a 90-min incubation with sec-butylamine, and nucleic acid synthesis was reduced to about 50% of the control value at this time. sec-Butylamine inhibited the incorporation of labeled carbon from [¹⁴C]glucose into the protein fraction of the hyphae to a greater degree than ¹⁴C derived from labeled proline, lysine, or leucine. These observations suggested that sec-butylamine interfered primarily with the intermediary metabolism of glucose rather than inhibiting a later stage of macromolecule synthesis. Hyphae incubated with [¹⁴C]glucose and sec-butylamine accumulated pyruvic acid to a level seven times greater than in control hyphae. Furthermore, sec-butylamine strongly inhibited ¹⁴CO₂ evolution from hyphae metabolizing [¹⁴C]pyruvate whereas CO₂ derived from acetate or glucose after a 45-min incubation was only slightly reduced by sec-butylamine. These observations implicate pyruvate oxidation as the primary site of sec-butylamine action in young hyphae of P. digitatum.     

High-pressure liquid chromatographic determination of sec-butylamine residues in potatoes

Fluorescamine, 4-phenylspiro[furan-2(3H), 1′(3′H)-isobenzofuran]-3,3′-dione, derivatives of primary aliphatic amines were separated by high-pressure liquid chromatography using a reverse-phase system with fluorescence detection. This technique was applied to the determination of residues of the fungicide sec-butylamine in potato tubers; the limit of detection was 0.36 pmol, equivalent to a residue of 0.1 mg kg^?1 in potato samples. A second amine, phenethylamine, was identified in extracts from artificially rotted potato flesh but this did not interfere with the analysis of sec-butylamine residues.     

The mechanism of fungistatic action of sec-butylamine: II. The effect of sec-butylamine on pyruvate oxidation by mitochondria of Penicillium digitatum and on the pyruvate dehydrogenase complex

sec-Butylamine at 5 mM inhibited the oxidation of pyruvate by mitochondria isolated from hyphae of Penicillium digitalum, but had little effect on the oxidation of citrate, isocitrate, succinate, malate, acetyl-coenzyme A, or reduced nicotinamide adenine dinucleotide. sec-Butylamine did not interfere with oxidative phosphorylation, as evidenced by similar $\text{P}\text{O}$ ratios in treated and control mitochondria. The pyruvate dehydrogenase complex (EC 1.2.4.1) isolated from young hyphae of P. digitatum was inhibited strongly by 20 mM sec-butylamine, whereas other tricarboxylic acid cycle enzymes were only slightly affected at most. Inhibition of the pyruvate dehydrogenase complex by sec-butylamine was competitive with respect to pyruvate. The K_i for sec-butylamine in the reaction was 1.38 × 10^?2M, and the K_m for pyruvate was 2.28 × 10^?4M. These observations and other evidence derived from studies with intact hyphae support the hypothesis that the pyruvate dehydrogenase complex is the primary site of the fungistatic action of sec-butylamine.     

The ultrastructure of germinating conidia of Penicillium digitatum inhibited by sec-butylamine

Electron micrographs of germinating conidia of P. digitatum showed osmiophilic inclusions scattered in the ground plasm or located within membrane-bound vacuoles where they were associated with various membranes and “myelin-like” structures. Cytochemical studies suggested that the osmiophilic inclusions within the vacuoles contained phospholipids and proteins. These phospholipoprotein inclusions may provide precursors needed for the assembly of membrane-bound organelles formed during germination. In conidia germinated in the presence of fungistatic concentrations of sec-butylamine (SBA) the vacuoles enlarged and the phospholipoprotein inclusions decreased in size and appeared to disintegrate and give rise to membrane-bound vesicles. SBA may deprive the conidium of materials normally utilized for membrane formation by the developing organelles.     

Ozone for control of post-harvest decay of table grapes caused byRhizopus stolonifer

The effect of ozone on post-harvest decay of table grapes was studied both with regard to its effectiveness and its possible mode of action. Ozone concentrations fell rapidly upon contact with organic matter and the amount which reacted with grape berries and the microflora on their surface was about 0.1mgg^-1, when supplied at a rate of 8mgmin^-1for 20min. The dose applied could be increased by longer periods of exposure, but symptoms of toxicity were observed on certain cultivars. The number of colony forming units (cfu) of fungi, yeasts and bacteria naturally present on the berry surface was considerably reduced by a 20min exposure to ozone. Ozone treatments significantly reduced the extent of berry decay caused by fungi following cold storage and increased shelf-life. A significant decrease in decay was observed in berries that were treated with ozone either before or after being inoculated withRhizopus stolonifer.This finding indicates that, in addition to its sterilizing effect, ozone also induced resistance to post-harvest decay development. The phytoalexins resveratrol and pterostilbene were elicited by ozone treatments, at levels similar to those produced by uv-c irradiation. Resveratrol accumulated in greater quantities than pterostilbene. Inoculation withR. stoloniferin addition to ozone treatment, raised the levels of both stilbenes even more. Exposing berries to ozone was almost as effective as SO₂fumigation for the control of storage decay caused byR. stoloniferand no deleterious effects were observed on the appearance of the grape cluster. Ozone treatments can therefore be considered as a possible substitute for SO₂fumigation for the control of post-harvest fungal decay.     

Effect of fenpropimorph and imazalil on sterol biosynthesis in penicillium italicum

Fenpropimorph was found to be highly active against Penicillium italicum (EC₅₀ 0.01/μg ml^?1). Conidia of P. italicum, treated with low concentrations of fenpropimorph, swelled in size and showed distorted germ tubes. During the initial stages of mycelial growth, fenpropimorph had little or no effect on the dry weight increase, which became strongly inhibited within 24 h after addition of the toxicant (0.05, 0.1 and 0.2 μg ml^?1). Irregular deposition of β–1,3 and β–1, 4 polysaccharides, probably chitin, was observed after treatment with fenpropimorph or imazalil. Fenpropimorph (0.05 and 0.2 μ ml^?1) caused the accumulation of a major demethyl-sterol that was different from ergosterol. It was identified as ergosta-8, 14, 24(28)-trien-3β-ol by mass, infrared, ultraviolet and proton nuclear magnetic resonance, spectrometric procedures. At both concentrations, the accumulation was already detected after incubation for 2 h. In contrast, imazalil (0.1 μg ml^?1) caused the accumulation of several methyl- and dimethyl-sterols which were tentatively identified as eburicol (24-methylene-24, 25-dihydrolanosterol), 4, 14α-dimethylergosta-8, 24(28)-dien-3-one, 14α-methylergosta-8, 24(28)-dien-3-one and obtusifoliol (4, 14α-dimethylergosta-8, 24(28)-dien-3α-ol). The accumulation of ergosta-8, 14,24(28)-trien-3β-ol indicates inhibition of the Δ¹⁴-reductase in P. italicum in a similar manner to that found previously in Ustilago maydis.     

Contamination of fresh and ensiled maize by multiple penicillium mycotoxins

Toxins produced by Penicillium species are reported in maize silage and have been associated with health problems in cattle. Our objectives were to evaluate the prevalence and dynamics of patulin (PAT), mycophenolic acid (MPA), cyclopiazonic acid (CPA), and roquefortine C (ROC) in fresh and ensiled maize. To achieve these objectives we developed a high-performance liquid chromatography method coupled with mass spectrometry to detect all four toxins simultaneously in silage. In addition we collected weather data, information on agronomic practices, and silage fermentation characteristics for each study site. Silage was collected at harvest and after ensiling in 2001 and 2002 from 30 Pennsylvania dairies. The average concentration of toxins (range in parentheses) was: PAT 0.08 microg/g (0.01 to 1.21), MPA 0.16 microg/g (0.02 to 1.30), CPA 0.12 microg/g (0.02 to 1.43), and ROC 0.38 microg/g (0.01 to 5.71). ROC was the most frequently detected toxin (60%), followed by MPA (42%), CPA (37%), and PAT (23%). Of 120 samples tested, 15% contained no detectible levels of toxin, 25% were contaminated with one toxin, 32% with two, 18% with three, and 10% with all four toxins. All four mycotoxins were found in freshly harvested material, contradicting the belief that Penicillium toxin formation occurs exclusively during storage. We observed that weather conditions during specific growth stages of the crop affected the final concentration of toxins in freshly harvested maize. In ensiled material, PAT levels were affected by concentrations of propionic and isobutyric acids. Based on our data, Penicillium mycotoxins can form while the crop is in the field and after ensiling, suggesting that preventative measures should begin prior to ensiling.     

The metabolism of fosetyl-aluminium and the evolution of residue levels in oranges and tangerines

The metabolism of fosetyl-aluminium to phosphonic acid in the peel of citrus fruit has been studied by gas chromatography using a nitrogen-phosphorus detector and also with a flame photometric detector. At the same time the phosphonic acid residues remaining in the peel and the pulp of citrus fruit, as a result of the application of aqueous solutions of fosetyl-aluminium at 2.5 g litre^?1 to trees prior to harvest was determined.     

The advantage of tbz + iprodione treatment for control of gray mold decay of celery caused by the heterogenic spore population ofBotrytis cinerea in Israel

A combined TBZ — iprodione treatment was more effective in inhibiting growthin vitro ofBotrytis cinerea isolates obtained from decayed celery than either of the fungicides applied separately. This was exhibited for both TBZ-resistant and TBZ-sensitive isolates. TBZ at 500 (μg ml^-1 plus iprodione at 1000 μg ml^-1 reduced celery decay beyond the reduction obtained by each fungicide alone. When applied prior to inoculation, the combined treatment prevented decay by the TBZ-sensitive/iprodione-resistant isolates and reduced initial decay by the TBZ-resistant/iprodione-sensitive isolates to 3–10% of the level without treatment. Under natural infection conditions iprodione showed better decay control than TBZ, and at 1500 μg ml^-1 it reduced initial decay during prolonged storage to 3% of the no-treatment level. Although TBZ (500 μg ml^-1) or iprodione (1000 μg ml^-1) applied separately reduced decay significantly, the combination of lower concentrations of each fungicide was sufficient to eliminate decay development almost totally. The combined treatment also inhibited decay bySclerotinia sclerotiorum, which contributed 3% of the total soft rot in stored celery.     

Rate, placement and timing of aldicarb applications to control Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), in oranges

BACKGROUND: The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is a cosmopolitan insect pest of citrus and vectors the bacterium Candidatus Liberibacter asiaticus, a suspected causal organism of citrus greening or ‘huanglongbing’ disease. Aldicarb 150 g kg^?1 GR (Temik^® 15 G) was evaluated at three rates, two placements and three timings for ACP control in orange trees. RESULTS: Application of aldicarb at 5.6, 2.8 and 1.4 kg AI ha^?1 in March 2006 reduced adults by 58–66%, 45–46% and 25–37% respectively compared with untreated controls in two separate trials. No difference was observed in placement (one versus two sides of the tree) or tree size (8 years old versus 12 years old). Application at 5.6 kg ha^?1 in January 2007 reduced adults by 86% and shoot infestation by 77% in spring, and was generally better than the November and especially February applications. Even more striking results were evident on adults caged on treated plants for 25 days in March. Spiders and ladybeetles were equally abundant in treated and untreated trees. CONCLUSION: Aldicarb application at 5.6 kg ha^?1 to the bed side of mature citrus trees 2–3 months before spring growth can suppress ACP through spring without a direct effect on principal psyllid natural enemies. Copyright © 2008 Society of Chemical Industry     

Acidified peel of litchi fruits selects for postharvestPenicillium decay

Litchi fruits are fumigated after harvest with sulfur dioxide (SO₂) to prevent their rapid browning. SO₂ blocks enzymatic activity but bleaches the fruits and, if this process is followed by dipping the fruit in dilute hydrochloric acid, the appealing red color is regained. Hot water brushing (HWB) is among the alternative methods that were developed to replace the use of SO₂. HWB reduced fungal population size on the surface of the fruit peel after treatment but did not eliminate fruit infection after storage. Whereas untreated fruits were infected with a variety of fungal species,Penicillium sp. was the only fungus that developed on the pericarp after storage in fruits that had been dipped in 1.5M HCl. Fruit treated by HWB followed by handling and storage under sterile conditions suffered greater decay than fruit stored under non-sterile conditions but with more ventilation. APenicillium sp. isolated from litchi grew well in liquid medium acidified to the pH range reported for SO₂ and HCl-treated litchi fruits. Morphological analysis identified fungal isolates asP. aurantiogriseum. Internal transcribed spacer sequence analysis of five isolates suggested a sequence similarity toP. commune. Our data support the hypothesis that dipping litchi fruit in hydrochloric acid eliminates infection by common opportunistic fungi and selects forPenicillium species that tolerate low pH. http://www.phytoparasitica.org posting April 30, 2004.     

秸秆腐熟剂菌种筛选及鉴定

为了快速降解秸秆,本试验从土壤中筛选出一株产纤维素酶的酵母菌,编号为J-3.通过形态学特征、生理生化特征及26S rDNA序列分析鉴定菌株J-3为胶红酵母菌(Rhodotorula mucilaginosa).经发酵条件优化产纤维素酶可以达到39.35U/mL,符合《农用微生物菌剂》GB20287-2006国家标准.为...     

Effect of temperature and leaf wetness duration on infection of sweet oranges by Asiatic citrus canker

Asiatic citrus canker, caused by Xanthomonas smithii ssp. citri , formerly X. axonopodis pv. citri , is one of the most serious phytosanitary problems in Brazilian citrus crops. Experiments were conducted under controlled conditions to assess the influence of temperature and leaf wetness duration on infection and subsequent symptom development of citrus canker in sweet orange cvs Hamlin, Natal, Pera and Valencia. The quantified variables were incubation period, disease incidence, disease severity, mean lesion density and mean lesion size at temperatures of 12, 15, 20, 25, 30, 35, 40 and 42°C, and leaf wetness durations of 0, 4, 8, 12, 16, 20 and 24 h. Symptoms did not develop at 42°C. A generalized beta function showed a good fit to the temperature data, severity being highest in the range 30–35°C. The relationship between citrus canker severity and leaf wetness duration was explained by a monomolecular model, with the greatest severity occurring at 24 h of leaf wetness, with 4 h of wetness being the minimum duration sufficient to cause 100% incidence at optimal temperatures of 25–35°C. Mean lesion density behaved similarly to disease severity in relation to temperature variation and leaf wetness duration. A combined monomolecular-beta generalized model fitted disease severity, mean lesion density or lesion size as a function of both temperature and duration of leaf wetness. The estimated minimum and maximum temperatures for the occurrence of disease were 12°C and 40°C, respectively.     

Surfactants for the control of apple mildew

Reduction of the overwintering powdery mildew fungus Podosphaera leucotricha resulted from one application of a variety of surfactants (at 50 g/litre a.i.) to apple trees during the dormant season. Examples of non-ionic, anionic and cationic materials were tested and some gave promising control. Successful treatments delayed bud burst during the following spring and caused some loss in fruit yield.     

Formulations for the control of dutch elm disease

Solutions of methyl benzimidazol-2-ylcarbamate (carbendazim, formerly MBC) in dilute hydrochloric, orthophosphoric or hypophosphorous acid and in dilute hydrochloric acid containing methanol (“Lignasan”) and aqueous solutions of 2-(thiazol-4-yl) benzimidazole (thiabendazole) hypophosphite, 8-hydroxyquinoline (oxine)hydro-chloride and dodecylguanidine acetate (dodine), were tested in the field for their efficacy in controlling Dutch elm disease caused by Ceratocystis ulmi. All gave some degree of control although some were phytotoxic, the benzimidazoles being the most active compounds tested. The anion and the concentration of acid present in the formulation may have some effect on retention of active material within the crown of the tree.     

Cross protection for the control of virus diseases

Control of ZYMV has been demonstrated in field trials at Wellesbourne and in Jersey by sap-inoculating courgette seedlings with a mild strain of the virus, prior to aphid-transmission of the severe strain. ZYMV has been found to be present in the testa and inner chlorenchyma tissue of the seed coat, but not in the cotyledon or embryo of mature courgette seeds.     

Observations on the potential of microwaves for weed control

Field tests of a prototype microwave‐based weed killer machine were conducted on Abutilon theophrasti, Panicum miliaceum, lucerne and oilseed rape pure stands. The approach can be considered a thermal weed control method, the microwave radiation causing dielectric heating of plant tissue water that eventually kills the plant. The method could overcome the limitations of other thermal methods, such as fire risk with flaming or the heavy loads required for hot water treatments. Species were effectively controlled by microwave irradiation, but their sensitivity and the evolution of damage symptoms over time differed. Lucerne showed no sigmoidal response and was the least affected by the treatment, while a log‐logistic curve expressed the dose–response relationships of the other species quite well. The estimated microwave dose for a 90% dry weight reduction ranged from 1015 kJ m^?2 in A. theophrasti to 3433 kJ m^?2 in P. miliaceum. Energy cost evaluation indicated that increased efficiency is required for this technique to compete with other thermal methods. Microwave efficiency could be increased by a flux configuration that minimizes soil penetration and maximizes absorption by plants, which, in turn, depends on plant growth form.     

大豆拟茎点种腐病菌的分离与鉴定

本文对美国进境大豆及其中混杂的豆杆上发现的拟茎点属（Phomopsis sp．）进行了形态学和分子生物学鉴定。无论从形态学还是从分子生物学角度都支持将该检测种鉴定为Phomopsis longicolla。该种病原真菌是在大豆上发生的危险性病原真菌，国内过去未见报道。     

Fumigant doses for the control of Trogoderma granarium1

The diapausing larvae of Trogoderma granarium are among the most tolerant of all storage pests to fumigation and effective dosage schedules are required. For this purpose, the toxicity of methyl bromide (MeBr), phosphine, methyl chloroform (MC), ethylene dichloride (EDC) and chlorobromomethane, and of mixtures of MeBr and MC, EDC and MC, and phosphine and MeBr, to diapausing larvae of the pest was examined at 15, 20 or 25°C. At 15 and 20°C the tolerance of larvae was high and very high dosages of MeBr or liquid fumigants, or long exposures to phosphine, were required for total kill. At 25°C larvae were more susceptible to phosphine but retained a high tolerance of other fumigants. A mixture of 2% MeBr in MC showed evidence of potentiation at 25°C and achieved control of larvae at moderate dosages.     

Acephate implants for control of the pine processionary caterpillar

Acephate (Orthene) cartridges spaced 14 cm apart along the stem circumference, implanted in November as a preventive treatment in 18-year-old pine trees, resulted in 100% mortality of young larvae of the pine processionary caterpillar,Thaumetopoea wilkinsoni Tams. A spacing of 10 cm was required to achieve 100% mortality of 3rd and 4th instar larvae. When implants were done in January, a spacing of 6 cm for control of the 4th and 5th instar larvae was needed.