Imazalil residue loading and green mould control in citrus packhouses

Imazalil (IMZ) is commonly applied in South African citrus packhouses for the control of green mould, caused by Penicillium digitatum, yet the disease still causes significant postharvest losses. The maximum residue limit (MRL) for IMZ on citrus fruit is 5 μg g⁻¹, whereas 2-3 μg g⁻¹ is a biologically effective residue level that should at least inhibit green mould sporulation. Standard compliance auditing of residue levels of citrus fruit, however, indicate that fruit from the majority of packhouses have residues of ≈1 μg g⁻¹. Poor disease control from insufficient residue loading might further be compounded by the presence of IMZ-resistant isolates of P. digitatum in packhouses. This study was conducted to assess the current status of IMZ application in South African packhouses, to determine the adequate residue levels needed to control green mould and inhibit its sporulation using both IMZ sensitive and resistant isolates, to investigate IMZ application methods and resultant residue levels in commercial citrus packhouses, and to study optimisation of modes of IMZ application in citrus packhouses. Factors studied were IMZ concentration, application type (spray vs. dip and drench), exposure time, solution temperature and pH, as well as curative and protective control of P. digitatum. The packhouse survey showed that the majority of packhouses applied IMZ in a sulphate salt formulation through a fungicide dip tank, and loaded an IMZ residue of ≈1 μg g⁻¹. In dip applications, IMZ had excellent curative and protective activity against Penicillium isolates sensitive to IMZ. However, curative control of IMZ resistant isolates was substantially reduced and protective control was lost, even at twice the recommended concentration, nor was sporulation inhibited. The use of sodium bicarbonate (2%) buffered imazalil sulphate solutions at pH ±8, compared with pH ±3 of the unbuffered solutions, markedly increased IMZ residue loading on Navel and Valencia oranges and improved curative and protective control of IMZ resistant isolates. Exposure time did not affect IMZ residue loading in IMZ sulphate solutions at pH 3, although the MRL was exceeded after 45 s exposure in pH 8 solutions. Imazalil applied through spray or drench application improved residue loading, but green mould control was less effective than after dip application.     

真空处理对青霉菌的抑制及吸湿性能影响研究

接种青霉菌橡胶木在温度15 ℃、绝对真空度7 mmHg(即933 Pa)下,分别处理0、12、24、36、48、60、72 h。经霉菌稀释计数观察青霉菌数量,傅里叶变换红外光谱、热重分析仪和动态水蒸气吸附分析仪分析橡胶木表面化学结构、热稳定性与吸湿解吸性能。结果表明,真空处理可明显抑制青霉菌生长,处理60 h,青霉菌几乎完全被杀死;接菌橡胶木与真空处理接菌橡胶木,其纤维素、半纤维素和木质素均有所降解,橡胶木表面亲水基团增加,表面亲水性增强,且后者易炭化,热稳定性能稍有降低;侵染青霉菌的橡胶木吸湿与解吸性能有所增大,真空处理后更趋明显。该研究可为真空处理抑制木材霉菌生长技术提供理论参考。     

Untargeted Metabolomics Approach for the Discovery of Environment-Related Pyran-2-Ones Chemodiversity in a Marine-Sourced Penicillium restrictum

Very little is known about chemical interactions between fungi and their mollusc host within marine environments. Here, we investigated the metabolome of a Penicillium restrictum MMS417 strain isolated from the blue mussel Mytilus edulis collected on the Loire estuary, France. Following the OSMAC approach with the use of 14 culture media, the effect of salinity and of a mussel-derived medium on the metabolic expression were analysed using HPLC-UV/DAD-HRMS/MS. An untargeted metabolomics study was performed using principal component analysis (PCA), orthogonal projection to latent structure discriminant analysis (O-PLSDA) and molecular networking (MN). It highlighted some compounds belonging to sterols, macrolides and pyran-2-ones, which were specifically induced in marine conditions. In particular, a high chemical diversity of pyran-2-ones was found to be related to the presence of mussel extract in the culture medium. Mass spectrometry (MS)- and UV-guided purification resulted in the isolation of five new natural fungal pyran-2-one derivatives—5,6-dihydro-6S-hydroxymethyl-4-methoxy-2H-pyran-2-one (1), (6S, 1’R, 2’S)-LL-P880β (3), 5,6-dihydro-4-methoxy-6S-(1’S, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (4), 4-methoxy-6-(1’R, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (6) and 4-methoxy-2H-pyran-2-one (7)—together with the known (6S, 1’S, 2’S)-LL-P880β (2), (1’R, 2’S)-LL-P880γ (5), 5,6-dihydro-4-methoxy-2H-pyran-2-one (8), (6S, 1’S, 2’R)-LL-P880β (9), (6S, 1’S)-pestalotin (10), 1’R-dehydropestalotin (11) and 6-pentyl-4-methoxy-2H-pyran-2-one (12) from the mussel-derived culture medium extract. The structures of 1-12 were determined by 1D- and 2D-MMR experiments as well as high-resolution tandem MS, ECD and DP4 calculations. Some of these compounds were evaluated for their cytotoxic, antibacterial, antileishmanial and in-silico PTP1B inhibitory activities. These results illustrate the utility in using host-derived media for the discovery of new natural products.     

胡椒调味油加工工艺条件的研究

以胡椒粉和大豆色拉油为主要原料制备风味独特的食用调味油,采用正交试验分析胡椒原料粒度、浸提温度以及质量比对胡椒调味油产品品质的影响,确定最佳工艺条件:以40目胡椒粉为原料,浸提温度为130℃,质量比(胡椒:大豆色拉油)为1:6(g/g).     

鹅源草酸青霉发酵鸡粪工艺的研究

为了利用微生物发酵鸡粪生产高质量的有机肥料,为鸡粪资源的合理利用和减少环境污染提供有效的解决途径。本文以从五龙鹅肠道分离的草酸青霉为发酵菌株,通过对鹅源草酸青霉发酵鸡粪生产生物肥料和减少环境污染的研究,来探讨发酵鸡粪的工艺条件。结果表明：利用草酸青霉为发酵菌株发酵鸡粪,使鸡粪中的氮、无机磷、有机质的含量分别提高12.13%、74.7%、15.6%;粗纤维的含量降低39.15%;氨气浓度降低了55.12%;发酵后的鸡粪大肠杆菌菌值为0.04,明显高于未发酵鸡粪（0.0004）;寄生虫卵死亡率为96%。通过正交优化试验确定的发酵鸡粪的工艺为：氮增幅最大时的发酵时间为60h,料水比为1：2,料层厚度为45cm,发酵温度为32℃;无机磷增幅最大时的发酵时间为60h,料水比为1：2,料层厚度为35cm,发酵温度为32℃;氨气浓度降低幅度最大时的发酵时间为60h,料水比为1：3,料层厚度为35cm,发酵温度为32℃。     

Dry mycelium of<Emphasis Type="Italic">Penicillium chrysogenum</Emphasis> protects cucumber and tomato plants against the root-knot nematode<Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

Incorporation into soil of dry mycelium ofPenicillium chrysogenum, a waste product of the pharmacological industry, enhanced plant growth and reduced root galling caused by the root-knot nematodeMeloidogyne javanica in cucumber and tomato plants. Incorporation into sandy loam soil in pots of dry mycelium at a concentration of 0.25% (w/w) resulted in complete protection of cucumber plants from the nematode. The number of juveniles recovered from soils containing dry mycelium was greatly reduced even at a concentration of 0.1% (w/w). In microplot studies conducted at two sites in two seasons, with three or four doses, dry mycelium caused a dose-dependent reduction in root galling index (GI) and promotion of plant growth of cucumber and tomato plants. Inin vitro studies, the water extract of dry mycelium immobilized nematode juveniles and reduced the egg hatching rate, but these effects were partly reversible after a rinse in water. Soil-drenching of cucumber and tomato seedlings with water extract of dry mycelium did not reduce GI or number of root-invading juveniles. The results show that dry mycelium promotes plant growth and protects plants against nematode infection. Protection, however, does not operatevia induced resistance. http://www.phytoparasitica.org posting April 6, 2003.     

Modes of Action of Pantoea agglomerans CPA-2, an Antagonist of Postharvest Pathogens on Fruits

Pantoea agglomerans CPA-2 is an effective antagonist against the postharvest pathogens Penicillium digitatum and Penicillium italicum on citrus fruits but its mode of action is unknown. Possible mechanisms studied in this work were antibiosis, induced resistance, competition and production of chitinolytic enzymes. P. agglomerans CPA-2 was unable to produce antibiotics or chitinolytic enzymes under the conditions tested. Induction of resistance by P. agglomerans CPA-2 was studied in oranges by measuring phenylalanine ammonia lyase and peroxidase enzyme activity in the orange peel at different time points after inoculation with the antagonist and/or the pathogen. No significant augmentation of enzyme activity after inoculation of oranges with P. agglomerans CPA-2 in the presence or absence of the pathogen was observed. P. agglomerans was effective only when it is in close contact with the pathogens. Competition for nutrients was studied using tissue culture plates with cylinder inserts, which allowed competition for nutrients to be studied without competition for space since physical contact between pathogen and antagonist was avoided. The presence of P. agglomerans in the tissue culture wells clearly decreased the germination of Penicillium conidia present in the cylinder when diluted orange peel extract or diluted potato dextrose broth was the nutrient source. Germination of Penicillium conidia, however, was almost completely inhibited when pathogen and antagonist were in physical contact. These results indicate that competition for nutrients is one of the modes of action of P. agglomerans CPA-2, but that physical contact between pathogen and antagonist is important for effective control.     

江西为害食用菌3种霉菌的生物学特性及综合防治技术

本文报道了江西为害食用菌的3种霉菌的生物学特性及其综合防治技术。它们是绿色木霉Trichoderma viride Persoon、产黄青霉Penicillium chrysogenum Thom、构巢曲霉Aspergillus nidulans (Eidam) Went。研究了这3种霉菌的生长特性、发生规律、传播途径、生活条件及其与几种食用菌菌丝的生长关系,并且提出了防治方法,为有效地控制食用菌中的霉菌生长提供了理论依据。     

Evaluation of food additives and low-toxicity compounds as alternative chemicals for the control of Penicillium digitatum and Penicillium italicum on citrus fruit

The effectiveness of low-toxicity chemicals as possible alternatives to synthetic fungicides for the control of post-harvest green and blue moulds of citrus was evaluated. A preliminary selection of chemicals, mostly common food additives, was made through in vivo primary screenings with oranges artificially inoculated with Penicillium digitatum or P italicum. Selected compounds and mixtures were tested as heated solutions in small-scale trials. Immersion of artificially inoculated oranges or lemons for 120 s in solutions at 40.6 degrees C and natural pH of potassium sorbate (0.2 M), sodium benzoate (0.2 M) or mixtures (0.1 + 0.1 M) of potassium sorbate with sodium benzoate, sodium propionate or sodium acetate were the most effective organic acid salts tested and reduced green mould by 70-80% after 7 days of storage at 20 degrees C. The mixtures did not significantly enhance the effectiveness of potassium sorbate or sodium benzoate alone. These solutions were as effective as sodium carbonate or calcium polysulphide treatments and, in general, they were more effective on lemons than on oranges. Satisfactory control of green and blue moulds was obtained by dipping oranges for 150 s in solutions of sodium molybdate (24.2 mM) or ammonium molybdate (1.0 mM) at 48 or 53 degrees C, but not at 20 degrees C. At 53 degrees C, however, the effectiveness of hot water was not enhanced by either molybdate. Molybdenum salts at higher concentrations were phytotoxic and stained the fruit. At non-phytotoxic concentrations, the effectiveness of these solutions was more influenced by temperature than by concentration. In general, the inhibitory effects of all compounds tested were not fungicidal but fungistatic and not very persistent. In conclusion, potassium sorbate, sodium benzoate and ammonium molybdate, among the wide range of chemicals tested, were superior for the control of post-harvest Penicillium decay of citrus fruit.     

Effect of the antagonist Candida sake on apple surface microflora during cold and ambient (shelf life) storage

This study examined the impact of the application of a biocontrol yeast, Candida sake CPA-1 (3 × 10⁶ colony forming units (cfu) ml^-1) on the resident microbial populations just prior to harvest, during 7 months cold storage and subsequent ambient shelf-life in two seasons on apples untreated or treated with a preharvest pesticide programme. The changes in populations of the antagonistic yeast ( C. sake) were also monitored. Generally, application of the antagonist had little effect on the total bacterial populations which remained very low both prior to harvest and subsequently during cold storage. White yeasts were predominant on the apples during the experimental period, with lower populations of pink yeasts. When apples were removed after 7 months to ambient conditions the yeast populations increased quickly but those apples treated with C. sake had significantly less white yeasts than untreated controls. The dominant filamentous fungi isolated were Cladosporium, Alternaria and Penicillium spp. Penicillium spp. which is responsible for major postharvest diseases was seldom isolated at preharvest but it became important during later cold storage and shelf life period. Populations of Cladosporium and Penicillium were significantly reduced by the C. sake treatment when removed from cold storage during the ambient shelf-life. In contrast, the Alternaria spp. were unaffected by the antagonist. The actual populations of C. sake applied decreased significantly immediately after application (24 h). However, they subsequently increased to a maximum after one month cold storage (10³ cfu g^-1), and populations increased again under ambient shelf-life conditions. The C. sake populations, and the resident microbial populations, were unaffected by preharvest fungicide applications. This study demonstrates that preharvest application of an antagonistic yeast such as C. sake has an impact on the resident mycoflora both during 7 months cold storage and during ambient shelf-life storage.