Revisiting the use of Iprodione and Trichoderma in the integrated management of onion white rot

The biocontrol properties of Trichoderma species are well documented, but their effectiveness in antagonism of the problematic Sclerotium cepivorum, the causal agent of white rot in Allium species, appears limited with reports of significant control only relating to deliberately-mutated strains of Trichoderma. Our previous studies have indicated the possibility of using selected naturally-occurring strains of the antagonist in the suppression of other diseases; now in vitro and controlled environment in vivo studies have indicated that a degree of control of Onion White Rot is possible, and that the selected antagonist strains can be used in integrated treatments with Iprodione to good effect. The possible value of such treatments is considered in light of other approaches to the suppression of this continuing problem.     

Differentiation of the causal pathogen of onion white rot Sclerotium cepivorum isolates by using the APIZYM system

The APIZYM system of detection of enzymes was proven to be useful in the differentiation of 15 European and Egyptian isolates of S. cepivorum, the incitant of onion white rot. The tested isolates produced alkaline phosphatase, esterase (c4), esterase lipase (c8), leucine arylamidase, valinearylamidase, trypsine, α-chymatrypsin, acid phosphatase, naphthol-AS-B1-phosphohydrolase, ß-galactosidase, ß-glucutronidase, α-glucosidase, ß-glucosidase and N-acetyl-ß-glucosaminidase and did not produced lipase (c14), crystine arylamidase, trypsine, ß-glucutronidase, α-mannosidase and α-fucosidase. According to enzyme activity, isolates can be divided into four groups (G). The differences between groups were in the activity of the enzymes α-chymotrypsin and α-glucosidase. The tested European isolates and the Egyptian isolates No.6 of the pathogen were in G1 and G2; however the rest of the Egyptian isolates were in G3 and G4.     

In vitro effect of fermented maize slurry and ethanol extracts of five plants against Sclerotium rolfsii Sacc. of tomato (Lycopersicon esculentum Mill.)

Sclerotium rolfsii Sacc. is an important fungal pathogen affecting the production of tomato (Lycopersicon esculentum Mill.). Conventional control methods using chemicals are expensive, which constitute environmental hazards. This has necessitated the search for alternatives in botanicals. Ethanol and supernatant solution of fermented maize slurry extracts from Ocimum gratissimum, Cymbopogon citratus, Xylopia aethiopica, Aframomum melegueta and Allium sativum were evaluated. In vitro test was carried out at 1, 2, 3, 4 and 5% concentrations against the pathogen. Mycelial growth was significantly reduced in the following descending order: O. gratissimum?>?A. melegueta?>?A. sativum?>?X. aethiopica?>?C. citratus. Supernatant solution extracts of O. gratissimum and A. melegueta at 5% concentration gave highest mycelial reduction (0.00?mm), respectively, while O. gratissimum ethanol extract recorded highest mycelial reduction (0.23?mm) among the ethanol extracts. Supernatant solution of fermented maize slurry extracts of O. gratissimum and A. melegueta at 5% concentration was more effective in reducing mycelial growth of S. rolfsii.     

Molecular characterization of European and Egyptian isolates of Sclerotium cepivorum,the incitant of onion white rot

Abstract

The tested European and Egyptian isolates of Sclerotium cepivorum were able to infect Giza 6 onion cultivar causing white rot disease with a different degrees of disease severity (ranging from sever to weak). The pattern of esterase isozymes produced by the tested isolates of the pathogen showed two main bands (arrows) which were different in density. Such differences in density of bands were present in every run and therefore appear to be indicators for differences among the tested isolates. Analysis of the protein pattern of the tested isolates of the pathogen indicated that the tested isolates had major proteins of a molecular weight of 52, 36, 23 and 16 kDa. Variation between isolates was detected by presence of bands of low molecular weight. Isolate Nos. 1, 4, 5, 7, 8, 9, 10 and 13 had a band at 17 kDa, whereas isolate Nos. 2, 3, 6, 11, 12, 14, and 15 had a band at 20 kDa. Using RAPD analysis to evaluate the genetic diversity of the tested isolates indicated that the tested field population of the pathogen was genetically heterogeneous but shared a number of common bands with molecular weights ranging from 650 to 2500 bp. Based on the DNA banding pattern the tested isolates can be assigned to seven genetically different groups. All tested isolates produced a band at 2500 bp except isolate No. 7. No correlation was exibited between patterns esterase isozmes, protein and DNA patterns of S. cepivorum isolates and their virulence or geographical origin.     

应用RAPD标记对斜生褐孔菌菌株亲缘关系的分析

采用RAPD技术对采集不同地区的8个斜生褐孔菌野生菌核分离得到的菌株亲缘关系进行研究,获得了斜生褐孔菌不同菌株的DNA指纹图谱。结果显示:12个引物共扩增出167条带,其中101条为多态性带,多态性比率为60.5%,同一培养时期的各菌株间RAPD图谱表明菌株间存在一定的种内及地理来源差异。若以遗传距离0.508为结合线,可将供试菌株划分为三大类,BCX01、BCX02归为一类,JL01、JL02、JL03、JL04、JL05归为一类,HLJ01单独聚为一类。     

Influence of the nitrogen source on the production and rheological properties of scleroglucan produced by Sclerotium rolfsii ATCC 201126

Scleroglucan production by Sclerotium rolfsii ATCC 201126 has been studied using nitrate or ammonium as nitrogen sources at several concentrations. In all the experiments carried out, both growth and production were modelled by an unstructured kinetic model using logistic and Luedeking–Piret equations for describing growth and production, respectively. The kinetic parameters for growth ( and Y_XN) and for production ( and ) were obtained by fitting the data to the model using the single-response non-linear regression technique by means of the algorithm of Marquardt coupled to a fourth-order Runge–Kutta algorithm. Biomass and scleroglucan production were higher when nitrate was the nitrogen source. Rheological properties of scleroglucan produced using nitrate as nitrogen source were studied and rheological parameters calculated, revealing similarity between this biopolymer and commercial scleroglucan.     

Immunolocalization and functional role of Sclerotium rolfsii lectin in development of fungus by interaction with its endogenous receptor

Many fungi are known to secrete lectins, but their functional roles are not clearly understood. Sclerotium rolfsii, a soilborne plant pathogenic fungus capable of forming fruiting bodies called sclerotial bodies, secrete a cell wall-associated Thomsen-Friedenreich antigen-specific lectin. To understand the functional role of this lectin, we examined its occurrence and expression during development of the fungus. Furthermore, putative endogenous receptors of the lectin were examined to substantiate the functional role of the lectin. Immunolocalization studies using FITC-labeled lectin antibodies revealed discrete distribution of lectin sites at the branching points of the developing mycelia and uniformly occurring lectin sites on the mature sclerotial bodies. During development of the fungus the lectin is expressed in small amounts on the vegetative mycelia and reaching very high levels in mature sclerotial bodies with a sudden spurt in secretion at the maturation stage. Capping of the lectin sites on the sclerotial bodies by lectin antibodies or haptens inhibit strongly the germination of these bodies, indicating functional significance of the lectin. At the maturation stage the lectin interacts with the cell wall-associated putative endogenous receptor leading to the aggregation of mycelium to form sclerotial bodies. The lectin-receptor complex probably acts as signaling molecule in the germination process of sclerotial bodies. Using biotinylated lectin, the receptors were identified by determining the specific lectin binding to lipid components, extracted from sclerotial bodies, and separated on thin-layer chromatograms. Preliminary characterization studies indicated that the receptors are glycosphingolipids and resemble inositolphosphoceramides. These findings together demonstrate the importance of lectin-receptor interactions to explain hitherto speculated functional role of the lectins and also the glycosphingolipids of fungi.     

Lipofuscins and sclerotial differentiation in phytopathogenic fungi

Lipofuscins of lipidic and proteinaceous origin were identified by their excitation and emission spectra in phytopathogenic fungal representatives of different sclerotial differentiation types. Lipofuscin pigments in Sclerotium rolfsii, Rhizoctonia solani, Sclerotinia minor and Sclerotinia sclerotiorum showed similar excitation and emission maxima (ex-em 330–450, 330–450, 330–470 and 3307–470 nm, respectively). Sclerotial differentiation of these fungi was proceeded by a 4.2, 2.5, 2.7, 2.5 and 6, 2.9, 3.8, 3.1 fold increase of lipofuscin accumulation (per lipid and protein content), per respective fungus, as compared to their undifferentiated stage. Lipofuscin levels were higher in older than in younger mycelia and this phenomenon was more profound in S. rolfsii. Since lipofuscins are considered as indicators of oxidative stress, these data are in accordance with the hypothesis that suggests oxidative stress to be a common underlying factor in sclerotial differentiation of sclerotia-forming filamentous phytopathogenic fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

A new microbial consortia containing entomopathogenic fungus,Beauveria bassiana and plant growth promoting rhizobacteria,Pseudomonas fluorescens for simultaneous management of leafminers and collar rot disease in groundnut

The virulence of 20 isolates of Beauveria bassiana (Balsamo) Vuillemin to larvae of the leafminer, Aproaerema modicella, was tested in the laboratory. Leafminer larvae were sprayed with a standard concentration of 1×10⁸ condia/mL of each B. bassiana isolate. All the B. bassiana isolates tested were pathogenic to A. modicella and the mortality varied between 16.7 and 68.9%. Beauveria bassiana isolate B2 was found to be the most virulent followed by isolate B4 which resulted in 59% mortality. Beauveria isolate B2 was selected for dose–response mortality studies with four different doses (1×10², 1×10⁴, 1×10⁶ and 1×10⁸ conidia/mL). Among the various doses tested, 1×10⁸ conidia/mL produced the highest mortality (70.7%). In addition, morphogenesis of the insect pest in all stages, larval, pupal and adult was greatly affected due to fungal infection. Further, B. bassiana isolate B2 and two Pseudomonas fluorescens strains, TDK1 and Pf1 were tested alone and in combination for suppression of groundnut leafminer and collar rot disease and promotion of plant growth and yield both under glasshouse and field conditions. The mixture of B. bassiana and P. fluorescens strains significantly reduced the leafminer and collar rot disease incidences when applied as talc-based formulation through seed, soil and foliar application under glasshouse and field conditions.