Studies on critical analysis of factors influencing improved production of protoplasts from Trichoderma reesei mycelium
TL;DR: The most suitable conditions for protoplasting were as follows: age of the organism in slant, 3 days; mycelium age, 20 h; volume of lytic enzymes, 190 ml;Mycelial weight (dry equivalent), 1.66 g; time of contact with lytic enzyme, 2 h; temperature of protoplasts, 30°C; phosphate buffer, 25 m m , pH 6.5; KCl as osmotic stabilizer, 0.7 m
About: This article is published in Enzyme and Microbial Technology.The article was published on 1992-03-01. It has received 43 citations till now. The article focuses on the topics: Trichoderma longibrachiatum & Mycelium.
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TL;DR: The use of high concentrations of chitin and ammonium sulphate and exclusion of peptone and urea from the medium resulted in the production of higher level of the enzyme, which was maximized to the level of 0.197 U.
Abstract: Statistical design was used to determine the optimal levels of medium components, the optimal initial pH of the enzyme production medium, the temperature of fermentation, age of the organism in the slant growth and the age of the inoculum for the production of chitinase in shake flask fermentations. The use of high concentrations of chitin and ammonium sulphate and exclusion of peptone and urea from the medium resulted in the production of higher level of the enzyme. The optimal concentrations of the medium components were 12.5 kg/m3 and 4.2 kg/m3 for the chitin and ammonium sulphate respectively. The effect of the addition of peptone and urea to the optimized medium was studied. The optimal values of initial pH and temperature were 5.6 and 28 °C respectively. The optimal age of the slant and the inoculum were found to be 105 h and 43 h respectively. The highest level of chitinase before optimization of the above variables was 0.054 U which was maximized to the level of 0.197 U.
38 citations
TL;DR: A temperature of 30°C and an initial pH of 4.7 were found to be optimal for β-1,3-glucanase production from T. harzianum in both surface culture and submerged culture processes.
38 citations
TL;DR: The results showed that the combined application of A. oligospora and SA provided the best nematode control and the activity of the enzymes and phenolic compounds increased in comparison with the control.
Abstract: Root-knot nematodes (RKN) (Meloidogyne spp.) are economically the most important pathogens of agricultural products. The aim of the present study was to control Meloidogyne javanica by using Arthrobotrys oligospora and salicylic acid (SA) and to analyse the kinetics of enzymes, phenylalanine ammonia lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO) and phenolic compounds accumulation in the root system of tomato after inoculation with M. javanica, A. oligospora and SA. The ability of A. oligospora to produce extracellular proteases was also examined. In greenhouse studies, we used soil drenching of A. oligospora (106 spores/ml) and soil drenching or leaf spraying of SA (5 mM) in six-leaf stage, separately and in combination. Experiments were performed in a completely randomised design. The efficiencies of treatments were appraised by using diameter of galls, number of galls per plant, number of egg masses per plant, number of eggs per egg mass, root and foliage fresh weight. The results showed that ...
34 citations
Cites methods from "Studies on critical analysis of fac..."
...The reaction was carried out as described by Kumari and Panda (1992) with four replicates....
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TL;DR: All the stages involved in fusion of protoplasts and some of the applications of protoplast fusion technique in fungal systems are discussed.
Abstract: Protoplast fusion is a non-specific recombination technique used for transfer of cytosolic organelles including genetic material. The process involves cell wall breakdown, regeneration of protoplasts, chemofusion and electrofusion. This review article discusses all the stages involved in fusion of protoplasts and some of the applications of protoplast fusion technique in fungal systems.
32 citations
TL;DR: The results have indicated that T. aureoviride URM 5158 is an effective biocontrol agent against cassava root rot caused by F. solani, because it presented competitive antagonist capability in vitro, the highest chitinase production, and reduced the cassavaRoot rot severity.
Abstract: Trichoderma has been used to manage a large number of pathogens, but there is a gap in the mechanisms used by these biocontrol agents regarding the physiological response of cassava plants (Manihot esculenta) when it is subjected to cassava root rot. The aims of this study were to investigate the antagonist activity of ten Trichoderma isolates against Fusarium solani on potato dextrose Agar (PDA), to quantify the chitinase production, to select and test in vivo the best isolate from each experiment and to assess the physiological response of cassava to the production of oxidative enzyme complex production (ascorbate peroxidase, catalase, peroxidase and polyphenol oxidase). All Trichoderma isolates have shown competitive capability against F. solani, and Trichoderma hamatum URM 6656 showed the highest inhibition of pathogen growth (88.91%). All isolates have shown chitinase activity, but Trichoderma aureoviride URM 5158 produced the highest amount of chitinase. T. hamatum URM 6656 and T. aureoviride URM 5158 were selected to be applied in vivo. The two Trichoderma strains reduced 64 and 60% of the disease severity in the shoot and 82 and 84% in the root. Cassava plants infected with Trichoderma have shown the highest peroxidase and ascorbate peroxidase production. Our results have indicated that T. aureoviride URM 5158 is an effective biocontrol agent against cassava root rot caused by F. solani, because it presented competitive antagonist capability in vitro, the highest chitinase production, and reduced the cassava root rot severity. The application of T. aureoviride has led to the maximum enzyme activity of reactive oxygen species group in cassava plants.
29 citations
References
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TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
225,085 citations
25,389 citations
TL;DR: The protoplast Fusion and the Hybridization of Fungal Species and Transformation in Fungi by Using Protoplasts by using Enzymatic and Nonenzymatic methods are studied.
Abstract: INTRODUCTION 21 ISOLATION OF PROTOPLASTS AND THEIR P ROPERTIES 22 Enzymatic Methods for Protoplast Isolation 22 Nonenzymatic Procedures for Protoplast Release 26 Properties of Protopiasts 27 WALL REGENERATION AND REVERSION OF PROTOPLASTS 28 Frequency of Protoplast Reversion 28 Morphology of Protoplast Reversion 29 Ultrastructure and Composition of the Regenerated Wall ..... 30 Biochemical Aspects oj Wall Biogenesis 31 PROTOPLAST FUSION AND GENETIC MANIPULATION 33 Protoplast Fusion and the Hybridization of Fungal Species 33 Transformation in Fungi by Using Protoplasts . .. ........ 36 CONCLUDING REMARKS 36
187 citations
TL;DR: Heterokaryon formation, at high frequency, after induced intraspecific fusion between protoplasts of auxotrophs of Penicillium chryosogiwim, P .
Abstract: Although protoplast fusion is of current interest because of its possibilities in pure and applied genetics, only a few reports exist on intraspecific fusion between fungal protoplasts and subsequent selection of heterokaryons. Naturally-occurring fusion between protoplasts of Geotrichum candidurn (Ferenczy, Kevei & Zsolt, I 974) and Aspergillz4s nidulans (Ferenczy, Kevei & Szegedi, I 975) has been reported. Binding & Weber (1974) described fusion between protoplasts of Phycomyces blakc~slceanus induced by seawater or calcium ions at high pH, but the frequency of heterokaryon formation was low in all cases. We here report heterokaryon formation, at high frequency, after induced intraspecific fusion between protoplasts of auxotrophs of Penicillium chryosogiwim, P . putulurn, P. roquejortii, A. nidulans, A . niger and Cephalosporium acrernonium, and fusion between protoplasts of P. chrysogenu~?z and P. rzotutunz auxotrophs, using solutions containing polyethylene glycol (PEG) mol.wt. 6000.
157 citations
TL;DR: All four in vitro methods of estimating the percentage viability of a cell suspension gave similar results with a recently harvested cell suspension but three of them grossly overestimated the proportion of viable cells in a suspension which had been stored at 4 °C for 11 days.
134 citations