Formation and Regeneration of Protoplasts from Conidiobolus lamprauges

Ishikawa, Fumihiko; Ōishi, Kunio

doi:10.1099/00221287-131-12-3311

Cited by 5 publications

(4 citation statements)

References 15 publications

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“…This is due to the fact that crude filtrate contains cocktail of different glycosyl hydrolases which synergistically acting on A. niger, favors rapid hydrolysis of fungal cell wall and endorse protoplasts formation. Our result was also comparable with other reports (20,21). One of the most important prerequisite features of protoplasts as genetic tool is their regeneration into mycelial form.…”

Section: Thermodynamic Viability Studysupporting

confidence: 95%

Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation

Halder

Maity

Jana

et al. 2014

Journal of Bioscience and Bioengineering

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Section: Thermodynamic Viability Studysupporting

confidence: 95%

Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation

Halder

Maity

Jana

et al. 2014

Journal of Bioscience and Bioengineering

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“…commune and C . lamprauges (de Vries & Wessels, 1975;Ishikawa & Oishi, 1985). On the other hand, 17 % of the sedimenting protoplasts from Scl.…”

Section: Activators and Inhibitors Of 13-p-glucan Synthase Activitymentioning

confidence: 99%

1,3- -Glucan synthase in cell-free extracts from mycelium and protoplasts of Sclerotium glucanicum

Kottutz

Rapp

1990

Journal of General Microbiology

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Conditions for obtaining stable protoplasts from Sclerotium glucanicum and their reversion to hyphal growth were determined. 1,3+Glucan synthase activity was detected in particulate enzyme fractions from mycelium and protoplasts of Scl. glucanicum. UDP-[U-14C]glucose was linearly incorporated into a p-glucan for about 20 min at 25 OC. Optimum pH and temperature values, as well as thermal stabilities of the 1,3-P-glucan synthase activity, were determined. High concentrations of EDTA were inhibitory. Enzyme activity was stimulated by ATP and GTP. The apparent K,,, value for UDP-glucose was 0.54 mM. The reaction product was characterized as 1,3-pglucan by 13C N M R spectroscopy and hydrolysis products of an exo-1,3-P-glucanase.

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“…Ramos and Garcia -Acha (1975) studied regeneration of Trichoderma viride protoplasts in winge medium and observed that a normal germ-tube began to form from protoplast and normal mycelium was formed and this new regenerated mycelia showed same characteristics as those of the original mycelia of Trichoderma viride. This pattern of regeneration of protoplasts in Trichoderma is in line with our findings of pattern of regenerating protoplasts in both strains A and R. The same regeneration pattern was observed byIshikawa and Oishi (1985) in the protoplasts of Conidiobolus lamprauges in soft agar.…”

supporting

confidence: 91%

Preparation and fusion of protoplasts from the phytopathogenic fungus Sclerotium rolfsii (Sacc.)

Sikandar¹

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1.11. Epidemiology 1.12. Control of Disease 1.13. Aims of Thesis 2. Materials and Methods 2.1. Organisms and Growth conditions 2.2. Preparation of semi-permeable cellophane membrane 2.3. Pretreatment of mycelia 2.4. Digestion mixtures 2.4.1. Osmotic stabilizers 2.4.2. Lytic enzymes 2.5. Protoplast formation 2.6. Determination of protoplast number 2.7. Purification of protoplast 2.8. Determination of protoplast size 2.9. Regeneration of protoplasts 2.9.1. Regeneration on solid medium 2.9.2. Regeneration in liquid medium 2.10. Fusion of protoplast 2.11. Study of stability of fusants 2.12. Growth rate of fusants 2.13. Pathogenic properties of fusants in green house pot experiment 2.13.1. Inoculum preparation 2.13.2. Soil preparation 2.13.3. Selecetion of host crops 66 2.13.4. Seed soaking and planting 66 2.13.5. Study of growth parameters of tomato, carrot and beans 67 2.13.5.1 Growth parameters studied 67 2.13.6. Disease incidence and mortality 68 2.13.7. Chlorophyll fluorescence 68 2.13.8. Estimation of chlorophyll a, b and carotenoids 69 2.14. Statistical analysis 70 3. Results 71 3.1.Strains A and R of Sclerotium rolfsii 71 3.2. Optimization of protoplast yields from Sclerotium rolfsii 71 3.2.1. Effect of pretreatment of mycelia 71 3.2.2. Effect of the osmotic stabilizers on protoplast yields and morphology74 3.2.2.1. Effect of different molarities of MgSO 4 on protoplast yields 76 3.2.3. Effect of different lytic enzymes on protoplast yields 76 3.3. Measurement of size of protoplast 3.4. Mechanism of protoplast release from the hyphae of Sclerotium rolfsii 3.5. Regeneration of protoplast into hyphae 3.6. Fusion of protoplasts of strains A and R 3.6.1. Recovery of the fusants 88 3.6.2. Morphological characters of fusants 3.6.3. Stability of the fusants 92 3.6.4. Growth rate of fusants 92 3.7. Assessment of pathogenicity of fusants and parent strains A and R of Sclerotium roflii 94 3.8. Physiological parameters 3.8.1.Chlorophyll fluorescence 3.8.2. Estimation of chlorophyll a, b and carotenoids 4. Discussion 4.1. Investigation into optimization for protoplast yields 4.3. Protoplast release mechanism 4.4. Protoplast regeneration 4.5. Protoplast fusion 4.6. Properties of fusants 4.7. Pathogenicity of fusants 5. References

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Formation and Regeneration of Protoplasts from Conidiobolus lamprauges

Cited by 5 publications

References 15 publications

Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation

Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation

1,3- -Glucan synthase in cell-free extracts from mycelium and protoplasts of Sclerotium glucanicum

Preparation and fusion of protoplasts from the phytopathogenic fungus Sclerotium rolfsii (Sacc.)

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