Topic
Protoplast
About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.
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TL;DR: Maximal yields of protoplasts were affected by the nature and the molarity of the osmotic stabilizer used, the pH of the lytic medium, period of lytic digestion and age and amount of mycelium used in the digestion mixture.
Abstract: Factors affecting the release of protoplasts from mycelium of Aspergillus flavus Link, A. nidulans (Eidam) Wint., Penicillium chrysogenum Thom, Fusarium culmorum (W. G. Smith) Sacc. and Neurospora crassa Shear & Dodge, using a Streptomyces lytic enzyme preparation, were investigated. Maximal yields of protoplasts were affected by the nature and the molarity of the osmotic stabilizer used, the pH of the lytic medium, period of lytic digestion and age and amount of mycelium used in the digestion mixture.
39 citations
TL;DR: Somatic hybridization between triploid and diploid bananas was attempted by using protoplast electrofusion and nurse culture techniques, which showed different ploidy levels of the somatic hybrids.
Abstract: Somatic hybridization between triploid and diploid bananas was attempted by using protoplast electrofusion and nurse culture techniques. Protoplasts from embryogenic cell suspensions of 'Maca' (Musa sp. AAB group) were fused with protoplasts from nonembryogenic calli of`Lidi' (Musa sp. AA group). Direct somatic embryogenesis was observed when the fusion-treated protoplasts were cultured with rice nurse cells (Oryza sativa L. A-58 line). Somatic hybrids were identified by using random amplified polymorphic DNA (RAPD) markers. Forty-four primers were tested for polymorphism between the two parental varieties, and eleven of them showed polymorphism. Among the 13 regenerated plants from one experiment, 11 (85%) were identified as somatic hybrids by the presence of both parent bands, and 2 showed only the 'Lidi'-band patterns. From the other experiment, 5 of 11 regenerated plantlets (45%) were identified as somatic hybrids. Flow cytometric ploidy analyses showed different ploidy levels of the somatic hybrids.
39 citations
TL;DR: It is proposed that polyethylene glycol (PEG)-mediated DNA transformation of protoplasts relies on the uptake of DNA via tension-dependent endocytotic vesiculation, and it is argued that such vediculation is also involved in protoplast fusion.
39 citations
TL;DR: The isolate LP10 that was isolated from soil collected from fuel station was the most active isolate in lipase production and identified as Streptomyces exfoliates LP10, which was the best lipase producer compared to its parents.
Abstract: Lipases (triacyl glycerol acyl hydrolase) catalyze hydrolysis and syntheses of ester formed from glycerol and long-chain fatty acids. They have many industrial applications, especially in food and detergent industries. Out of 33 bacterial isolates, a group of 20 bacterial isolates produced lipase enzyme on tributyrin agar and Tween 80 agar media. In liquid medium, the lipase activity was ranged from 1.5 to 6.9 IU/ml. Among the evaluated bacteria, the isolate LP10 that was isolated from soil collected from fuel station. It was the most active isolate in lipase production (6.9 IU/ml). Using morphological, physiological and biochemical studies, it was identified as an isolate belonging to the genus Streptomycesand identified as Streptomyces exfoliates LP10. Identification was confirmed using 16S rDNA analysis. Growth of the selected bacterium in medium containing tributyrin and Tween 60 at initial pH 6 in addition to incubation at 37°C for three days yielded the maximum lipase production. The molecular weight of the purified enzyme was 60 kDa, determined using gel electrophoresis. Improvement of lipase production was carried out between Streptomyces exfoliates LP10 and Streptomyces niveus using protoplast fusion. Five fusants were obtained. Fusant LP3 was the best lipase producer (3 times higher) compared to its parents.
Key words: Lipase, Streptomyces exfoliates, tributyrin, protoplast fusion, 16S rDNA.
39 citations
TL;DR: The amount and quality of proteins extracted from the protoplast cells were much higher than that from the mycelial cells and no streaking or tailing was observed in the protoplasts, suggesting that effective extraction of intracellular proteins from protoplast might help suppress degradation of proteins during this process.
Abstract: Since filamentous fungi including basidiomycetous fungi possess an exceptionally robust cell wall as in microorganisms, effective extraction of intracellular proteins is a key step for fungal proteomic studies. To overcome the experimental obstacle caused by cell walls, we utilized fungal protoplasts, prepared from the brown-rot basidiomycete, Tyromyces palustris. The amount and quality of proteins extracted from the protoplast cells were much higher than that from the mycelial cells. Quantitative comparisons of proteome maps prepared from mycelial and protoplast cells indicated protein spots with a wider range of molecular weights and pIs in the protoplast sample. Furthermore, no streaking or tailing was observed in the protoplasts, suggesting that effective extraction of intracellular proteins from protoplasts might help suppress degradation of proteins during this process. In addition to the efficiency of protein extraction, simple and efficient subcellular fractionation was also achieved using protoplast cells.
39 citations