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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01 Jan 1994
TL;DR: An introcuction to optical microscopy for plant cell biology with the use of fluorescent probes for studies of living plant cells and general and enzyme histochemistry.
Abstract: 1.: An introcuction to optical microscopy for plant cell biology. 2.: The use of fluorescent probes for studies of living plant cells. 3.: General and enzyme histochemistry. 4.: Electron microscopy. 5.: In situ hybridization of RNA. 6.: DNA-DNA in situ hybridization - methods for light microscopy. 7.: Immunochemistry for light and electron microscopy. 8.: Plant protoplast techniques. 9.: Chemical analysis of the components of the primary cell wall. 10.: Immunofluorescent techniques for analysis of the cytoskeletan. 11.: Isolation of endo- and plasma membranes. 12.: Protein transport into intact chloroplasts and isolated thylakoids. 13.: Ion-selective micro-electrodes. 14.: Microsampling and measurements of solutes in single cells
204 citations
TL;DR: The response of protoplasts isolated from aleurone layers of barley to internally and externally applied hormone was analyzed and suggested that the site of perception of GA3 and ABA in the barley Aleurone protoplast is on the external face of the plasma membrane.
Abstract: The response of protoplasts isolated from aleurone layers of barley (Hordeum vulgare L. cv Himalaya) to internally and externally applied hormone was analyzed to localize the site of perception of the hormonal signal. Protoplasts responded to externally applied gibberellic acid (GA3) with increased synthesis and secretion of [alpha]-amylase, transient expression of the glucuronidase reporter gene fused to the hormone-responsive elements of the [alpha]-amylase promoter, and the vacuolation typical of GA3-treated aleurone cells. When up to 250 [mu]M GA3 was microinjected into the protoplast cytoplasm, none of these responses were observed. This did not reflect damage to the protoplasts during the microinjection procedure, since microinjected protoplasts remained responsive to externally applied hormone. Nor did it reflect loss of microinjected GA3 from the protoplast, since 50% of microinjected [3H]GA20 was retained by protoplasts for at least 24 h. Externally applied abscisic acid (ABA) could reverse the stimulation of [alpha]-amylase synthesis and secretion, whereas microinjecting up to 250 [mu]M ABA was ineffective at antagonizing the stimulatory effect of GA3. These results suggest that the site of perception of GA3 and ABA in the barley aleurone protoplast is on the external face of the plasma membrane.
204 citations
TL;DR: High frequencies of fusion were induced between isolated tobacco leaf protoplasts when they were incubated at 37 °C in the presence of 0.05 M CaCl2 in 0.4 ᴍ mannitol at pH 10.5 and had no irreversible, deleterious effects on the protoplast.
Abstract: High frequencies of fusion were induced between isolated tobacco leaf protoplasts when they were incubated at 37 °C in the presence of 0.05 M CaCl₂ in 0.4 ᴍ mannitol at pH 10.5. Subsequent to the fusion treatment the protoplasts were washed and cultured in a suitable medium and within two weeks, actively dividing cell colonies were observed. The fusion treatment had no irreversible, deleterious effects on the protoplasts
203 citations
TL;DR: Improved the thermotolerance and ethanol tolerance of an industrial yeast strain SM-3 by genome shuffling while simultaneously enhancing the ethanol productivity.
Abstract: Genome shuffling is a powerful strategy for rapid engineering of microbial strains for desirable industrial phenotypes. Here we improved the thermotolerance and ethanol tolerance of an industrial yeast strain SM-3 by genome shuffling while simultaneously enhancing the ethanol productivity. The starting population was generated by protoplast ultraviolet irradiation and then subjected for the recursive protoplast fusion. The positive colonies from the library, created by fusing the inactivated protoplasts were screened for growth at 35, 40, 45, 50 and 55°C on YPD-agar plates containing different concentrations of ethanol. Characterization of all mutants and wild-type strain in the shake-flask indicated the compatibility of three phenotypes of thermotolerance, ethanol tolerance and ethanol yields enhancement. After three rounds of genome shuffling, the best performing strain, F34, which could grow on plate cultures up to 55°C, was obtained. It was found capable of completely utilizing 20% (w/v) glucose at 45–48°C, producing 9.95% (w/v) ethanol, and tolerating 25% (v/v) ethanol stress.
197 citations
TL;DR: There is a disappearance of these masses, an increase in cell wall volume, and a return to smooth contour by the protoplast surface, paralleled by a return of the Golgi apparatus to an unhypertrophied state.
194 citations