Academic literature on the topic 'Protoplasts'

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Journal articles on the topic "Protoplasts"

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Han, Jong-Eun, Han-Sol Lee, Hyoshin Lee, Hyunwoo Cho, and So-Young Park. "Embryogenic Stem Cell Identity after Protoplast Isolation from Daucus carota and Recovery of Regeneration Ability through Protoplast Culture." International Journal of Molecular Sciences 23, no. 19 (September 30, 2022): 11556. http://dx.doi.org/10.3390/ijms231911556.

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Protoplasts are single cells isolated from tissues or organs and are considered a suitable system for cell studies in plants. Embryogenic cells are totipotent stem cells, but their regeneration ability decreases or becomes lost altogether with extension of the culture period. In this study, we isolated and cultured EC-derived protoplasts (EC-pts) from carrots and compared them with non-EC-derived protoplasts (NEC-pts) with respect to their totipotency. The protoplast isolation conditions were optimized, and the EC-pts and NEC-pts were characterized by their cell size and types. Both types of protoplasts were then embedded using the alginate layer (TAL) method, and the resulting EC-pt-TALs and NEC-pt-TALs were cultured for further regeneration. The expression of the EC-specific genes SERK1, WUS, BBM, LEC1, and DRN was analyzed to confirm whether EC identity was maintained after protoplast isolation. The protoplast isolation efficiency for EC-pts was 2.4-fold higher than for NEC-pts (3.5 × 106 protoplasts·g−1 FW). In the EC-pt group, protoplasts < 20 µm accounted for 58% of the total protoplasts, whereas in the NEC-pt group, small protoplasts accounted for only 26%. In protoplast culture, the number of protoplasts that divided was 2.6-fold higher for EC-pts than for NEC-pts (7.7 × 104 protoplasts·g−1 FW), with a high number of plants regenerated for EC-pt-TALs, whereas no plants were induced by NEC-pt-TAL. Five times more plants were regenerated from EC-pts than from ECs. Regarding the expression of EC-specific genes, WUS and SERK1 expression increased 12-fold, and LEC1 and BBM expression increased 3.6–6.4-fold in isolated protoplasts compared with ECs prior to protoplast isolation (control). These results reveal that the protoplast isolation process did not affect the embryogenic cell identity; rather, it increased the plant regeneration rate, confirming that EC-derived protoplast culture may be an efficient system for increasing the regeneration ability of old EC cultures through the elimination of old and inactivate cells. EC-derived protoplasts may also represent an efficient single-cell system for application in new breeding technologies such as genome editing.
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Perera, Srini C., and Peggy Ozias-Akins. "Regeneration from Sweetpotato Protoplasts and Assessment of Growth Conditions for Flow-sorting of Fusion Mixtures." Journal of the American Society for Horticultural Science 116, no. 5 (September 1991): 917–22. http://dx.doi.org/10.21273/jashs.116.5.917.

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Petiole protoplasts of the sweetpotato [Ipomoea batatas (L.) Lam.] cultivars Red Jewel and Georgia Jet formed cell walls within 24 hours and divided in 2 to 3 days. Pretreating enzyme solutions with activated charcoal increased the viability and division frequency of protoplasts. Culture of protoplast-donor plants in a medium containing STS did not affect plant growth, protoplasm yield, or viability, but did increase the division frequency. Culture of protoplasts for 24 hours in a medium containing DB, a cell wall synthesis inhibitor, or staining of protoplasts with FDA did not significantly affect division frequency. The division frequency of protoplasts cultured in liquid medium was significantly higher than that of protoplasts cultured in agarose-solidified medium. Cell cycle analysis of petioles and freshly isolated protoplasts showed that the latter has a significantly higher proportion of nuclei in G1 phase. Protoplasts did not initiate DNA synthesis or mitosis within the first 24 hours of culture. Low-frequency regeneration of shoots from protoplast-derived callus was accomplished on MS medium containing 1.0 mg ldnetin/liter when preceded by MS medium modified to contain only (in mg·liter-1) 800 NH4NO3, 1400 KNO3, 0.5 2,4-D, 0.5 kinetin, and 1.0 ABA. Roots produced from protoplast-derived callus formed adventitious shoots after 4 weeks on MS medium containing 2% sucrose, 0.02 mg kinetin/liter and 0.2% Gelrite. Secondary shoot formation from regenerated roots will be a more effective means of obtaining plants from protoplasts than direct shoot regeneration from callus. Chemical names used: silver thiosulfate (STS): 2.6-dichlorobenzonitrile (DB); fluorescein diacetate (FDA): 2.4-diacetate (FDA); 2.4 dichlorophenoxyacetic acid (2,4-D); abscisic acid (ABA).
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Struck, C., R. Rohringer, and R. Heitefuss. "Isolation and lectin-binding properties of barley epidermal and mesophyll protoplasts." Canadian Journal of Botany 72, no. 11 (November 1, 1994): 1688–91. http://dx.doi.org/10.1139/b94-207.

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Protoplasts from primary leaves of barley (Hordeum vulgare L.) were obtained by enzymatic digestion and fractionated by discontinuous density gradient centrifugation to yield highly enriched fractions of mesophyll and epidermal protoplasts. A characterization of both protoplast types resulted in a clear differentiation of the outer protoplast surfaces. The protoplasts were examined for affinity to various lectins by agglutination tests and by labeling with lectin – fluorescein isothiocyanate conjugates. Both types of protoplasts agglutinated with soybean lectin. Fluorescein isothiocyanate-labeled soybean lectin was uniformly distributed on the protoplast surface. Mesophyll protoplasts, but not protoplasts from the epidermis, were agglutinated by Concanavalin A. Both types of protoplasts exhibited fluorescence labeling with Concanavalin A – fluorescein isothiocyanate conjugate. This label often showed a patchy distribution on the protoplast surface. Tetragonolobus lectin and β-D-glucosyl Yariv artificial antigen agglutinated mesophyll but not epidermal protoplasts. One of three tested monoclonal antibodies with specificity for arabinogalactans had affinity to the surface of mesophyll and epidermal protoplasts. Key words: agglutination, arabinogalactan protein, cell surface, epidermal protoplasts, fluorescence labeling, lectin.
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Sinha, Anupam, Andrew C. Wetten, and P. D. S. Caligari. "Effect of biotic factors on the isolation of Lupinus albus protoplasts." Australian Journal of Botany 51, no. 1 (2003): 103. http://dx.doi.org/10.1071/bt01104.

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Several tissue types of Lupinus albus L. were investigated as sources for the isolation of protoplasts. Cotyledons from in vitro seedlings were found to yield the highest number of protoplasts compared with leaves, hypocotyls and roots. A combination of the protoplast isolation enzymes, cellulase and Pectolyase Y23, was capable of releasing the highest number of protoplasts compared with a combination of cellulase and Macerase. Protoplast yield increased with increasing cotyledon age but was accompanied by a progressive decline in protoplast viability. The optimal combination of protoplast yield and viability occurred when the protoplasts were isolated from 14- to 18-day-old cotyledons. The ratio between the volume of enzyme solution and the tissue biomass did not affect the protoplast production significantly. This is the first report of the isolation of protoplasts from a lupin cotyledon and, following the procedure described in this paper, an average yield of 1.2 × 106 protoplasts per gram of fresh tissue was obtainable.
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Qiu, Quan-Sheng, Ze-Zhou Wang, Nang Zhang, Qi-Gui Cai, and Rong-Xi Jiang. "Aquaporins in the plasma membrane of leaf callus protoplasts of Actinidia deliciosa var. deliciosa cv. Hayward." Functional Plant Biology 27, no. 1 (2000): 71. http://dx.doi.org/10.1071/pp99033.

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The water transport activity of Actinidia deliciosa protoplasts was determined using a cell imaging system. Results showed that the protoplast volume increased swiftly when placed in a hypoton-ic medium, and also increased with an increase in medium osmotic gradients. The osmotic water permeability coefficient (Pf) values were 0.118 × 10–3, 0.121 × 10–3, and 0.133 × 10–3 cm s–1 when the osmotic gradients were 75, 100, and 125 mosmol, respectively. The water transport activity of protoplas-ts could be inhibited by HgCl2 and stimulated by amphotericin B. Moreover, ZnCl2 and ZnSO4 had a significant inhibitory effect on the water transport activity of the protoplasts. Our results indicate that the Actinidia deliciosa protoplasts had properties typical of aquaporins, suggesting that aquaporins were present at the plasma membrane.
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Ishikawa, Francine Hiromi, Quélen de Lima Barcelos, Elaine Aparecida de Souza, and Eustáquio Souza Dias. "Factors affecting the production and regeneration of protoplasts from Colletotrichum lindemuthianum." Ciência e Agrotecnologia 34, no. 1 (February 2010): 74–79. http://dx.doi.org/10.1590/s1413-70542010000100009.

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The present work reports factors affecting the production and regeneration of protoplasts from Colletotrichum lindemuthianum. The usefulness of protoplast isolation is relevant for many different applications and has been principally used in procedures involving genetic manipulation. Osmotic stabilizers, lytic enzymes, incubation time and mycelial age were evaluated in terms of their effects on protoplast yield. The optimal condition for protoplast production included the incubation of young mycelia (48 h) in 0.6 mol l-1 NaCl as the osmotic stabilizer, with 30 mg ml-1 Lysing Enzymes from Trichoderma harzianum for 3 h of incubation. In these conditions protoplasts production was higher than 10(6) protoplatos ml-1 in the digestion mixture, number suitable enough for experiments of transformation in fungi. Sucrose concentrations of 1.2 mol l-1 and 1 mol l-1 were the most suitable osmotic stabilizers for the regeneration after 48 h, with rates of 16.35% and 14.54%, respectively. This study produced an efficient method for protoplast production and reverted them into a typical mycelial morphology using a Colletotrichum lindemuthianum LV115 isolate.
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Tusa, N., J. W. Grosser, and F. G. Gmitter. "Plant Regeneration of `Valencia' Sweet Orange, `Femminello' Lemon, and the Interspecific Somatic Hybrid following Protoplasm Fusion." Journal of the American Society for Horticultural Science 115, no. 6 (November 1990): 1043–46. http://dx.doi.org/10.21273/jashs.115.6.1043.

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Protoplasm culture following the chemical fusion of `Valencia' sweet orange [Citrus sinensis (L.) Osb.] protoplasts, isolated from an embryogenic suspension culture, with `Femminello' lemon [Citrus limon (L.) Burro. f.] leaf protoplasts resulted in the regeneration of an interspecific allotetraploid somatic hybrid plant, two autotetraploid lemon plants, and diploid plants from both parents. The regeneration of plants from lemon leaf protoplasts is an example of protoplast-to-plant regeneration from non-nucellus-derived tissue for Citrus. Regenerated plants were classified according to leaf morphology, chromosome number, and analyses of phosphohexose isomerase (PHI), peroxidase (PER), and 6-phosphoglucose dehydrogenase (PGD) zymograms. The somatic hybrid plant was vigorous, with leaves morphologically intermediate to the parents. The tetraploid lemon plants were similar to diploids, although less vigorous and with thicker leaves. The tetraploid lemon and somatic hybrid plants, if fertile, could be used in interploid sexual crosses to breed triploid seedless lemon cultivars with tolerance of mal secco disease from sweet orange. Further investigation of plant regeneration from leaf protoplasts could increase the number of totipotent Citrus clones amenable to somatic hybridization and genetic transformation experiments.
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Leinhos, Volker, and Rodney Arthur Savidge. "Isolation of protoplasts from developing xylem of Pinusbanksiana and Pinusstrobus." Canadian Journal of Forest Research 23, no. 3 (March 1, 1993): 343–48. http://dx.doi.org/10.1139/x93-050.

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Protoplasts were isolated from developing xylem of Pinusbanksiana Lamb, and Pinusstrobus L. by incubating freshly harvested tissue in a cellulose–pectinase mixture having mannitol as osmoticum. Protoplasts were then purified using a discontinuous sucrose–mannitol gradient. More than 70% of the isolated protoplasts were of small diameter (12–27 μm) and had dense cytoplasm and many small vacuoles, suggesting that they originated from ray cells. Larger protoplasts constituted about 25% of the protoplast population; these contained single large vacuoles and only parietal cytoplasm, suggesting that they originated from fusiform cells. Using combined gas chromatography–mass spectrometry, coniferin was confirmed to be present in protoplast preparations. By high-performance liquid chromatography (258 nm UV detection), coniferin was readily detected in protoplasts and in extracts of developing xylem from both species. On a fresh-weight basis, coniferin occurred at 1.0–1.6 mM in protoplasts. In late June, coniferin in developing xylem could be accounted for totally by protoplast coniferin content. In late July, protoplasts contained 93 and 61% of the coniferin content in developing xylem of P. strobus and P. banksiana, respectively.
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Saxena, Praveen K., and John King. "Optimal conditions for the isolation and culture of protoplasts from a cell suspension culture of an isoleucine–valine-requiring auxotroph of Datura innoxia." Canadian Journal of Botany 65, no. 8 (August 1, 1987): 1736–40. http://dx.doi.org/10.1139/b87-237.

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Conditions have been standardized for obtaining high yields of viable protoplasts from cell suspension cultures of an isoleucine–valine-requiring auxotroph (IV-1) of Datura innoxia P. Mill. Isolation of protoplasts critically required the use of a salt solution, containing sodium chloride and potassium chloride (0.125 M each), as osmotic stabilizers. The yield, viability, and divisional activity of the protoplasts isolated with mannitol or sucrose were poor. Protoplast-releasing enzymes used to isolate IV-1 protoplasts could be used twice without any loss in the yield or viability of the protoplasts. Cultured protoplasts developed cell walls and underwent sustained divisions in a modified Murashige and Skoog medium enriched with organic acids. Pretreatment of isolated protoplasts with glycine (0.1 M) and calcium chloride (0.05 M) prior to culture increased the frequency of cell colony formation. Protoplast-derived cells developed calli on transfer to agar-solidified medium.
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Riyadi, Imron. "Isolasi Protoplas Tanaman Kacang Panjang secara Enzimatis." Buletin Plasma Nutfah 12, no. 2 (October 6, 2016): 62. http://dx.doi.org/10.21082/blpn.v12n2.2006.p62-68.

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<p>The technique, kind and concentration of enzyme that were appro-priate and optimum affected the isolation process and rendement result of plant protoplasts. A research was conducted to enhance the protoplast rendements of long bean (Vigna sinensis, L.) that was isolated by enzyme Cellulase RS and Macerozyme R-10 as single and combination in a solution. Concentrations of enzyme were used as much as 2.0-3.0% w/v for Cellulase RS and 0.4-0.6% w/v for Macerozyme R-10. Those solutions contain mannitol 25 mM as osmotycum. Isolation process was done on shaker with 50 rpm (rotation per minute) speed in dark room for 3 hours. Results show that C3 treatment (concentration of Cellulase RS enzyme as much as 3.0% w/v) yielded protoplasts density 17.40 x 105 protoplasts/ g fresh weight of mesophyl and M2 treatment (concentration of Macerozyme R-10 enzyme as much as 0.5% w/v) resulted 17.46 x 105 protoplasts/g. As a whole, the best treat-ment was achieved by C2M2 (combination between Cellulase RS as much as 2.5% and Macerozyme R-10 enzyme as much as 0.5% w/v) which resulted protoplasts density 32.67 x 105 protoplasts/g fresh weight of mesophyl</p><p> </p><p><strong>Abstrak</strong></p><p>Teknik, jenis, dan konsentrasi enzim yang tepat dan optimum berpengaruh dalam proses isolasi dan hasil rendemen protoplas tanaman. Penelitian ini bertujuan untuk meningkatkan rendemen protoplas kacang panjang (Vigna sinensis L.) yang diisolasi dengan enzim Cellulase RS dan Macerozyme R-10 secara individu dan penggabungan dua enzim dalam satu larutan. Konsentrasi enzim yang digunakan adalah 2,0-3,0% b/v untuk Cellulase RS dan 0,4-0,6% b/v untuk Macerozyme R-10. Zat osmotikum yang digunakan adalah mannitol 25 mM. Proses isolasi dilakukan di atas gyotoric shaker dengan kecepatan 50 ppm (putaran per menit) dalam kondisi gelap selama 3 jam. Hasil penelitian menunjukkan bahwa perlakuan C3 (konsentrasi enzim Cellulase RS 3,0% b/v) menghasilkan densitas 17,40 x 105 protoplas/g dan perlakuan M2 (konsentrasi enzim Macerozyme R-10 0,5% b/v) menghasilkan densitas 17,46 x 105 protoplas/g berat segar mesofil daun. Secara keseluruhan, perlakuan terbaik dicapai oleh C2M2 (konsentrasi enzim Cellulase RS 2,5% dan enzim Macerozyme R-10 0,5% b/v) yang menghasilkan densitas 32,67 x 105 protoplas/g berat segar mesofil daun.</p>
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Dissertations / Theses on the topic "Protoplasts"

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Lynch, Paul Thomas. "The uptake of fungal protoplasts by plant protoplasts." Thesis, University of Liverpool, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.328131.

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Mullins, P. J. "Protoplasts from Phytophthora." Thesis, University of Liverpool, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.381366.

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Lawrence, W. A. "Microinjection of tobacco protoplasts." Thesis, University of East Anglia, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.372559.

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Attree, S. M. "Properties of Pteridium protoplasts." Thesis, University of Manchester, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.378009.

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Sheard, J. P. "Glucose uptake by pea mesophyll protoplasts." Thesis, University of East Anglia, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.235210.

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Hansen, Christine S. "Construction of galactose assimilating, carotenoid producing yeasts by protoplast fusion." Thesis, University of British Columbia, 1988. http://hdl.handle.net/2429/27935.

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Protoplasts were prepared from two yeast strains P. rhodozyma (ATCC 24202) and K. fragilis (ATCC 8455). Protoplasts prepared from P. rhodozyma were facilitated by prior growth of the cells in a media containing S-(2-aminoethyl)-L-cysteine. Protoplasts from these two yeast genera were fused either by the use of electrofusion or polyethylene glycol treatment. Stable carotenoid producing cell lines were selected by growth at 30°C on yeast nitrogen base plus galactose. Selected single fusants display taxonomic characteristics common to both genera with a cellular morphology and a carotenoid composition similar to that of P. rhodozyma.
Land and Food Systems, Faculty of
Graduate
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Souza, Bárbara Lizandra Perini de. "Fusão de protoplastos entre Penicillium echinulatum e Trichoderma harzianum para obtenção de variabilidade visando a produção de celulases." reponame:Repositório Institucional da UCS, 2007. https://repositorio.ucs.br/handle/11338/881.

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O estudo de fungos celulolíticos tem-se mostrado relevante, tendo em vista o interesse econômico do complexo celulases, especialmente na indústria têxtil e, mais recentemente, para propósitos energéticos. No presente trabalho, a fusão de protoplastos foi utilizada para combinar genótipos de mutantes parcialmente desreprimidos para produção de celulases de Penicillium echinulatum (9A02S1B9) e richoderma harzianum (AS5CH3), utilizando a técnica do doador morto, buscando-se obter recombinantes com maior produção de celulases. Nesta estratégia, ambas as linhagens tiveram seu micélio tratado com Glucanex 0,01 g/mL, para quebra da parede celular. Os protoplastos resultantes da linhagem portadora de marca de resistência ao benomil (9A02S1B9) foram inativados por calor (técnica do doador morto) de 60oC antes da etapa de fusão, a qual após foi induzida por PEG4000 e Ca2+, com protoplastos da linhagem sensível ao benomil (AS5CH3). A partir de um produto de fusão, foram selecionados 24 sub-clones, após estratégias de estabilização e seleção para precocidade e eficiência na formação de halo de hidrólise de celulose em placas de Petri. Os produtos de fusão apresentaram morfologia e esporulação semelhantes a um dos parentais, sendo treze semelhantes à Penicillium, nove semelhantes à Trichoderma e dois mostrando formas alteradas. Os produtos de fusão que segregaram para morfologia de T. harzianum apresentaram a característica de resistência ao benomil, sendo capazes de crescer e esporular em meios contendo até 100 μg/mL deste inibidor. A morfologia, o perfil de bandas, obtidos por RAPD, e o padrão de secreção de celulases dos produtos de fusão foram sempre mais semelhantes a um dos parentais. Os clones apresentaram variação quanto ao halo de hidrólise de celulose em placas de Petri e na atividade sobre papel filtro FPAases, -glicosidase ou endoglicanase, quando crescidas em cultivo submerso ou em estado sólido. Desta variabilidade, verificaram-se aumentos significativos para algumas das linhagens em relação aos parentais. A aplicação da metodologia de fusão de protoplastos para obter recombinantes entre P. echinulatum e T. harzianum, empregando a técnica do doador morto, mostrou-se adequada na geração de variabilidade para produção de celulases.
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The study of cellulolytic fungi has proved to be important, considering economic interest of the cellulase complex, especially in the textile industry and, more recently, for energy purposes. In this work, the protoplast fusion was used to combine genotypes of mutants partially non repressed for cellulases production of Penicillium echinulatum (9A02S1B9) and Trichoderma harzianum (AS5CH3) using the technique dead donor, intending to obtain recombinants with higher cellulases production. In this strategy, both strains had their mycelium treated with Glucanex  0,01 g/mL, to lyse the cell wall. The protoplast obtained from the benomyl-resistant (9A02S1B9) were heat-inactivated (technique of dead donor) at 60ºC, before the step of fusion, induced by PEG4000 and Ca2+, with protoplast of the sensitive-benomyl strain (AS5CH3). Twenty four sub-clones were selected from one fusion product, after stabilization and selection strategies for precocity and efficiency in the formation clearing zones of by cellulose hydrolysis in Petri plates. The fusion products showed similar morphology and sporulation to one of parents, thirteen similar to Penicillium, nine similar to Trichoderma and two showed altered forms. The fusion products which segregate to the morphology of T. harzianum resistance to benomyl, being able to grow and sporulate in media containing up to 100 μg/mL of this inhibitor. The morphology, the profile of bands, obtained by RAPD, and the pattern of cellulase secretion by fusion products were ever more similar to one of parents. The fusants presented variation in the halo of cellulose hydrolysis in Petri plates, and in the activity on filter paper (FPAases), - glicosidase or endoglicanase, when grown submerged cultivation or solid state. From this variability, significant improvement was verified for some of the parental strains. The application of the protoplast fusion methodology to obtain recombinant between P. echinulatum and T. harzianum, using the technique of dead donor, has proved to be adequate to generate variability in the production of cellulases.
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Bourne, N. "The regeneration and transformation of Bacillus protoplasts." Thesis, Cardiff University, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376554.

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Newell, Jane Marie. "Vacuole development in evacuolated oat mesophyll protoplasts." Thesis, University of Southampton, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.295919.

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Ward, Kenneth Glenn. "Microinjection and regeneration of tobacco and potato protoplasts." Thesis, University of Cambridge, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.303971.

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Books on the topic "Protoplasts"

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1927-, Pilet Paul-Emile, ed. The Physiological properties of plant protoplasts. Berlin: Springer-Verlag, 1985.

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Bengochea, Teresa, and John H. Dodds. Plant Protoplasts. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6.

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C, Fowke L., and Constabel F, eds. Plant protoplasts. Boca Raton, FL: CRC Press, 1985.

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J, Reinert, and Binding H. 1939-, eds. Differentiation of protoplasts and of transformed plant cells. Berlin: Springer-Verlag, 1986.

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Lawrence, Wendy Ann. Microinjection of tobacco protoplasts. Norwich: University of East Anglia, 1986.

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I͡Akovenko, K. N. Protoplasty mikroorganizmov. Minsk: "Nauka i tekhnika", 1985.

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Bajaj, Y.P.S., 1936-, ed. Plant protoplasts and genetic engineering. Berlin: Springer-Verlag, 1993.

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Bajaj, Y. P. S., ed. Plant Protoplasts and Genetic Engineering I. Berlin, Heidelberg: Springer Berlin Heidelberg, 1989. http://dx.doi.org/10.1007/978-3-642-73614-8.

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Pilet, Paul-Emile, ed. The Physiological Properties of Plant Protoplasts. Berlin, Heidelberg: Springer Berlin Heidelberg, 1985. http://dx.doi.org/10.1007/978-3-642-70144-3.

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Bajaj, Y. P. S., ed. Plant Protoplasts and Genetic Engineering III. Berlin, Heidelberg: Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-78006-6.

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Book chapters on the topic "Protoplasts"

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Tanaka, Ichiro. "Haploid protoplasts: pollen protoplasts." In In Vitro Haploid Production in Higher Plants, 287–307. Dordrecht: Springer Netherlands, 1996. http://dx.doi.org/10.1007/978-94-017-0477-9_13.

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Jones, M. G. K. "Cereal Protoplasts." In Cereal Tissue and Cell Culture, 204–30. Dordrecht: Springer Netherlands, 1985. http://dx.doi.org/10.1007/978-94-009-5133-4_7.

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Peberdy, J. F. "Fungal Protoplasts." In Genetics and Biotechnology, 49–60. Berlin, Heidelberg: Springer Berlin Heidelberg, 1995. http://dx.doi.org/10.1007/978-3-662-10364-7_4.

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David, A. "Conifer Protoplasts." In Cell and Tissue Culture in Forestry, 2–15. Dordrecht: Springer Netherlands, 1987. http://dx.doi.org/10.1007/978-94-009-4484-8_2.

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Bengochea, Teresa, and John H. Dodds. "Introduction." In Plant Protoplasts, 1–2. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_1.

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Bengochea, Teresa, and John H. Dodds. "Isolation and culture." In Plant Protoplasts, 3–27. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_2.

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Bengochea, Teresa, and John H. Dodds. "Regeneration of plants." In Plant Protoplasts, 28–43. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_3.

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Bengochea, Teresa, and John H. Dodds. "Protoplast fusion." In Plant Protoplasts, 44–58. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_4.

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Bengochea, Teresa, and John H. Dodds. "Protoplasts as physiological tools." In Plant Protoplasts, 59–66. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_5.

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Bengochea, Teresa, and John H. Dodds. "Uptake of foreign materials." In Plant Protoplasts, 67–76. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_6.

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Conference papers on the topic "Protoplasts"

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Stevanović, Katarina, Tanja Pajić, Aleksandar Krmpot, Mihailo Rabasović, Milan Žižić, Miroslav Živić, and Nataša V. Todorović. "Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts." In 2nd International Conference on Chemo and Bioinformatics. Institute for Information Technologies, University of Kragujevac, 2023. http://dx.doi.org/10.46793/iccbi23.221s.

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The success of patch-clamp giga-ohm seal formation on filamentous fungi Phycomyces blakesleeanus protoplasts was investigated to evaluate their usefulness in ion channel studies on filamentous fungi. Protoplasts were obtained by laser-mediated nanosurgery of the cell wall stained with Calcofluor White. To enable a successful seal formation, it is critical to prevent cell wall regeneration. Since wall integrity responses in fungi involve kinase-dependent pathways, we aimed to sufficiently reduce intracellular ATP availability using the respiratory inhibitor azide. The effect of azide on phosphate metabolites of Phycomyces blakesleeanus, as determined by 31P NMR spectroscopy, was a reduction in intracellular ATP accompanied by a decrease in long chain polyphosphates. Subsequently, all seal formation measurements were performed in the presence of azide, and protoplast viability was confirmed by cytoplasmic streaming. The success of seal formation depends on the size of the protoplasts, as larger protoplasts are more prone to successful seal formation. It was also found that the laser power used in nanosurgery could influence the success of seal formation, as higher values were associated with a lower success rate. The protoplasts released by laser nanosurgery produced by our newly developed method are a good model system for patch clamp on filamentous fungal membranes because their plasma membrane can easily form high-quality seals with the patch pipette.
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Fang, Kefeng, Ping Lu, and Tongquan Yu. "Concanavalin Agglutinin Induced Adhesion of Sexual Protoplasts of Tobacco." In 2010 4th International Conference on Bioinformatics and Biomedical Engineering (iCBBE 2010). IEEE, 2010. http://dx.doi.org/10.1109/icbbe.2010.5517502.

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Kaler, K. V. I. S., and A. K. C. Tai. "Dynamic (active feedback controlled) dielectrophoretic levitation of Canola protoplasts." In Proceedings of the Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE, 1988. http://dx.doi.org/10.1109/iembs.1988.94510.

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Zhang, Gaina. "Plant regeneration from mesophyll protoplasts of Radix Gentianae Macrophyllae." In 2011 International Conference on Remote Sensing, Environment and Transportation Engineering (RSETE). IEEE, 2011. http://dx.doi.org/10.1109/rsete.2011.5966163.

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Tarasenko, V. I., T. A. Bolotova, M. V. Koulintchenko, and Y. M. Konstantinov. "STUDY OF DNA IMPORT INTO MITOCHONDRIA IN VIVO USING ARABIDOPSIS PROTOPLASTS." In The All-Russian Scientific Conference with International Participation and Schools of Young Scientists "Mechanisms of resistance of plants and microorganisms to unfavorable environmental". SIPPB SB RAS, 2018. http://dx.doi.org/10.31255/978-5-94797-319-8-1385-1387.

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"606 BGRS/SB-2022 StCDF1 gene editing in wild potato protoplasts." In Bioinformatics of Genome Regulation and Structure/Systems Biology (BGRS/SB-2022) :. Institute of Cytology and Genetics, the Siberian Branch of the Russian Academy of Sciences, 2022. http://dx.doi.org/10.18699/sbb-2022-342.

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Sokolovsky, Sergei G. "Laser monitoring of cytoplasm Ca2+ concentration in etiolated oat protoplasts." In Laser Applications in Life Sciences: 5th International Conference, edited by Pavel A. Apanasevich, Nikolai I. Koroteev, Sergei G. Kruglik, and Victor N. Zadkov. SPIE, 1995. http://dx.doi.org/10.1117/12.197413.

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Hu Jing, Mao Hanpin, Ma Wanzheng, and Han Lvhua. "Optimum parameters of preparation of Epidermal Cell Protoplasts within the Allium cepa L." In 2011 International Conference on New Technology of Agricultural Engineering (ICAE). IEEE, 2011. http://dx.doi.org/10.1109/icae.2011.5943939.

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Nishimoto, K., K. Taguchi, and K. Aritoshi. "Cell Culture and Trapping of Yeast Protoplasts Using Au Thin-Film Dielectrophoresis Chip." In 2015 International Conference on Electrical, Automation and Mechanical Engineering. Paris, France: Atlantis Press, 2015. http://dx.doi.org/10.2991/eame-15.2015.18.

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Mastuti, Retno, and Mufidatur Rosyidah. "In vitro enzymatic isolation of protoplasts from tissues of the medicinal plant Physalis angulata L." In THE 9TH INTERNATIONAL CONFERENCE ON GLOBAL RESOURCE CONSERVATION (ICGRC) AND AJI FROM RITSUMEIKAN UNIVERSITY. Author(s), 2018. http://dx.doi.org/10.1063/1.5061838.

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Reports on the topic "Protoplasts"

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Steponkus, P. L. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Office of Scientific and Technical Information (OSTI), January 1991. http://dx.doi.org/10.2172/7302592.

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Steponkus, P. L. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Office of Scientific and Technical Information (OSTI), January 1993. http://dx.doi.org/10.2172/6551768.

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Steponkus, P. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Office of Scientific and Technical Information (OSTI), January 1990. http://dx.doi.org/10.2172/7190629.

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Sink, Ken, Shamay Izhar, and Abraham Nachmias. Asymmetric Somatic Hybridization: Developing a Gene Transfer System for Solanaceous Vegetable Crops. United States Department of Agriculture, February 1996. http://dx.doi.org/10.32747/1996.7613010.bard.

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Highly asymmetric somatic hybrid plants were obtained by PEG/DMSO fusion of gamma irradiated (100, 250, 7500 and 1000 Gy) protoplasts of a (KmR-) interspecific hybrid Lycopersicon esculentum x L. pennellii (EP) with protoplasts of eggplant (E). Somatic hybrid calli were selected based on kanamycin resistance and verified by PCR of the NptII gene, RAPD's and Southern's using potato rDNA pTHG2 probes. Flow cytometry indicated all hybrid calli that did not regenerate shoots were 5-9n. Three asymmetric plants regenerated only from callus close to 4n and such calli oly occurred when EP received 100 Gy. The asymmetric plants had eggplant morphology and regenerated from one hybrid callus with 6.29 average size tomato chromosomes. Limited amounts of EP DNA were found in the three somatic hybrid plants H18-1 to -3 by dot-blot hybridization with probe pTHG2, to be equivalent to 6.23, 5.41, and 5.95 % EP, respectively. RFLP analysis of Lycopersicon esculentum and L. pennellii specific chromosomes revealed that only fragments of 8 to 10 out of the 24 EP chromosomes are present in the asymmetric plants. Transgenic plants 2-3, 2-4 and 10-3 were found resistant to verticillium; suggesting successful transfer of the Ve complex from S. torvum to eggplant.
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Steponkus, P. L. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Progress report, May 16, 1992--January 9, 1993. Office of Scientific and Technical Information (OSTI), May 1993. http://dx.doi.org/10.2172/10148698.

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Steponkus, P. L. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. [Annual report], May 16, 1993--January 29, 1994. Office of Scientific and Technical Information (OSTI), June 1994. http://dx.doi.org/10.2172/10154320.

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Author, Not Given. (Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts): Progress report, 16 May 1988--9 January 1989. Office of Scientific and Technical Information (OSTI), January 1989. http://dx.doi.org/10.2172/6171109.

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Steponkus, P. L. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Summary progress report, May 16, 1987--June 1, 1991. Office of Scientific and Technical Information (OSTI), December 1991. http://dx.doi.org/10.2172/10166751.

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Dawson, William O., and Moshe Bar-Joseph. Creating an Ally from an Adversary: Genetic Manipulation of Citrus Tristeza. United States Department of Agriculture, January 2004. http://dx.doi.org/10.32747/2004.7586540.bard.

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Citrus is one of the major agricultural crops common to Israel and the United States, important in terms of nutrition, foreign exchange, and employment. The economy of both citrus industries have been chronically plagued by diseases caused by Citrus tristeza virus (CTV). The short term solution until virus-resistant plants can be used is the use of mild strain cross-protection. We are custom designing "ideal" protecting viruses to immunize trees against severe isolates of CTV by purposely inoculating existing endangered trees and new plantings to be propagated as infected (protected) citrus budwood. We crossed the substantial technological hurdles necessary to accomplish this task which included developing an infectious cDNA clone which allows in vitro manipulation of the virus and methods to then infect citrus plants. We created a series of hybrids between decline-inducing and mild CTV strains, tested them in protoplasts, and are amplifying them to inoculate citrus trees for evaluation and mapping of disease determinants. We also extended this developed technology to begin engineering transient expression vectors based on CTV as tools for genetic improvement of tree crops, in this case citrus. Because of the long periods between genetic transformation and the ultimate assay of mature tree characteristics, there is a great need for an effective system that allows the expression or suppression of target genes in fruiting plants. Virus-based vectors will greatly expedite progress in citrus genetic improvement. We characterized several components of the virus that provides necessary information for designing virus-based vectors. We characterized the requirements of the 3 ’-nontranslated replication promoter and two 3 ’-ORF subgenomic (sg) mRNA controller elements. We discovered a novel type of 5’-terminal sgRNAs and characterized the cis-acting control element that also functions as a strong promoter of a 3 ’-sgRNA. We showed that the p23 gene controls negative-stranded RNA synthesis and expression of 3 ’ genes. We identified which genes are required for infection of plants, which are host range determinants, and which are not needed for plant infection. We continued the characterization of native dRNA populations and showed the presence of five different classes including class III dRNAs that consists of infectious and self-replicating molecules and class V dRNAs that contain all of the 3 ’ ORFs, along with class IV dRNAs that retain non-contiguous internal sequences. We have constructed and tested in protoplasts a series of expression vectors that will be described in this proposal.
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Lapidot, Moshe, Linda Hanley-Bowdoin, Jane E. Polston, and Moshe Reuveni. Geminivirus-resistant Tomato Plants: Combining Transgenic and Conventional Strategies for Multi-viral Resistance. United States Department of Agriculture, December 2010. http://dx.doi.org/10.32747/2010.7592639.bard.

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Begomoviruses, which constitute one genus of the Geminiviridae family, are single-stranded DNA viruses that infect many dicotyledonous crops important to large agricultural industries as well as to subsistence growers. Although all begomoviruses are transmitted by whiteflies (Bemisia tabaci), they have proven difficult to manage even with heavy insecticide applications. The begomovirus, Tomato yellow leaf curl virus (TYLCV), has been a problem in tomato production in Israel since the 1950s and in the United States since 1997. Approximately 89 begomoviruses have now been reported to infect tomato. Crop losses due to begomoviruses such as TYLCV and Tomato mottle virus (ToMoV), are limiting factors in tomato cultivation in Israel, the U.S., and many tomato-growing regions throughout the world. To overcome these limitations, we proposed a two-step strategy that combines transgenic and conventional resistance in order to develop tomato plants that are resistant to multiple begomoviruses. In the first step, we have developed transgenic tomato plants expressing trans-dominant interfering mutants Rep and C3 from TYLCV and ToMoV, and tested whether these plants are resistant to infection by these two viruses. In the second step we have tested whether pyramiding transgenic and conventional resistance is superior to either strategy alone. The specific objectives of the proposal were: 1. Design and test trans-dominant interfering constructs for TYLCV and ToMoV Rep and C3 in transient replication interference assays. 2. Generate and test transgenic tomato plants expressing mutant Rep and C3 in resistance assays. 3. Generate and test conventional resistant lines that also express mutant Rep and C3. Two viral replication interfering constructs, expressing the trans-dominant interfering mutants Rep and C3, were designed and constructed during this project. One construct, pNSB1630 was based on TYLCV sequences and the other, pNSB1682, based on ToMoV sequences. The TYLCV transformation construct was tested in a protoplasts replication assay, and was found to inhibit TYLCV replication. The ToMoV transformation construct is yet to be tested in a protoplast assay. Both transformation vectors, pNSB1630 and pNSB1682, were used to transform four different tomato lines, and generate transgenic plants. The tomato lines used for transformation were: FL7613, MM, TY172, TY199. FL7613 and MM are susceptible to both TYLCV and ToMoV. TY172 and TY199 are breeding lines developed at Volcani Center. TY172 is resistant to TYLCV but susceptible to ToMoV, while TY199 is resistant to both TYLCV and ToMoV. When transgenic T1 plants expressing the pNSB1630 constructed were screened for TYLCV resistance, it was found that these plants showed very low level of TYLCV resistance, if any. However, some of these lines showed high level of resistance to ToMoV. Only five transgenic T1 lines expressing the pNSB1682 construct were tested (so far) for resistance to ToMoV. It was found that all five lines express very high level of resistance to ToMoV. Although we haven’t finished (yet) the screen of all the transgenic lines, it is already clear that we were able to successfully combine genetic resistance for TYLCV with transgenic resistance to ToMoV.
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