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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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Journal ArticleDOI
TL;DR: Transformation efficiency, plant regeneration of transformed root clones, protoplast yield and ploidy stability were the highest as compared to the other genotypes, and the application of these transformed plants as marker lines in gene mapping and gene expression studies is indicated.
Abstract: Agrobacterium transformation of stem internodes of four monohaploid (839-79, 849-7, 851-23, 855-1) and two diploid (M9 and HH260) potato genotypes using hairy root-inducing single (LBA 1020, LBA 9365, LBA 9402) and binary (LBA 1060KG) vectors is reported. Various media and successive culture steps were tested for plant regeneration from different transformed root clones. The fate of introduced genetic markers in root clones and regenerated plants (hairy root phenotype, hormone autotrophy, opine production, kanamycin resistance, β-glucuronidase activity), the ploidy stability and protoplast yield were analysed. The transformation efficiency of stem internodes (hairy root production) and the regeneration capacity of the transformed root clones greatly differed within and between the various potato genotypes. The regenerated plants obtained after transformation with both types of vectors often showed the absence of one or more genetic markers. However, transformation with the binary Agrobacterium vector generally resulted in the stable presence of the opines in all transformed root clones and most regenerated plants. In HH260, transformation efficiency, plant regeneration of transformed root clones, protoplast yield and ploidy stability were the highest as compared to the other genotypes. The application of these transformed plants as marker lines in gene mapping and gene expression studies is indicated.

31 citations

Journal ArticleDOI
TL;DR: Large secretory cells are found in the pericarp ofOkra capsules and it is suggested that these cells may play a major role in the formation of okra mucilage.
Abstract: Large secretory cells are found in the pericarp of okra capsules. In early developmental stages these cells are conspicuous because of the great quantity of Golgi apparatus-derived, PAS positive substance stored between the protoplast and the cell wall. It is suggested that these cells may play a major role in the formation of okra mucilage.

31 citations

Patent
18 Jun 1991
TL;DR: In this paper, the embryogenic cell aggregates (type II callus) were used to isolate totipotent protoplasts after subcultivation of the callus as cell suspension culture.
Abstract: Starting from immature embryos on hormone-free nutrient media there is formation, at the stem base of the germ plant, of an auxin-autotrophic, embryogenic callus which retains its embryogenesis ability on subcultivation on hormone-free medium over a lengthy period. Besides fully developed embryos, also produced under suitable culture conditions (6-9% sucrose in the medium) are adventitious embryos. When the sucrose content is reduced to 2-3% and 2,4-dichlorophenoxyacetic acid is added there is formation of soft, granular calli which consist of embryogenic cell aggregates (type II callus). It is possible to isolate totipotent protoplasts after subcultivation of the type II callus as cell suspension culture. The maize plants according to the invention are regenerated from these protoplasts.

31 citations

Journal ArticleDOI
22 Jan 1998-Planta
TL;DR: In this paper, the authors measured net H+ and Ca2+ fluxes from individual corn coleoptile protoplasts using a non-invasive microelectrode technique for ion flux measurements.
Abstract: The ability to measure directly individual protoplast ion fluxes is a valuable addition to patch clamp and other techniques when using protoplasts to study membrane transporters. Before interpreting observations on protoplasts in terms of behaviour of intact cells and tissues, some methodological questions should be addressed. These include effects of space and time variations of transporter activities over the membrane, the osmotic dependence of specific ion transporters and the effect of the regenerating cell wall. In this study net H+ and Ca2+ fluxes were measured from individual corn (Zea mays L.) coleoptile protoplasts using a non-invasive microelectrode technique for ion flux measurements. For Ca2+, the flux distribution was almost symmetrical, ranging ±30 nmol · m−2 · s−1 around zero. For H+ it was skewed towards efflux ranging from −100 to +10 nmol · m−2 · s−1. The distribution of H+ fluxes through the protoplast surface was a complex mosaic which changed with time, sometimes showing oscillations. These flux variations with time and position around the surface, apparently driven by endogenous mechanisms, may be relevant to protoplast pH homeostasis. When the new cell wall was partially regenerated on the next day, the correlation between H+ and Ca2+ fluxes increased, which is consistent with the weak-acid Donnan-Manning model of cell wall ion exchange.

31 citations

Journal ArticleDOI
TL;DR: Growth and development of the protoplasts were significantly affected by osmolarity, nitrogen source, plating density and culture medium dilution; however, a low frequency of plantlet recovery from protoplast-derived somatic embryos was observed.
Abstract: Protoplasts were isolated from embryogenic suspension cultures derived from avocado (Persea americana Mill.) zygotic embryos and nucellus in an enzyme digestion solution consisting of 1% cellulase Onozuka RS, 1% Macerase R10, 0.2% Pectolyase Y-23, 0.7 M mannitol. 24.5 mM CaCl2, 0.92 mM NaH2PO4 and 6.25 2-[N-morpholino]ethanesulfonic acid (1.5 ml) mixed with 0.7 M MS–8P (2.5 ml). MS-8P medium consisted of Murashige and Skoog salts without NH4NO3, 1 mg l–1 thiamine HCl, 100 mg l–1 myo-inositol, 3.1 g l–1 glutamine and 8P organic addenda. Medium osmolarity was adjusted with 0.15 M sucrose and 0–0.55 M mannitol. Protoplast yields of 3.5×106 protoplasts g–1 were obtained. Growth and development of the protoplasts were significantly affected by osmolarity, nitrogen source, plating density and culture medium dilution. Under optimum conditions, proembryos developed directly from embryogenic protoplasts and subsequently into somatic embryos. Optimum conditions for somatic embryo development included the culture of protoplasts at a density of 0.8–1.6×105 ml–1 in 0.4 M MS–8P for 2–3 weeks, followed by subculture in 0.15 M MS–8P at a diluted density of 20–40× for 1 month in darkness to obtain somatic embryos. Mature somatic embryos were recovered on semisolid medium; however, a low frequency of plantlet recovery (≤1%) from protoplast-derived somatic embryos was observed.

31 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
202358
2022153
202160
202060
201978
201855