Cytoplast formation and enrichment from mesophyll tissues of Nicotiana spp.

Archer, Edward; Landgren, Craig R.; Bonnett, Howard T.

doi:10.1016/0304-4211(82)90175-4

Cited by 22 publications

(6 citation statements)

References 10 publications

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“…The difference between cell lines may have arisen because protoplasts of Lp9A may be more susceptible than Lp6JJ protoplasts to a suboptimal osmolality of the medium. On the S/M gradient, the protoplasts banded quickly at the interphase between the 18.2% mannitol and the 33% sucrose layers and were thus surrounded by a medium with an osmolality of >1200 mOsmol kg ~ H20, which perhaps caused a plasmolysis-like effect as has been described for larger cells (Bradley 1978;Gw6zdz & Waliszewska 1979;Archer et al 1982;Wichers et al 1984). Alternatively, different cell lines may show different sensitivities to Percoll, a factor which was found to influence detrimentally cytoplast viability and yield from Beta protoplasts.…”

Section: Discussionmentioning

confidence: 95%

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High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation

Ark¹,

Hall²,

Creemers-Molenaar³

et al. 1992

Plant Cell Tiss Organ Cult

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Methods for the isolation of cytoplasts from suspension culture-derived protoplasts of the monocot Lolium perenne (perennial ryegrass) and the dicot Beta vulgaris (sugarbeet) have been determined. After comparing a range of gradients it was found that a discontinuous sucrose/mannitol gradient gave the highest cytoplast yields for both species tested: of the protoplasts loaded onto the gradient, for Beta >30% and for Lolium up to 45% could be recovered as cytoplasts. Sufficient protoplasts could be loaded onto the gradient to produce suitable numbers of cytoplasts for use in asymmetric somatic hybridisation experiments. Cytoplasts could be isolated from several suspension cultures of different ages. The cytoplast fraction was recovered from the upper part of the gradient in all cases and was only slightly contaminated (2-8%) with protoplasts. Lolium cytoplasts were small, evacuolate cells with granular cytoplasm. In contrast, Beta cytoplasts were larger and predominantly vacuolate. Both contained mitochondria as determined using fluorescence staining.

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Section: Discussionmentioning

confidence: 95%

“…The first method involves the use of media with high osmotic values prior to, or during protoplast isolation (Bradley 1978;Gw6zdz & Waliszewska 1979;Archer et al 1982;Wichers et al 1984). The high osmotic value causes rapid plasmolysis which can lead to intracellular budding and the formation of more than one protoplast per cell.…”

Section: Introductionmentioning

confidence: 99%

High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation

Ark¹,

Hall²,

Creemers-Molenaar³

et al. 1992

Plant Cell Tiss Organ Cult

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“…Especially noteworthy are those involved with various aspects of photosynthesis, such as polypeptides, thylakoid membranes, large subunit of RUBCase, Photosystem II proteins, m-RNA and t-RNA, chlorophyll synthesis, and translation proteins. Considerable re search is still needed to modify donor protoplasts that lack a nucleus (1) and have the chloroplasts and/or mitochondria inhibited or in activated so that they can be used more widely as natural biological entities for the transfer of discrete cell components for plant mod ification.…”

Section: Discussionmentioning

confidence: 99%

Protoplast Fusion for Plant Improvement

Sink

1984

horts

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Most horticultural plant breeders probably are skeptical of the “potential for crop improvement by genetic manipulation through the use of protoplasts,” which has been professed optimistically in review papers and at genetic engineering conferences and meetings (45). This attitude may be warranted to some degree (43). Certainly, the ability to manipulate quantitatively inherited traits will be very difficult to achieve. Exogenous DNA may be introduced into protoplasts, but the subsequent integration, phenotypic expression, and sexual transmission of the new DNA are prerequisite to any possible exploitation by plant breeders. Significant advances have been and are being made presently, however, toward the realization of somatic cell genetic manipulations by the use of protoplasts. These efforts should increase genetic diversity of available germplasm and provide more efficient means for handling specific traits at various steps in plant breeding.

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“…Elongated cells, such as palisade cells and cells from suspension culture, have been frequently used as starting material to obtain cytoplasts [1,6,23]. This technique is supplemented with high speed centrifugation, which forcibly removes nuclei from the protoplasts [23].…”

Section: Isolationmentioning

confidence: 99%

“…Anucleate plant cell systems have been developed for use in somatic cell hybridization as carriers of cytoplasmic traits, such as male sterility [1,6,23]. Anucleate algal cells of Micrasterias and Acetabularia have been shown to be valuable in physiological studies concerning the role of the nucleus on cellular activities in development [5] and anucleate cells of higher plants have been used in studies concerning nuclear influence on chloroplast metabolism [29].…”

Section: Introductionmentioning

confidence: 99%

Isolation and culture of anucleate protoplasts from cotton fiber; assessment of viability and analysis of regenerated wall polymers

Gould

Palmer

Dugger

1986

Plant Cell Tiss Organ Cult

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Procedures were developed for the isolation and culture of an anucleate protoplast system from cotton fibers actively undergoing secondary wall synthesis. Because the fibers at this stage are elongated single cells (30#m X 1-2 cm), most of the cellular vesicles released in the process of isolation are anucleate. After purification, the protoplast population was nuclei-free. When transferred to culture medium, the anucleate protoplasts (cytoplasts) synthesized starch, hydrolyzed fluorescene diacetate for up to 9 days and formed cell wall material for at least 7 days. The composition of the regenerated cell walls was dependent upon the substrate supplied in the medium: ~3-1,3-1inked glucans were predominantly synthesized when l mM UDP[14C]glucose was supplied; /3-1,4-1inked glucans were predominantly synthesized when 1 mM [14C]-glucose was supplied. Thus the composition of the regenerated cell wails formed by the anucleate protoplasts was similar to the secondary cell wall synthesized by intact cotton fibers under the same culture conditions.

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Cytoplast formation and enrichment from mesophyll tissues of Nicotiana spp.

Cited by 22 publications

References 10 publications

High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation

High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation

Protoplast Fusion for Plant Improvement

Isolation and culture of anucleate protoplasts from cotton fiber; assessment of viability and analysis of regenerated wall polymers

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