Isolation and characterization of cytoplasts and miniprotoplasts derived from protoplasts of cultured cells

Lörz, Horst; Paszkowski, Jerzy; Dierks-Ventling, Christa; Potrykus, Ingo

doi:10.1111/j.1399-3054.1981.tb04517.x

Cited by 70 publications

(18 citation statements)

References 18 publications

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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%

“…This technique is supplemented with high speed centrifugation, which forcibly removes nuclei from the protoplasts [23]. The procedure reported here utilizes these approaches.…”

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%

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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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Section: Isolationmentioning

confidence: 99%

“…This technique is supplemented with high speed centrifugation, which forcibly removes nuclei from the protoplasts [23]. The procedure reported here utilizes these approaches.…”

Section: Isolationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…It is also possible to remove vacuoles before microinjection but regeneration and division may be decreased [52].…”

Section: Cotton Researchmentioning

confidence: 99%

Science Behind Cotton Transformation

Rao¹,

Ali²,

Khan³

et al. 2016

Cotton Research

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The introduction of foreign genes into plant has made possible to bring out desired traits into crop of our own interest. With the advancement in cell biology, regeneration of plants from single cell and advent of different procedures for gene transformation to the plants have opened new avenues for the efficient and applicable implementation of biotechnology for the modifications of desired crop characteristics. Identifications and isolation of different genes for various traits from different organisms have made possible to get the crop plants with modified characters. Over time improvement has been made in transformation technology depending upon the crop of interest. The efficiency of plant transformation has been increased with advances in plant transformation vectors and methodologies, which resulted in the improvement of crops. A detailed discussion on advanced techniques for genetic modification of plants with their handy use and limitation has been focused in this chapter.

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“…8 Miniprotoplasts are protoplasts from which large vacuoles have been removed by centrifugation. 9,10 When miniprotoplasts were cultured, the genesis of vacuoles and cell wall occurred. 11 As vacuoles were formed, the activities of hydrolytic enzymes and vacuolar H + -PPase in cells increased, and vacuoles formed de novo could be stained with neutral red.…”

Section: Introductionmentioning

confidence: 99%

A Novel type of Autophagy Occurs together with Vacuole Genesis in Miniprotoplasts Prepared from Tobacco Culture Cells

Yano¹,

Hattori²,

Moriyasu³

2007

Autophagy

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KeY wordsautophagy, cysteine protease inhibitor, miniprotoplast, protoplast, tobacco culture cells (BY-2), vacuole genesis AbbreviAtions MS medium Murashige and Skoog culture medium AcKnowledgeMentsWe would like to thank Dr. Tetsuro Mimura and Dr. Alison Hills for critical reading of the manuscript. AbstrActMature plant cells have large vacuoles. But how these vacuoles are formed has not been fully understood. It has been reported that autophagy is involved in the genesis of plant vacuoles. Thus we examined whether autophagy occurs in the vacuole genesis of a plant cell model called miniprotoplasts, in which preexising large vacuoles have been removed. We prepared miniprotoplasts from tobacco culture cells (BY-2) and observed the formation of vacuoles by light and electron microscopy. The miniprotoplasts had few vacuoles immediately after preparation, but had large vacuoles after 1 to 2 d. When the cysteine protease inhibitor E-64c or E-64d was added to culture media, almost all vacuoles formed contained materials of cytoplasmic origin. This result suggests that autophagy occurs together with the genesis of the vacuoles in miniprotoplasts. 3-Methyladenine and phosphatidylinositol 3-kinase inhibitors such as wortmannin and LY294002, all of which block starvation-induced autophagy in tobacco culture cells and constitutive autophagy in Arabidopsis root cells, did not affect the autophagy in miniprotoplasts. Thus the form of autophagy in miniprotoplasts is probably different from the form of autophagy that arises as a result of sucrose starvation and constitutive autophagy in root tip cells. The causal connection between autophagy and vacuole genesis in miniprotoplasts was not clarified in this study.

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Isolation and characterization of cytoplasts and miniprotoplasts derived from protoplasts of cultured cells

Cited by 70 publications

References 18 publications

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

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

Science Behind Cotton Transformation

A Novel type of Autophagy Occurs together with Vacuole Genesis in Miniprotoplasts Prepared from Tobacco Culture Cells

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