Ultrastructural characterization and three-dimensional reconstruction of isolated maize (Zea mays L.) egg cell protoplasts

Faure, J. E.; Mogensen, H. Lloyd; Kranz, Erhard; Digonnet, C.; Dumas, Christian

doi:10.1007/bf01403723

Cited by 55 publications

(35 citation statements)

References 27 publications

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“…A pear shape, together with a large vacuolated central nucleus, allowed for recognition of the egg cell in situ. The typical, granular-structured cytoplasm of the egg cells corresponded to the features earlier reported for maize (Faure et al 1992) and wheat (You and Jensen 1985). Egg cells could be isolated through a combination of enzymatic maceration and mechanical manipulation of the dissected ovules, comparable to the technique that was developed for maize (Kranz et al 1991;Kranz and Lörz 1993), or by purely mechanical isolation.…”

supporting

confidence: 57%

Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench.)

Woo¹,

Adachi²,

Jong³

et al. 1999

Can. J. Plant Sci.

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. 1999. Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench.). Can. J. Plant Sci. 79: 593-595. In vitro fertilization could be useful to overcome the problem of self-incompatibility of common buckwheat. Therefore, experiments were conducted to isolate viable protoplasts from egg cells for use in vitro fertilization. Protoplasts from viable egg cells of common buckwheat were isolated by enzyme treatment followed by mechanical isolation. Incubation of ovules for up to 4 h allowed the boundary wall of the egg cells to become partially digested and the protoplasts of the egg cells came out of the micropylar or chalazal end after squashing with a mean isolation frequency of approximately 30% (6 out of 20 ovules). The egg cell protoplasts were larger than those of the synergids.

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supporting

confidence: 57%

Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench.)

Woo¹,

Adachi²,

Jong³

et al. 1999

Can. J. Plant Sci.

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“…Highly viable protoplasts of the unfertilized egg cell were isolated by micromanipulation from enzymatically digested ovules (Kranz et al, 1991;Faure et al, 1992). The protoplasts could be electrofused to isolated sperm cells with a high frequency (Kranz et al, 1991;Faure et al, 1993), and the fusion products were regenerated to plants by cocultivation with a nonmorphogenic maize cell suspension (Kranz and Lorz, 1993).…”

Section: Introductionmentioning

confidence: 99%

Regeneration of Fertile Barley Plants from Mechanically Isolated Protoplasts of the Fertilized Egg Cell.

et al. 1994

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A simple procedure is described for the mechanical isolation of protoplasts of unfertilized and fertilized barley egg cells from dissected ovules. Viable protoplasts were isolated from m75% of the dissected ovules. Unfertilized protoplasts did not divide, whereas almost all fertilized protoplasts developed into microcalli. These degenerated when grown in medium only. When cocultivated with barley microspores undergoing microspore embryogenesis, the protoplasts of the fertilized egg cells developed into embryo-like structures that gave rise to fully fertile plants. On average, 75% of cocultivated protoplasts of fertilized egg cells developed into embryo-like structures. Fully fertile plants were regenerated from 4 0 % of the embryo-like structures. The isolation-regeneration techniques may be largely genotype independent, because similar frequencies were obtained in two different barley varieties with very different performance in anther and microspore culture. Protoplasts of unfertilized and fertilized eggs of wheat were isolated by the same procedure, and a fully fertile wheat plant was regenerated by cocultivation with barley microspores.

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“…Single-cell TEM was applied for observation of a protoplast by using an automatic micromanipulation system and ultra-low gelling temperature agarose No. 9 (Faure et al, 1992). As shown in Figure 3b, an elongated fiber forming protoplast of Larix was transferred efficiently to an agarose No.…”

Section: Observation By Lcsmmentioning

confidence: 87%

“…Recently, some callose fibers were also found in protoplast cultures of several tree mangrove species, without high salt concentrations in the medium, and their callose component was confirmed by an enzymatic method (Kurita et al, 2008). micromanipulation techniques developed for selection of fused protoplasts (Sasamoto et al, 2000), we improved the single cell TEM technique (Faure et al, 1992) for observing the site of (sub)-fibrils formation.…”

Section: Introductionmentioning

confidence: 99%

Laser Confocal Scanning Microscopy and Transmission Electron Microscopy to Visualize the Site of Callose Fiber Elongation on a Single Conifer Protoplast Selected With a Micromanipulator

Oyanagi¹,

Kurita²,

Fukumoto³

et al. 2014

JPS

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We have developed a new method to observe the site of big and long spiral callose fiber elongation from a protoplast in liquid culture medium. Protoplasts of embryogenic cells of a conifer, Larix leptolepis, were cultured in the NH 4 NO 3 -free Murashige and Skoog's medium containing 50 mM MgCl 2 and 6% sucrose in a well in a 96-or 24-well culture plates. The protoplasts with elongated callose fibers were selected before fixation with glutaraldehyde, by picking up using a micromanipulator after electric treatment (DC 3 kV/cm) in the medium containing Alexafluor 488 phalloidin. Under laser confocal scanning microscopy (LCSM), two sites in the cell were stained clearly. One site was the nucleus and the other was the plasma membrane from which fibers elongated. Single cell transmission electron microscopy (TEM) was developed for observation of the microstructure at the site of fiber elongation on a single protoplast. A single protoplast which had a fiber was selected using a micromanipulator and transferred to an agarose bead at a low gelling temperature. The cells were fixed with cold glutaraldehyde and processed for TEM analysis. Elongated thin fibrils with vesicle-like structures could be observed by TEM.

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Ultrastructural characterization and three-dimensional reconstruction of isolated maize (Zea mays L.) egg cell protoplasts

Cited by 55 publications

References 27 publications

Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench.)

Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench.)

Regeneration of Fertile Barley Plants from Mechanically Isolated Protoplasts of the Fertilized Egg Cell.

Laser Confocal Scanning Microscopy and Transmission Electron Microscopy to Visualize the Site of Callose Fiber Elongation on a Single Conifer Protoplast Selected With a Micromanipulator

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