Cell-Wall Regeneration by Protoplasts Isolated from Higher Plants

Willison, J. H. M.; Klein, Anita S.

doi:10.1007/978-1-4684-1116-4_4

Cited by 27 publications

(7 citation statements)

References 57 publications

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“…The nakedness of their surfaces makes them particularly amenable to microscopical examination (see 17) and to experimental manipulation with chemical probes.…”

Section: Discussionmentioning

confidence: 99%

“…Plant protoplasts are valuable experimental systems for the study of cell-wall secretion (see reviews: 14,17): they permit the examination ofthe process from a unique wall-less starting point; they eliminate cellular interactions which complicate tissue experimental systems; and the lack of wall around the initial isolated protoplast makes it feasible to attempt to distinguish the interrelated processes of the biosynthesis, the secretion and the assembly of individual cell wall components.…”

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confidence: 99%

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Suberin Production by Isolated Tomato Fruit Protoplasts

Rao¹,

Willison²,

Ratnayake³

1984

Plant Physiol.

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The multilamellar wall secreted by protoplasts isolated from locule tissue of tomato (Lycopersico scuklntum L.) fruit was purified, nd an extract was obtained after depolymerization with BF3-methanol. Analysis of this extract using thin layer chromatography demonstrated the presence of fatty acid methyl esters, fatty alcohols, dicarboxylic add dimethyl esters, and w-hydroxy acid methyl esters. These components were quantified using an Iatroscan thin layer chromatography-flame ionization detection system. The different chain lengths in each group were identified and quantified using gas chromatography. The results clearly indicated the presence of suberin.Plant protoplasts are valuable experimental systems for the study of cell-wall secretion (see reviews: 14, 17): they permit the examination ofthe process from a unique wall-less starting point; they eliminate cellular interactions which complicate tissue experimental systems; and the lack of wall around the initial isolated protoplast makes it feasible to attempt to distinguish the interrelated processes of the biosynthesis, the secretion and the assembly of individual cell wall components.Fruit locule tissue protoplasts of tomato synthesize two types of envelope within 24 h of culture; some of these protoplasts make a wall having a pectocellulosic appearance, others make a wall which appears multilamellated in thin sections examined electron microscopically (13). After making the mlw2 some of the protoplasts secrete a second envelope of the pectocellulosic type (13,16 The protoplast band which formed at the top ofthe gradient was collected in glass Petri dishes and suspended in Murashige and Skoog culture medium (12) with 20 ml/l coconut milk. The dishes were sealed with Parafilm and stored in dark at room temperature (20-25°C). In general, the protoplast isolation and the culture procedures followed those of Gamborg and Wetter (4).For electron microscopy, 7-d-old protoplasts were harvested and fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer and similarly buffered 2% OSO4. They were embedded in TAAB embedding resin (TAAB Laboratories, Reading, U.K.), and sections were poststained with uranyl acetate and lead citrate.Preparations of the mlw material were obtained from protoplasts after 7 d culture, using a well-established procedure for the isolation of suberized lamellae (3). In this procedure, protoplasts were rinsed in distilled H20, ground with pestle and mortar, treated with pectinase and cellulase, then rinsed in water again. Soluble lipids wee extracted from the mlw isolate, and the polymer was depolymerized with BF3-methanol, again using procedures of Dean and Kolattukudy (3). Extracts were prepared from tomato periderm wound callus by the same methods.TLC was performed on Adsorbosil-5 Prekotes (Applied Science Laboratories Inc.) of dimensions 20 x 20 cm, activated at 1 1OC for 30 min. Separation of the lipid fractions was effected in the solvent system: hexane/diethyl ether/acetic acid (85/15/ 1, v/v/v). After spraying with 2', 7'-dich...

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“…The nakedness of their surfaces makes them particularly amenable to microscopical examination (see 17) and to experimental manipulation with chemical probes.…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Suberin Production by Isolated Tomato Fruit Protoplasts

Rao¹,

Willison²,

Ratnayake³

1984

Plant Physiol.

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“…So it can be assumed that protoplasts require, like elongating cells, delivery of membrane for cell enlargement. Moreover, protoplasts in general exhibit a high rate of cell wall synthesis and require, like elongating cells, transport vesicles for delivery of cell wall compounds to the cell surface (Willison and Klein, 1982). Notably, the most frequently detected size of exo-and endocytotic events reflect interaction of vesicles with the plasma membrane that have a diameter of 70-100 nm (Figure 2).…”

Section: Exo-and Endocytosis In Coleoptile Protoplasts 119mentioning

confidence: 99%

Unitary exocytotic and endocytotic events in Zea mays L. coleoptile protoplasts

Thiel¹,

Kreft²,

Zorec³

1998

The Plant Journal

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SummaryCell-attached capacitance measurements were used to detect unitary exocytotic and endocytotic events in protoplasts from Zea mays L. coleoptiles with a high-frequency lock-in amplifier. A background noise level of around 70 aF permitted the detection of interacting vesicles with a diameter as small as 60 nm. The distribution of discrete on-steps (exocytosis) was similar to that of off-steps (endocytosis) with a modal values at about 200 aF for both events. From the frequency of endocytotic events (1.5 ⍨ 1.4 min -1 ) it is estimated that 200 min are required for the whole membrane to be ingested in a non-stimulated protoplast.

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“…1, 2) and scanning electron microscopy (Burgess and Linstead 1976;Burgess et al 1977 a, b), freeze etching (Grout 1975;Willison and Klein 1982) and high resolution replica techniques (Williamson et al 1977). The quantity of cell wall material present on the surface of protoplasts has an important effect on the frequency of protoplast fusion (Weber et al 1976).…”

Section: Ultmstructure Of Isolated Protoplastsmentioning

confidence: 97%

Ultrastructural Studies of Plant Protoplast Fusion

Fowke

1989

Plant Protoplasts and Genetic Engineering I

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Cell-Wall Regeneration by Protoplasts Isolated from Higher Plants

Cited by 27 publications

References 57 publications

Suberin Production by Isolated Tomato Fruit Protoplasts

Suberin Production by Isolated Tomato Fruit Protoplasts

Unitary exocytotic and endocytotic events in Zea mays L. coleoptile protoplasts

Ultrastructural Studies of Plant Protoplast Fusion

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