Isolation of Plasma Membrane from Ryegrass (Lolium multiflorum) Endosperm Protoplasts

Schibeci, Angelo

doi:10.1007/978-3-642-70144-3_6

Cited by 3 publications

(2 citation statements)

References 37 publications

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“…Determination of the pH of sugar solutions over time showed that galactose caused a smaller pH change than sucrose (data not shown), suggesting that galactose could provide more stable pH conditions. Furthermore, Schibeci (25) suggested that galactose molecules are common components of plant plasma membranes, and that galactose-binding proteins are useful for stabilization of the plasma membrane.…”

Section: Resultsmentioning

confidence: 99%

Improvement of Longevity and Viability of Sperm Cells Isolated from Pollen of Zea mays L.

Zhang¹,

Williams²,

Campenot³

et al. 1992

Plant Physiol.

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Our previous studies showed that the common maize (Zea mays L.) sperm isolation medium (Brewbaker and Kwack salts in 0.44 M sucrose without buffering) caused cell lysis in vitro. In an attempt to remedy this situation, 6 sugars, 10 buffers, 5 pH values, and 3 membrane protective agents were screened to improve longevity and viability of isolated Zea mays sperm cells as estimated by hemacytometry and flow cytometry. Use of 0.55 M galactose in the isolation solution increased sperm yield by 2.5-fold compared with sucrose, and suspension of isolated sperm cells in the galactose solution gave the best longevity among the six sugars. Buffering the galactose solution with 2 mm 2-(N-morpholino)ethanesulfonic acid significantly improved longevity, whereas other buffers had no effect or decreased the longevity and/or viability. Among the five pH values tested (5.0, 6.0, 6.7, 7.0, and 8.0), pH 6.7 appeared to be optimal for maintenance of both longevity and viability.Screening of membrane protectants showed that cysteine caused a rapid decrease in cell viability and increased lysis, whereas dithiothreitol increased the cell numbers but lowered their viability.Addition of 0.1% bovine serum albumin increased cell numbers and viability, and about 70% of the cells remained viable after 72 h of suspension. Cell longevity and viability were also improved in 0.44 M sucrose when the solution was conditioned with 2-(Nmorpholino)ethanesulfonic acid and bovine serum albumin. Use of 2-(N-morpholino)ethanesulfonic acid and bovine serum albumin in the isolation and suspension medium significantly improved the viability and longevity of sperm cells isolated from Zea mays pollen.

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Section: Resultsmentioning

confidence: 99%

Improvement of Longevity and Viability of Sperm Cells Isolated from Pollen of Zea mays L.

Zhang¹,

Williams²,

Campenot³

et al. 1992

Plant Physiol.

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“…In order to obtain additional evidence that the 75 kD antiserum was directed against epitopes present on the plasma membrane, an immunofluorescence assay was used. Because animal IgG fractions were shown to bind to plant protoplast surfaces- (21,26,28) cumvented. Figure 4 demonstrates that the 75 kD antiserum was effective as a protoplast surface label.…”

Section: Methodsmentioning

confidence: 99%

Plant Plasma Membrane Proteins

Grimes

Breidenbach²

1987

Plant Physiol.

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A major 75 kD protein group from the tomato plasma membrane was semipurified on polyacrylamide gels and used to raise a rabbit antiserum. The resulting antiserum recognized a single 75 kilodalton band from phase partitioned tomato plasma membrane (from both sispension cells and mature, green fruit) after resolution on one-dimensional polyacrylamide gels. Two-dimensional polyacrylamide gel analysis of proteins from tomato plasma membrane showed that the 75 kilodalton antiserum recognized a group of proteins ranging from 63.1 to 88.2 kilodaltons (mean = 75.6 kilodaltons) and with isoelectric point values ranging from 5.7 to 6.3. No other spots were visible on the two-dimensional blots. This antiserum was shown to bind protoplast surface epitopes by indirect immunofluorescence. The presence of this protein group in both monocotyledonous and dicotyledonous plants was established by immunoblotting the tomato 75 kilodalton antiserum against proteins obtained from plasma membrane-enriched fractions from corn roots and soybean roots. The data suggest that this 75 kilodalton protein group is a major proteinaceous component of the plant plasma membrane.

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Plasma Membrane Isolation

Sandelius

Morré

1990

The Plant Plasma Membrane

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Isolation of Plasma Membrane from Ryegrass (Lolium multiflorum) Endosperm Protoplasts

Cited by 3 publications

References 37 publications

Improvement of Longevity and Viability of Sperm Cells Isolated from Pollen of Zea mays L.

Improvement of Longevity and Viability of Sperm Cells Isolated from Pollen of Zea mays L.

Plant Plasma Membrane Proteins

Plasma Membrane Isolation

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