The uses of isolated protoplasts in plant genetics

Abstract

SUMMARYProtoplasts may be isolated from a wide range of fungal and green plant species. In many of these, protoplasts can be induced to regenerate, and to give either plants directly, or cell cultures, which may in turn subsequently give rise to plants. Fusion of protoplasts can be induced by a variety of agents, and it may be possible to select hybrid cell lines or plants following treatment of mixtures of protoplasts. Hybrid selection procedures include direct visual selection using fusion partners which ale morphologicaily distinct, and various procedures which involve the use of complementing mutant strains.Somatic hybridisation following protoplast fusion has led to the selection of novel hybrids, not obtainable by sexual hybridisation, and has made possible the genetic analysis of sexually sterile strains. It also provides a unique method of studying cytoplasmic inheritance in higher plants, and allows the construction of strains having new combinations of nuclear and cytoplasmic genomes.Isolated protoplasts are likely to be excellent material for mutagenic treatment and because they take up a wide range of material, they are also valuable in a variety of genetic techniques, aimed at the transfer of specific genetic information into plant species. Protoplasts have been utilised in the experiments leading to transformation in Saccharomyces cerevisiae.

The Promise of Using Protoplasts in Grapevine Research

“…The successful removal of the cellulose matrix surrounding the cell allows for these membrane-bound protoplasts to be used for many applications that are otherwise not possible when the cell wall acts as a physical barrier to the external environment. The use of protoplasts in such experiments have generated an encompassing experimental system for cellular biologists, providing an understanding of plant membrane biology, the structure and chemistry of the plasma membrane, the cytoplasmic organelles associated with the plasma membrane, the uptake of macromolecules and membrane transport into the protoplasts, primary and secondary metabolism through idioblasts formation, and organelle isolation such as the nuclei, chloroplast and vacuoles (Cove et al, 1979;Cocking et al, 1985;Eeckhaut et al, 2013). Although these protoplasting applications have been shown to be useful in a range of plant species, a limiting factor remains the phenomenon of recalcitrance in many other plant species, such as grapevine.…”

Effect of Enzyme Treatments on Protoplast Isolation from Leaves of Vetiver (Vetiveria spp.)

“…It is often found more effective to apply a combination of enzymes, such as cellulase, hemicellulase and pectinase, to obtain optimum isolation. Cellulase and hemicellulase generally used to break the plant cell wall, while pectinase used for the separation of cell aggregates (Cove, 1979;Sija et al, 2016). The success of protoplast isolation depends on several factors that affect the release of protoplast in plants, such as the concentration and combination of enzyme, duration of enzyme incubation, pH and osmoticum of the enzyme solution, temperature and the extent of cell wall thickening (Chamani et al, 2012).…”

Auxin Conjugation by Tobacco Mesophyll Protoplasts

“…Protoplasts are a key tool in the genetic manipulation of plants (6). However, inducing the division of mesophyll protoplasts in suitable media is stilf a challenge for numerous species of agronomic interest (22).…”