Fungal Protoplasts
In the 40 years since the first reported isolation of protoplasts from filamentous or mycelial fungi (Peberdy 1971), these structures have become central to research in several key areas of fungal biology. The switch in focus in genetics from the classica
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PEBERDY 1
CONTENTS I. II. Ill. IV.
A.
B. V.
A. B. C. D. VI.
Introduction ............................. . Protoplast Isolation ....................... . Protoplast Regeneration ................... . Protoplasts and Fungal Genetics ............ . Protoplast Fusion ......................... . Transformation .......................... . Protoplasts as Tools in Biochemistry and Cell Biology ......................... . Cell Wall Formation and Regeneration ...... . Cell Biology ............................. . Protein Secretion Metabolic Activities of Protoplasts .......... . Conclusions ............................. . References .............................. .
I. Introduction
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yields for many fungi, which in turn has reawakened the interest of researchers addressing cell biological, physiological, and biochemical questions and led to the adoption of protoplasts as experimental tools wherever relevant. Research involving protoplasts has been reviewed several times over the past 20 years, both generally and more specifically in the genetic context. For a historical perspective the reader is invited to consult reviews by Hocart and Peberdy (1990), Peberdy (1979a, 1983, 1987a, 1989), Peberdy and Ferenczy (1985), Peberdy and Hocart (1987). The aim of this chapter is to update progress in several areas, including protoplast isolation and fusion, and to consider in more depth the recent studies on cell biology, physiology, and biochemistry.
In the 40 years since the first reported isolation of protoplasts from filamentous or mycelial fungi (Peberdy 1971), these structures have become II. Protoplast Isolation central to research in several key areas of fungal biology. The switch in focus in genetics from the The list of fungi that have been used for proclassical to a molecular approach is the major toplast isolation grows continuously. Some of the factor. Two genetic techniques, protoplast fusion more recent examples are given in Table 1. Some and transformation, together with electrophoretic of the fungi listed have been reported on pretechniques to determine the molecular karyotype of fungi are all dependent on the effective use of viously, and reference is made to an alternative and generally improved procedure. Many new protoplasts. As a consequence, other aspects of species are listed for the first time and these research that employ protoplasts have benefited include mycobiontic fungi isolated from lichens more generally. At the beginning, protoplasts and mycorrhizal fungi. Interest in these species held high curiosity value but their scientific reflects the growing desire in exploiting fungi in potential was not fully realized. However, as the many areas of biotechnology. desire to extend the protoplast isolation technique The basic technology for protoplast isolato more and more fungi has occurred, cortion is very simple. The fungal material, cells, respondingly more attention has been given to mycelium, or spores, is suspended in a solution improving this essential process
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