The development of a protoplast fusion technique for oxytetracyclineproducing streptomyces rimosus strains, and its evaluation for the application for a breeding programme, has been described. Protoplast isolation and fusion protoplasts are cells without the cell wall. Application of the protoplast fusion technique to engineer. Protoplasts can be generated by stripping the cell wall from plant, bacterial, or fungal cells by mechanical, chemical or enzymatic means. Pdf protoplast fusion technology for somatic hybridisation in. The application of this technique to citrus would be of great value. Protoplast fusion and somatic hybridization in vitro. These workers compared the effectiveness of conventional techniques of obtaining nuclear fusion between strains of c. The highest regeneration frequency was obtained on the ms hormone free medium. Share your knowledge share your word file share your pdf file share your ppt file. Updated protoplast preparations for gene transfection 109 protoplast isolation protocol is based on the method developed by sheen and colleagues sheen, j. Archaic term meaning the first individual of a type or race. Us6569680b2 efficient method of protoplast culture. Somatic fusion, also called protoplast fusion, is a type of genetic modification in plants by which two distinct species of plants are fused together to form a new hybrid plant with the.
A cell of a plant, fungus, bacterium, or archaeon from which the cell wall has been removed, leaving the protoplasm and plasma membrane. Protoplast fusion is a wellestablished technique that is applicable to an array of organisms including bacteria and both lower and higher eukaryotes. Hybridization studies based on the use of crossing and the protoplast fusion technique revealed the relationship between the various taxa of the yeast genus schizosaccharomyces lindner. Transfer of resistance to xanthomonas campestris pv. Protoplast fusion may be used to produce interspecific or even intergeneric hybrids. Limitations, advantages and applications of somatic hybridization. Protoplast fusion technique has a great potential for genetic analysis and for strain. The technique is used for brassica species, for forcing chicory, and in the near future probably also for rapeseed. This technique is used to create hybrids between plant species that cannot be crossed sexually.
Protoplast fusion techniques in fungi springerlink. Brassica improvement, protoplast fusion, cytoplasmic hybrids, mitochondrial genome recombination, cytoplasmic male sterilityfertilityrestorer systems, manipulation of flower morphology introduction protoplast fusion technique involves the fusion of. This technique in the future will be one of the most frequently used research tools for tissue. The process involves cell wall breakdown, regeneration of. However, the application ofthis technique is limited by the composition. Protoplast fusion technology and its biotechnological. It may allow the exchange of dna between unrelated. Impacts of banning protoplast fusion on the range of. Protoplast fusion becomes an important tool of gene manipulation because it breakdown the barriers to genetic exchange imposed by conventional mating systems. Moreover, the fusion of protoplasts of genetically different lines or species has also been possible. Protoplast fusion and somatic hybridization with the development of techniques for enzymatic isolation of protoplasts and subsequent regeneration, a new tool of genetic manipulation of plants has now become available.
Protoplast fusion is important in biochemical andgenetic engineering studies, butmanydetails regarding its mechanismare notknown. Increasing the variability oflycopersicon peruvianum mill. Pdf protoplasts are the cells of which cell walls are removed and cytoplasmic membrane is the outermost layer in such. They are the biologically active and most significant material of cells. The effect of high ph and calcium on tobacco leaf protoplast fusion. Agricultural applications of plant protoplast fusion. Plant protoplast fusion protoplast fusion has many uses in plant research.
In this way the protoplasts are retained and compressed by the flow of liquid. Plant protoplasts can be fused and the fusion products cultured to produce somatic hybrid plants. Protoplast fusion an overview sciencedirect topics. Isolation and fusion of protoplasts from mesophyll cells.
Isolation and fusion of protoplasts from mesophyll cells of dendrobium pompadour kamnoon kanchanapooma, saowarath jantarob and duangkae rakchadb a department of biology, faculty of science, prince of songkla university, hatyai, songkhla 90112, thailand. Fusion of protoplasts of potato and tomato, and production of hybrid plant pomato. Nootjaree tudses1, siripong premjet,2,3 and duangporn premjet1,3. Pdf the success of interspecific breeding between phaseolus vulgaris l. Transfer of mitochondria by protoplast fusion in saccharomyces. The application of protoplast fusion techniques micro.
Application of the protoplast fusion technique to engineer a recombinant microorganism to more efficiently degrade chlorophenols a,honglei chen, b, yu liu,a a,cfangong kong,a dlucian a. Protoplast fusion induced by the application of electric shock 18. Enzymatic hydrolysis, microalgae, cell wall, protoplast fusion, strain improvement. By this technique occasional fusion of protoplast has been observed. Plant protoplast fusion by electromanipulation sciencedirect.
If by contrast cellfusion allows for a free mixing of the cy. Protoplast fusion is a nonspecific recombination technique used for transfer of cytosolic organelles including genetic material. An alginatelayer technique for culture of brassica oleracea l. The beadtype culture technique where segments of protoplast containing gels are floated in larger volumes of liquid culture medium and aerated on a rotary shaker has considerably improved the plating efficiency in many systems and has enabled sustained development in systems where protoplasts did not previously divide.
Production of somatic hybrid plants through protoplast fusion in. Fusion of proplast incudes three stages 1 agglutination 2 membrane fusion 3rounding off of the fused protoplast carlson et al 1972 first time fused the leaf protoplasts of nicotina gluca and n. Protoplast fusion would continue to be an existing area of research in modern biotechnology 11. This technique has been used to produce germplasm previously unavailable to. Get a printable copy pdf file of the complete article 582k. Often the presence of polyethylene glycol or electric fields will enhance the frequency of fusion. An alginatelayer technique for culture of brassica. Protoplast fusion and growth of hybrid cells 219 table 6. The effect of peg muting solution on fusion of protoplasts derived from cultured soybean cells and vicia haastana leaves eluting solution s percentage of heterokaryocytes to total surviving protoplasts and to soybean alone b,o. In this study we describe the use of protoplast fusion techniques within the genus phaseolus, as an alternative to. Pdf protoplast fusion technology and its biotechnological. This technique is known as electro fusion of protoplasts.
In the fusion unit the dna from the component cells may undergo recombination. Membranefluidity may affect the rate ofprotoplast fusion. Plant protoplasts for cell fusion and direct dna uptake. This lecture explains about the protoplast fusion technique also known as somatic cell fusion which allows us to produce plant cell hybrids with increased drought tolerance and immunity. Plant protoplast fusion and growth of intergeneric hybrid. Limitations, advantages and applications of somatic.
Free fulltext pdf articles from hundreds of disciplines, all in one place. From a series of intra and interspecific crosses as well as protoplast fusions, we concluded that a revision of the genus is required. Establishment of method for protoplast fusion with pegmediated between jatropha curcas l. Read this article to learn about the importance, isolation techniques, culture and regeneration of protoplasts and also learn about sub protoplasts sub protoplasts is divided into three types.
Protoplast fusion is a wellestablished technique that is applicable to an array of. Application of the protoplast fusion technique to engineer a recombinant microorganism to more efficiently degrade chlorophenols. In protoplast technology,two genetically different protoplast isolated from the somatic cells and are. Recently, mild electrical stimulation is being used to fuse protoplasts. Procedure for successful somatic hybridization is as below. Protoplast protoplasts are the cells of which cell walls are removed and cytoplasmic membrane is the outermost layer in such cells. Protoplast fusion technology vary protoplast from speciesgenera, fusion to create new genotypes, plant regeneration organogenesissomatic embryogenesis, isolated in large quantities, protoplasts must be. Protoplast fusion would continued to be an existing area of research in modern biotechnology.
Isolated protoplasts are devoid of cell walls which make them easy tools for undergoing fusions in vitro. Protoplast definition of protoplast by the free dictionary. Several protoplastbased genetic engineering techniques emerged, including protoplast fusion, dna microinjection, electroporation, and polyethylene glycol pegmediated gene transfer. What importance does protoplast fusion have as a breeding method at present. A demonstration of the use of protoplast fusion for an industrial fungus is provided by the work of hamlyn and ball 1979 on the cephalosporin producer, c. During fusion, two or more protoplasts come in contact and adhere with one another either spontaneously or in presence of fusion inducing chemicals. Protoplast fusion will help in the combination of mitochondria and chloroplasts to result in a unique nuclearcytoplasmic genetic combination. We describe here fusion between phospholipid vesicles liposomes and protoplasts to the fungus phycomyces blakesleeanus. The plant cell walls acts as a physical barrier and protects cytoplasm from microbial. The invention a novel and efficient method for protoplast culture comprising the steps of isolating the protoplasts from a cell suspension, mixing the protoplasts with equal volumes of alginate solution, placing 4050. A technique by which two protoplasts are fused to form hybrid cells that can grow into mature hybrid organisms.
Honglei chen, a,b, yu liu, a fangong kong, a lucian a. Sikandar hayat1, christos christias2 1postdoctoral fellow alberta research council, vegreville, alberta, canada. The relation between the composition of the phospholipid molecular species in a cell membrane and the velocity of protoplast fusion was studied using cells cultured at a low temperature 10 c. Protoplast fusion article about protoplast fusion by the. Electrofusion, that is, cell fusion in an electric field, has become of great interest as a powerful technique for the fusion of plant, animal, and microbial cells as it is simpler, more rapid, and less harmful to the cells than the conventional polyethylene glycol peg method and is available for mass production of fusion. After adhesion, membranes of protoplasts fuse in some localised areas and, eventually, the cytoplasm of the two protoplasts intermingle. When cell wall is mechanically or enzymatically removed the isolated protoplast is known as naked plant cell on which most of recent researches are based. Lucia, a,c and xiaojing feng d recombinant strain xz61 was constructed by the protoplast fusion technique with the goal of endowing it with the ability to efficiently degrade pentachlorophenol. Lucia, and xiaojing feng recombinant strain xz61 was constructed by the protoplast fusion.
The fusion of protoplasts in microalgae constitutes a tool for strain improvement because it allows both intra and interspecific genetic recombination, resulting in organisms with new or improved. Protoplast fusion and somatic hybridization biotechnology. Wepreviously proposed that membranefluidity is determined bythe phase transition ofthe phospholipid molecularspecies 3, 17. An electrical field of low strength 10 kv m1 gives rise to dielectrophoretic dipole generation within the protoplast suspension. Protoplast fusion technology was employed to reconstruct fusants from this fungus. A bacterial cell from which the rigid cell wall has. Strain improvement of pleurotus species by protoplast fusion. Pdf microalgae protoplasts isolation and fusion for.