In this review we focus on recent progress in protoplast regeneration, symmetric and asymmetric hybridization and novel technology developments. Disadvantages of Protoplast Fusion During the mechanical method of isolation of protoplasts: 1. It yields a very small amount of protoplasts. achievements and limitations of protoplast research pdf ppt on achievements of india in space research, achievement limitation of protoplast.

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Spontaneously fused protoplasts, however, cannot regenerate into whole plants, except undergoing a few cell divisions. The somatic hybridization involves three aspects.

To be addressed then is the extent to which sexual incompatibility limits the production of other useful hybrids. Figure 1 shows a general model of the different steps of protoplast regeneration, compiling achievenents most promising research areas for further research in the near future. Plant protoplasts are of immense utility in somatic plant cell genetic manipulations and improvement of crops.

Biotechnology in forest tree improvement with special references to developing countries

In some cases, asymmetric fusions were realized without fragmentation treatment Li et al. This method was first used for the fusion of tobacco protoplasts, and is now in use for other plants also. Some of the genetic traits in certain plants are cytoplasmically controlled. As the formation of heterokaryon occurs during hybridization, the nuclei can be stimulated to segregate so that one protoplast contributes achievementx the cytoplasm while the other contributes nucleus alone or both nucleus and cytoplasm.

The heterokaryons are bigger and green in colour while the parental protoplasts are either small or colourless.

During the last decade, interest in protoplast research was renewed, partly due to public antagonism toward genetically modified organisms.


Asymmetric hybrids may be regarded as cybrids but for the introgressed genes. A PDF file should load here.

Progress in plant protoplast research

Asymmetric hybrids have abnormal or wide variations in the chromosome number than the exact total of two species. After the fusion process, the protoplast population consists of a heterogenous mixture of un-fused chloroplasts, homokaryons and heterokaryons Fig.

A selected list of interspecific hybrids produced through protoplast fusion along with the number of chromosomes in the hybrids is given in Table Petunia parodii protoplasts species B form small colonies, but are resistant to actinomycin D. It is suggested that the high amount of secondary metabolites is quenching ROS w ang et al.

Poor regeneration from protoplasts Tree taxa for which plant regeneration from protoplasts has been reported include Eucalyptus, Populus alba, P. Cytoplasmic male sterile cybrids of rice have been produced and used in hybrid seed production Drew Differences in the status of protoplasts actively dividing or quiescent from the two species of plants result in formation of asymmetric hybrids.

SSR markers are suitable tools to prove recombination through the occurrence of new bands and the disappearance of others Guo et al. Molecular tools based on DNA analysis have known significant progress over the last years and are now routineously used for characterization of somatic hybrids. Many a times, production of full-pledged hybrids through fusion of protoplasts of distantly related higher plant species is rather difficult due to instability of the two dissimilar genomes in a common cytoplasm.

Also, it transfers both mono- and polygenic traits Thieme et al. Although somatic hybridization is a novel approach in plant biotechnology, there are research focusing on the subject matter for reducing the effect of the limitations.

Freshly isolated protoplasts can be fused by induction. The theoretical potential of flow cytometry for cell sorting is extended, but practical obstacles such as the preservation of a stable osmotic potential and the efficient recuperation of sorted protoplast populations are omnipresent and have so far inhibited flow cytometrical hybrid selection on a wide scale. This may be due to production of 3n hybrids by fusing 2n of one species with n of another species. This method is useful for the selection hybrids of two plant species, if one of them is sensitive to a drug.


Promising as it may be, it has its shortcomings in establishing recombination events and should for that aim be combined with sequencing.

The Zeiss CombiSystem combines MicroTweezers for optical trapping and particle positioning with laser beam microdissection and subsequent laser directed transport in a collection vessel. There are no barriers of incompatibility at interspecific, inter-generic or even at inter-kingdom levels for the protoplast fusion. To this end, transcriptome and proteome analyses should be implemented further. Several generations may be required to eliminate the undesirable genes. As mentioned before, protoplast isolation generates significant levels of stress and influences polyamine and ROS biosynthesis, both of which may negatively interfere with subsequent regeneration Cassells and Curry ; Papadakis et al.

Autotetraploid plant regeneration by indirect somatic embryogenesis from leaf Several interspecific and inter-generic hybrids with disease resistance have been created.

This treatment renders the protoplasts inactive and non-dividing, but they are efficient donors of cytoplasmic constituents when fused with recipient protoplasts. Regeneration of new species and improved culture techniques opened new horizons for practical breeding in a number of crops.