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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Chromosome rejection as a consequence of the distant relationship between both partners is very likely Guo 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. Importance, Isolation, Culture and Regeneration.

Somatic hybrids that ans the complete nuclear complements of both parents are generally rare Xiaand this fact has drawn more attention to the evaluation of chromosome number and structure stability following fusion. This survey does not aim to cite all protoplast related research, but to provide an overview of the most innovative developments and insights. Another advantage of protoplast embedding may be an improved signaling cascade before the first cytokinesis.

Somatic Hybridization: Aspects, Applications and Limitations

Zhang Protolast Sorghum bicolor is one of the most important crops. The parental protoplasts cannot grow on nitrate medium while the hybrid protoplasts can grow Fig.

Drug sensitivity technique was originally developed by Power et al for the selection of hybrids of Petunia sp. Enucleated protoplasts can be isolated by high-speed centrifugation. This is a question and answer forum for students, teachers and general visitors for exchanging articles, answers and notes.

They positively influenced protoplast viability and regeneration Kakoniova et al. The same techniques can also be applied for sorting and selection of heterokaryons.

It is cultivated globally achivements only. Regeneration research would greatly benefit from more detailed studies on the effects of isolation and early culture on model plants antioxidant mechanisms.

Some workers use flow cytometry and achivements cell sorting techniques for the analysis of plant protoplasts while their viability is maintained. In fusogen or electro-induced fusions, about one third of the fusions occur between more than two protoplasts. Next generation sequencing enables fairly cheap high quality nucleic acid sequence data obtention of complete genomes in a short period of time, and will therefore definitely contribute to somatic hybrid genome screening and stability studies in the near future.


This is of particular interest where a useful gene is to be introduced to a commercial variety from a wild relative, and offers a rapid alternative to traditional backcrossing programmes.

Progress in plant protoplast research.

For most tissues from tree species, regeneration from protoplasts remains problematical. It is suggested that the achhievements amount of secondary metabolites is quenching ROS w ang et al. Somatic hybridization has opened new possibilities for the in vitro genetic manipulation of plants to improve the crops. Petunia parodii protoplasts species B form small colonies, but are resistant to actinomycin D.

The same was observed when untreated Arabidopsis thaliana protoplasts were fused with Uv treated Bupleurum protoplasts w ang et al. Two species of nitrate reductase deficiency— one protooplast to lack of apoenzyme nia-type mutant and the other due to lack of molybdenum cofactor cnx- type mutant are known.

Many recent developments in molecular biology have improved the understanding of genetic constitution of somatic plant hybrids. Several interspecific and inter-generic hybrids with disease resistance have been created. The isoenzymes are often variable within the same plant. The importance of protoplast sources and embedding systems is discussed.

Achievements and limitations of protoplast research pdf

Changes in the production of reactive oxygen species ROS and reactive nitrogen species were studied in more achhievements in cucumber protoplasts Petrivalsky et al. In spite of continuing good progress in the field, the early expectations of protoplast fusion are yet to be realised, major limitations remain to be overcome, and most of the research is still empirical.

The different levels of as well ROS and antioxidant enzymes and scavengers in Citrus callus and mesophyll are suggested to play a key role in defining the regeneration potential of protoplasts of both pprotoplast types Xu et al.

Theoretically, protoplasts are totipotent, achievemnts that they have the capability to dedifferentiate, re-enter the cell cycle, go through repeated mitotic divisions and then proliferate or regenerate into various organs.

This allowed them to report the phenomenon of massive mitochondrial fusion that within 24 h led to a near-complete mixing of the mitochondrial population.

Somatic hybridizations and cybridizations in Citrus resulted in rootstocks resistant to biotic and abiotic constraints and in increased yield and fruit quality Dambier et al.


The hybridity must be established only from euploid and not from aneuploid hybrids. For this purpose, the vegetative and floral characters are considered. This is difficult to assess. The major limitation of this method is the requirement of specialized and costly equipment.

In the research of w ang et al. Apart from meiotic abnormalities, chromosome fragment deletion and rearrangements also occur in sterile somatic hybrids Iovene et al. High resolution melting can become a standard for mtDNA and cpDNA screening, as, through combination with a PCR reaction, it can outcompete laborious and costly sequencing analysis. Some of the genetic traits in certain plants are cytoplasmically controlled. Lagging chromosomes could contribute to the formation of small pollen grains that are linked to sterility Guo et al.

A diagrammatic representation of protoplast fusion is depicted in Fig. To this end, transcriptome and proteome analyses should be implemented further. There are several fusion-inducing agents which are collectively referred to as fusogens e. During protoplast isolation, endophytic bacteria can be released and subsequently cocultured Klocke et al.

To this end, a further rationalization of protoplast regeneration is indispensable, and we expect that regeneration efforts will continue to steadily drop out of trial and error based experimentations. ISSR analysis showed that the level of intergenomic recombination can be increased by reducing ploidy level of Solanum hybrids, through androgenesis, by tetrasomic inheritance Toppino et al.

Most hybrids are fertile and indeed contain some partial resistance against these parasites. The simulation model of growth and cell divisions for the root apex with an It can create different homokaryon or heterokaryon types, as well as alloplasmic hybrids cybrids Xia In essence, protoplast fusion comprises removal of cell walls and then the amalgamation of cell contents.

For fertility, chromosome addition meaning that the number of chromosomes in the hybrid equals the sum of the chromosomes in the fusion partners is not absolutely required. Alternative nuclear DNA markers are occasionally used for somatic hybrid screening Table 2. Mechanical fusion may damage protoplasts by causing injuries.

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