Protoplast Fusion in Banana (Musa spp.): Comparison of Chemical (PEG: Polyethylene Glycol) and Electrical Procedure

Assani, Akym; Chabane, Djamila; Haïcour, R.; Bakry, Frédéric; Wenzel, G.; Foroughi‐Wehr, B.

doi:10.1007/s11240-005-4633-9

Cited by 33 publications

(24 citation statements)

References 25 publications

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“…In the cell alignment experiment, most cells (Z95%) were attracted toward the edge of the microelectrode one by one and most cells were aligned as cell-cell twins. Compared with traditional electrofusion method [24,25], the probability of forming cell-cell twins is much higher (42-68%), which is essential for high fusion efficiency.…”

Section: Cell Alignmentmentioning

confidence: 99%

A high‐throughput dielectrophoresis‐based cell electrofusion microfluidic device

Yang

Yin

et al. 2011

Electrophoresis

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A high-throughput cell electrofusion microfluidic chip has been designed, fabricated on a silicon-on-insulator wafer and tested for in vitro cell fusion under a low applied voltage. The developed chip consists of six individual straight microchannels with a 40-μm thickness conductive highly doped Si layer as the microchannel wall. In each microchannel, there are 75 pairs of counter protruding microelectrodes, between which the cell electrofusion is performed. The entire highly doped Si layer is covered by a 2-μm thickness aluminum film to maintain a consistent electric field between different protruding microelectrode pairs. A 150-nm thickness SiO₂ film is subsequently deposited on the top face of each protruding microelectrode for better biocompatibility. Owing to the short distance between two counter protruding microelectrodes, a high electric field can be generated for cell electrofusion with a low voltage imposed across the electrodes. Both mammalian cells and plant protoplasts were used to test the cell electrofusion. About 42-68% cells were aligned to form cell-cell pairs by the dielectrophoretic force. After cell alignment, cell pairs were fused to form hybrid cells under the control of cell electroporation and electrofusion signals. The averaged fusion efficiency in the paired cells is above 40% (the highest was about 60%), which is much higher than the traditional polyethylene glycol method (<5%) and traditional electrofusion methods (∼12%). An individual cell electrofusion process could be completed within 10 min, indicating a capability of high throughput.

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Section: Cell Alignmentmentioning

confidence: 99%

A high‐throughput dielectrophoresis‐based cell electrofusion microfluidic device

Yang

Yin

et al. 2011

Electrophoresis

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“…through nonconventional strategies, i.e. protoplast culture and fusion, mutagenesis, somaclonal variation and genetic transformation (Ganapathi et al, 2001;Matsumoto et al, 2002;Assani et al, 2005;Houllou-Kido et al, 2005;Kumar et al, 2005;Pei et al, 2005). In addition, the incorporation of biotechnological procedures in breeding programs depends upon effi cient and reliable plant regeneration protocols.…”

Section: Introductionmentioning

confidence: 99%

Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

Morais-Lino

Santos-Serejo

Silva

et al. 2008

Pesq. agropec. bras.

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-The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male fl owers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

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“…7.1 Fusion of gentian protoplasts. a Embryogenic cell suspension of G. cruciata and (b) its "white" protoplasts, c shoot culture of G. tibetica and (d) "green" leaf mesophyll protoplasts, e "pearl chain" formation in AC electric field mixture of "white" and "green" protoplasts after DC pulse, f arrows indicate newly formed heterokaryons, g a single heterokaryon before and h after cytoplasmic mixing, i an increase in heterokaryon cell volume 1 week after fusion, j asymmetric heterokaryon division after 2 weeks in culture, k a multicellular hybrid aggregate after 3 weeks of culture, l granular embryogenic callus, m regenerating somatic embryo, n embryo conversion into plantlets, o acclimatized somatic hybrid plant Table 7.3 Combinations of species used in studies on gentian somatic hybridization and the results of particular protoplast fusions Chemical fusion can sometimes be more effective than electrofusion (Assani et al 2005). However, the frequency of heterokaryons obtained with the use of PEG for the same 3 gentian species combinations ranged from 2.42 % to only 3.65 % (Table 7. 3; Ładyżynski et al 2006).…”

Section: Conditions Of Protoplast Fusionmentioning

confidence: 99%

Protoplast Culture and Somatic Cell Hybridization of Gentians

Tomiczak

Mikuła

Rybczyński

2015

The Gentianaceae - Volume 2: Biotechnology and Applications

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During the last three decades, less than fifteen papers have described the results of scientific investigations in the field of gentian protoplast technology and somatic hybridization. Despite rather limited research already done on this subject, several important goals have been achieved. Protoplast-to-plant systems have been developed either for leading ornamental species or for specific medicinal plants. Two major protoplast sources were evaluated in gentians, namely differentiated leaf mesophyll cells and undifferentiated callus/cell suspensions. Plant regeneration proceeded by the two different pathways of shoot organogenesis or somatic embryogenesis. Some examples of somaclonal variation at the ploidy level were demonstrated within the pool of protoplast-derived regenerants. Totipotency exhibited by gentian protoplasts was exploited to create three different somatic hybrid combinations: intergeneric Swertia mussotii (+) Bupleurum scorzonerifolium, and interspecific Gentiana kurroo (+) G. cruciata and G. cruciata (+) G. tibetica. IntroductionPlant cell totipotency as postulated by Schleiden and Schwann in the middle of the nineteenth century is the basis of modern plant biotechnology (Vasil 2008). Based on the ability of a living single plant cell to dedifferentiate and to convert into other cell types, it is possible to obtain a completely new plant from a cell and even from its protoplast. Consequently, plant regeneration from single protoplasts underlies the genetic manipulation technologies of somatic hybridization and direct genetic transformation by DNA uptake. However, the prerequisite for practical application of these technologies is the development of efficient and reproducible protoplast-to-plant systems for the species of interest.

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Protoplast Fusion in Banana (Musa spp.): Comparison of Chemical (PEG: Polyethylene Glycol) and Electrical Procedure

Cited by 33 publications

References 25 publications

A high‐throughput dielectrophoresis‐based cell electrofusion microfluidic device

A high‐throughput dielectrophoresis‐based cell electrofusion microfluidic device

Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

Protoplast Culture and Somatic Cell Hybridization of Gentians

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