Modulation and direction of the electrofusion response in plant protoplasts

Tempelaar, M. J.; Duyst, A.; Vlas, S.y. De; Król, Grzegorz; Symmonds, C.; Jones, Michael G. K.

doi:10.1016/0168-9452(87)90136-1

Cited by 39 publications

(9 citation statements)

References 16 publications

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“…The observed bursting of mesophyll protoplasts in the electric field could limit the recovery of fusants in mesophyll (+) mesophyll protoplast combinations. Tempelaar et al (1987) proposed an addition of I mM Ca 2+ ions in order to improve the protoplast integrity and to enhance the fusion yield. In our case, however, the presence of divalent calcium cations caused the overheating of protoplast mixture and, consequently, the death of the protoplasts.…”

Section: Discussionmentioning

confidence: 99%

Electrofusion of protoplasts from Solanum tuberosum, S. nigrum and S. bulbocastanum

et al. 2001

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Leaf protoplasts of two wild species, Solanum nigrum var. gigantea (S. ngr gig) and S. bulbocastanum Dun. (S. blb), were electrofused with leaf protoplasts of two diploid potato clones, H-8105 and ZEL-1136, respectively, in order to confer the late blight-resistance from the wild species to the cultivated potato. The S. ngr gig mesophyll (+) H-8105 mesophyll combination resulted in regenerants of mostly normal ngr phenotype. Two regenerants from this combination were proved to be true hybrids by RAPD analysis but they rooted poorely in vitro and did not survive the transfer to soil. The S. ngrgig (+) H-8105 fusion combination was also performed with H-8105 cell suspension derived protoplasts enabling an easy identification of interspecific fusants on basis of their intermediate morphology. From the S. ngrgig mesophyll (+) H-8105 cultured cell combination, many abnormal shoots were regenerated. The two lines which survived had normal ngr phenotype but the presence of tube rosum (tbr) genome in those regenerants was not confirmed by RAPD analysis. No plants with tbr phenotype were obtained from both of S. ngr gig (+) H-8105 combinations. On the contrary, when S. bib mesophyll protoplasts were electrofused with ZEL-1136 mesophyll protoplasts, all regenerated plants had tbr phenotype, indicating much lower morphogenetic potential of S. bulbocastanum in comparison with that of S. nigrum vat. gigantea. However, the hybridity of those regenerants has not been confirmed by RAPD analysis with two different primers. The efficiency of the applied fusion procedure and analysis of the regenerants is discussed.

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Section: Discussionmentioning

confidence: 99%

Electrofusion of protoplasts from Solanum tuberosum, S. nigrum and S. bulbocastanum

et al. 2001

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“…Protoplasts isolated from leaves are generally found to fuse at lower field strengths than protoplasts isolated from roots or suspension cultures (Tempelaar and Jones, 1985;Tempelaar et al, 1987), and, under optimal conditions, fusion efficiency is generally higher for leaf protoplasts (Tempelaar and Jones, 1985;Tempelaar et al, 1987). However, the DC pulse settings that give optimal fusion can vary depending on cell source.…”

Section: Factors Affecting Protoplast Electrofusionmentioning

confidence: 99%

Electrofusion of Plant Protoplasts and the Production of Somatic Hybrids

Bates

1992

Guide to Electroporation and Electrofusion

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“…Although 100 #m CaC12 in the electrofusion buffer stabilized grape protoplasts, calcium did not enhance the fusion of small vacuoles with the plasmalemma of grape protoplasts. In other electrofusion studies [9,18], calcium (up to 1 mM CaC12) usually doubled the number of protoplast fusions that occurred.…”

Section: Electrofusion Of Small Vacuoles and Protoplastsmentioning

confidence: 99%

PEG-enhanced, electric field-induced fusion of tonoplast and plasmalemma of grape protoplasts

Lackney

Spanswick

Hirasuna

et al. 1990

Plant Cell Tiss Organ Cult

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Cultured grape cells accumulate anthocyanins in vacuoles rather than secreting them into the nutrient medium. Therefore, grape cells that contain tonoplast segments in their plasmalemma should be capable of excreting anthocyanins rather than sequestering them in their vacuoles. In initial attempts to construct such 'novel' cells, small vacuoles were fused with the plasmalemma of cultured plant cells. Protoplasts were isolated from grape calluses that produce and accumulate anthocyanins. Small vacuoles were formed by gently rupturing vacuoles isolated from grape protoplasts. Although small vacuoles and protoplasts became aligned in an AC field, the tonoplast and plasmalemma did not readily fuse when subjected to 3 DC pulses of 1200 V cm 1 for 50/~s each. Changes in the intensity, number and/or duration of the DC pulses had no effect on the fusion process. When 1.0% polyethylene glycol was added to the electrofusion buffer, however, small vacuoles and protoplasts fused within a few minutes after the DC pulses were applied. These 'novel' grape cells remained viable for several hours.

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Modulation and direction of the electrofusion response in plant protoplasts

Cited by 39 publications

References 16 publications

Electrofusion of protoplasts from Solanum tuberosum, S. nigrum and S. bulbocastanum

Electrofusion of protoplasts from Solanum tuberosum, S. nigrum and S. bulbocastanum

Electrofusion of Plant Protoplasts and the Production of Somatic Hybrids

PEG-enhanced, electric field-induced fusion of tonoplast and plasmalemma of grape protoplasts

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