Plant regeneration from the protoplasts of Solanum tuberosum, S. nigrum and S. bulbocastanum

Szczerbakowa, Anna; Borkowska, M.; Wielgat, Bernard

doi:10.1007/s11738-000-0001-x

Cited by 9 publications

(7 citation statements)

References 20 publications

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“…As shown previously (Szczerbakowa et al 2000), the used accession ofS. bulbocastanum, P1243512, did not regenerate from the leaf protoplasts under the applied culture conditions: the formed calli turned brown and inevitably died without regeneration.…”

Section: Discussionsupporting

confidence: 56%

Interspecific somatic hybrids Solanum bulbocastanum (+) S. tuberosum H-8105

2003

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A b s t r a c tlnterspecific somatic hybrids between a dihaploid potato clone H-8105 susceptible to Phytophthora infi, stans (Mont.) de Bary and a resistant diploid tuberizing species Solanum bulbocastanum were generated and analysed. Only ten regenerants displaying the intermediate morphology with dominating characteristics of the wild parent (simple leaves, anthocyanin pigmentation) were produced in 15 weeks after a single PEG-mediated fusion event. The RAPD patterns confirmed the hybridity of all of them. The hybrids rooted poorly and grew slowly in vitro. The cytological analysis revealed a high degree of aneuploidy in the hybrids with morphological and growth anomalies in vitro, while the morphologically normal hybrids were tetraploids. All the S. bulbocastanum (+) H-8105 hybrids were unstable in culture and three of them were consequently lost during three years of propagation in vitro. The possible reasons for instability of somatic hybrids between the distantly related species are discussed.

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Section: Discussionsupporting

confidence: 56%

Interspecific somatic hybrids Solanum bulbocastanum (+) S. tuberosum H-8105

2003

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“…Plant regeneration from the protoplasts of fusion partners was previously described in details (Szczerbakowa et al 2000). The parental protoplasts responded to culture conditions differently: the regeneration (shoot formation) was achieved for mesophyll protoplasts of both potato protoclones and S. nigrum var.…”

Section: Regeneration Of the Parental Protoclonesmentioning

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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“…The applied technique of protoplast isolation and regeneration was performed as described previously (Szczerbakowa et al 2000). Parental protoplasts in a 1:1 mixture at final density of 10 6 cm -3 were fused in a medium consisting of 25% PEG 6,000, 0.1 M Ca(NO 3 ) 2 , 0.3 M mannitol and 10% DMSO, adjusted to pH 9.0.…”

Section: Protoplast Isolation Fusion and Regenerationmentioning

confidence: 99%

Nuclear DNA content and chromosome number in somatic hybrid allopolyploids of Solanum

Szczerbakowa

Tarwacka

Śliwińska

et al. 2011

Plant Cell Tiss Organ Cult

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An attempt was made to change the proportion of the parental genomes in interspecific hybrids Solanum nigrum ? S. tuberosum (ngr ? tbr) by means of repeated protoplast fusion. In order to enlarge the potato input into the hybrid genome, the protoplasts of two ngr ? tbr hybrids of different ploidy (7x and 8x) were fused with the protoplasts of two different diploid potato clones in three combinations. Protoclonal variability was studied in three populations of new ngr ? tbr allopolyploids maintained in vitro. The absolute nuclear DNA content (2C) was measured using flow cytometry to estimate the ploidy of the hybrids. The ploidy level of the selected clones was verified by chromosome counts in root meristems. The newly synthesized allopolyploids (75 clones) showed only a small gain in nuclear DNA content above the mean value determined for the parents, instead of the expected addition of an entire diploid potato genome to the combined parental ngr ? tbr genome. An increase in nuclear DNA was observed mostly in the clones having the 7x hybrid as a parent (75% of allopolyploids from two combinations). When the 8x hybrid was used as a parent, only two allopolyploids (5%) exhibited a significantly increased nuclear DNA content. The 8x level of ngr ? tbr allopolyploids was shown to be stable and was only occasionally exceeded. Somatic hybrids ngr ? tbr offer a model system for studying the molecular mechanism(s) and processes involved in stabilization and establishment of the synthetic Solanum allopolyploids.

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Plant regeneration from the protoplasts of Solanum tuberosum, S. nigrum and S. bulbocastanum

Cited by 9 publications

References 20 publications

Interspecific somatic hybrids Solanum bulbocastanum (+) S. tuberosum H-8105

Interspecific somatic hybrids Solanum bulbocastanum (+) S. tuberosum H-8105

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

Nuclear DNA content and chromosome number in somatic hybrid allopolyploids of Solanum

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