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  • 1
    Electronic Resource
    Electronic Resource
    Species-specific sequences in the genus Solanum: identification, characterization, and application to study somatic hybrids of S. brevidens and S. tuberosum (1990)
    Springer
    Theoretical and applied genetics 80 (1990), S. 693-698 
    Details
    ISSN: 1432-2242
    Keywords: Potato ; Solanum brevidens ; Somatic hybrids ; Species-specific probes ; Squash blots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To aid in the identification and analysis of somatic hybrids between potato (Solanum tuberosum, dihaploid line PDH 40) and the non tuber-bearing wild species S. brevidens, a series of species-specific repetitive DNA sequences have been isolated. This was accomplished by making libraries of HaeIII-digested total DNA of S. tuberosum and S. brevidens, by cloning fragments into the SmaI site of plasmid pUC18 and transforming them into E. coli (JM83). The S. brevidens library consisted of 1,000 recombinant clones, and that of S. tuberosum, 700. Nitrocellulose filters with recombinant clones were hybridised to nick-translated total DNA of S. brevidens and also S. tuberosum, and, following autoradiography, clones that hybridised strongly to the DNA of only one of the species were chosen. Two highly repeated S. brevidens clones (pSB1, 400 bp and pSB7,210 bp), one highcopy-number s. tuberosum clone (pST10, 200 bp) and one low-copy-number sequence of S. tuberosum (pST3, 1.5 kbp) were selected for further analysis by Southern hybridisation to digested total DNA. Clone pSB7 gave a ladder pattern on hybridisation to EcoR1-digested total DNA of S. brevidens, with signals at multiples of 200 bp DNA. Using these probes it was possible to verify the hybridity of putative hybrids of dihaploid S. tuberosum and S.brevidens, and to confirm by Southern analysis and by slot blots the parental genome dosage of hexaploid hybrids (two s. brevidens: one S. tuberosum, and vice-versa). The S. tuberosum-specific probe, pSTIO, hybridised with DNA of three other tuber-bearing wild species (S. hjertingii, S. capsicibaccatum and S. berthaultii). A squash-blot procedure was developed using the probes that would allow early identification of somatic hybrid callus. There are a number of useful applications of such species-specific probes in the identification and analysis of somatic hybrids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00224231
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  • 2
    Electronic Resource
    Electronic Resource
    Resistance to potato leaf roll virus and potato virus Y in somatic hybrids between dihaploid Solanum tuberosum and S. brevidens (1988)
    Springer
    Theoretical and applied genetics 76 (1988), S. 113-117 
    Details
    ISSN: 1432-2242
    Keywords: Potato ; Solanum brevidens ; somatic hybrid ; PLRV ; PVY
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Many somatic fusion hybrids have been produced between a dihaploid potato Solanum tuberosum and the sexually-incompatible wild species S. brevidens using both chemical and electrical fusion techniques. S. brevidens was resistant to both potato leaf roll virus (PLRV) and potato virus Y (PVY), the viruses being either at low (PLRV) or undetectable (PVY) concentrations as determined by enzyme-linked immunosorbent assay (ELISA). The S. tuberosum parent was susceptible to both viruses. A wide range of resistance, expressed as a decrease in virus concentration to both viruses was found amongst fusion hybrids, four of which were especially resistant. The practicality of introducing virus resistance from S. brevidens into cultivated potatoes by somatic hybridisation is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00288840
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  • 3
    Electronic Resource
    Electronic Resource
    Production of somatic hybrids by electrofusion in Solanum (1988)
    Springer
    Theoretical and applied genetics 76 (1988), S. 260-266 
    Details
    ISSN: 1432-2242
    Keywords: Solanum tuberosum ; Solanum brevidens ; electrofusion ; somatic hybrids ; cytology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Conditions are described for large scale electrofusion of mesophyll protoplasts of dihaploid S. tuberosum with those of diploid S. brevidens. Overall fusion frequencies of 20%–30% were achieved, and following fusion, large numbers of protoplast-derived calli were obtained. Putative somatic hybrid plants were selected from the regenerated shoots by examining their morphological characteristics. Twenty-one somatic hybrids were confirmed by isoenzyme analysis and six somatic hybrids were further confirmed by Southern hybridization. Tetraploid hybrids were obtained, but cytogenetic studies indicated that more of the regenerated hybrids were hexaploid than had previously been found following chemical fusion of the same partners. Some advantages of electrofusion over chemical fusion are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00257854
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  • 4
    Electronic Resource
    Electronic Resource
    Field assessment of dihaploid Solatium tuberosum and S. brevidens somatic hybrids (1988)
    Springer
    Theoretical and applied genetics 76 (1988), S. 880-886 
    Details
    ISSN: 1432-2242
    Keywords: Solanum tuberosum ; Solanum brevidens ; Somatic hybrids ; Planned release ; Field trial
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Following both chemical and electrical fusion of protoplasts of a dihaploid line of potato (Solanum tuberosum), (PDH40), with those of the wild species, Solanum brevidens, 11 and 40 somatic hybrid plants, respectively were obtained. Fifteen of these somatic hybrid genotypes and the two parents were studied further in a small field trial to assess field performance and phenotypic variability. In the UK, somatic hybrid plants are classified as genetically engineered organisms, and the UK Advisory Committee on Genetic Manipulation have imposed various restrictions on field experiments. Examination of the somatic hybrids in the field showed extensive phenotypic variability, and no two genotypes were identical. Some of the variation reflected changes in chromosome numbers, but other factors were also involved. Half the somatic hybrid genotypes produced tubers in the field, although the tubers were smaller and differed morphologically from those of PDH40. The results of the study suggest that the extent of somaclonal variation manifested in somatic hybrids is greater than that found in protoplast-derived plants of potato. The implications of this observation and the current regulations concerning field experiments of somatic hybrid plants in the UK are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00273676
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  • 5
    Electronic Resource
    Electronic Resource
    Molecular, cytogenetic and morphological characterization of somatic hybrids of dihaploid Solanum tuberosum and diploid S. brevidens (1989)
    Springer
    Theoretical and applied genetics 78 (1989), S. 696-704 
    Details
    ISSN: 1432-2242
    Keywords: Potato ; Solanum brevidens ; Somatic hybrids ; Characterization (molecular, cytological, morphological)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Fifty-eight somatic hybrid plants, produced both by chemical (11) and electrical fusion (47) of protoplasts of dihaploid Solanum tuberosum and S. brevidens, have been analysed by molecular, cytological and morphological methods. The potentially useful euploid plants constituted 34% of the total, of which 20% were tetraploid and 14% hexaploid; the remainder were aneuploid at the tetraploid, hexaploid and octoploid levels. Analysis of chloroplast DNA showed that 55% of hybrids contained chloroplasts from S. brevidens and 45% from S. tuberosum. Hexaploids, the products of three protoplasts fusing together, were analyzed with specific DNA probes, and this revealed that nuclear genome dosages could be either 2∶1 S. tuberosum∶S. brevidens, or vice-versa. Chloroplast types of hexaploids were not influenced by nuclear genome dosage, and all six possible combinations of genome dosage and chloroplast types were found amongst tetraploids and hexaploids. To examine the morphology of the hybrid population and its possible relation to the chromosome number and chloroplast DNA type, 18 morphological characteristics were measured on greenhouse-grown plants and analyzed by principal component and canonical variate analyses. Both analyses showed that nuclear ploidy has the most prominent influence on the overall morphology of the hybrids. Differential parental genome expression in the morphology of the hybrids is discussed. These results provide useful data on the range of genetic combinations that can be expected to occur amongst somatic hybrid plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00262566
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  • 6
    Electronic Resource
    Electronic Resource
    Production of asymmetric hybrids between Solanum tuberosum and irradiated S. brevidens (1993)
    Springer
    Theoretical and applied genetics 85 (1993), S. 729-734 
    Details
    ISSN: 1432-2242
    Keywords: Protoplast fusion ; Gamma irradiation ; Partial genome transfer ; Solanum tuberosum ; Solanum brevidens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Asymmetric somatic hybrids were obtained by fusion of Solanum tuberosum (PDH40) protoplasts with 300- or 500-Gy irradiated protoplasts of S. brevidens. These radiation doses were sufficient to prevent the growth of the S. brevidens protoplasts. Putative hybrids were selected on the basis of phenotype from regenerated shoots and identified with a S. brevidens-specific probe. From these, 31 asymmetric hybrids were confirmed by morphological characteristics, isoenzyme patterns and RFLP analysis. The morphology of the asymmetric hybrids was intermediate between that of S. tuberosum and symmetric hybrids of both species (obtained without irradiation treatment). Chromosome counts from 17 asymmetric hybrids showed that the chromosome number of the hybrids ranged from 31 to 64. The asymmetric hybrids probably had one or two genome complements (i.e. either 24 or 48 chromosomes) from S. tuberosum and 7–22 chromosomes from S. brevidens. There was no clear correlation between the radiation dose and the degree of elimination of the S. brevidens genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00225012
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