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  • 1985-1989  (22)
  • 1955-1959  (10)
  • 1890-1899
  • 1820-1829
  • gene transfer
  • somaclonal variation
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 42 (1989), S. 155-162 
    ISSN: 1573-5060
    Keywords: Rubus laciniatus ; thornless blackberry ; brambles ; tissue culture ; periclinal chimera ; in vitro ; ex vitro ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary ‘Thornless Evergreen’ (TE) blackberry (Rubus laciniatus Willd.) is a periclinal chimera in which the epidermis has mutated to thornlessness (designated ‘t’), while the internal portions of the plant remain genetically thorny (tTT). Over 300 ex vitro plants (from shoot tip cultures of TE) were established in the field and observed over two fruiting seasons for various vegetative traits and fruiting characteristics. Both parental types and variants were included in the field plantings. Plants were evaluated for growth habit, vigor, flowering, and fruiting traits. Continuous variation for growth habit, vigor, flower number, and fertility was observed. Some of the variation could be explained in terms of chimeral segregation (pure thornless vs. chimeral types). Tissue culture-induced (somaclonal) variation was assumed to be the source of most of the other variation.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 40 (1989), S. 75-81 
    ISSN: 1573-5060
    Keywords: Brassica juncea ; Indian mustard ; plant generation ; somaclones ; yield ; oil content ; dwarf type ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Multiple shoot formation in cotyledonary callus of Indian mustard (Brassica juncea cv. Prakash) was induced on modified MS media supplemented with high cytokinin (kinetin or zeatin) and low IAA concentrations. Complete plants were obtained on prolonged incubation of shoots on the same medium. 6-Benzyladenine alone or in combination with IAA or NAA did not support plantlet regeneration. A total of 71 plants were transferred to greenhouse. The seed, however, could be collected from 37 plants only. The seed was sown in the field to evaluate the material for somaclonal variation in R1 generation. Data were recorded for yield, plant height, number of primary branches, siliqua number, 1,000 seed weight and oil content. Somaclonal lines showed tremendous amount of variation for all the characters studied. A number of plants in this generation showed significantly higher yield and/or other improved agriculturally important characteristics as compared to the control. A line with dwarf plant type was also identified. A number of plants were selected from this generation and carried forward to R2 generation. Most of these lines bred true in R2 generation. The material seems to be very promising for future breeding programmes.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Fish physiology and biochemistry 7 (1989), S. 409-413 
    ISSN: 1573-5168
    Keywords: gene transfer ; transgenic fish ; growth hormone ; antifreeze polypeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Successful production of transgenic fish by gene transfer technology is a very important breakthrough in the techniques of genetic manipulation in animals. This will have an impact of an unprecedented scale in fish biology, aquaculture and mariculture. This is a summary of the workshop on the Transgenic Fish presented at this Symposium. The Workshop discussed the current knowledge, experimental difficulties and related topics of the transgenic fish. It recommended further research on better gene constructs, methods development, safety containment and the closer collaboration of researchers of different disciplines.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 19 (1989), S. 159-165 
    ISSN: 1573-5044
    Keywords: cultivars ; roots ; shoots ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four tronchuda (Brassica oleracea var. tronchuda Bailey) cultivars were tested for their ability to regenerate in vitro on Murashige & Skoog (MS) medium supplemented with 3 different combinations of α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). Explants were either axillary bud-free whole cotyledons or hypocotyls from 7-day-old darkgrown seedlings. The ability to regenerate varied by cultivars, explants and the concentration of growth regulators. Hypocotyl explants of all 4 cultivars, and cotyledon explants of 2 cultivars, developed plantlets within 4 weeks. Hypocotyl explants produced more shoots than cotyledons. Cotyledon explants produced more roots than hypocotyls. Best shoot regeneration was on MS medium supplemented with 2 mgl-1 BAP and 0.1 mgl-1 NAA. ‘Portuguesa’ produced the most shoots. Some regenerants varied in leaf shape and phyllotaxy.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-0603
    Keywords: Agrobacterium ; gene transfer ; somatic embryos ; walnut ; β-glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Somatic embryos have been successfully used as a target tissue for transformation and regeneration of transgenic walnut plants. Walnut somatic embryos, initiated originally from developing zygotic embryos, proliferate numerous secondary embryos from single cells in the epidermal layer. These single cells in intact somatic embryos are susceptible to transformation by genetically engineeredAgrobacterium tumefaciens and provide a means to regenerate nonchimeric transgenic plants. This gene transfer system has been made more efficient using, a) vector plasmids containing two marker genes encoding β-glucuronidase (GUS) and aminoglycoside phosphotransferase (APH(3′)II) and B) a more virulent strain ofAgrobacterium. This system should be applicable to any crop that undergoes repetitive embryogenesis from singleAgrobacterium-susceptible cells.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-5044
    Keywords: Oryza sativa L. ; somaclonal variation ; grain and quality characters ; protein content
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Variations in some grain and quality characters in progenies of regenerated rice plants were studied. Grain length and weight decreased significantly, yet gel consistency increased. Variations in these quantitative characters of all cultivars studied were consistent, showing the tendencies of the variations. Grain protein contents of the somaclones were higher in one cultivar. Variability of most traits was increased by combining low-dosage radiation and tissue culture.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0730-2312
    Keywords: tumorigenicity ; metastasis ; ha-ras ; oncogene ; gene transfer ; skin cancer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: High-molecular-weight genomic DNA isolated from a human cutaneous squamous cell carcinoma (AS) was assayed for its ability to induce tumorigenic transformation of NIH 3T3 cells. Subcutaneous injection of NIH 3T3 cells cotransfected with DNAs from AS tumor and pSV2-neo plasmid not only induced tumors at the site of injection, but also metastasized spontaneously to the lungs in 100% of nude mice injected. DNA isolated from a representative primary tumor and a metastasis was again used in a second round of transfection. Injection of secondary transfectants into nude mice again resulted in induction of both subcutaneous tumors and spontaneous long metastases. Southern blot hybridization with ras-specific probes revealed that DNA from both primary tumors and metastases induced by AS tumor DNA contained highly amplified Ha-ras oncogene. Furthermore, DNAs from secondary tumors and metastases induced by DNA from a primary tumor and a metastasis also contained similar highly amplified Ha-ras oncogene. These results suggest that the amplified Ha-ras oncogene may be responsible for induction of both tumorigenic and metastatic phenotypes in NIH 3T3 cells transfected with DNA from AS tumor.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Potato research 30 (1987), S. 371-380 
    ISSN: 1871-4528
    Keywords: dihaploids ; protoclones ; protoplasts ; Solanum tuberosum ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Zusammenfassung Die Verwendung dihaploider Kartoffelklone (2n=2x=24) in Züchtungsprogrammen, vor allem die Rückkehr zum tetraploiden Grad, wird sehr oft durch deren fehlende Fertilität verhindert. Dieses Problem dürfte am elegantesten durch somatische Fusion der Dihaploiden überwunden werden. Als Ergebnis einer solchen Prozedur würden sich, ausser der Paarung beider Ploidiestufen, die Addition qualitativer und quantitativer Merkmale ergeben. Grösstes Hindernis für die Anwendung der Protoplastenfusion ist das Fehlen eines brauchbaren Selektionssystems für die Separierung von homo- und heterokaryotischen Fusionsprodukten. In der hier beschriebenen Methode wurde der sehr kräftige Wuchs einiger Kalli für die Vorselektion einiger vermuteter Hybriden verwendet. Nach Anwendung der Polyethylenglykol-Fusions-methode (PEG, Tabelle 1) konnten von fünf selektierten dihaploiden Klonen (P1–P5) grosse Zahlen von Kalli und Pflanzen regeneriert werden, obwohl die PEG-Behandlung einen negativen Einfluss auf die Regenerationsrate hatte (Tabelle 2). Insgesamt wurden nach PEG-Behandlung 115 Kalli als vermutliche Hybriden, ihrer extremen Wuchsleistung entsprechend, selektiert. Tabelle 3 vergleicht die Ploidiestufen dieser selektierten Klone mit der von unbehandelten Elternklonen. Wegen der somaklonalen Variation wurden auch von nicht fusionierten Protoplasten viele tetraploide Klone gefunden. Ihre Zahl war allerdings signifikant kleiner, und unter den nicht PEG-behandelten Protoplasten waren immer einige diploide vorhanden. Die Tabellen 4 und 5 zeigen die Merkmale von 9 und 10 selektierten Klonen (Hy 1–10) in vitro und in vivo für Sprosslänge, Zahl der Nodien, Blattfläche, Blattform, Zahl der Wurzeln und allgemeiner Wachstumsleistung. In allen Fällen waren die gemessenen Parameter bei den selektierten Klonen signifikant grösser als bei den Kontrollen. Folglich kann das stärkere Wachstum der selektierten Klone nicht nur mit somaklonaler Variation erklärt werden. Es ist ein starkes Indiz für die Hybridnatur. Das Isoenzym-Muster der Esterasen unterstreicht diese Schlussfolgerung. Den Ergebnissen zufolge ist es möglich, somatische Hybriden anhand ihrer hybriden Vitalität vorzuselektieren. Dies sollte die Möglichkeit zur Entdeckung somatischer Hybriden in ausreichender Häufigkeit für praktische Züchtungsprogramme erhöhen.
    Abstract: Résumé L'utilisation de clônes dihaploïdes (2n=2x=24) dans les programmes d'hybridation, et particulièrement le retour au niveau tétraploïde, est entravée par leur manque de fertilité. Ce problème pourrait être maitrisé élégamment par la fusion somatique de dihaploïdes. D'un tel procédé résulterait de plus l'héritage de caractères qualitatifs et quantitatifs apportés par le doublement de ploïdie. Le principal obstacle pour utiliser la fusion de protoplastes est l'absence d'un système de sélection approprié pour la séparation des homo et hétérokaryotes produits par la fusion. Par la méthode décrite dans cet article la grande vigueur de croissance de quelques cals a été mise à profit pour la présélection des présumés hybrides. Après l'adaptation du procédé de fusion au polyéthylène-glycol (PEG, tableau 1) sur cinq clônes dihaploïdes sélectionnés (P1–P5) un grand nombre de cals et de plantes pourrait être régénéré, bien que le traitement PEG ait une influence négative sur le taux de régénération (tableau 2). Au total 115 cals étaient sélectionnés après le traitement comme de présumés hybrides, en raison de leur extrème vigueur. Le tableau 3 compare les niveaux de ploïdie de ces clônes sélectionnés avec ceux des parents non traités. La variation somatique de protoplastes non fusionnés est également trouvée dans de nombreaux clônes tétraploïdes, leur nombre est cependant significativement plus petit, et parmi les protoclônes régénérés de protoplastes non traités quelques diploïdes sont toujours présents. Les tableaux 4 et 5 montrent les caractéristiques de 9 et 10 clônes sélectionnés (Hy 1–10) in vitro et in vivo, respectivement pour la longueur de pouses, le nombre de noeuds, la surface et la forme des feuilles, le nombre de racines et la vigueur générale. Dans tous les cas, les paramètres mesurés ont des valeurs significativement plus élevées dans les clônes sélectionnés que dans les témoins. En conséquence, leur vigoureuse croissance ne peut être expliquée seulement par la variation somatique mais elle constitue une indication sur la vigueur hybride. L'analyse des estérases souligne cette conclusion (figure 1). Ces résultats montrent qu'il est possible de présélectionner des hybrides somatiques par leur vigueur, ce qui augmente les possibilités de les détecter en quantité suffisante dans les programmes d'hybridation.
    Notes: Summary Tetraploid potato plants were regenerated after polyethylene-glycol-induced protoplast fusion between dihaploids. Hybrid vigour of the regenerated calli was used for preselection of fusion products. Nearly all the selected vigorous clones possessed chromosome counts at the tetraploid level. Fusion products were compared to the parental material to auto-fused plants of and to three protoclones expressing different degrees of somaclonal variation. The selected clones, where grown in vitro in growth rooms and in pots in the glasshouse, showed increased vigour compared to their parents, to auto-fused and to 4x protoclones. Plants of clones from very vigorous calli, when assessed by height, the number of nodes per plant, leaf morphology and tuber production, showed hybrid vigour. The hypothesis that superior clones result from heterokaryons after protoplast fusion or that they arise from other in vitro events such as somaclonal variation is discussed.
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  • 9
    ISSN: 1573-5028
    Keywords: Bradyrhizobium ; gene transfer ; nitrogen fixation ; plasmid ; root-nodule symbiosis ; vector construction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A non-essential DNA region carrying two different repeated sequences (RSβ3 and RSα9) adjacent to a nitrogen fixation (nif) gene cluster has been identified previously in Bradyrhizobium japonicum strain 110. In closely related B. japonicum strains a similar genomic arrangement was found. We constructed a mobilizable plasmid vector carrying RSβ3 and RSα9, and a kanamycin resistance cassette (nptII gene) plus suitable cloning sites inserted between the two repeated sequences. Using this vector (pRJ1035), stable integration of a lacZ gene fusion into the B. japonicum genomic RS region was achieved. The resulting strain yielded more than 10-fold higher β-galactosidase activity in soybean root nodules as compared to a B. japonicum strain carrying the same lacZ fusion on a pRK290-based plasmid.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 9 (1987), S. 189-196 
    ISSN: 1573-5044
    Keywords: cereals ; genetic variability ; plant tissue culture ; somaclonal variation ; Sorghum bicolor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sorghum bicolor (L.) Moench, plants were regenerated from 4 to 5 month old callus cultures originally derived from seedling explants. Somaclonal variation was examined in the SC2 and SC3 generations of eight different clones plus the non-tissue cultured parent line. Characters examined included the area of the third leaf, height, tiller number, total shoot weight, seed number, grain yield, days to flowering, and chlorophyll content in the SC2 generation. Some plants were phenotypically different from the parent line in the SC2 generation. All clones were similar to the parent in third leaf area and chlorophyll content. Three clones had significant height reduction and six had significantly higher total plant matter production. Seven of the clones had significant increases in number of secondary tillers, and all had significant decreases in days to flowering. Only three clones had significant increases in grain yield and seed number; however, all the clones had smaller seeds. In subsequent studies of SC2 and SC3 generations of three somaclones, characters examined included height, tiller number, total shoot weight, and days to flower. In one of the somaclones none of the original differences in characters was maintained in two subsequent growing seasons in the SC2 or SC3 generation. The other two somaclones maintained the increased tiller number in the next growing season in both the SC2 and SC3 generations. In one of the somaclones plant height reduction occurred in the SC2 and SC3 generations. All other traits were not maintained.
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  • 11
    ISSN: 1573-5060
    Keywords: Solanum tuberosum ; potato ; amino acids ; amino acid analogue resistance ; frost tolerance ; plant regeneration ; proline ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A number of previously selected hydroxyproline (hyp) resistant cell lines of a diploid potato (Solanum tuberosum L., clone H2578, 2n=2x=24) could be regenerated into plants which were further analysed. Hyp resistance, although lower than in the originally selected calli, was still present in regenerated shoots and in callus initiated from these shoots and it was not lost upon (mini)tuber propagation. Regenerated shoots showed a wide range of phenotypic variation. The chromosome number, analysed in 4 clones, appeared to be hypotetraploid (44 or 45). Tuber-propagated regenerants generally showed increased frost tolerance both at the plant and the cell level. In leaves this seemed to be associated with increased levels of both proline and total amino acid content. However, in callus only the total amino acid content but not proline was still elevated. In the wild type the frost tolerance of the tubers appeared to be lower than that of the leaves. Between tubers of wild type and the hyp resistant regenerants no differences in frost tolerance were found, although proline and total amino acid content tended to be higher in tubers from the regenerants.
    Type of Medium: Electronic Resource
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 8 (1987), S. 209-216 
    ISSN: 1573-5028
    Keywords: hairy root ; gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic engineering of legumes and other important dicotyledonous plants is limited because of the difficulty of regenerating plants via cell culture. Since a considerable number of crop plants can be regenerated only from root culture, the introduction of foreign genes into Agrobacterium rhizogenes-induced hairy roots may expand the list of crop plants that could be genetically engineered. Here we report genetic transformation of alfalfa (Medicago sativa L.), a valuable forage legume, using a virulent strain of Agrobacterium rhizogenes containing, in addition to its Ri-plasmid, a binary vector containing a nopaline synthase gene. Plant cells transformed by this vector can be easily identified by their ability to produce nopaline. Transformed alfalfa plants were recovered from A. rhizogenes-induced hairy roots. These transgenic plants were characterized by normal leaf morphology and stem growth but a root system that was shallow and more extensive than normal. These plants were also fertile, set seeds upon self-pollination and outcrossing. Nopaline was detected in R1 progeny. Southern blot analysis confirmed the presence of multiple copies of T-DNAs from the Riplasmid in the plant genome in addition to the vector T-DNA.
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  • 13
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; crown gall ; somaclonal variation ; T-DNA rearrangements ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract After three years of apparent stability in tissue culture, the single cell derived shooty crown gall line sNT1.013 produced a revertant shoot which had switched from non-rooting (Rod+) and octopine synthesizing (Ocs+) to Rod- Ocs-, indicating that in this revertant TL-DNA genes 4 (causing the Rod+ trait) and gene 3 (causing the Ocs+ trait) had been inactivated. Southern blots revealed that the inactivation of these T-DNA genes was the result of a considerable rearrangement of DNA sequences, accompanied by deletions and possibly also by DNA amplifications. This study for the first time unambiguously proves that foreign genes which have been introduced via Agrobacterium tumefaciens can, at a low frequency, be inactivated after T-DNA integration because of reorganization of T-DNA sequences during tissue culture. This can be considered as an event of somaclonal variation.
    Type of Medium: Electronic Resource
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 8 (1987), S. 461-469 
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; binary vectors ; gene transfer ; kanamycin resistance ; legume transformation ; Trifolium repens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A system was established for introducing cloned genes into white clover (Trifolium repens L.). A high regeneration white clover genotype was transformed with binary Agrobacterium vectors containing a chimaeric gene which confers kanamycin resistance. Transformed kanamycin resistant callus was obtained by culturing Agrobacterium inoculated stolon internode segments on selective medium. The kanamycin resistance phenotype was stable in cells and in regenerated shoots. Transformation was confirmed by the expression of an unselected gene, nopaline synthase in selected cells and transgenic shoots and by the detection of neomycin phosphotransferase II enzymatic activity in kanamycin resistant cells. Integration of vector DNA sequences into plant DNA was demonstrated by Southern blot hybridisation.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 36 (1987), S. 659-665 
    ISSN: 1573-5060
    Keywords: Hordeum vulgare ; barley ; regeneration ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 42 (1986), S. 1128-1137 
    ISSN: 1420-9071
    Keywords: Cell fusion ; cell hybrids ; gene transfer ; human genome ; mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 5 (1986), S. 104-107 
    ISSN: 1432-203X
    Keywords: Plant tissue culture ; somaclonal variation ; mutable allele ; transposable element ; Medicago sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A white-flowered mutant (“WFM”) was regenerated from tissue culture of a purple-flowered plant of tetraploid alfalfa (Medicago sativa L.). When WFM was recultured, many regenerated plants (〉20%) were purple-flowered. Genetic analysis established that a functional allele, C2, of a locus required for anthocyanin pigmentation was in the simplex condition (C2c2c2c2) in the donor genotype when it mutated to an unstable recessive (“mutable”) allele, c2-m4, which is carried by WFM. Tissue culture experiments demonstrated that c2-m4 reverts to function at a high frequency in vitro. Results indicate that reversion occurs early in culture and may be the result of a genome shock associated with callus formation. Reversion also occurs in planta, but at a much lower frequency than in vitro. The c2-m4 allele is transmitted to progeny which revert in tissue culture. Revertant alleles, like the progenitor allele, are stable and are sexually transmitted. The action of a transposable element which is especially active in vitro is suggested.
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 5 (1986), S. 108-110 
    ISSN: 1432-203X
    Keywords: Plant tissue culture ; somaclonal variation ; mutable allele ; transposable element ; Medicago sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An unstable recessive (“mutable”) allele, c2-m4, of a locus required for anthocyanin pigmentation in alfalfa (Medicago sativa L.) reverts to a stable functional state at high frequency in vitro. It was previously established that a white-flowered mutant (“WFM”) and a white-flowered progeny of WFM (“WHGW3”) each carry the unstable allele. More than 20% of plants regenerated from tissue cultures of WFM and WHGW3 are revertant. It is here established that most nonrevertant plants regenerated from cultures of WFM and WHGW3 are stabilized in the recessive condition. Reculture of nonrevertants of WFM and WHGW3 indicated that there are three classes of nonrevertants: (i) Nonrevertants which revert in vitro at a high frequency typical of WFM; (ii) Nonrevertants which revert upon reculture but at significantly lower frequencies than WFM; and (iii) Nonrevertants which do not revert upon reculture. These observations are discussed in terms of transposable element action in vitro.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 6 (1986), S. 167-172 
    ISSN: 1573-5044
    Keywords: cape primrose ; Gesneriaceae ; anther culture ; somaclonal variation ; caulogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regeneration of plants by caulogenesis of anther derived callus of Streptocarpus X hybridus ‘Concorde’ was obtained on MS basal medium with 2.0 mg/l NAA and 0.5 mg/l BAP. All regenerated plants were diploid and of somatic cell origin as determined by chromosome counts, flower color and morphological similarity to the anther donor plant, and flower color segregation among progeny of anther-derived plants. Variation expressed as deformed and dwarfed transient morphology was observed in 2 anther-derived plants. Variation was also observed among progenies of 5 anther-derived plants from the same anther donor plant for days to flowering, flowers per peduncle, peduncle length, and leaf area. Additional morphological variation including transient leaf variegation among seedlings and twisted or inverted leaves on mature plants was also observed among progeny of anther-derived plants.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 34 (1985), S. 525-537 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; bread wheat ; somaclonal variation ; chromosomal aberrations ; wheat breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transmission of somaclonal variation was studied in callus derived (SC1) plants of wheat, Triticum aestivum L. Second (SC2), third (SC3) and fourth (SC4) generations were compared with those obtained from embryo culture (E1-E4). SC2 generation had significantly lower grain yield (−37.3%), reduced thousand kernel weight (−7.8%) and shorter height (−2.1%) than that of E2, and included 5.7% seed-sterile and 15.0% partially seed-sterile plants. SC3 generation showed reduced yield (−12.6%), lower thousand kernel weight (−4.9%) and shorter plant height (−1.9%) than E3 generation. SC4 progeny of a short height, partially sterile SC1 variant included aberrant plants: a haploid, few aneuploids, mixoploids, some showing multiploidy, spindle abnormalities, multivalents, bridges and fragments, and tillers having supernumerary spikes with branched rachis. Somaclonal variation resulted from genome instability in callus, and likely involved transposable elements.
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  • 21
    ISSN: 1573-5060
    Keywords: Lotus corniculatus ; birdsfoot trefoil ; tissue culture ; somaclonal variation ; alteration stabiliy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Plants of Lotus corniculatus regenerated from single genotype-derived calli showed for several agronomic and morphological traits a variation similar to that present in the original population. Comparisons of the results obtained under field conditions during the 2nd and 3rd year from plant regeneration show: a) the maintenance of significant variance within callus-derived population for several traits examined; b) the total disappearance of ‘minus variants’ indicating a time-related decrease of depressive in vitro effects.
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  • 22
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    Plant cell, tissue and organ culture 4 (1985), S. 159-169 
    ISSN: 1573-5044
    Keywords: Plant tissue culture ; callus ; regeneration ; somaclonal variation ; isozymes ; chromosome instability ; celery ; Apium graveolens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetically marked tissues of celery (Apium graveolens) were employed to contrast genetic and chromosomal stability in serially bulk-transferred callus and regenerated plants. After six months in culture, 84% of the callus cells were karologically indistinguishable from normal, while the remainder exhibited chromosome loss and/or fusion. All of 50 clones derived from this tissue expressed the control phenotype with respect to heterozygous isozyme markers. Of 95 plants regenerated from the same tissue, 94 were phenotypically indistinguishable from the original explant donor, and cytogenetic analyses revealed the presence in 4.3% of an accessory chromosome, while the remainder were normal diploids. Analysis of the selfed progeny of these regenerated plants revealed the presence of a new recessive mutation causing abnormal leaf morphology at a frequency of 1.8%. Only one of 40 cells in 12-month-old callus tissue was karyologically indistinguishable from normal, the remainder consisting primarily of hypodiploids. The observation that all 50 clones were phenotypically heterozygous was statistically inconsistent with the hypothesis that hypodiploidy was associated with random complete chromosome loss. The culture had, at this point, lost the ability to regenerate. It is speculated that embryogenic cloning of celery may be suitable under certain circumstances for direct field establishment, but that levels of new genetic variation are sufficiently high to preclude its use for seed production.
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  • 23
    ISSN: 1573-5060
    Keywords: meristem ; shoot apex ; ballistic microtargeting ; gene transfer ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The classical approach of gene transfer to a given plant species delivers the foreign gene to transformable cells and then puts the effort into generating plants. This approach is very difficult in many important crop plants, including cereals, and the results of regeneration are very genotype-dependent. In contrast, we use regenerable cells and try to transform them. Shoot apical meristems provide a tissue which regenerates in situ a fertile plant for most given genotypes or species. Transformation of meristem cells may lead to transgenic sectors in chimeras. These sectors may contribute to the gametes and, thus, to transgenic offspring, which then should be homohistonts and not sectorial chimeras like their parents. Our model plant for these studies is wheat. Microtargeting is a ballistic approach which is particularly suitable for the controlled delivery of microprojectiles to meristem cells in situ (Sautter et al., 1991). We summarize in this paper our experience with ballistic microtargeting of transgenes to wheat shoot apical meristem cells in situ.
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  • 24
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    Euphytica 85 (1955), S. 295-302 
    ISSN: 1573-5060
    Keywords: tissue culture ; somaclonal variation ; plant breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Somaclonal variation is a tool that can be used by plant breeders. The review examines where this tool can be applied most effectively and the factors that limit or improve its chances of success. The main factors that influence the variation generated from tissue culture are (1) the degree of departure from organised growth, (2) the genotype, (3) growth regulators and (4) tissue source. Despite an increasing understanding of how these factors work it is still not possible to predict the outcome of a somaclonal breeding programme. New varieties have been produced by somaclonal variation, but in a large number of cases improved variants have not been selected because (1) the variation was all negative, (2) positive changes were also altered in negative ways, (3) the changes were not novel, or (4) the changes were not stable after selfing or crossing. Somaclonal variation is cheaper than other methods of genetic manipulation. At the present time, it is also more universally applicable and does not require ‘containment’ procedures. It has been most successful in crops with limited genetic systems and/or narrow genetic bases, where it can provide a rapid source of variability for crop improvement.
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  • 25
    ISSN: 1573-5060
    Keywords: doubled haploids ; micropropagation ; mutant cultivars ; mutation techniques ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Conventional mutation techniques have often been used to improve yield, quality, disease and pest resistance in crops, or to increase the attractiveness of flowers and ornamental plants. More than 1700 mutant varieties involving 154 plant species have been officially released. In some economically important crops, e.g. barley, durum wheat and cotton, mutant varieties occupy the majority of cultivated areas in many countries. Mutation techniques have become one of the major tools in the breeding of ornamentals such as alstroemeria, begonia, chrysanthemum, carnation, dahlia and streptocarpus. The use of in vitro techniques such as anther culture, shoot organogenesis, somatic embryogenesis and protoplast fusion can overcome some of the limitations in the application of mutation techniques in both seed and vegetatively propagated crops. In vitro culture in combination with induced mutations can speed up breeding programmes, from the generation of variability, through selection, to multiplication of the desired genotypes. The expression of induced mutations in the pure homozygote obtained through microspore, anther or ovary culture, can enhance the rapid recovery of the desired traits. In some vegetatively propagated species, mutations in combination with in vitro culture technique, may be the only method of improving an existing cultivar. Currently, many molecular studies rely on the induction and identification of mutants in ‘model species’ for construction and subsequent saturation of genetic maps, understanding of developmental genetics and elucidation of biochemical pathways. Once identified and isolated, the genes that encode agronomically-important features can be either introduced directly into crop plants or used as probes to search for similar genes in crop species. It seems most likely that the recent developments based on these technologies will soon provide improved methods for selection of desired mutants.
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  • 26
    ISSN: 1573-5060
    Keywords: gene transfer ; Hordeum vulgare ; neomycin phosphotransferase II ; particle bombardment ; transgenic barley
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS). Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed. Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.
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  • 27
    ISSN: 1573-5060
    Keywords: aluminium toxicity ; soil acidity ; somaclonal variation ; sorghum ; Sorghum bicolor (L.) Moench ; tissue culture ; salt stress ; drought stress ; variants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sorghum bicolor (L.) Moench is generally quite sensitive to salt and acid (high aluminium) soil stresses, but quite tolerant of drought stress. As with any stress phenomenon, intra-specific variability exists within the genus. In vitro cell selection and somaclonal variation offer an alternative to traditional breeding methodology for generating improved breeding lines for hybrid development. A field selection protocol was developed for the three soil stresses and inter-stress evaluations were conducted in an effort to find multiple, stress-tolerant genotypes. The acid soil-drought stress, super-tolerant selections were located by the R7 generation when exposed to a combined aluminium-drought stress field environment and when the regeneration population (number of regenerated lines from one callus source) was maintained at 15,000 plants or higher. A variant frequency of 0.1 to 0.2% for stress tolerance and acceptable agronomic traits among the surviving somaclones, provided an adequate number of phenotypes with desirable agronomic characteristics and a high level of soil stress tolerance. Subsequent research verified that the stress-tolerant regenerants had superior acid soil and drought stress tolerance to that of the donor parents, that their yield capabilities under stress were superior to their parents, and that their stress tolerance attributes were transferred in hybrid combinations. In vitro selection was not effective in increasing the number of field stress survivors. In fact, superior germplasms were developed from non-stressed callus or salt-stressed callus. In vitro selection reduced regeneration frequency and subsequent survival of plants under field stress. In vitro-stressed regenerants should be subjected only to non-stressed environments to maintain population numbers for field selection and thereafter should be subjected to stress environments during later (R5+) generations. The optimal strategy for the exploitation of somaclonal variation may be through short-term cell culture (〈 12 months) with no attempt at in vitro selection.
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  • 28
    ISSN: 1573-5060
    Keywords: callus culture ; organogenesis ; pea ; Pisum sativum ; somaclonal variation ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The possibility of producing agronomically-useful somaclones via organogenesis and somatic embryogenesis from callus cultures of pea (Pisum sativum L.) was studied. Organogenic calli were induced from immature leaflets on MSB medium with NAA and BAP. Embryogenic calli were derived either from immature zygotic embryos (using 2,4-D) or from shoot apices (using picloram) of aseptically-germinated seedlings. The seed progenies (T1 to T3-generation) of primary regenerants were grown in field conditions and their phenotypic variation was evaluated and compared with control, non-tissue culture-derived plant material. In addition, electrophoretic analyses of selected isoenzyme systems and total proteins have been done. The results do not show dramatic changes in qualitative and quantitative traits. The evaluation of at least two future generations (T4, T5) is planned.
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  • 29
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    Euphytica 85 (1955), S. 323-327 
    ISSN: 1573-5060
    Keywords: Brassica napus ; fatty acids ; gas chromatography ; Lunaria annua ; protoplast regeneration ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A programme of research was designed to investigate methods for the modification of the fatty acid profiles of high performance lines of oilseed rape (Brassica napus L.) in an attempt to produce lines with enhanced levels of industrially useful fatty acids. The methodology employed to achieve these objectives was based on the exploitation of somaclonal or protoclonal variation, and targeted somatic hybridization using wild cruciferous germplasm as fusion partners. A range of somaclonal lines was produced from shoot regeneration protocols. These lines underwent replicated, randomised glasshouse trials for morphological assessment followed by gas chromatographic analysis to monitor any changes in fatty acid profile. It was found that a small number of lines exhibited potentially useful changes in oleic acid and polyunsaturated fatty acid content. Protoplast regeneration and electrofusion protocols for a range of winter oilseed rape lines were developed, and methods for the isolation and fusion of protoplasts of the wild crucifer Lunaria annua (chosen for its high nervonic acid content) established.
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  • 30
    ISSN: 1573-5060
    Keywords: Linum usitatissimum ; linseed ; mutation breeding ; somaclonal variation ; fatty acids ; genetic engineering
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In the early 1980s the phenomenon of somaclonal variation induced by cell culture was exploited to produce genetic variation in linseed. The linseed variety Andro, derived from the widely grown Canadian variety McGregor, was selected in saline culture and was released for production in Canada. ‘Andro’ possesses traits very different from its parent, such as increased seedling vigour and tolerance to heat stress. Additional stable somaclonal variation in characters such as yield, days to maturity, seed weight and oil content were subsequently induced in ‘McGregor’. However, despite extensive screening of the somaclonal variants, no significant variation in the fatty acid profile was found. Chemical mutagenesis using ethyl methanesulphonate was, however, succesful in modifying the fatty acid profile of McGregor. Initial screening of M2 seed by the thiobarbituric acid colourimetric procedure was followed by gas chromatography to select half-seeds with atypical fatty acid profiles. Two independent, partially dominant genes were identified that were responsible for reducing the linolenic acid (18 : 3) from 50% to 2% while increasing linoleic acid (18 : 2) to 70%. A single, partially dominant gene, inherited independently of the linolenic acid genes, increased palmitic acid (16 : 0) from 7% to 30% and palmitoleic acid (16 : 1) from trace amounts to 4%. Agrobacterium-mediated transformation of linseed has also been successful. Herbicide tolerance genes for glyphosate, sulfonylurea and phosphinothricin have been incorporated into Canadian varieties. Commercially useful levels of tolerance to sulfonylurea herbicides have been achieved with no adverse agronomic affect. It is expected that a transgenic variety containing this resistance will be registered for commercial production in Canada in 1994. Standard breeding techniques, the application of antisense technology and the overexpression of fatty acid synthesis genes are being used to further modify the fatty acid profile of linseed, as well as for the transfer of abiotic stress-related genes identified in bromegrass.
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  • 31
    ISSN: 1573-5060
    Keywords: Vicia narbonensis ; gene transfer ; gene expression ; seeds ; 2S albumin ; methionine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Epicotyl explants were co-cultivated with Agrobacterium tumefaciens EHA101 to transfer a chimeric 2S albumin gene construct carried in the binary Ti plasmid vectors pGSGLUC1 or pGA472 into the grain legume Vicia narbonensis. This gene encoding the sulphur-rich Brazil nut albumin was under the control of either the CaMV 35S promoter which permits gene expression in all organs, or the Vicia faba legumin B4 promoter which elicits seed-specific gene expression. After callus formation and selection for kanamycin resistance, somatic embryos were induced which, in the case of transformation with the vector pGSGLUC1, were screened for GUS activity. Embryos that produced GUS were in addition analysed for 2S albumin formation. Selected transgenic embryos were cloned by multiple shoot regeneration. Rooted and fertile plants were obtained by grafting transgenic shoots on the appropriate seedlings. R1 and R2 generations were raised and analysed for GUS as well as 2S albumin gene expression. Expression of the 35S promoter/2S albumin gene fusion took place in all organs of the transgenic plants including the cotyledons of seeds, whereas seed-specific gene expression was found in transformants with the legumin promoter/2S albumin gene fusion. The 2S albumin accumulated in the 2S protein fraction of transgenic seeds and its primary translation product was processed into the 9 and 3 kDa polypeptide chains. The foreign protein was localised in the protein bodies of the grain legume. Analysis of the R2 plants indicated Mendelian inheritance of the 2S albumin gene. In homozygous V. narbonensis plants the amounts of 2S albumin were twice that present in the corresponding heterozygous plants. Whereas only low level formation of the foreign protein was achieved if the gene was under the control of the 35S promoter, approximately 3.0% of the soluble seed protein was 2S albumin if seed-specific gene expression was directed by the legumin B4 promoter. Some of these transformants exhibited a three-fold increase in the methionine content of the salt-soluble protein fraction extracted from seeds.
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  • 32
    ISSN: 1573-5060
    Keywords: gene transfer ; crop species ; particle bombardment ; transgenic plants ; cereals ; legumes ; woody plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The limiting component in the creation of transgenic crops has been the lack of effective means to introduce foreign genes into elite germplasm. However, the development of novel direct DNA transfer methodology, by-passing limitations imposed by Agrobacterium-host specificity and cell culture constraints, has allowed the engineering of almost all major crops, including formerly recalcitrant cereals, legumes and woody species. The creation of transgenic rice, wheat, maize, barley, oat, soybean, phaseolus, peanut, poplar, spruce, cotton and others, in an efficient and in some cases, variety-independent fashion, is a significant step towards the routine application of recombinant DNA methodology to the improvement of most important agronomic crops. In this review we will focus on key elements and advantages of particle bombardment technology in order to evaluate its impact on the accelerated commercialization of products based on agricultural biotechnology and its utility in studying basic plant developmental processes and function through transgenesis. Fundamental differences between conventional gene transfer methods, utilizing Agrobacterium vectors or protoplast/suspension cultures, and particle bombardment will be discussed in depth.
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