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  • 11
    Electronic Resource
    Electronic Resource
    Analysis of organelle genomes in a somatic hybrid derived from cytoplasmic male-sterile Brassica oleracea and atrazine-resistant B. campestris (1987)
    Springer
    Theoretical and applied genetics 74 (1987), S. 303-309 
    Details
    ISSN: 1432-2242
    Keywords: Somatic hybridization ; Atrazine resistance ; Mitochondrial recombinants ; Brassica ; Cytoplasmic male sterility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An atrazine-resistant, male-fertile Brassica napus plant was synthesized by fusion of protoplasts from the diploid species B. oleracea and B. campestris. Leaf protoplasts from B. oleracea var. italica carrying the Ogura male-sterile cytoplasm derived from Raphanus sativus were fused with etiolated hypocotyl protoplasts of atrazine-resistant B. campestris. The selection procedure was based on the inability of B. campestris protoplasts to regenerate in the media used, and the reduction of light-induced growth of B. oleracea tissue by atrazine. A somatic hybrid plant that differed in morphology from both B. oleracea and B. campestris was regenerated on medium containing 50 μM atrazine. Its chromosome number was 36–38, approximately that of B. napus. Furthermore, nuclear ribosomal DNA from this hybrid was a mixture of both parental rDNAs. Southern blot analyses of chloroplast DNA and an assay involving tetrazolium blue indicated that the hybrid contained atrazine-resistant B. campestris chloroplasts. The hybrid's mitochondrial genome was recombinant, containing fragments unique to each parent, as well as novel fragments carrying putative crossover points. Although the plant was female-sterile, it was successfully used to pollinate B. napus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00274711
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  • 12
    Electronic Resource
    Electronic Resource
    Synthesis of male sterile, triazine-resistant Brassica napus by somatic hybridization between cytoplasmic male sterile B. oleracea and atrazine-resistant B. campestris (1989)
    Springer
    Theoretical and applied genetics 78 (1989), S. 445-455 
    Details
    ISSN: 1432-2242
    Keywords: Brassica napus ; Protoplast fusion ; Triazine resistance ; Cytoplasmic male sterility ; Mitochondrial DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Fusion of leaf protoplasts from an inbred line of Brassica oleracea ssp. botrytis (cauliflower, n=9) carrying the Ogura (R1) male sterile cytoplasm with hypocotyl protoplasts of B. campestris ssp. oleifera (cv “Candle”, n=10) carrying an atrazine-resistant (ATR) cytoplasm resulted in the production of synthetic B. napus (n=19). Thirty-four somatic hybrids were produced; they were characterized for morphology, phosphoglucose isomerase isoenzymes, ribosomal DNA hybridization patterns, chromosome numbers, and organelle composition. All somatic hybrids carried atrazine-resistant chloroplasts derived from B. campestris. The mitochondrial genomes in 19 hybrids were examined by restriction endonuclease and Southern blot analyses. Twelve of the 19 hybrids contained mitochondria showing novel DNA restriction patterns; of these 12 hybrids, 5 were male sterile and 7 were male fertile. The remaining hybrids contained mitochondrial DNA that was identical to that of the ATR parent and all were male fertile.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00265310
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  • 13
    Electronic Resource
    Electronic Resource
    A comparison of cytoplasmic revertants to fertility from different CMS-S maize sources (1988)
    Springer
    Theoretical and applied genetics 76 (1988), S. 609-618 
    Details
    ISSN: 1432-2242
    Keywords: S-type cytoplasmic male sterility ; Reversion to fertility ; Mitochondrial genome organization ; Recombination ; Maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial genome organizations of a number of independent culture-derived fertile CMS-S revertants with the nuclear genotype W182BN were compared to spontaneous field revertants with the genotypes WF9, M825/Oh07 and 38-11. Regions of the genome around sequences homologous to the terminal repeats of the linear S1 and S2 episomes characteristic of CMS-S mitochondria were used as hybridization probes on Southern blots of BamHI and SalI digested mitochondrial DNA. The results obtained suggest that the nuclear, not the cytoplasmic, genotype of the parent plant affects the type of novel mitochondrial DNA organization found in the revertant. The DNA reorganization during reversion from CMS-S in tissue culture appears to be similar to that observed in spontaneous revertants obtained during the normal plant life-cycle. Unlike the situation for reversion from CMS-T, no common DNA sequence or reading frame appeared to be lost or disrupted in revertants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00260916
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  • 14
    Electronic Resource
    Electronic Resource
    Preparation and flow cytometric analysis of metaphase chromosomes of tomato (1991)
    Springer
    Theoretical and applied genetics 82 (1991), S. 101-111 
    Details
    ISSN: 1432-2242
    Keywords: Tomato chromosome suspension ; Flow cytometry ; Univariate analysis ; Bivariate analysis ; Chromosome sorting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A procedure for the preparation of tomato chromosome suspensions suitable for flow cytometric analysis is described. Rapidly growing cell suspension cultures of Lycopersicon esculentum cv VFNT cherry and L. pennellii LA716 were treated with colchicine to enrich for metaphase chromosomes. Metaphase indices between 20 and 35% were routinely obtained when cultures were exposed to 0.1% colchicine for 15–18 h after 2 days of subculture. Mitotic cells were isolated by brief treatment with cell wall digesting enzymes in a medium with low osmolarity (∼325 mOsm/kg of H52O). The low osmolarity medium was needed to avoid the chromosome clumping and decondensation seen in standard media. Suspensions of intact chromosomes were prepared by lysing swollen protoplasts in various buffers (MgSO4, polyamines, hexylene glycol, or KCl-propidium iodide) similar in contents to the buffers used to isolate mammalian chromosomes. For univariate flow cytometric analysis, chromosome suspensions were stained with a fluorescent DNA-binding stain (propidium iodide, Hoechst 33258, mithramycin, or chromomycin A3) and analyzed using an EPICS flow cytometer (Profile Analyzer or 753). Peaks for the chromosomes, chromatids, clumps of chromosomes, nuclei, and fluorescent debris were seen on a histogram of log of fluorescence intensity, and were confirmed by microscopic examination of the objects collected by flow-sorting. Chromosome suspensions prepared in MgSO4 buffer have the highest frequency of intact chromosomes and the least fluorescent cellular debris. Peaks similar to theoretical univariate flow karyotypes of tomato chromosomes were seen on the observed univariate flow karyotypes, but were not as well resolved. Bivariate flow analysis of tomato chromosome suspension using double-stain combination, Hoechst 33258 and chromomycin A3, and two laser beams showed better resolution of some chromosomes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00231283
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  • 15
    Electronic Resource
    Electronic Resource
    Organellar segregation, rearrangement and recombination in protoplast fusion-derived Brassica oleracea calli (1993)
    Springer
    Theoretical and applied genetics 85 (1993), S. 761-769 
    Details
    ISSN: 1432-2242
    Keywords: Organelle segregation ; Mitochondrial recombination ; Brassica oleracea ; Ogura cytoplasmic male sterility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cauliflower protoplasts were fused to determine the effect of protoplast source and pretreatment on organellar segregation in fusion products. Mitochondrial and chloroplast type were determined for over 250 calli from eight fusions between iodoacetate-treated or γ-irradiated leaf or hypocotyl protoplasts with fertile or Ogura cytoplasms. Organelles in fusion-derived calli were identified with five mitochondrial probes and one chloroplast probe. Mitochondrial and chloroplast segregation were independent but biased. Most calli had B. oleracea chloroplasts, but more calli had Ogura mitochondria than B. oleracea ones. Neither protoplast source nor pretreatment alone affected organelle segregation. However, iodoacetate treatment of hypocotyl protoplasts reduced their mitochondrial contribution to the fusion products although it did not affect chloroplast segregation. Over half of the calli had mitochondrial genomes distinct from those of either fusion partner; many of these contained the complete mitochondrial genome of one partner along with some mitochondrial DNA from the other. Out of 258 calli, 83 showed evidence of mitochondrial recombination, most commonly by formation of a novel 11-kb PstI fragment near the atp9 region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00225017
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  • 16
    Electronic Resource
    Electronic Resource
    Synthesis of high erucic acid rapeseed (Brassica napus L.) somatic hybrids with improved agronomic characters (1995)
    Springer
    Theoretical and applied genetics 91 (1995), S. 1129-1136 
    Details
    ISSN: 1432-2242
    Keywords: Brassica napus ; somatic hybrids ; protoplast fusion ; Erucic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Novel Brassica napus somatic hybrids have been created through protoplast fusion of B. oleracea var. botrytis and B. rapa var. oleifera genotypes selected for high erucic acid (22:1) content in the seed oil. Fifty amphidiploids (aacc) and one putative hexaploid (aacccc) hybrid were recovered in one fusion experiment. Conversely, only one amphidiploid and numerous regenerates with higher DNA contents were produced in a similar fusion using a different B. rapa partner. Hybridity was confirmed by morphology, isozyme expression, flow cytometry, and DNA hybridization. Analysis of organellar DNA revealed a distinct bias toward the inheritance of chloroplasts from the B. rapa (aa) genome. All amphidiploids set self-pollinated seed. A erucic acid content as high as 57.4% was found in the seed oil of one regenerated plant. Fatty acid composition was stable in the R1 generation and was coupled with increased female fertility. Other novel agronomic characters in the hybrids recovered include large seed size, lodging resistance, and non-shattering seed pods.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00223931
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  • 17
    Electronic Resource
    Electronic Resource
    Transfer of resistance to Xanthomonas campestris pv campestris into Brassica oleracea L. by protoplast fusion (1995)
    Springer
    Theoretical and applied genetics 91 (1995), S. 1293-1300 
    Details
    ISSN: 1432-2242
    Keywords: Somatic hybridization ; Black rot ; Disease resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Black rot caused by the bacterium Xanthomonas campestris pv campestris is one of the most serious diseases of Brassica oleracea. Since sources of resistance to the disease within B. oleracea are insufficient and control means are limited, the development of resistant breeding lines is extremely desirable. Certain lines of B. napus contain very high resistance controlled by a dominant gene, but crossing the two species sexually is very difficult. Therefore, somatic hybrids were produced by protoplast fusion between rapid cycling B. oleracea and a B. napus line highly resistant to X. campestris pv campestris. Hybrid identity was confirmed by morphological studies, flow cytometric estimation of nuclear DNA content, and analysis of random amplified polymorphic DNA (RAPD). Inoculations with the pathogen identified four somatic hybrids with high resistance. The resistant hybrid plants were fertile and set seed when selfed or crossed reciprocally to the bridge line ‘15’ (Quazi 1988). Direct crosses to B. oleracea were unsuccessful, but embryo rescue facilitated the production of a first-backcross generation. The BC1 plants were resistant to the pathogen. Progeny from the crosses to ‘line 15’ were all susceptible. Embryo rescue techniques were not obligatory for the development of a second-backcross generation, and several resistant BC2 plants were obtained.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00220944
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  • 18
    Electronic Resource
    Electronic Resource
    Camalexin induction in intertribal somatic hybrids between Camelina sativa and rapid-cycling Brassica oleracea (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 164-170 
    Details
    ISSN: 1432-2242
    Keywords: Key words Intertribal somatic hybrids ; Brassica ; Camelina sativa ; Alternaria ; Camalexin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Camelina sativa, a wild relative of Brassica crops, is virtually immune to blackspot disease caused by Alternaria brassicicola. Intertribal somatic hybrids were produced between C. sativa and rapid-cycling Brassica oleracea as a step toward the transfer of resistance to this disease into Brassica vegetable crops. The plants recovered were confirmed as somatic hybrids by flow cytometry and RAPD analysis. All hybrids showed a morphology intermediate between the two parents. Rooted plants grew in soil up to 4–5 weeks, and some produced sterile flowers. Two of three hybrids tested showed a high level of resistance to  A. brassicicola. Resistance was correlated with the induction of high levels of the phytoalexin camalexin 48 h after inoculation, as in the resistant Camelina fusion partner. In contrast, susceptible somatic hybrids produced much lower levels of camalexin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051053
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  • 19
    Electronic Resource
    Electronic Resource
    Rearrangement, amplification, and assortment of mitochondrial DNA molecules in cultured cells of Brassica campestris (1989)
    Springer
    Theoretical and applied genetics 77 (1989), S. 17-25 
    Details
    ISSN: 1432-2242
    Keywords: Brassica ; Callus culture ; Mitochondrial DNA ; Genomic variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We compared Brassica campestris mitochondrial and chloroplast DNAs from whole plants and from a 2-year-old cell culture. No differences were observed in the chloroplast DNAs (cpDNAs), whereas the culture mitochondrial DNA (mtDNA) was extensively altered. Hybridization analysis revealed that the alterations are due entirely to rearrangement. At least two inversions and one large duplication are found in the culture mtDNA. The duplication element is shown to have the usual properties of a plant mtDNA high frequency “recombination repeat”. The culture mtDNA exists as a complex heterogeneous population of rearranged and unrearranged molecules. Some of the culture-associated rearranged molecules are present in low levels in native plant tissue and appear to have sorted out and amplified in the culture. Other mtDNA rearrangements may have occurred de novo. In addition to alterations of the main mitochondrial genome, an 11.3 kb linear mtDNA plasmid present in whole plants is absent from the culture. Contrary to findings in cultured cells of other plants, small circular mtDNA molecules were not detected in the B. campestris cell culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00292310
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  • 20
    Electronic Resource
    Electronic Resource
    Atrazine-resistant cytoplasmic male-sterile-nigra broccoli obtained by protoplast fusion between cytoplasmic male-sterile Brassica oleracea and atrazine-resistant Brassica campestris (1991)
    Springer
    Theoretical and applied genetics 83 (1991), S. 201-208 
    Details
    ISSN: 1432-2242
    Keywords: Brassica ; Atrazine resistance ; Cytoplasmic male sterility ; Protoplast fusion ; CMS-nigra
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplast fusion was used to combine the cytoplasmic traits of atrazine resistance and male sterility in Brassica oleracea var. italica (broccoli). Leaf protoplasts from broccoli with the petaloid B. nigra type of cytoplasmic male sterility were fused with hypocotyl protoplasts from an atrazine-resistant biotype of B. campestris var. oleifera cv Candle (oilseed rape). A total of 19 colonies regenerated shoots, all of which were broccolilike in phenotype, i.e., lacked trichomes. Four shoots, all from one colony, were atrazine resistant, surviving and growing in the presence of 25 μM atrazine. A leaf piece assay also confirmed that they were atrazine resistant. Molecular analysis showed that they contain chloroplasts from the atrazine-resistant B. campestris parent and mitochondria from the B. nigra parent. No recombination or rearrangement of the mitochondrial genomes in the fusion products was detected. These four plants and their progeny all showed the petaloid B. nigra type of male sterility.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00226252
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