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  • 1
    Electronic Resource
    Electronic Resource
    Organelle assortment and mitochondrial DNA rearrangements in Brassica somatic hybrids and cybrids (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 85 (1992), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Protoplast fusion permits manipulations of organelle genomes not readily achieved by other in vitro procedures or sexual crosses. Although considerable information is now available about the fate of chloroplasts and mitochondria in fusion products of various genera, many additional questions about factors affecting organelles after fusion remain to be answered. Brassica species are particularly favorable materials for such studies. Organelle assortment, mitochondrial DNA (mtDNA) recombination, and plant phenotypes observed after fusion of protoplasts from cytoplasmic male sterile B. oleracea with protoplasts from B. campestris, B. oleracea or B. napus are described. The somatic hybrids and cybrids obtained at Cornell have been used for detailed studies of recombinant mtDNA, including correlation of a specific mtDNA region with the ogura type of cytoplasmic male sterility, and have provided plant materials for possible use in hybrid breeding programs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1992.tb04742.x
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  • 2
    Electronic Resource
    Electronic Resource
    Novel mitochondrial genomes in Brassica napus somatic hybrids (1992)
    Springer
    Current genetics 22 (1992), S. 243-249 
    Details
    ISSN: 1432-0983
    Keywords: Brassica ; Somatic hybrids ; Cytoplasmic male sterility ; Mitochondrial DNA rearrangement
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial genomes of nine male-fertile and two Ogura cytoplasmic male-sterile (cms) Brassica napus somatic hybrids were probed with 46 mitochondrial DNA fragments. The distribution of information obtained from each fusion partner was not random. Several regions, including the coxI gene and a major recombination repeat sequence, were always derived from the Brassica campestris fusion partner, and some regions were always derived from the Ogura mitochondrial genome. Novel fragments occurred in seven distinct regions. Some of the rearrangement breakpoints were located near the evolutionary breakpoints relating the mitochondrial genomes of the Brassica species. The sizes of the mitochondrial genomes in the somatic hybrids ranged from 224.8 to 285.3 kb. A direct correlation between a specific gene and the cms phenotype was not observed; however, a possible cms-associated region was identified. It corresponds to a region that was identified through analysis of fertile revertants from a cms B. napus cybrid.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351732
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  • 3
    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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  • 4
    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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  • 5
    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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  • 6
    Electronic Resource
    Electronic Resource
    Insect-resistant plants with improved horticultural traits from interspecific potato hybrids grown in vitro (1990)
    Springer
    Theoretical and applied genetics 80 (1990), S. 95-104 
    Details
    ISSN: 1432-2242
    Keywords: Solanum tuberosum ; Solanum berthaultii ; Insect resistance ; Introgression ; Potato somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plants were regenerated from petiole calli of interspecific hybrids of Solanum tuberosum x S. berthaultii, an insect-resistant wild species. Callus culture was used to generate genetic changes to overcome the restricted recombination between the two genomes. Two plants out of 58 (3.5%) from calli of hybrid J114-1 showed stable and heritable differences from the hybrid over two cycles of evaluations in the field. Replicated trials were conducted in 1987 and 1988, using two populations of plants propagated by nodal cuttings from the original regenerates maintained in vitro. One regenerate showed insect resistance and increased marketable yield (approximately two fold) in the field. The other had higher levels of phenolic exudate in one of the two types of foliar trichomes associated with the insect resistance mechanism. Some desirable changes were discernible only in sexual progeny of regenerates, not in the regenerates themselves. In a backcross to S. tuberosum, 7 of 14 (50%) regenerates from hybrid F743-4 showed more progeny (up to 15-fold) with improved trichome traits and horticultural characteristics than the original hybrid. The variations were not associated with changes in ploidy. Fifteen plants obtained from these crosses are currently being incorporated into breeding lines. These results suggest that a period of callus culture followed by plant regeneration may aid in the introgression of desirable traits from wild species into crop plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00224021
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  • 7
    Electronic Resource
    Electronic Resource
    Improved protoplast culture and stability of cytoplasmic traits in plants regenerated from leaf protoplasts of cauliflower (Brassica oteracea ssp.botrytis) (1990)
    Springer
    Plant cell, tissue and organ culture 21 (1990), S. 227-236 
    Details
    ISSN: 1573-5044
    Keywords: Brassica oleracea ; protoplasts ; regeneration ; cytoplasmic traits ; mitochondrial DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nearly 1000 plants have been regenerated from leaf protoplasts of two cauliflower (Brassica oleracea ssp.botrytis) alloplasmic inbred lines. One line (7642A) carried the Ogura (R1) cms cytoplasm derived from radish; the other line (7642B) carried a normalBrassica cytoplasm and was the fertile maintainer for the cms line. The majority of regenerated plants displayed normal vegetative morphology; they formed normal cauliflower heads and retained the floral characteristics of seed-grown plants from which they were derived. We found no change in either male sterility or in the low temperature-induced chlorosis associated with the 7642A line. Mitochondrial DNA analysis by hybridization with five cloned mtDNA probes revealed no apparent alteration in 75 regenerated plants of both lines. These results indicate that cytoplasmic traits inBrassica oleracea are stable after one cycle of in vitro culture and regeneration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00047615
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