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  • 1
    Electronic Resource
    Electronic Resource
    Quick Identification of the Members of the Glutamine Synthetase Gene Family from Sunflower by Simultaneous Amplification of cDNA with Degenerate Primers (1998)
    Springer
    Biologia plantarum 41 (1998), S. 339-355 
    Details
    ISSN: 1573-8264
    Keywords: chloroplastic isoforms ; cytosolic isoforms ; Helianthus annuus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A single degenerate glutamine synthetase (GS)-specific primer was used to amplify the 3′ end of cDNAs derived from different GS genes that are expressed in leaves and roots of sunflower (Helianthus annuus L. cv. Peredovic). Four types of GS cDNA (I, II, III and IV) were simultaneously amplified from leaves and five types (I, II, V, VI, VII) from roots with a minimum investment of time and experimental work. cDNAs II, III and IV encode chloroplastic isoforms as deduced by the presence of chloroplastic GS-specific features in their sequences. The rest of cDNAs codifies cytosolic isoforms. Using cDNA-specific probes and primers, homologous sequences to all GS cDNAs amplified from cv. Peredovic, except to cDNAs III and IV, were detected in the inbred line R41. This result strongly suggests that the three cDNAs for chloroplastic isoform are allelic sequences from the same locus, and since cDNA type IV contains sequences derived from cDNAs II and III, it indicates a recombinational origin. The results presented are consistent with the existence of a GS gene family in sunflower with at least five members. Four of them, named ggs1.1 to ggs1.4, codify for the cytosolic isoforms (cDNAs I, V, VI and VII). A fifth member, named ggs2, from which three allelic sequences (cDNAs II, III and IV) have been cloned, encodes the chloroplastic isoform.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1001825706687
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  • 2
    Electronic Resource
    Electronic Resource
    Molecular and cytogenetic characterization of repetitive DNA sequences from Lolium and Festuca: applications in the analysis of Festulolium hybrids (1992)
    Springer
    Theoretical and applied genetics 84 (1992), S. 145-154 
    Details
    ISSN: 1432-2242
    Keywords: Lolium multiflorum ; Festuca arundinacea ; Repetitive DNA sequences ; In situ hybridization ; Festulolium hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A set of species-specific repetitive DNA sequences was isolated from Lolium multiflorum and Festuca arundinacea. The degree of their species specificity as well as possible homologies among them were determined by dot-blot hybridization analysis. In order to understand the genomic organization of representative Lolium and Festuca-specific repetitive DNA sequences, we performed Southern blot hybridization and in situ hybridization to metaphase chromosomes. Southern blot hybridization analysis of eight different repetitive DNA sequences of L. multiflorum and one of F. arundinacea indicated either tandem and clustered arrangements of partially dispersed localization in their respective genomes. Some of these sequences, e.g. LMB3, showed a similar genomic organization in F. arundinacea and F. pratensis, but a slightly different organization and degree of redundancy in L. multiflorum. Clones sequences varied in size between 100 bp and 1.2 kb. Estimated copy number in the corresponding haploid genomes varied between 300 and 2×104. Sequence analysis of the highly species-specific sequences from plasmids pLMH2 and pLMB4 (L. multiflorum specific) and from pFAH1 (F. arundinacea specific) revealed some internal repeats without higher order. No homologies between the sequences or to other repetitive sequences were observed. In situ hybridization with these latter sequences to metaphase chromosomes from L. multiflorum, F. arundinacea and from symmetric sexual Festulolium hybrid revealed their relatively even distribution in the corresponding genomes. The in situ hybridization thus also allowed a clearcut simple identification of parental chromosomes in the Festulolium hybrid. The potential use of these species-specific clones as hybridization probes in quantitative dot-blot analysis of the genomic make-up of Festulolium (sexual and somatic) hybrids is also demonstrated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00223994
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  • 3
    Electronic Resource
    Electronic Resource
    CMS system inNicotiana: flower development, patterns of mitochondrial DNA and mitochondrial gene expression (1992)
    Springer
    Sexual plant reproduction 5 (1992), S. 13-26 
    Details
    ISSN: 1432-2145
    Keywords: Nicotiana ; Cytoplasmic male sterility ; Flower morphogenesis ; mtDNA organization ; In organello protein synthesis ; Mitochondrial transcripts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A detailed description of in planta floral ontogeny based on scanning electron microscopy, developmental histology and morphology is presented for three different alloplasmic gene-cytoplasmic male-sterile (CMS)Nicotiana tabacum isonuclear lines with cytoplasms ofN. bigelovii, N. debneyi and N. suaveolens and compared to the corresponding nuclear donorN. tabacum genotype. This allowed the precise determination of the developmental stages affected in the mutant forms as well as a thorough phenotypic characterization of them. The organization of the mitochondrial genome and expression of mitochondrial DNA (mtDNA) was investigated in the three different alloplasmic CMS tobacco analogs and compared to the corresponding malefertile parentalNicotiana species. Southern hybridizations of total cellular DNA and mtDNA from the different sets of lines with probes specific for mitochondrial genes coding for cytochrome oxidase subunits I, II and III, apocytochrome b, ATPase subunits α and 9 as well as 18S-5S ribosomal RNA indicated that: (a) mtDNA organization is different between mitochondrial genomes of fertile and sterile lines but identical in two different fertile tobacco lines; however genetic similarity among different mitochondrial genome types can be revealed by restriction fragment patterns; (b) although several differences were detected between the male-sterile and male-fertile plants, most of these were related to the origin of the mitochondria (cytoplasm donorNicotiana species); (c) identical mtDNA rearrangements — distinct to the cytoplasm donor — occur in cytoplasmic malesterile tobacco analogs bearing cytoplasm fromN. bigelovii in two differentN. tabacum nuclear backgrounds, indicating that mitochondrial genome structure inNicotiana is altered by substitution of the nuclear back-ground, since (d) inter- and intraspecific mitochondrial genome diversity among differentNicotiana species and the corresponding alloplasmic CMS tobacco analogs can be determined by hybridization with mtDNA specific probes. Analysis of in organello translation products in the three CMS-systems described confirmed the presence of variant proteins synthesized by mitochondria from CMS and male-fertileNicotiana isonuclear lines. In addition, differences due to the origin of both the nucleus and the cytoplasm, which involve changes in the presence or size of particular polypeptides, are apparent. A common feature of all three systems — including two different nuclear backgrounds — is the enhanced synthesis of a 31-kDa polypeptide in the strictly isonuclear CMS lines compared to the male-fertile tobacco. In addition, organellar gene expression was studied by Northern blot analysis of transcripts homologous to mitochondrial gene probes, revealing variant mtRNA species associated with some CMS lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00714554
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  • 4
    Electronic Resource
    Electronic Resource
    Intergeneric symmetric and asymmetric somatic hybridization in Festuca and Lolium (1955)
    Springer
    Euphytica 85 (1955), S. 235-245 
    Details
    ISSN: 1573-5060
    Keywords: intergeneric somatic hybrids ; forage grasses ; fescue ; Festuca arundinacea ; F. rubra ; ryegrasses ; Lolium multiflorum ; L. perenne ; Alopecurus pratensis ; species-specific repetitive DNA sequences
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated. Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with ≤250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes. The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred. Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023952
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  • 5
    Electronic Resource
    Electronic Resource
    Xanthine accumulation and vacuolization inChlamydomonas reinhardtii cells (1995)
    Springer
    Protoplasma 186 (1995), S. 93-98 
    Details
    ISSN: 1615-6102
    Keywords: Chlamydomonas ; Accumulation ; Purines ; Vacuole ; Xanthine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Utilization of xanthine as the sole nitrogen source for growth byChlamydomonas reinhardtii cells involved the formation of a transient, intracellular pool of xanthine. Up to 20% of the total xanthine supplied to the medium was not assimilated after uptake but stored in the cells at concentrations that exceeded xanthine solubility in water. At the subcellular level, a massive accumulation of starch grains in the chloroplast and the appearance of many vacuoles in the cytoplasm distinguished xanthine-grown from ammonium-grown cells. Starch accumulation, but not development of vacuoles, was also observed in N-starved cells. Uptake experiments with radio-labelled xanthine showed that this accumulates only in the cytoplasm, most probably inside vacuoles. The electron-dense material observed in vacuoles of xanthine-grown cells suggests that the intracellular xanthine is in part solid xanthine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01276941
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  • 6
    Electronic Resource
    Electronic Resource
    Sunflower cytosolic-glutamine-synthetase genes showing similar organ-specificity patterns exhibit very different expression intensities by reverse-transcription polymerase chain reaction and Northern analysis (1999)
    Springer
    Protoplasma 207 (1999), S. 154-157 
    Details
    ISSN: 1615-6102
    Keywords: Glutamine synthetase gene family ; Differential expression ; Sunflower ; Reverse-transcription polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four genes coding for the cytosolic isoforms of glutamine synthetase (GS; EC 6.3.2.1) were recently identified in sunflower. The levels of transcripts from those genes have been analyzed in selected organs where ammonium is being produced by different processes that might differentially regulate the genes' expression. Genes exhibiting consistently high (ggs1.1 and−1.4), low (ggs1.2), and very low (ggs1.3) expression have been found. However, the four cytosolic-GS genes were expressed according to very similar patterns of organ specificity suggesting a low degree of specialization within this gene family in sunflower.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01282995
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