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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 263 (2000), S. 925-933 
    ISSN: 1617-4623
    Keywords: Key words S-like RNase ; Phosphate starvation ; Senescence ; RFLP ; Almond
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA for an S-like RNase (RNase PD2) has been isolated from a pistil cDNA library of Prunus dulcis cv. Ferragnés. The cDNA encodes an acidic protein of 226 amino acid residues with a molecular weight of 25 kDa. A potential N-glycosylation site is present at the N-terminus in RNase PD2. A signal peptide of 23 amino acid residues and a transmembrane domain are predicted. The two active-site histidines present in enzymes of the T2/S RNase superfamily were detected in RNase PD2. Its amino acid sequence shows 71.2% similarity to RNS1 of Arabidopsis and RNase T2 of chickpea, respectively. Northern blotting and RT-PCR analyses indicate that PD2 is expressed predominantly in petals, pistils of open flowers and leaves of the almond tree. Analyses of shoots cultured in vitro suggested that the expression of RNase PD2 is associated with phosphate starvation. Southern analysis detected two sequences related to RNase PD2 in the P. dulcis genome. RFLP analysis showed that S-like RNase genes are polymorphic in different almond cultivars. The PD2 gene sequence was amplified by PCR and two introns were shown to interrupt the coding region. Based on sequence analysis, we have defined three classes of S-like RNase genes, with the PD2 RNase gene representing a distinct class. The significance of the structural divergence of S-like RNase genes is further discussed.
    Type of Medium: Electronic Resource
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  • 2
    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
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