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  • 1
    ISSN: 1617-4623
    Keywords: Barley ; Physical mapping ; Recombination ; Powdery mildew
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Variants of the pulsed-field gel electrophoresis technique were used in conjunction with two-dimensional DNA gel electrophoresis (2-DDGE) to determine the ratio of physical to genetic distance in two genetically defined intervals on barley chromosome 1H.2-DDGE analysis demonstrated that two loci that define a 0.3 cM interval, as determined by hybridization with BCD249, reside on a single 450-kbMluI fragment. This result indicates a maximum ratio of physical to genetic distance in this interval of 1500 kb/cM as compared to 3.7–4.2 Mb/cM for the barley genome as a whole. High molecular weight (HMW) DNA restricted withNotI and probed sequentially with MWG068 and BCD249 yield diffuse bands at approximately 2.8 Mb and 3.0 Mb in the C.I. 16151 and C.I. 16155 parental lines, respectively. These results suggest the maximum ratio of physical to genetic distance in the interval defined by these probes is 7.8 Mb/cM. unique HMW DNA restriction fragment length polymorphisms (RFLP) were attributed to the presence of recombination breakpoints. Data from the recombination breakpoint analysis were used to estimate a ratio of physical to genetic distance of 2.5 Mb/cM in theXbcd249.2-Xmwg068 interval and 0.465 Mb/cM in theXbcd249.1-Xbcd249.2 interval. Both physical linkage and recombination breakpoint analysis indicate theXbcd249.1-Xbcd249.2 interval is approximately five-fold smaller, physically, than theXbcd249.2-Xmwg068 interval.
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  • 2
    ISSN: 1432-2242
    Keywords: Key words Barley ; BAC library ; P-loop genes ; Resistance-gene analog (RGA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Modern cultivated barley is an important cereal crop with an estimated genome size of 5000 Mb. To develop the resources for positional cloning and structural genomic analyses in barley, we constructed a bacterial artificial chromosome (BAC) library for the cultivar Morex using the cloning enzyme HindIII. The library contains 313344 clones (816 384-well plates). A random sampling of 504 clones indicated an average insert size of 106 kbp (range=30–195 kbp) and 3.4% empty vectors. Screening the colony filters for chloroplast DNA content indicated an exceptionally low 1.5% contamination with chloroplast DNA. Thus, the library provides 6.3 haploid genome equivalents allowing a 〉99% probability of recovering any specific sequence of interest. High-density filters were gridded robotically using a Genetix Q-BOT in a 4×4 double-spotted array on 22.5-cm2 filters. Each set of 17 filters allows the entire library to be screened with 18432 clones represented per filter. Screening the library with 40 single copy probes identified an average 6.4 clones per probe, with a range of 1–13 clones per probe. A set of resistance-gene analog (RGA) sequences identified 121 RGA-containing BAC clones representing 20 different regions of the genome with an average of 6.1 clones per locus. Additional screening of the library with a P-loop disease resistance primer probe identified 459 positive BAC clones. These data indicate that this library is a valuable resource for structural genomic applications in barley.
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  • 3
    ISSN: 1432-2242
    Keywords: Maize ; Cytoplasmic male sterility ; Fertility restorer loci ; Linkage mapping ; Complementary genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract There are three major groups of cytoplasmic male-sterile cytoplasms in maize; C (Charrua), S (USDA), and T (Texas). These cytoplasms can be classified by the unique nuclear genes that suppress the male-sterility effects of these cytoplasms and restore pollen fertility. Typically, plants that carry Texas (T) cytoplasm are male fertile only if they carry dominant alleles at two unlinked nuclear restorer loci,rf1 andrf2. To facilitate analysis of T-cytoplasm-mediated male sterility and fertility restoration, we have mappedrf1 andrf2 relative to closely-linked RFLP markers using five populations. Therf1 locus and/or linked visible markers were mapped in four populations; therf2 locus was mapped in two of the populations. Data from the individual populations were joined with the aid of JoinMap software. The resulting consensus maps placerf1 between umc97 and umc92 on chromosome 3 andrf2 between umc153 andsus1 on chromosome 9. Markers that flank therf1 andrf2 loci have been used to identify alleles atrf1 andrf2 in segregating populations. These analyses demonstrate the possibility of tracking separate fertility restorer loci that contribute to a single phenotype.
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  • 4
    ISSN: 1432-2242
    Keywords: Genetics ; Disease resistance ; Monocots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An F2 oat population was produced by crossing the diploid (n=7) species Avena strigosa (CI 3815) with A. wiestii (CI 1994), resistant and susceptible, respectively, to 40 isolates of Puccinia coronata, the causal agent of crown rust. Eighty-eight F2 individuals were used to construct an RFLP linkage map representing the A genome of cultivated hexaploid oat. Two hundred and eight RFLP loci have been placed into 10 linkage groups. This map covers 2416 cM, with an average of 12 cM between RFLP loci. Eighty-eight F3 lines, derived from F2 individuals used to construct the map, were screened for resistance to 9 isolates of P. coronata. One locus, Pca, was found to confer a dominant resistance phenotype to isolates 203, 258, 263, 264B, 290, 298, 325A, and 345. Pca also conferred resistance to isolate 276; however, an unlinked second gene may also be involved.
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  • 5
    ISSN: 1617-4623
    Keywords: Cochliobolus heterostrophus race T disease ; Maize cytoplasmic male sterility ; Mitochondria ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We asked whether the mitochondrial T-urf13 gene, associated with the male sterility phenotype of T cytoplasm in maize, can be expressed in Saccharomyces cerevisiae and whether this expression can mimic the effects observed in maize. We introduced the universal code equivalent of the T-urf13 gene into the S. cerevisiae nucleus by transformation and directed its translation product into mitochondria by means of a fusion with the targeting presequence from Neurospora crassa ATPase subunit 9. We show that expression of the universal code equivalent of the T-urf13 gene in the yeast nucleus does indeed mimic its effects in maize: respiratory growth of yeast is inhibited, respiration-deficient cytoplasmic mutants accumulate and NADH oxidation of isolated mitochondria is uncoupled. All these effects are observed only if the mitochondrial targeting peptide and methomyl or HmT toxin are present.
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  • 6
    ISSN: 1617-4623
    Keywords: Key words Barley ; Physical mapping ; Recombination ; Powdery mildew
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Variants of the pulsed-field gel electrophoresis technique were used in conjunction with two-dimensional DNA gel electrophoresis (2-DDGE) to determine the ratio of physical to genetic distance in two genetically defined intervals on barley chromosome 1H. 2-DDGE analysis demonstrated that two loci that define a 0.3 cM interval, as determined by hybridization with BCD249, reside on a single 450-kb MluI fragment. This result indicates a maximum ratio of physical to genetic distance in this interval of 1500 kb/cM as compared to 3.7–4.2 Mb/cM for the barley genome as a whole. High molecular weight (HMW) DNA restricted with NotI and probed sequentially with MWG068 and BCD249 yield diffuse bands at approximately 2.8 Mb and 3.0 Mb in the C.I. 16151 and C.I. 16155 parental lines, respectively. These results suggest the maximum ratio of physical to genetic distance in the interval defined by these probes is 7.8 Mb/cM. Unique HMW DNA restriction fragment length polymorphisms (RFLP) were attributed to the presence of recombination breakpoints. Data from the recombination breakpoint analysis were used to estimate a ratio of physical to genetic distance of 2.5 Mb/cM in the Xbcd249.2-Xmwg068 interval and 0.465 Mb/cM in the Xbcd249.1-Xbcd249.2 interval. Both physical linkage and recombination breakpoint analysis indicate the Xbcd249.1-Xbcd249.2 interval is approximately five-fold smaller, physically, than the Xbcd249.2-Xmwg068 interval.
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  • 7
    ISSN: 1617-4623
    Keywords: Maize ; Cytoplasmic male sterility ; Mitochondrial DNA ; Nuclear-mitochondrial interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transcripts of the maize mitochondrial genes atp6, urf13-T and ORF25 were examined by Northern analysis from five normal (N), Texas male sterile (T), and T male sterile cytoplasm lines restored to fertility. A 5 kb DNA region containing the atp6 promoter is duplicated 5′ to urf13-T and ORF25 in T cytoplasm. DNA sequence differences in the 5 kb repeat detected between N and T cytoplasms may be related to atp6 transcriptional differences between the two cytoplasms. Nuclear restorer genes directed an RNA processing event that apparently decreased the abundance of mature urf13-T transcripts, which may correlate with the observed reduction of the 13 kDa gene product in restored lines. Fertility restoration had no effect on the transcription of atp6. Nuclear background, distinct from fertility restoration, also affected the abundance of urf13-T transcripts and directed the synthesis of an ORF25-specific transcript.
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Anaesthesia 17 (1962), S. 0 
    ISSN: 1365-2044
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Anaesthesia 17 (1962), S. 0 
    ISSN: 1365-2044
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
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  • 10
    ISSN: 1573-5028
    Keywords: cytoplasmic male sterility ; disease susceptibility ; Cochliobolus heterostrophus ; Phyllosticta maydis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Polyspecific antibody to a 17 amino acid synthetic peptide from the maize T-cytoplasm urf13-T mitochondrial open reading frame immunoprecipitated a 13 kD polypeptide from 35S-methionine incorporations of T cytoplasm maize. Male-fertile, toxin-insensitive mutants in which the urf13-T sequence is deleted do not synthesize the 13 kD polypeptide. A mutant designated T-4, which carries a 5 bp insertion and a premature stop codon, synthesizes a truncated polypeptide, corresponding to an open reading frame of 8.3 kD. Thus the 13 kD polypeptide is trunctated or absent in mutants expressing male fertility and toxin insensitivity in T-cytoplasm maize.
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