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  • 1
    ISSN: 1871-4528
    Keywords: PSTV detection ; DNA-RNA probes ; RNA transcription ; nick-translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary 32P-labelled probes prepared by nick-translation, two methods (A and B) of labelling PSTV cDNA in M13 phage DNA, and RNA transcription were compared for their sensitivity in detecting PSTV. Nick-translated and method B probes were the least sensitive. The minimum concentration of PSTV detected by these probes was 100 pg. Method A probe detected 10–20 pg of PSTV whereas RNA transcripts were capable of detecting as little as 0.33 pg of PSTV. RNA transcripts are easy to prepare, and their use in hybridization analysis do not cause background reactions. Sometimes low non-specific signals are produced but they can be eliminated by washing membranes with RNase A. RNA probes are now used for routine detection of PSTV in tuber flesh and sprouts, leaves and true seed of potato at the International Potato Center (CIP).
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1572-994X
    Keywords: viroids ; apscaviroids ; phylogenetic analysis ; RNA quasi-species ; subviral RNAs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Field-grown citrus trees often harbor complex mixtures of 4–5 different viroid species, and the presence of citrus viroid III (CVd-III) has been shown to reduce the rate of tree growth without inducing disease. To more fully define the structure of its quasi-species, we have examined nine citrus viroid complexes for the presence of previously undescribed sequence variants of CVd-III. Analysis of 86 full-length cDNAs generated from these nine viroid complexes by RT-PCR revealed the presence of 20 new CVd-III variants. Chain lengths ranged from 293–297 nucleotides, and sequence changes were confined largely to the lower portions of the central conserved region and variable domain. The previously described variants CVd-IIIa (297 nt) and CVd-IIIb (294 nt) were clearly predominant, but phylogenetic analysis indicated that certain isolates may contain representatives of two additional fitness peaks. At least one group of CVd-III variants appears to have arisen as a result of RNA recombination. Populations recovered from diseased/declining trees were the most diverse, but even dwarfing isolates originating from old line Shamouti trees showed considerable variability.
    Type of Medium: Electronic Resource
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