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  • 1
    Electronic Resource
    Electronic Resource
    ‘Universal’ primers for PCR-sequencing of grass chloroplastic acetyl-CoA carboxylase domains involved in resistance to herbicides (2005)
    Oxford, UK : Blackwell Science Ltd
    Weed research 45 (2005), S. 0 
    Details
    ISSN: 1365-3180
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Primers were designed to amplify two regions involved in sensitivity to herbicides inhibiting the plastidic acetyl-CoA carboxylase (ACCase) from grasses (Poaceae). The first primer pair amplified a 551-bp amplicon containing a variable Ile/Leu codon at position 1781 in Alopecurus myosuroides sequence. The second primer pair amplified a 406-bp amplicon containing four variable codons (Trp/Cys, Ile/Asn, Asp/Gly, Gly/Ala) at positions 2027, 2041, 2078 and 2096, respectively, in A. myosuroides sequence. Both primer pairs amplified the targeted fragments from genes encoding plastidic ACCases, but not from the very similar genes encoding cytosolic ACCases. Clear DNA sequences were obtained from fresh or dried plant material from the field, and from 29 various grass species. Sequences revealed that the gene encoding plastidic ACCase in Poa annua and Festuca rubra contained a Leu1781 codon, in agreement with both species being inherently tolerant to herbicides inhibiting ACCase. Sequencing confirmed the hybrid origin of P. annua. Compared with ACCase enzyme assay, polymerase chain reaction is faster, can be performed from a single plant and suppresses the need for radioactive experiments. It can be completed with basic molecular biology laboratory equipment. It is the tool of choice for diagnosing resistance caused by alteration(s) of the plastidic ACCase.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3180.2005.00467.x
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  • 2
    Electronic Resource
    Electronic Resource
    A single polymerase chain reaction-based assay for simultaneous detection of two mutations conferring resistance to tubulin-binding herbicides in Setaria viridis (2005)
    Oxford, UK : Blackwell Science Ltd
    Weed research 45 (2005), S. 0 
    Details
    ISSN: 1365-3180
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A simple method based upon polymerase chain reaction (PCR) was developed to detect mutant alleles of the gene encoding α2-tubulin, which confers recessive resistance to tubulin-binding herbicides in Setaria viridis. Multiplex, bidirectional allele-specific PCR (Mbi-PASA) was shown to specifically and reliably detect the presence of all sensitive (Leu136, Thr239) and resistant (Phe136, Ile239) α2-tubulin alleles in a single reaction. Double-blind analysis of 2000 S. viridis seedlings using seed bioassay and Mbi-PASA confirmed that the presence of two mutant α2-tubulin alleles in a seedling was always associated with cross-resistance to dinitroaniline and benzoic acid herbicides, sensitivity to a benzamide herbicide, and hypersensitivity to carbamate herbicides. No other resistance mechanism was detected in the S. viridis populations screened. Successful Mbi-PASA genotyping was achieved with fresh and dried plant fragments from the field. Compared with bioassays, Mbi-PASA is faster and more robust, removing the need for live plant material. It is the only way of detecting recessive resistance before resistant plants occur in a field. Mbi-PASA can be performed with basic molecular biology laboratory equipment, and is suitable for high-throughput genotyping adaptation. It is the tool of choice for resistance diagnosis in such cases where only a few recessive, target-derived, genes control resistance to herbicides.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3180.2005.00446.x
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  • 3
    Electronic Resource
    Electronic Resource
    PCR cloning and detection of point mutations in the eburicol 14a-demethylase (CYP51) gene from Erysiphe graminis f. sp. hordei, a “recalcitrant” fungus (1998)
    Springer
    Current genetics 34 (1998), S. 399-403 
    Details
    ISSN: 1432-0983
    Keywords: Key wordsErysiphe graminis ; Polymerase chain reaction ; Cytochrome P450 ; Fungicide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Molecular studies of some micro-organisms are hampered by the difficulty of obtaining sufficient amounts of nucleic acids. A cloning strategy based on PCR has therefore been used to clone the eburicol 14α-demethylase (CYP51) gene of the obligate fungus Erysiphe graminis f. sp. hordei (Egh) using minute amounts of genomic DNA. The CYP51 gene encodes the enzymatic target of a major group of fungicides. Sequencing CYP51 from different Egh isolates revealed the occurrence of two alleles for this gene. An allele-specific PCR assay was developed to detect each CYP51 allele.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002940050413
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    Paper (German National Licenses)
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