ALS gene proline (197) mutations confer ALS herbicide resistance in eight separated wild radish (Raphanus raphanistrum) populations

Qin Yu, X.Q. Zhang, A. Hashem, Michael Walsh, Stephen Powles

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    89 Citations (Web of Science)

    Abstract

    The biochemical and molecular basis of resistance to acetolactate synthase (ALS)-inhibiting herbicides was investigated in eight resistant (R) and three susceptible (S) wild radish populations. In vitro enzyme assays revealed an ALS herbicide-resistant ALS enzyme in all R populations. ALS enzyme extracted from the shoots of all eight R populations was highly resistant to the ALS-inhibiting sulfinnylurea herbicide chlorsulfuron (20- to 160-fold) and the triazolopyrimidine herbicide metosulam (10- to 46-fold) and moderately resistant to metsulfuron (three to eightfold). There was little or no cross-resistance to the imidazolinone herbicides imazapyr and imazethapyr. The ALS gene fragment covering potential mutation sites in these populations was amplified, sequenced, and compared. All eight R populations had point mutations in the codon for the proline residue in Domain A. However, the point mutations varied and encoded four different amino acid substitutions: histidine, threonine, alanine, and serine. No nucleotide difference in the DNA sequence of Domains C and D resulting in amino acid substitutions was observed between the R and S populations examined. In addition, a three- to fivefold higher ALS-specific activity was consistently observed in all R populations compared with S populations, whereas Northern blot analysis detected a similar level of ALS mRNA, suggesting a possible translational-posttranslational regulation of the enzyme. It is concluded that selection pressure from chlorsulfuron on eight separate wild radish populations has resulted in target site mutation at the same proline residue in the ALS gene. Higher ALS activity also may play a role in the resistance level.
    Original languageEnglish
    Pages (from-to)831-838
    JournalWeed Science
    Volume51
    Issue number6
    DOIs
    Publication statusPublished - 2003

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