A chicory (Cichorium intybus L. var. Witloof) line, previously selected in vitro for tolerance to the sulfonylurea herbicide chlorsulfuron and regenerated into resistant fertile plants showing a 1500 to 2000 higher resistance than wild-type, was characterized in order to understand the biochemical basis of the resistance. Transcription, specific activity level and properties of acetolactate synthase (ALS, EC 4.1.3.18), the first enzyme in branched-chain amino acid synthesis and the target of sulfonylurea and imidazolinone herbicides, were evaluated in plant tissues and compared with those of a sensitive control. Whilst Northern blot analysis detected similar levels of ALS-mRNA, a significant difference was found in ALS specific activity. However, its higher level in sensitive plants, could not account for the selected tolerance. When ALS activity from wild-type and resistant plants was measured in the presence of increasing chlorsulfuron concentrations, a striking variation in enzyme sensitivity was found. In the two cases 50%-inhibition was achieved at 21 nmol/L and 12 μmol/L (R = 570), respectively, suggesting that the tolerant line could owe its resistance to a mutational event resulting in a decreased affinity of the target enzyme. Mutant ALS displayed cross-tolerance to the imidazolinone herbicide imazamethabenz, a 125-fold higher level of which was required to cause 50 %-inhibition of enzyme activity. Herbicide tolerance was accompanied also by a reduced sensitivity to the regulative feedback inhibition by leucine and valine, with concentrations causing 50%-inhibition about 10 and 3-fold higher, respectively, than those found for the wild-type enzyme.
Biochemical characterization of chlorsulfuron resistance in cichorium intybus L. var. Witloof
FORLANI, Giuseppe;
1997
Abstract
A chicory (Cichorium intybus L. var. Witloof) line, previously selected in vitro for tolerance to the sulfonylurea herbicide chlorsulfuron and regenerated into resistant fertile plants showing a 1500 to 2000 higher resistance than wild-type, was characterized in order to understand the biochemical basis of the resistance. Transcription, specific activity level and properties of acetolactate synthase (ALS, EC 4.1.3.18), the first enzyme in branched-chain amino acid synthesis and the target of sulfonylurea and imidazolinone herbicides, were evaluated in plant tissues and compared with those of a sensitive control. Whilst Northern blot analysis detected similar levels of ALS-mRNA, a significant difference was found in ALS specific activity. However, its higher level in sensitive plants, could not account for the selected tolerance. When ALS activity from wild-type and resistant plants was measured in the presence of increasing chlorsulfuron concentrations, a striking variation in enzyme sensitivity was found. In the two cases 50%-inhibition was achieved at 21 nmol/L and 12 μmol/L (R = 570), respectively, suggesting that the tolerant line could owe its resistance to a mutational event resulting in a decreased affinity of the target enzyme. Mutant ALS displayed cross-tolerance to the imidazolinone herbicide imazamethabenz, a 125-fold higher level of which was required to cause 50 %-inhibition of enzyme activity. Herbicide tolerance was accompanied also by a reduced sensitivity to the regulative feedback inhibition by leucine and valine, with concentrations causing 50%-inhibition about 10 and 3-fold higher, respectively, than those found for the wild-type enzyme.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
