ABSTRACT
Triazolopyrimidine sulfanilides are a class of highly active herbicides whose primary target is acetolactate synthase. Spontaneous mutants of tobacco (Nicotiana tabacum) (KS-43) and cotton (Gossypium hirsutum) (PS-3 and DO-2) resistant to triazolopyrimidine sulfonanilide were selected in tissue culture. Acetolactate synthase partially purified from the three mutants were 80- to 1000-fold less sensitive to inhibition by the compound compared with the corresponding wild-type enzyme. The mutants also varied in the cross-resistance pattern to other acetolactate synthase inhibiting herbicides in the sulfonylurea, imidazolinone, and pyrimidyl-oxy-benzoate chemical families. Thus, acetolactate synthase from KS-43, PS-3, and DO-2 cultures have different mutations. The affinities for pyruvate, thiamine pyrophosphate, as well as the activity of the mutant enzymes were found to be comparable to the corresponding wild-type enzymes. However, the enzyme from PS-3 was highly resistant to feedback inhibition by valine and leucine. In contrast, acetolactate synthase from KS-43 and DO-2 were inhibited by valine and leucine to nearly the same extent as the wild-type enzymes. Also, PS-3 cultures accumulated much higher levels of the branched chain amino acids compared to the wild-type cotton culture. The mutation in the PS-3 enzyme has therefore rendered it insensitive to feedback regulation by valine and leucine.
ABSTRACT
[(14)C]Atrazine (2-chloro-4-[ethylamino]-6-[isopropylamino]-s-triazine) and [(14)C]glyphosate (N-[phosphonomethyl]glycine) were xylem fed to sunflower shoots at 100 micromolar for 1 hour in the light, then placed in the dark at 100% relative humidity for 1, 4, 7, or 10 hours. The distribution of atrazine and glyphosate between shoot parts, in the leaves, and between the aoplast and symplast of the leaf was determined. The apoplastic concentrations and distribution patterns of atrazine and glyphosate in the leaves were evaluated using a pressure dehydration technique, our results were compared to the previously reported distribution patterns of the naturally occurring apoplastic leaf solutes, and the apoplastic dye PTS (trisodium 3-hydroxy-5,8,10-pyrenetrisulfonate). The pattern of atrazine and glyphosate distribution in the shoot, and between the leaf apoplast and symplast, was found to reflect the potential of these herbicides to enter the shoot symplast. The results of this study are discussed with respect to current theories of xenobiotic transport in plants, and have been found to be consistent with the intermediate permeability hypothesis for xenobiotic transport.
ABSTRACT
A simple, repeatable, and accurate method is described for the collection of apoplastic and membrane-filtered symplastic sap fractions, and for the determination of the origin of these fractions within the leaf. The apoplastic distribution patterns of the naturally occurring apoplastic leaf solutes, and the apoplastic dye PTS (trisodium 3-hydroxy-5, 8, 10-pyrenetrisulfonate) were compared. Aliquots of sap were expressed from detached sunflower leaves in a pressure chamber over intervals of 0.02 to 0.04 megapascal. Three distinct fractions were detected in the expressed sap volume. These were successively released and identified as a petiole-midrib fraction, a minor vein-cell wall fraction, and a mixed fraction consisting of a contribution from the minor vein-cell wall with an increasing proportion of membrane-filtered cell sap.
