Physiological consequences of mutation for ALS-inhibitor resistance

• Published in: Weed science 1999-08, Vol.47 (4), p.383-392
• Main Authors: Eberlein, Charlotte V., Guttieri, Mary J., Berger, Philip H., Fellman, John K., Mallory-Smith, Carol A., Thill, Donn C., Baerg, Roger J., Belknap, William R.
• Format: Article
• Language: English
• Subjects: acetohydroxybutyric acid; acetolactate synthase; acetolactic acid; Alleles; amino acid metabolism; amino acid sequences; Amino acids; backcrossing; binding sites; Branched chain amino acids; butyrates; chemical constituents of plants; enzyme activity; enzyme inhibitors; Experiment design; Experimental replication; gene expression; Generally accepted auditing standards; Genetic mutation; Herbicide resistance; Herbicides; histidine; inhibition; isoleucine; Lactuca sativa; Lactuca serriola; leaves; leucine; molecular sequence data; mutants; mutation; nucleotide sequences; Physiology, Chemistry, and Biochemistry; Polymerase chain reaction; proline; pyruvic acid; seeds; stability; substrates; sulfonylurea; valine
• ISSN: 0043-1745; 1550-2759
• DOI: 10.1017/S0043174500091967

Biochemical and physiological effects of target site resistance to herbicides inhibiting acetolactate synthase (ALS) were evaluated using sulfonylurea-resistant (R) and -susceptible (S) near isonuclear Lactuca sativa ‘Bibb’ lines derived by backcrossing the resistance allele from Lactuca serriola L. into L. sativa. Sequence data suggest that resistance in L. sativa is conferred by a single-point mutation that encodes a proline197 to histidine substitution in Domain A of the ALS protein; this is the same substitution observed in R L. serriola. K mapp (pyruvate) values for ALS isolated from R and S L. sativa were 7.3 and 11.1 mM, respectively, suggesting that the resistance allele did not alter the pyruvate binding domain on the ALS enzyme. Both R and S ALS had greater affinity for 2-oxobutyrate than for pyruvate at the second substrate site. Ratios of acetohydroxybutyrate: acetolactate produced by R ALS across a range of 2-oxobutyrate concentrations were similar to acetohydroxybutyrate: acetolactate ratios produced by S ALS. Specific activity of ALS from R L. sativa was 46% of the specific activity from S L. sativa, suggesting that the resistance allele has detrimental effects on enzyme function, expression, or stability. ALS activity from R plants was less sensitive to feedback inhibition by valine, leucine, and isoleucine than ALS from S plants. Valine, leucine, and isoleucine concentrations were about 1.5 times higher in R seed than in S seed on a per gram of seed basis, and concentrations of valine and leucine were 1.3 and 1.6 times higher, respectively, in R leaves than in S leaves. Findings suggest that the mutation for resistance results in altered regulation of branched-chain amino acid synthesis.