Biotypes of field bindweed that vary in sensitivity to glyphosate were studied to determine the physiological or biochemical bases for these differences. Studies con ducted using whole plants and in vitro cultured shoots identified several potentially important differences between the most tolerant (biotype 4) and most susceptible (biotype 1) biotypes. Biotype 4 plants had greater 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS) activity and higher concentrations of phenolic compounds, indicating greater shikimate pathway activity than biotype 1. This may reflect a higher growth ability of biotype 4, as observed in shoot cultures. 5-Enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity in whole plants increased in all parts of biotype 4 by 7 d after a glyphosate treatment of 1.1 kg ae ha−1, whereas activities in biotype 1 plants did not increase at any time. However, this may not be the only mechanism of glyphosate tolerance because EPSPS activity in cultured shoots of both biotypes increased equally in response to glyphosate, even though biotype 4 shoots were able to survive and grow on a glyphosate-containing medium that inhibited growth of biotype 1 shoots. We propose that multiple mechanisms operating at a cellular/metabolic level combine to enable biotype 4 to tolerate higher glyphosate rates than biotype 1.

Glyphosate is a broad-spectrum herbicide that has been used extensively for more than 20 yr. The first glyphosate-resistant weed biotype appeared in 1996; it involved a rigid ryegrass population from Australia that exhibited an LD50 value approximately 10-fold higher than that of sensitive biotypes. We have characterized gene expression levels and glyphosate sensitivity of 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS), the target enzyme for glyphosate inhibition, in sensitive and resistant lines derived from this population. Restriction fragment length polymorphism analyses were also performed to examine the distribution of EPSPS gene variants and the gene copy number. A two- to threefold increase in basal EPSPS messenger RNA (mRNA) and enzyme activity levels was observed in the most resistant lines analyzed; however, differences among lines in the sensitivity of EPSPS to glyphosate were not apparent. Induction of EPSPS was observed within 48 h after application of 1.5 kg ae ha−1 of glyphosate. This was reflected in elevated levels of both EPSPS mRNA and enzyme activity. Similarly, 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase mRNA levels increased after glyphosate treatment; however, basal and induced transcript levels were comparable for sensitive and resistant lines in this case. The restriction fragment length polymorphism analyses showed no evidence for gene amplification or cosegregation of a specific EPSPS gene variant with glyphosate resistance. EPSPS expression in lines exhibiting an intermediate level of resistance was indistinguishable from that in glyphosate-sensitive lines, suggesting that the mechanism could, at least in part, be non–target-based.