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Recent increases in glyphosate use in perennial crops of California, USA, are hypothesized to have led to an increase in selection and evolution of resistance to the herbicide in Conyza canadensis populations. To gain insight into the evolutionary origins and spread of resistance and to inform glyphosate resistance management strategies, we investigated the geographical distribution of glyphosate resistance in C. canadensis across and surrounding the Central Valley, its spatial relationship to groundwater protection areas (GWPA), and the genetic diversity and population structure and history using microsatellite markers. Frequencies of resistant individuals in 42 sampled populations were positively correlated with the size of GWPA within counties. Analyses of population genetic structure also supported spread of resistance in these areas. Bayesian clustering and approximate Bayesian computation (ABC) analyses revealed multiple independent origins of resistance within the Central Valley. Based on parameter estimation in the ABC analyses, resistant genotypes underwent expansion after glyphosate use began in agriculture, but many years before it was detected. Thus, diversity in weed control practices prior to herbicide regulation in GWPA probably kept resistance frequencies low. Regionally coordinated efforts to reduce seed dispersal and selection pressure are needed to manage glyphosate resistance in C. canadensis.

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Reduced control of some glyphosate-resistant hairy fleabane populations with paraquat has raised concerns about evolved multiple resistance to both glyphosate and paraquat. The objective of this study was to confirm the presence of multiple-resistant (glyphosate and paraquat) hairy fleabane populations in California. A series of dose-response experiments was conducted to evaluate the effect of glyphosate and paraquat in a known susceptible (S) and putative multiple-resistant (R) population of hairy fleabane. The greenhouse experiments were conducted during summer, fall, and winter under controlled temperature and natural light conditions. Multiple-resistant hairy fleabane was identified; however, the level of resistance to glyphosate varied substantially among seasons. During the summer, the glyphosate rate required to reduce growth by 50% (GR50) for the R population was 0.94 kg ae ha−1, 5.2-fold more than for the S population. In the fall and winter experiments, however, the R population response to glyphosate was similar to the S population with a GR50of 0.22 kg ae ha−1or less. Multiple-resistant plants were controlled in the fall and winter at rates that did not control the same population during summer. GR50of paraquat varied among seasons (0.94, 0.24, and 0.07 kg ai ha−1during summer, fall, and winter, respectively); however, plant mortality was more consistent. This is the first reported case of glyphosate–paraquat resistance in hairy fleabane and the multiple-resistant population could pose a significant challenge to annual no-till and perennial cropping systems in California. Further research on the mechanisms of resistance and the physiological factors underlying the seasonally variable response to glyphosate is needed.

Glyphosate-resistant weeds are an increasing problem in perennial cropping systems in the Central Valley of California, USA. To elucidate the evolutionary origins and spatial spread of resistance, we investigated the geographical distribution of glyphosate resistance and the population genetic diversity and structure of Conyza bonariensis and compared the results with previously studied C. canadensis. Thirty-five populations from orchards and vineyards across the Central Valley were sampled. Population genetic structure was assessed using microsatellite markers. Population-level resistance was assessed in glasshouse screening of plants grown from field-collected seed. Bayesian clustering and analyses of multilocus genotypes indicated multiple origins of resistance, as observed in C. canadensis. Pairwise F ST analysis detected spatial spread of resistance in the south of the Central Valley, also similar to C. canadensis. The results strongly indicate that the southern valley was an environment markedly more suitable than the northern valley for resistance spread and that spread in Conyza species was driven by increased uniformity of strong selection in the southern valley, due to recent regulation on herbicides other than glyphosate. Accordingly, resistant C. canadensis individuals occurred at high frequencies only in the southern valley, but interestingly resistant C. bonariensis occurred at high frequencies throughout the valley. Expression of resistance showed varying degrees of plasticity in C. bonariensis. The lower selfing rate and substantially greater genotypic diversity in C. bonariensis, relative to C. canadensis, indicate greater evolutionary potential over shorter time periods. Interspecific hybridisation was detected, but its role in resistance evolution remains unclear.

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