Glyphosate-resistant Individuals Research Articles

Dynamics of Glyphosate Resistance in Lolium perenne: Pollen-Mediated Gene Flow under Field Conditions in Argentina

The genus Lolium has shown a high tendency to evolve resistance to herbicides. Considering the high level of allogamy of L. perenne and the importance of the wind as an agent for pollen dispersion, the aim of this work was to assess and quantify the effective glyphosate-resistant gene flow by pollen in L. perenne under field conditions. Pollen-mediated gene flow from glyphosate-resistant to -susceptible plants was studied using a donor–receptor design. The results indicate that the effective gene flow reached trap plants distanced ≤35 m downwind from the source of pollen. The progeny of receptor plants growing 0 m from the pollen donors showed 13% of individuals with low glyphosate sensitivity. This represents a seven-fold increase in the frequency of glyphosate-resistant individuals found in the progeny of plants isolated during the flowering period (negative control). Similarly, the progenies of plants grown at 15 and 25 m from the source of glyphosate resistance showed an increase in the frequency of resistant plants by about four- and two-fold, respectively. When the receptor plants were located at >35 m, no glyphosate-resistant plants were detected in their progenies. Management should aim for prevention or delay of flowering of glyphosate-resistant plants.

Distribution of Conyza sp. in Orchards of California and Response to Glyphosate and Paraquat

Resistance to glyphosate in hairy fleabane and horseweed is a problem in orchards and vineyards in California. Population genetic analyses suggest that glyphosate resistance evolved multiple times in both species, but it is unknown if resistance to other herbicides is also present. Two approaches of research were undertaken to further evaluate herbicide resistance in Conyza sp. in the perennial crop systems of California. In the initial study, the distribution of Conyza sp. in the Central Valley, using a semistructured field survey, was coupled with evaluation of the presence and level of glyphosate resistance in plants grown from field-collected seed. In a subsequent study, single-seed descendants representing distinct genetic groups were self-pollinated in the greenhouse and these accessions were evaluated for response to glyphosate and paraquat. Conyza sp. were commonly found throughout the Central Valley and glyphosate-resistant individuals were confirmed in all field collections of both species. The level of glyphosate resistance among field collections varied from 5- to 21-fold compared with 50% glyphosate resistance (GR50) of the susceptible, with exception of one region with a GR50 similar to the susceptible. When self-pollinated accessions from different genetic groups were screened, the level of glyphosate resistance, on the basis of GR50 values, ranged from 1.7- to 42.5-fold in hairy fleabane, and 5.9- to 40.3-fold in horseweed. Three accessions of hairy fleabane from different genetic groups were also resistant to paraquat (40.1- to 352.5-fold). One glyphosate-resistant horseweed accession was resistant to paraquat (322.8-fold), which is the first confirmed case in California. All paraquat-resistant accessions of Conyza sp. identified so far have also been resistant to glyphosate, probably because glyphosate resistance is already widespread in the state. Because glyphosate and paraquat resistances are found across a wide geographical range and in accessions from distinct genetic groups, multiple resistant Conyza sp. likely developed independently several times in California.

Evolution of a double amino acid substitution in the 5-enolpyruvylshikimate-3-phosphate synthase in Eleusine indica conferring high-level glyphosate resistance.

Glyphosate is the most important and widely used herbicide in world agriculture. Intensive glyphosate selection has resulted in the widespread evolution of glyphosate-resistant weed populations, threatening the sustainability of this valuable once-in-a-century agrochemical. Field-evolved glyphosate resistance due to known resistance mechanisms is generally low to modest. Here, working with a highly glyphosate-resistant Eleusine indica population, we identified a double amino acid substitution (T102I+P106S [TIPS]) in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene in glyphosate-resistant individuals. This TIPS mutation recreates the biotechnology-engineered commercial first generation glyphosate-tolerant EPSPS in corn (Zea mays) and now in other crops. In E. indica, the naturally evolved TIPS mutants are highly (more than 180-fold) resistant to glyphosate compared with the wild type and more resistant (more than 32-fold) than the previously known P106S mutants. The E. indica TIPS EPSPS showed very high-level (2,647-fold) in vitro resistance to glyphosate relative to the wild type and is more resistant (600-fold) than the P106S variant. The evolution of the TIPS mutation in crop fields under glyphosate selection is likely a sequential event, with the P106S mutation being selected first and fixed, followed by the T102I mutation to create the highly resistant TIPS EPSPS. The sequential evolution of the TIPS mutation endowing high-level glyphosate resistance is an important mechanism by which plants adapt to intense herbicide selection and a dramatic example of evolution in action.

Gene amplification of 5-enol-pyruvylshikimate-3-phosphate synthase in glyphosate-resistant Kochia scoparia.

Field-evolved resistance to the herbicide glyphosate is due to amplification of one of two EPSPS alleles, increasing transcription and protein with no splice variants or effects on other pathway genes. The widely used herbicide glyphosate inhibits the shikimate pathway enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Globally, the intensive use of glyphosate for weed control has selected for glyphosate resistance in 31 weed species. Populations of suspected glyphosate-resistant Kochia scoparia were collected from fields located in the US central Great Plains. Glyphosate dose response verified glyphosate resistance in nine populations. The mechanism of resistance to glyphosate was investigated using targeted sequencing, quantitative PCR, immunoblotting, and whole transcriptome de novo sequencing to characterize the sequence and expression of EPSPS. Sequence analysis showed no mutation of the EPSPS Pro106 codon in glyphosate-resistant K. scoparia, whereas EPSPS genomic copy number and transcript abundance were elevated three- to ten-fold in resistant individuals relative to susceptible individuals. Glyphosate-resistant individuals with increased relative EPSPS copy numbers had consistently lower shikimate accumulation in leaf disks treated with 100 μM glyphosate and EPSPS protein levels were higher in glyphosate-resistant individuals with increased gene copy number compared to glyphosate-susceptible individuals. RNA sequence analysis revealed seven nucleotide positions with two different expressed alleles in glyphosate-susceptible reads. However, one nucleotide at the seven positions was predominant in glyphosate-resistant sequences, suggesting that only one of two EPSPS alleles was amplified in glyphosate-resistant individuals. No alternatively spliced EPSPS transcripts were detected. Expression of five other genes in the chorismate pathway was unaffected in glyphosate-resistant individuals with increased EPSPS expression. These results indicate increased EPSPS expression is a mechanism for glyphosate resistance in these K. scoparia populations.

Inheritance of Glyphosate Resistance in Hairy Fleabane (Conyza bonariensis) from California

Inheritance of glyphosate resistance was investigated in hairy fleabane populations from California as part of providing the information needed to predict and manage resistance and to gain insight into resistance mechanism (or mechanisms) present in the populations. Three glyphosate-resistant individuals grown from seed collected from distinct sites near Fresno, CA, were crossed to individuals from the same susceptible population to create reciprocal F1populations. A single individual from each of the F1populations was used to create a backcross population with a susceptible maternal parent, and an F2population. Based on dose response analyses, reciprocal F1populations were not statistically different from each other, more similar to the resistant parent, and statistically different from the susceptible parent, consistent with nuclear control of the trait and dominance to incomplete dominance of resistance over susceptibility in all three crosses. Glyphosate resistance in two of the three crosses segregated in the backcross and the F2populations as a single-locus trait. In the remaining cross, the resistant parent had approximately half the resistance level as the other two resistant parents, and the segregation of glyphosate resistance in backcross and F2populations conformed to a two-locus model with resistance alleles acting additively and at least two copies of the allele required for expression of resistance. This two-locus model of the segregation of glyphosate resistance has not been reported previously. Variation in the pattern of inheritance and the level of resistance indicate that multiple resistance mechanisms may be present in hairy fleabane populations in California.

Survey of glyphosate-resistant kochia (Kochia scoparia L. Schrad.) in Alberta

Hall, L. M., Beckie, H. J., Low, R., Shirriff, S. W., Blackshaw, R. E., Kimmel, N. and Neeser, C. 2014. Survey of glyphosate-resistant kochia ( Kochia scoparia L. Schrad.) in Alberta. Can. J. Plant Sci. 94: 127-130. Glyphosate-resistant (GR) kochia was identified in Warner county in southern Alberta in 2011. To determine the scale of the distribution and frequency of GR kochia, a randomized stratified survey of more than 300 locations (one population per location) in southern Alberta was conducted in the fall of 2012. Mature plants were collected, seed separated, and F1 seedlings screened by spraying with glyphosate at 900 g a.e. ha-1 under greenhouse conditions. Screening confirmed 13 GR kochia sites: seven in Warner county, five in Vulcan county, and one in Taber county. The frequency of GR individuals in a population ranged from 0.3 to 98%. GR kochia were found in arid areas where chemical fallow is a significant component of the rotation. Economic and agronomic impact of this GR weed biotype is compou...

Survey of glyphosate-resistant kochia (Kochia scoparia L. Schrad.) in Alberta

Hall, L. M., Beckie, H. J., Low, R., Shirriff, S. W., Blackshaw, R. E., Kimmel, N. and Neeser, C. 2014. Survey of glyphosate-resistant kochia ( Kochia scoparia L. Schrad.) in Alberta. Can. J. Plant Sci. 94: 127–130. Glyphosate-resistant (GR) kochia was identified in Warner county in southern Alberta in 2011. To determine the scale of the distribution and frequency of GR kochia, a randomized stratified survey of more than 300 locations (one population per location) in southern Alberta was conducted in the fall of 2012. Mature plants were collected, seed separated, and F1 seedlings screened by spraying with glyphosate at 900 g a.e. ha−1 under greenhouse conditions. Screening confirmed 13 GR kochia sites: seven in Warner county, five in Vulcan county, and one in Taber county. The frequency of GR individuals in a population ranged from 0.3 to 98%. GR kochia were found in arid areas where chemical fallow is a significant component of the rotation. Economic and agronomic impact of this GR weed biotype is compounded because of multiple resistance to acetolactate synthase-inhibiting herbicides.