As cases of resistance to herbicides escalate worldwide, there is increasing demand from growers to test for weed resistance and learn how to manage it. Scientists have developed resistance-testing protocols for numerous herbicides and weed species. Growers need immediate answers and scientists are faced with the daunting task of testing an increasingly large number of samples across a variety of species and herbicides. Quick tests have been, and continue to be, developed to address this need, although classical tests are still the norm. Newer methods involve molecular techniques. Whereas the classical whole-plant assay tests for resistance regardless of the mechanism, many quick tests are limited by specificity to an herbicide, mode of action, or mechanism of resistance. Advancing knowledge in weed biology and genomics allows for refinements in sampling and testing protocols. Thus, approaches in resistance testing continue to diversify, which can confound the less experienced. We aim to help weed science practitioners resolve questions pertaining to the testing of herbicide resistance, starting with field surveys and sampling methods, herbicide screening methods, data analysis, and, finally, interpretation. More specifically, this article discusses approaches for sampling plants for resistance confirmation assays, provides brief overviews on the biological and statistical basis for designing and analyzing dose–response tests, and discusses alternative procedures for rapid resistance confirmation, including molecular-based assays. Resistance confirmation procedures often need to be slightly modified to suit a specific situation; thus, the general requirements as well as pros and cons of quick assays and DNA-based assays are contrasted. Ultimately, weed resistance testing research, as well as resistance management decisions arising from research, needs to be practical, feasible, and grounded in science-based methods.

Annual bluegrass is commonly controlled by acetolactate synthase (ALS)-inhibiting herbicides in managed turfgrass. An annual bluegrass population with suspected resistance to ALS-inhibiting herbicides was collected from Grand National Golf Course in Opelika, AL (GN population). Subsequent testing confirmed resistance of the GN population to foramsulfuron, trifloxysulfuron, bispyribac-sodium (bispyribac), and imazaquin when compared to a susceptible population collected locally at Auburn University (AU population). Sequencing of the ALS gene revealed a point mutation resulting in an amino acid substitution at Trp574. Cloning of the ALS gene surrounding the Trp574 region yielded two distinct ALS gene sequences: one producing Trp574 and one producing Leu574. Trp574 to Leu has been previously correlated with resistance to ALS-inhibiting herbicides. Both AU and GN gene sequences contained other similar silent and missense mutations. This research confirms resistance of annual bluegrass to ALS-inhibiting herbicides with Trp574 to Leu amino acid substitution being the most likely mode of resistance based on past literature.

Studies were conducted to determine the growth, fecundity, and competitive ability of an acetyl-CoA carboxylase (ACCase)–inhibitor resistant (R) sterile wild oat biotype compared with a susceptible (S) biotype. Seed germination studies indicated that there were no differences in seed germination and seedling vigor between R and S biotypes at any temperature regime. R and S biotypes were grown under noncompetitive and competitive arrangement in the greenhouse. Under noncompetitive greenhouse conditions, growth of the R biotype was similar to that of the S biotype on the basis of plant height, canopy area, and plant biomass. Seed production and weight of R and S plants were also at the same levels. Furthermore, relative competitiveness among the R and S sterile wild oat biotypes was investigated by means of replacement series experiments. The R and S biotypes were compared under seven mixture proportions (6 : 0, 5 : 1, 4 : 2, 3 : 3, 2 : 4, 1 : 5, and 0 : 6). No significant differences in competitive ability were observed between R and S biotypes on the basis of plant height, canopy area, or plant biomass. In most cases, relative crowding coefficient (RCC) values at 20, 60, and 100 d after transplanting (DAT) were close to one, indicating equal competitiveness between the R and S biotypes of wild oat used in this competitive study. However, in some cases, the RCC value was 1.31 for plant height, evident of a slight competitive advantage for the R biotype at 100 DAT. In general, ACCase-inhibitor R and S sterile wild oat biotypes were equally competitive, clearly without any growth penalty for R plants in either noncompetitive or competitive conditions.

Florida hydrilla populations have shown an alarming increase in resistance to fluridone, an herbicide used extensively for controlling invasive US hydrilla populations. A rapid PCR and sequencing method was developed to identify and screen hydrilla genomic DNA for three previously identified phytoene desaturase (pds) gene mutations that confer resistance to fluridone. Ninety hydrilla accessions were screened for fluridone resistant genotypes including 46 accessions from the US and 44 accessions from 15 other countries. In Florida, hydrilla from five of nine sites tested was heterozygous for wild-type and herbicide-resistant alleles. Additionally, a new resistant population was identified from Lake Seminole in Georgia, the first genetically confirmed strain of resistant hydrilla outside of Florida. All resistance-conferring mutations were located on the same homologous haplotype of US dioecious hydrilla. All other hydrilla samples tested possessed only wild type alleles, including monoecious strains that had been exposed to fluridone. Management implications are discussed.

Crops transformed to provide resistance to herbicides with two different mechanisms of action provide new opportunities for control of herbicide-resistant weeds. However, unexpected interactions may develop, especially for herbicides not generally used in tank-mixtures. The objectives of this study were to evaluate weed control and determine herbicide interactions and fluorescence responses with combinations of glyphosate and glufosinate on selected weeds prevalent in Michigan cropping systems. Field studies to determine herbicide interactions resulted in synergism only at 0.84 kg ae ha−1 of glyphosate and 0.47 kg ai ha−1 glufosinate in 2008. Early synergism (7 d after treatment [DAT]) was observed in the field at several combined rates for common lambsquarters and velvetleaf in 2009, and in the greenhouse for giant foxtail. Differences between years were perhaps due to the effect of environmental conditions on herbicide absorption and translocation. Antagonism was observed in the field in 2009 for velvetleaf, common lambsquarters, and giant foxtail especially at 840 g ae ha−1 glyphosate and 118 g ai ha−1 glufosinate, 28 DAT. Antagonism was also observed in the greenhouse for giant foxtail and Canada thistle, 28 DAT. Fluorescence measurements on Canada thistle in the greenhouse showed that glufosinate and glufosinate plus glyphosate acted rapidly to quench electron transport of photosystem II (PS II) system of photosynthesis, and the fluorescence characteristics of the glyphosate and glufosinate combinations were indistinguishable from glufosinate alone.

A suspected glyphosate-resistant (R) junglerice population was collected from a glyphosate-R corn field near Durham in northern California where glyphosate had been applied at least twice a year for over 6 yr. Based on the amount of glyphosate required to reduce growth by 50% (ED50), the R population was 6.6 times more R than the susceptible (S) standard population. Based on the glyphosate concentration that inhibits EPSPS by 50% based on shikimate accumulation (I50) in leaf discs, R plants were four times more R than S plants. By 3 d after treatment with 0.42 kg ae ha−1 glyphosate, the S population had accumulated approximately five times more shikimate than the R population. No differences in [14C]-glyphosate uptake and translocation were detected between R and S plants. However, partial sequencing of the EPSPS gene revealed a mutation in R plants causing a proline to serine change at EPSPS position 106 (P106S). Our results reveal the first case of a P106S target site mutation associated with glyphosate resistance in junglerice.

The biochemical basis of resistance to the acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicide diclofop-methyl was investigated in a resistant wild oat population (R1), which does not exhibit a resistant ACCase. Rates of foliar uptake and translocation of [14C]-diclofop were the same in the R1 vs. susceptible (S) populations. However, the level of phytotoxic diclofop acid was always found to be lower in the R1 vs. S plants, with a concomitant higher level (up to 1.7-fold) of nontoxic polar diclofop metabolites in R1 relative to the S plants. These results indicate that a non–target-site-based mechanism of enhanced rate of diclofop acid metabolism confers resistance in population R1. Moreover, the high-performance liquid chromotography elution profile of the major diclofop metabolites in R1 is similar to that of wheat, suggesting resistance in individuals of population R1 involves a wheat-like detoxification system mediated by cytochrome P450 monooxygenases. In addition, lower level of tissue diclofop acid was also observed using nonradioactive ultra-performance liquid chromatography–mass spectrometry analysis in resistant individuals of three other resistant wild oat populations (R2, R3, and R4) known to posses ACCase gene resistance mutations. These results establish that either one or at least two independent resistance mechanisms (target-site ACCase resistance mutations and non–target-site enhanced rates of herbicide metabolism) can be present in individual wild oat plants.

Field bindweed is extremely susceptible to aminocyclopyrachlor compared to other weed species. Laboratory studies were conducted to determine if absorption, translocation, and metabolism of aminocyclopyrachlor in field bindweed differs from other, less susceptible species. Field bindweed plants were treated with 3.3 kBq 14C-aminocyclopyrachlor by spotting a single leaf mid-way up the stem with 10 µl of herbicide solution. Plants were then harvested at set intervals over 192 h after treatment (HAT). Aminocyclopyrachlor absorption reached a maximum of 48.3% of the applied radioactivity by 48 HAT. A translocation pattern of herbicide movement from the treated leaf into other plant tissues emerged, revealing a nearly equal aminocyclopyrachlor distribution between the treated leaf, aboveground tissue, and belowground tissue of 13, 14, and 14% of the applied radioactivity by 192 HAT. Over the time-course, no soluble aminocyclopyrachlor metabolites were observed, but there was an increase in radioactivity recovered bound in the nonsoluble fraction. These results suggest that aminocyclopyrachlor has greater translocation to belowground plant tissue in field bindweed compared with results from other studies with other herbicides and other weed species, which could explain the increased level of control observed in the field. The lack of soluble metabolites also suggests that very little metabolism occurred over the 192 h time course.

Soybean plants exposed POST to 2,4-D can have reduced seed yield depending on the dose and time of exposure, but it is unclear how 2,4-D affects specific yield components. Objectives were to quantify soybean injury, characterize changes in seed yield and yield components of soybean plants exposed to 2,4-D, and determine if seed-yield loss can be estimated from visual assessment of crop injury. Ten rates (0, 0.1, 1.1, 11.2, 35, 70, 140, 280, 560, and 2,240 g ae ha−1) of 2,4-D were applied to Becks brand 342 NRR soybean at three soybean growth stages (V2, V5, or R2). The soybeans were planted near Lafayette, IN and Urbana, IL in 2009 and 2010 and near Fowler, IN in 2009. Twenty percent visual soybean injury was caused by 29 to 109 g ha−1 2,4-D at 14 d after treatment (DAT) and 109 to 245 g ha−1 at 28 DAT. Nonlinear regression models were fit to describe the effect of 2,4-D on seed yield and yield components of soybean. Seed yield was reduced by 5% from 87 to 116 g ha−1 and a 10% reduction was caused by 149 to 202 g ha−1 2,4-D at all application timings. The number of seeds m−2, pods m−2, reproductive nodes m−2, and nodes m−2 were the most sensitive yield components. Path analysis indicated that seeds m−2, pods m−2, main stem reproductive nodes m−2, and main stem nodes m−2 were the most influential yield components in seed-yield formation. Seed-yield loss was significant (P < 0.0001) and highly correlated (R2 = 0.95 to 0.99) to visual soybean injury ratings. A 10% seed-yield loss was caused by 35% soybean injury observed at 14 DAT, whereas a 10% seed-yield loss was a result of 40, 19, and 15% soybean injury observed at 28 DAT when soybean was exposed to 2,4-D at the V2, V5, and R2 growth stages, respectively.

Alligatorweed is well known for its potassium (K+)-accumulating capabilities and its strong resistance to undesired growth conditions. The results of this study revealed properties of K+ accumulation and its contribution to drought stress in alligatorweed. In addition, we attempted to characterize the molecular mechanisms of K+ accumulation in this plant. Alligatorweed plants showed a consistent increase in biomass in response to external K+ concentrations, ranging from micromolar levels up to 50 mmol L−1; K+ was also accumulated accordingly in the plants. The stem was the most K+-accumulating organ, accumulating up to 13% of the K+. Moreover, this K+ superaccumulation caused improved resistance to drought stress. The apparent K+ uptake by the roots showed a typical high-affinity property, and the Michaelis constant increased at higher rates of plant K+ in the starting materials. Furthermore, three putative, K+-uptake transporter complementary DNAs (cDNAs) were isolated from alligatorweed (ApKUP1, ApKUP2, and ApKUP3, respectively) using degenerated primers and rapid amplification of cDNA end techniques. The expression of ApKUP1 and ApKUP3 was predominately localized to the leaves, whereas ApKUP2 was expressed throughout the entire plant. The expression of ApKUP1 and ApKUP3 was stimulated in the stems and roots when K+ was depleted from the external medium. Moreover, ApKUP3 expression was enhanced in the stem in response to abscisic acid treatment and drought stress. In conclusion, our findings provide further insight into the mechanisms of K+ accumulation linked to K+ uptake in alligatorweed.

A segment of the debate surrounding the commercialization of genetically engineered (GE) crops, such as glyphosate-resistant (GR) crops, focuses on the theory that implementation of these traits is an extension of the intensification of agriculture that will further erode the biodiversity of agricultural landscapes. A large field-scale study was conducted in 2006 in the United States on 156 different field sites with a minimum 3-yr history of GR corn, cotton, or soybean in the cropping system. The impact of cropping system, crop rotation, frequency of using the GR crop trait, and several categorical variables on emerged weed density and diversity was analyzed. Species richness, evenness, Shannon's H′, proportion of forbs, erect growth habit, and C3 species diversity were all greater in agricultural sites that lacked crop rotation or were in a continuous GR crop system. Rotating between two GR crops (e.g., corn and soybean) or rotating to a non-GR crop resulted in less weed diversity than a continuous GR crop. The composition of the weed flora was more strongly related to location (geography) than any other parameter. The diversity of weed flora in agricultural sites with a history of GR crop production can be influenced by several factors relating to the specific method in which the GR trait is integrated (cropping system, crop rotation, GR trait rotation), the specific weed species, and the geographical location. The finding that fields with continuous GR crops demonstrated greater weed diversity is contrary to arguments opposing the use of GE crops. These results justify further research to clarify the complexities of crops grown with herbicide-resistance traits, or more broadly, GE crops, to provide a more complete characterization of their culture and local adaptation.

Greenhouse studies were conducted to evaluate the growth response of itchgrass to water stress. Itchgrass plants produced the greatest aboveground biomass and seeds at 75% of field capacity and these parameters at 50 and 100% of field capacity were similar. With further increase in water stress, seed production was sharply reduced, but itchgrass was still able to produce an average of 63 and 9 seeds plant−1 at 25 and 12.5% of field capacity, respectively. Itchgrass plants responded to increasing water stress with increased leaf weight ratio; it was 2.5 times greater at 12.5% of field capacity than at 100% of field capacity. In another study, compared with daily irrigation, intervals of 9 d between irrigations reduced aboveground biomass of itchgrass by 27% and 12-d intervals reduced aboveground biomass by 67%. Compared with the daily irrigation regime, itchgrass seed production was reduced by 61% at intervals of 12 d between irrigations; however, the weed plants produced a considerable number of seeds (153 seeds plant−1) at the 12-d intervals. The ability of itchgrass to produce biomass and seeds under water stressed conditions necessitates strategies that minimize weed survival while maximizing irrigation efficiency for the crop at the same time.

According to climate models, drier summers must be expected more frequently in Central Europe during the next decades, which may influence plant performance and competition in grassland. The overall source–sink relations in plants, especially allocation of solutes to above- and below-ground parts, may be affected by drought. To investigate solute export from a given leaf of broadleaf dock, a solution containing 57Co and 65Zn was introduced through a leaf flap. The export from this leaf was detected by analysing radionuclide contents in various plant parts. Less label was allocated to new leaves and more to roots under drought. The observed alterations of source–sink relations in broadleaf dock were reversible during a subsequent short period of rewatering. These findings suggest an increased resource allocation to roots under drought improving the functionality of the plants.

Smutgrass is an invasive, well-rooted perennial that has long been recognized as an aggressive weed throughout Florida and in the subtropical regions of the United States. Small smutgrass and giant smutgrass are the two predominant smutgrass varieties found in Florida. The native soil pH of Florida flatwoods is 4.5 to 5.0 which is lower than the level of soil pH recommended for optimum bahiagrass growth. Therefore, replacement series experiments were conducted in a greenhouse in 2010 and 2011 to compare the competitive ability of bahiagrass with each of the two varieties of smutgrass at three levels of soil pH (4.5, 5.5, and 6.5), two densities; 4 (low) and 8 (high) plants pot−1, and at five planting ratios of 100 : 0, 75 : 25, 50 : 50, 25 : 75, and 0 : 100. Relative competitive ability and aggressivity of giant smutgrass was greater than bahiagrass across all pH levels and densities, whereas relative competitive ability and aggressivity of bahiagrass was greater than small smutgrass in all pH levels and densities, except at pH 6.5. At pH 5.5, biomass accumulation of giant smutgrass was at least 73% higher than bahiagrass, whereas small smutgrass biomass was at least 46% lower than bahiagrass at equal planting ratios of both low and high densities. Differential responses were observed on bahiagrass competitive ability with small and giant smutgrass. Amending soil pH is not a likely option to increase the growth and competitive ability of bahiagrass over giant smutgrass. However, for small smutgrass, it is likely to increase the aggressivity of bahiagrass in bahiagrass–small smutgrass mixture, unless the soil pH is raised above 5.5.

Exotic invasive species are nonnative species that thrive outside of their native habitat, and while it is difficult to determine which exotic plants will become invasive, successful invaders often share a wide range of traits including high growth rate and reproductive output, vegetative reproduction, high population growth rates, early reproductive age, phenotypic and physiological plasticity, and high resource use efficiency. Here we report on the response of pampasgrass, an important exotic invasive plant of the western United States, to experimental variations in soil nitrogen (N) and water availability. Given its ability to invade a wide variety of ecosystems in southern California, we hypothesized that pampasgrass would have higher water and N use efficiency under conditions of low water and N availability but rapid growth and resource use under conditions of high water and N availability. Our data support this hypothesis and indicate that pampasgrass exhibited large variations in growth, carbon allocation, morphology, and N and phosphorus (P) nutrition to variations in N availability and water table depth. Many of these traits are highly correlated with invasive performance, and the high N and P use efficiency observed under low soil N (control) and water table, coupled with the large increase in physiological performance and resource use under high N and water table, indicate that pampasgrass is highly flexible to soil resource levels that are typical for coastal sage scrub and riparian ecosystems of southern California. Such flexibility in resource use could allow pampasgrass to persist in low-resource environments and expand as resource levels increase.

Purple nutsedge is among the most troublesome weeds of vegetables in the Southeast US and a substantial impediment in the search for methyl bromide alternatives. Greater understanding of the environmental cues that regulate tuber sprouting may assist in improved nutsedge management. Experiments were conducted to evaluate the effect of diurnal temperature variation on sprouting of purple nutsedge tubers. Two temperature regimes were evaluated: the first averaged 28 C, with daily fluctuations ranging from 0 to 19.5 C; the second temperature regime averaged 16 C, with daily fluctuations ranging from 0 to 18.5 C. When average temperature was 28 C, cumulative tuber sprouting ranged from 88 to 92%, with no detectable differences among diurnal fluctuations. The high average temperature in the first study may have negated any type of enforced sprouting suppression. However, when average temperature was lowered to 16 C (simulating early spring diurnal fluctuations under polyethylene mulch), there was a positive linear correlation between maximum tuber sprouting and temperature variation. With an average temperature of 16 C, the absence of temperature variation resulted in 52% purple nutsedge sprouting, while 87% sprouting occurred when daily temperature varied 18.5 C at the same average temperature. The use of various types of mulching material can affect average soil temperatures and diurnal variations, potentially shifting nutsedge emergence. Further studies are needed to determine if these data on tuber sprouting in response to alternating temperatures can facilitate more efficient weed management.

Weed growth and N assimilation usually increase with N application rate. With the increasing price of N fertilizer, a better understanding N assimilation by weeds is necessary to maximize economic return. Total plant yield is generally independent of population density, except when plants are very small or at very low population density. If plant yield is independent of population density, weed N assimilation may also be independent of population density. However, the effect of weed population density on N assimilation has not been thoroughly investigated. A 2011 controlled-environment study was established in East Lansing, MI, to evaluate the effect of weed population density and N application rate on growth and N assimilation by common lambsquarters and redroot pigweed. Study factors included four weed densities (1, 2, 4, and 8 plants pot−1), three N application rates (0, 67, and 134 kg N ha−1), and two weed species (redroot pigweed and common lambsquarters). Weeds were destructively harvested 3 wk after emergence, and shoot height, biomass, total N concentration, N use efficiency, and N assimilation were measured. Redroot pigweed was taller, had greater shoot biomass, and a greater shoot N assimilation than did common lambsquarters. With similar environmental conditions, redroot pigweed is expected to be more competitive than common lambsquarters. Shoot N assimilation increased with increasing weed population density, indicating that N assimilation was not independent of population density 3 wk after emergence because weeds were small or at low population density.

Glyphosate-resistant Palmer amaranth is a serious problem in southern cropping systems. Much phenotypic variation is observed in Palmer amaranth populations with respect to plant growth and development and susceptibility to herbicides. This may be related to levels of genetic diversity existing in populations. Knowledge of genetic diversity in populations of Palmer amaranth may be useful in understanding distribution and development of herbicide resistance. Research was conducted to assess genetic diversity among and within eight Palmer amaranth populations collected from North Carolina and Georgia using amplified fragment length polymorphism (AFLP) markers. Pair-wise genetic similarity (GS) values were found to be relatively low, averaging 0.34. The highest and the lowest GS between populations were 0.49 and 0.24, respectively, while the highest and the lowest GS within populations were 0.56 and 0.36, respectively. Cluster and principal coordinate (PCO) analyses grouped individuals mostly by population (localized geographic region) irrespective of response to glyphosate or gender of individuals. Analysis of molecular variance (AMOVA) results when populations were nested within states revealed significant variation among and within populations within states while variation among states was not significant. Variation among and within populations within state accounted for 19 and 77% of the total variation, respectively, while variation among states accounted for only 3% of the total variation. The within population contribution towards total variation was always higher than among states and among populations within states irrespective of response to glyphosate or gender of individuals. These results are significant in terms of efficacy of similar management approaches both in terms of chemical and biological control in different areas infested with Palmer amaranth.

A model that describes the emergence of ripgut brome was developed using a two-season data set from a no-tilled field in northeastern Spain. The relationship between cumulative emergence and hydrothermal time (HTT) was described by a sigmoid growth function (Chapman). HTT was calculated with a set of water potentials and temperatures, iteratively used, to determine the base water potential and base temperature. Emergence of ripgut brome was well described with a Chapman function. The newly-developed function was validated with four sets of data, two of them belonging to a third season in the same field and the other two coming from independent data from Southern Spain. The model also successfully described the emergence in different field management and tillage systems. This model may be useful for predicting ripgut brome emergence in winter cereal fields of semiarid Mediterranean regions.