Eclipta, widespread in tropical, subtropical, and temperate regions, is one of the main malignant broadleaf weeds and thrives in moist and dryland fields. Field rates of acetolactate synthase (ALS) inhibitors have failed to control eclipta in some farmlands in China. One ALS inhibitor–resistant population (R) collected from Jiangsu province in China was confirmed in the greenhouse in our preliminary work. Whole-plant assays revealed that this R population was highly resistant to four sulfonylureas (pyrazosulfuron-ethyl, 134-fold; bensulfuron-methyl, 172-fold; metsulfuron-methyl, 30-fold; and tribenuron-methyl, 195-fold), two triazolopyrimidines (pyroxsulam, 98-fold; penoxsulam, 30-fold), and one pyrimidinylthio-benzoate (bispyribac-sodium, 166-fold) and was moderately resistant to two imidazolinones (imazethapyr, 10-fold; imazapic, 19-fold). ALS enzyme-activity assays showed insensitivity of the ALS from the R population (resistance index values ranged from 12 to 293) to all of the above ALS inhibitors in vitro. Chromatograms from ALS gene sequence analysis detected a homozygous Pro-197-Ser amino acid substitution in the R population. These results confirmed that the Pro-197-Ser substitution results in broad-spectrum cross-resistance to ALS inhibitors in the eclipta R population. To our knowledge, this study is the first to report broad cross-resistance to ALS inhibitors in eclipta and to obtain the full-length ALS gene sequence.

Herbicide-resistant weeds pose a considerable threat to agriculture, but their resistance mechanisms are poorly understood. Differential gene expression analysis of a weed subjected to herbicide treatment is a key step toward more mechanistic studies. Such an analysis, often involving quantitative real-time PCR (qPCR), requires suitable reference genes as internal controls. In this study, we identified optimal reference genes in the noxious weed, Japanese foxtail. This weed has evolved resistance to acetyl-coenzyme A carboxylase (ACCase) inhibitors. We analyzed the stability of eight commonly used candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase [GAPDH]; ubiquitin [UBQ]; capsine phosphatase [CAP]; beta-tubulin [TUB]; eukaryotic initiation factor 4a [EIF4A]; elongation factor-1 alpha [EF1]; 18S ribosomal RNA [18S]; 25S ribosomal RNA [25S]) from root, stem, and leaf tissue of plants that were either resistant or sensitive to ACCase inhibitors, with or without herbicide stress, using qPCR. The results were further ranked and analyzed using geNorm, NormFinder, and BestKeeper software. These analyses identified EF1 and UBQ in roots, EF1, TUB, CAP, and 18S in stems, and EF1, GAPDH, and 18S in leaves as suitable references for qPCR normalization. We have identified a set of reference genes that can be used to study herbicide resistance mechanisms in Japanese foxtail.

The Enlist™ traits provide 2,4-D resistance in several crops. Though corn is naturally tolerant to 2,4-D, the engineered trait conferred by the aryloxyalkanoate dioxygenase-1 (AAD-1) enzyme provides enhanced 2,4-D tolerance and confers resistance to the graminicide herbicide family, the aryloxyphenoxypropionates. The objectives of this research were 2-fold: (1) measure and compare uptake, translocation, and metabolism of 2,4-D in Enlist™ (E, +AAD1) and non–AAD-1 transformed (NT, −AAD1) isogenic corn hybrids; and (2) and investigate the effect of glyphosate and/or the Enlist™ adjuvant system (ADJ) on these factors and corn injury. Uptake of radiolabeled 2,4-D acid applied alone in corn was not altered by the addition of ADJ when tank mixed at 24 h after application (HAA). By contrast, uptake of radiolabeled 2,4-D was significantly lower (69%) compared with 2,4-D plus ADJ (89%) at 24 HAA with a premixed formulation of 2,4-D choline plus glyphosate-dimethylamine (Enlist Duo™ herbicide [EDH]). Translocation of 2,4-D between the two corn hybrids was not different. E corn metabolized more 2,4-D (100% of absorbed) than NT corn (84%), and glyphosate did not alter 2,4-D metabolism. Furthermore, the metabolism of 2,4-D to nonphytotoxic dichlorophenol (DCP) and subsequent DCP-derived metabolites formed in E corn was examined. Injury to E corn is not typically observed in the field; however, injury symptoms were clearly evident in E corn (within 24 HAA) when formulated acetochlor was tank mixed with EDH, which correlated with an increase in 2,4-D uptake during this time period. In summary, the lack of injury in E corn following EDH applied alone may be attributed to a relatively low amount of 2,4-D uptake and the combination of natural and engineered 2,4-D metabolic pathways.

Fenoxaprop-P-ethyl, a phenoxy herbicide of the aryloxy–phenoxy–propionic acid group, had a strong control effect when applied POST to weedy rice in this study, with the effective concentrations of 294 μM and 218 μM of herbicide causing 50% inhibition (IC50) in plant height and fresh weight values, respectively. However, fenoxaprop-P-ethyl caused phytotoxicity in cultivated rice. Isoxadifen-ethyl, a widely used herbicide safener in rice, can decrease the phytotoxicity caused by fenoxaprop-P-ethyl. Owing to the extremely similar morphological features and physiological properties of weedy and cultivated rice, it is not practical to spray isoxadifen-ethyl directly on cultivated rice plants to safen them. Applying the safener directly to cultivated rice seeds may be a practical alternative method. To improve the biological activity of isoxadifen-ethyl seed treatments, novel compounds were designed by splicing other groups, including amines, amino acids, and 2- methoxy-5-nitrophenol sodium salt, to the parental structure of isoxadifen-ethyl. Through hydrolysis, acyl chlorination, acyl amination, and esterification, a series of isoxadifen-ethyl derivatives were synthesized and their structures were determined by mass spectrometry and 1H nuclear magnetic resonance spectroscopy. The biological activities of five of the isoxadifen-ethyl derivatives, which possessed recovery effects similar to isoxadifen-ethyl, were able to relieve herbicide phytotoxicity. In pot experiments, isoxadifen-ethyl showed almost no activity as a seed treatment, while three derivative compounds, when used independently as seed treatments, were able to prevent the damage caused by fenoxaprop-P-ethyl. The results will help to develop a new control method for weedy rice, thereby decreasing production costs and increasing farmers’ incomes.

Glyphosate and 2,4-D have been commonly used for control of common and giant ragweed before planting of corn and soybean in the midwestern United States. Because these herbicides are primarily applied in early spring, environmental factors such as temperature may influence their efficacy. The objectives of this study were to (1) evaluate the influence of temperature on the efficacy of 2,4-D or glyphosate for common and giant ragweed control and the level of glyphosate resistance and (2) determine the underlying physiological mechanisms (absorption and translocation). Glyphosate-susceptible (GS) and glyphosate-resistant (GR) common and giant ragweed biotypes from Nebraska were used for glyphosate dose–response studies, and GR biotypes were used for 2,4-D dose–response studies conducted at two temperatures (day/night [d/n]; low temperature [LT]: 20/11 C d/n; high temperature [HT]: 29/17 C d/n). Results indicate improved efficacy of 2,4-D or glyphosate at HT compared with LT for common and giant ragweed control regardless of susceptibility or resistance to glyphosate. The level of glyphosate resistance decreased in both the species at HT compared with LT, primarily due to more translocation at HT. More translocation of 2,4-D in GR common and giant ragweed at HT compared with LT at 96 h after treatment could be the reason for improved efficacy. Similarly, higher translocation in common ragweed and increased absorption and translocation in giant ragweed resulted in greater efficacy of glyphosate at HT compared with LT. It is concluded that the efficacy of 2,4-D or glyphosate for common and giant ragweed control can be improved if applied at warm temperatures (29/17 C d/n) due to increased absorption and/or translocation compared with applications during cooler temperatures (20/11 C d/n).

The effect of reduced light intensity on the growth and development of three common grass weeds, blackgrass, silky windgrass, and annual bluegrass, was studied. Two identical greenhouse experiments displaced in time were performed with six light levels aiming at 0%, 20%, 50%, 80%, 90%, and 95% shade corresponding to a mean daily light integral (DLI) of 12.4, 9.63, 7.13, 2.74, 0.95, and 0.69 mol m−2 d−1 in experiment 1 and 21.2, 18.0, 10.7, 3.71, 1.64, 1.20 mol m−2 d−1 in experiment 2. Climate screens of acrylic fabric were used to create the light levels. A DLI of 0.69 to 3.71 mol m−2 d−1 substantially reduced the plant height, the number of leaves, leaf chlorophyll content index, stomatal conductance, maximum photochemical efficiency of photosystem II, and dry matter of blackgrass. It also reduced plant height, the number of leaves, and dry matter and delayed flowering of windgrass and annual bluegrass. Annual bluegrass reacted most rapidly when light levels increased from the lowest levels by producing more leaves. DLI thresholds for blooming were estimated to be about 7.13 mol m−2 d−1 for windgrass and 1.64 mol m−2 d−1 for annual bluegrass. Annual bluegrass was able to bloom and sustain biomass even at a DLI of 1.64 mol m−2 d−1. This ability may contribute to an explanation of why annual bluegrass is among the most common weed species in highly competitive and well-fertilized crops even though it is much smaller than the two other grass species.

Timing of weed emergence and seed persistence in the soil influence the ability to implement timely and effective control practices. Emergence patterns and seed persistence of kochia populations were monitored in 2010 and 2011 at sites in Kansas, Colorado, Wyoming, Nebraska, and South Dakota. Weekly observations of emergence were initiated in March and continued until no new emergence occurred. Seed was harvested from each site, placed into 100-seed mesh packets, and buried at depths of 0, 2.5, and 10 cm in fall of 2010 and 2011. Packets were exhumed at 6-mo intervals over 2 yr. Viability of exhumed seeds was evaluated. Nonlinear mixed-effects Weibull models were fit to cumulative emergence (%) across growing degree days (GDD) and to viable seed (%) across burial time to describe their fixed and random effects across site-years. Final emergence densities varied among site-years and ranged from as few as 4 to almost 380,000 seedlings m−2. Across 11 site-years in Kansas, cumulative GDD needed for 10% emergence were 168, while across 6 site-years in Wyoming and Nebraska, only 90 GDD were needed; on the calendar, this date shifted from early to late March. The majority (>95%) of kochia seed did not persist for more than 2 yr. Remaining seed viability was generally >80% when seeds were exhumed within 6 mo after burial in March, and declined to <5% by October of the first year after burial. Burial did not appear to increase or decrease seed viability over time but placed seed in a position from which seedling emergence would not be possible. High seedling emergence that occurs very early in the spring emphasizes the need for fall or early spring PRE weed control such as tillage, herbicides, and cover crops, while continued emergence into midsummer emphasizes the need for extended periods of kochia management.

Locoweeds are plants of the genera Astragalus and Oxytropis (Fabaceae family) and are toxic to cattle, sheep, and horses. The toxic property of locoweeds is due to the alkaloid swainsonine (SWA), which is synthesized by an endophytic fungus Alternaria spp. section Undifilum. Although the endophyte–locoweed complex is often considered mutualistic, empirical evidence for benefits to host plants is lacking. This study: 1) compared the growth, photosynthesis, and leaf pigment and antioxidant concentrations between endophyte-infected and endophyte-free plants under well-watered and water-deficit conditions; and 2) measured SWA to determine whether SWA concentrations are attenuated by water deficit and leaf age. Locoweed species in this study were woolly loco and silky crazyweed. Endophyte-infected and endophyte-free (by removal of seed coat) seedlings, as confirmed by DNA analyses, were grown under greenhouse conditions for 6 mo, after which plants were subjected to three 12- to 15-d water-deficit periods that created sublethal drought conditions. Results suggest that the endophyte did not influence photosynthetic gas exchange and leaf pigment concentrations. Under well-watered conditions only, endophyte-infected woolly loco plants had lower shoot and root biomass and higher concentrations of α-tocopherol than endophyte-free plants. SWA analyses revealed taxon-specific effects of water deficit, with water deficit increasing SWA concentrations in young leaves of woolly loco but not affecting SWA concentration in silky crazyweed. These results suggest that the endophyte behaves as a parasite in woolly loco plants grown under optimal but not under water-limited conditions. Further, results indicate that drought conditions elevate the toxicity of woolly loco plants. Improved understanding of endophyte-locoweed interactions and factors influencing SWA levels will contribute to the development of livestock management strategies to predict toxicity in particular locoweed populations.

Weed management is a major constraint in organic cropping systems. In 2004, the Cornell Organic Vegetable Cropping Systems Experiment was established in central New York state using a split-plot randomized complete block design with two crop rotation entry points (split-plot factor). Four organic vegetable cropping systems that varied in cropping intensity and tillage (main plot factor) were compared: (1) intensive, (2) intermediate, (3) bio-extensive, and (4) ridge tillage. The basic crop rotation was cabbage, lettuce, potato, and winter squash, with additional short-season crops in the intensive system and with cover crops and fallow substituted for cabbage and potato in the bio-extensive system. In 2014, two uniformity trials were conducted in which oat and then a mixture of sorghum-sudangrass plus Japanese millet were grown uniformly over the entire experiment. Prior to sowing oat, soil samples were collected from each plot and an emergence bioassay was conducted to assess the soil weed seedbank. Crop biomass, weed density, and weed biomass were sampled in the uniformity crops. Soil weed seedbank density was three to four times greater in the intensive, intermediate, and ridge-tillage systems than in the bio-extensive system. The bio-extensive system also had lower weed density and weed biomass in the oat uniformity trial compared with the other three systems. Oat biomass did not differ between the cropping systems. Weed density and biomass in oat were also affected by the crop rotation entry point. Cropping system legacy effects on weed abundance and community composition were greater in the oat than in the sorghum-sudangrass plus Japanese millet uniformity trial. Our results illustrate the effects of different organic vegetable production practices on weed community structure and highlight the value of tilled fallow periods, cover crops, and prevention of weed seed rain for reducing weed populations.

The Harrington Seed Destructor (HSD), a novel weed control technology, has been highly effective in Australian cropping systems. To investigate its applicability to conditions in western Canada, stationary threshing was conducted to determine the impact of weed species, seed size, seed number, chaff load, and chaff type on efficacy of seed destruction. Control varied depending on species, with a range of 97.7% to 99.8%. Sieve-sized volunteer canola seed had a linear relationship of increasing control with increasing 1,000-seed weight. However, with greater than 98% control across all tested seed weights, it is unlikely that seed size alone will significantly influence control. Consistently high levels of control were observed at all tested seed densities (10 seeds to 1 million seeds). The response of weed seed control to chaff load was quadratic, but a narrow range of consistently high control (>97%) was again observed. Chaff type had a significant effect on weed seed control (98% to 98.6%); however, seed control values in canola chaff were likely confounded by a background presence of volunteer canola. Overall, the five parameters studied statistically influence control of weed seeds with the HSD. However, small differences between treatments are unlikely to affect the biological impact of the machine, which provides high levels of control for those weed seeds that can be introduced into the harvester.

Weeds are ubiquitous and economically damaging in southern U.S. rice systems. Barnyardgrass has consistently been one of the most prevalent and troublesome of these. Although most rice cultivars do not suppress weeds dramatically, certain Indica cultivars and commercial hybrids are known to suppress barnyardgrass aggressively in conventional, drill-seeded rice systems in the southern United States. A field study was conducted to determine the degree to which either reducing or increasing standard seeding rates would affect natural suppression of weeds by conventional inbred and weed-suppressive cultivars. Five cultivars were evaluated at three seeding rates (160 [low], 320 [medium; conventional recommendation for inbred cultivars], and 480 [high] seeds m−2) and two weed levels (weed-free and weedy). Cultivars included a conventional, non–weed suppressive long-grain, ‘Wells’; high-tillering weed-suppressive cultivars ‘PI312777,’ ‘Rondo,’ and ‘4612’ from Asia; and the commercial hybrid ‘XL723.’ Overall, PI 312777 produced the most tillers, whereas XL 723 exhibited the greatest midseason shoot biomass and the greatest weed suppression. Yields of PI 312777 and 4612, both of which are Indica cultivars considered to be good weed suppressors, changed minimally across all seeding rates when compared with the other cultivars and thus tolerated weeds at the low rate nearly as well as at the high rate. Such a tolerance to weeds might be useful in the maintenance of weed suppression at reduced rice-seeding rates and suggests that reduced seeding rates of PI 312777 and 4612 would be less risky for yield loss when compared with the other cultivars tested. Visual suppression ratings were positively correlated with rice yield within weed-infested plots, suggesting that yield performance under weed pressure might be a good indicator of weed-suppression ability of cultivars in these systems. In contrast with PI 312777 and 4612, yields of the conventional inbred cultivar and commercial hybrid appeared to benefit from the high seeding rate. Overall, moderate to high seeding rates are likely to be needed for consistent weed suppression for all of the cultivar types evaluated in this study.

Intercropping with functionally diverse crops can reduce the availability of resources that could otherwise be used by weeds. An experiment was conducted across 6 site-years in New York and Maryland in 2013 and 2014 to examine the effects of functional diversity and crop species richness on weed suppression. We compared four annual crop species that differed in stature and nitrogen acquisition traits: (1) pearl millet, (2) sorghum sudangrass, (3) cowpea, and (4) sunn hemp. Crops were seeded in monoculture and in three- and four-species mixtures using a replacement design in which monoculture seeding rates were divided by the number of species in the intercrop. Crop and weed biomass were sampled at ~45 and 90 d after planting. At the first sampling date, intercrops produced more crop biomass than monocultures in all but 1 site-year; however, weed biomass in intercrops was lower than monocultures in only 1 site-year. By the second sampling date, crop biomass was consistently greater in the intercrops than in the monocultures, and weed biomass was lower in the intercrops than in monocultures in 2 site-years. Although we observed several negative relationships between crop species richness and weed biomass, crop biomass was a more important factor than species richness for suppressing weeds. Despite the weak weed suppression from the two legumes compared with the two grasses, legume crops can provide other benefits, including increased forage quality, soil nitrogen for subsequent crops, and resources for pollinators if allowed to flower. On the other hand, if weed suppression is the top priority, our results suggest that monocultures of high biomass–producing grasses will provide more effective suppression at a lower seed cost than functionally diverse intercrops that include low biomass–producing legumes in warm-season intercrops.