The influence of environmental factors on germination and emergence of horseweed was examined in growth chamber experiments. Germination was highest (61%) under 24/20 C day/night temperature under light. Horseweed seed germination was observed under both light (13 h photoperiod) and complete darkness (24 h), but germination under continuous darkness was only 0 to 15% compared with 0 to 61% under light. All other experiments were conducted under 24/20 C and 13-h light conditions. Germination was 19 to 36% over a pH range from 4 to 10, with a trend toward higher germination under neutral-to-alkaline conditions. Horseweed germination was > 20% at < 40 mM NaCl concentration and lowest (4%) at 160 mM NaCl. These data suggest that even at high soil salinity conditions, horseweed can germinate. Germination of horseweed decreased from 25% to 2% as osmotic potential increased from 0 (distilled water) to −0.8 MPa, indicating that germination can still occur under moderate water stress conditions. Horseweed seedling emergence was at its maximum on the soil surface, and no seedlings emerged from seeds placed at a depth of 0.5 cm or higher.

An in vivo shikimate accumulation assay with excised leaf tissue was developed to provide a fast and reliable method for identifying glyphosate-resistant plants. The assay is based on glyphosate-induced accumulation of shikimate. There was a linear accumulation of shikimate in excised leaf discs of soybean and canola treated with 250 μM glyphosate for 48 h. The IC50 for the accumulation of shikimate in soybean and corn leaf discs was 34 and 87 μM, respectively. Leaf discs excised from glyphosate-resistant corn or soybean did not accumulate shikimate when treated with 500 μM glyphosate. Leaf discs taken from a number of field-grown plants accumulated shikimate in a glyphosate dose–dependent manner. The accumulation of shikimate was dependent on light and the age of the leaf from which the disc was taken. The assay worked either in 96-well microtiter plates or in vials, and it clearly differentiated between glyphosate-resistant and -susceptible crops in which the resistance is due to an alteration of the target site for glyphosate. The assay was simple and robust and has the potential to be used as a high throughput assay to detect glyphosate resistance in weeds.

Greenhouse and laboratory experiments were conducted to investigate mechanisms of glyphosate resistance in horseweed populations from Mississippi, Arkansas, Delaware, and Tennessee. A nondestructive leaf-dip bioassay was developed to confirm resistance and susceptibility in individual test plants. A single leaf was excised from each plant, and the petiole and bottom one-fourth of leaf was dipped in a 600 mg ae L−1 glyphosate solution for 2 d followed by visually estimating the injury on a scale of 0 to 10. Plants were classified as resistant (R) if the score was 2 to 3 and susceptible (S) if the score was 5 to 6. 14C-glyphosate solution was applied on the adaxial surface of a fully expanded leaf of the second whorl of four-whorl rosette plants. Plants were harvested 48 h after treatment and radioactivity was determined in treated leaf, other leaves, crown, and roots. Absorption of 14C-glyphosate was similar (47 to 54%) between R and S plants from within and among the four states, suggesting absorption is not involved in glyphosate resistance. The amount of radioactivity translocated from the treated leaf was reduced in R plants compared with S plants. The reduction in translocation of 14C-glyphosate ranged from 28% in Mississippi-R biotype to 48% in Delaware-R biotype compared with their respective S biotypes. Epicuticular wax mass ranged from 6 to 80 μg cm−2 among horseweed biotypes, with no differences between R and S biotypes within each state. Treating two leaves with glyphosate solution at the field use rate (0.84 kg ae ha−1) killed S plants but not R plants (38 to 58% control) regardless of state origin. These results suggest that a simple bioassay can be used to screen biotypes for suspected resistance and that reduced translocation of glyphosate plays a major role in glyphosate resistance in R biotypes of horseweed.

Field, laboratory, and greenhouse experiments were conducted to determine the seed production potential and effect of environmental factors on germination, emergence, and survival of texasweed. Texasweed produced an average of 893 seed per plant, and 90% were viable. Seed exhibited dormancy, and prechilling did not release dormancy. Percent germination ranged from 56% for seed subjected to no prechilling to 1% for seed prechilled at 5 C for 140 d. Seed remained viable during extended prechilling conditions, with 80% of seed viable after 140 d of prechilling. Texasweed seed germinated over a range of 20 to 40 C, with optimum germination (54%) occurring with a fluctuating 40/30 C temperature regime. Seed germinated with fluctuating 12-h light/dark and constant dark conditions. Texasweed seed germinated over a broad range of pH, osmotic potential, and salt concentrations. Seed germination was 31 to 62% over a pH range from 4 to 10. Germination of texasweed ranged from 9 to 56% as osmotic potential decreased from − 0.8 MPa to 0 (distilled water). Germination was greater than 52% at less than 40 mM NaCl concentrations and lowest (27%) at 160 mM NaCl. Texasweed seedlings emerged from soil depths as deep as 7.5 cm (7% emergence), but emergence was > 67% for seed placed on the soil surface or at a 1-cm depth. Texasweed seed did not germinate under saturated or flooded conditions, but seed survived flooding and germinated (23 to 25%) after flood removal. Texasweed seedlings 2.5 to 15 cm tall were not affected by emersion in 10-cm-deep flood for up to 14 d. These results suggest that texasweed seed is capable of germinating and surviving in a variety of climatic and edaphic conditions, and that flooding is not a viable management option for emerged plants of texasweed.

Field studies were conducted in 2005 and 2006 to determine the most effective chemical options within three individual herbicide-based burndown programs, glyphosate, paraquat and glufosinate, for controlling glyphosate-resistant horseweed in Mississippi. Burndown treatments were applied April 5, 2005 and March 15, 2006 to horseweed plants 15 to 30 cm in height. Glyphosate at 0.86 kg ae/ha alone provided 60 to 65% horseweed control 4 wk after treatment (WAT). Control 4 WAT ranged from 73 to 74% when the glyphosate rate was increased to 1.25 kg/ha. Glyphosate at 0.86 kg/ha applied in combination with 2,4-D at 0.84 kg ae/ha or dicamba at 0.28 ae/ha maximized control of horseweed (≥ 90%) 4 WAT and soybean yield. Horseweed control 4 WAT with paraquat alone at 0.84 kg ai/ha ranged from 55 to 63% and control did not improve by increasing the rate to 0.98 kg/ha. Addition of 2,4-D or dicamba to paraquat maximized horseweed control both years (78 to 89%), whereas soybean yield was maximized with addition of dicamba or metribuzin at 0.42 kg ai/ha. Glufosinate applied alone at 0.47 kg ai/ha resulted in at least 88% control of horseweed and maximized soybean yield. Results indicate that effective management of glyphosate-resistant horseweed can be obtained in glyphosate-resistant soybean in glyphosate-, paraquat-, and glufosinate-based preplant weed control programs.