Effect of New Auxin Herbicide Formulations on Control of Herbicide Resistant Weeds and on Microbial Activities in the Rhizosphere

Widespread distribution of glyphosate-resistant weeds in soybean-growing areas across Mississippi has economically affected soybean planting and follow-up crop management operations. New multiple herbicide-resistant crop (including soybean) technologies with associated formulations will soon be commercialized. The objectives of this research were to determine the efficacy of new 2,4-D + glyphosate and dicamba formulations on herbicide resistant weeds, and to determine the impact of the new 2,4-D + glyphosate formulation on microbial communities in the soybean rhizosphere involved in nutrient cycling. New 2,4-D + glyphosate and dicamba formulations registered for use on 2,4-D and dicamba-resistant soybean, respectively, adequately controlled glyphosate resistant and susceptible pigweeds (Palmer amaranth and tall waterhemp) and common ragweed. The 2,4-D + glyphosate formulation did not significantly impact soil microbial activities linked to nutrient cycling in the soybean rhizosphere. These results indicate these new 2,4-D + glyphosate and dicamba formulations can be effective in controlling glyphosate resistant and other herbicide resistant weeds while not having adverse effects on the activities of beneficial soil microorganisms.

Target Site-Based Resistance to ALS Inhibitors, Glyphosate, and PPO Inhibitors in an <i>Amaranthus palmeri</i>Accession from Mississippi

Extensive acceptance of glyphosate-resistant (GR) row crops coupled with the simultaneous increase in glyphosate usage has sped the evolution of glyphosate resistance in economically important weeds. GR Amaranthus palmeri populations are widespread across the state with some exhibiting multiple resistance to acetolactate synthase (ALS) inhibiting herbicides such as pyrithiobac. A GR and ALS inhibitor-resistant accession was also resistant to the protoporphyrinogen oxidase (PPO) inhibiting herbicide fomesafen. The PPO inhibitor resistance profile and multiple herbicide resistance mechanisms in this accession were investigated. In addition to fomesafen, resistance to postemergence applications of acifluorfen, lactofen, carfentrazone, and sulfentrazone was confirmed. There was no resistance to preemergence application of fomesafen, flumioxazin, or oxyfluorfen. Molecular analysis of the ALS gene indicated the presence of point mutations leading to single nucleotide substitutions at codons 197, 377, 574, and 653, resulting in proline-to-serine, arginine-to-glutamine, tryptophan-to-leucine, and serine-to-asparagine replacements, respectively. The resistant accession contained up to 87-fold more copies of the EPSPS gene compared to a susceptible accession. A mutation leading to a deletion of glycine at codon 210 (ΔG210) of PPO2 gene was also detected. These results indicate that the mechanism of resistance in the Palmer amaranth accession is target-site based, i.e., altered target site for ALS and PPO inhibitor resistance and gene amplification for glyphosate resistance.

Morphological Characterization of <i>Amaranthus palmeri</i>x <i>A. spinosus</i>Hybrids

The growth of clones of seven Amaranthus palmeri x A. spinosus hybrids was compared to type specimens of A. palmeri and A. spinosus. The hybrids came from the field where they were originally discovered and clones of the type specimens and hybrids were established under greenhouse conditions and used to compare growth rates. A. palmeri had the highest growth rate and A. spinosus the lowest growth rate based on height, node counts, and dry weight accumulation. A. palmeri also had the greatest number of days to flowering and A. spinosus the fewest. Hybrids had intermediary growth rates and days to flowering, but differed from each other with regard to sex identity. The hybrids were either dioecious like A. palmeri or, if monoecious, had patterns unlike A. spinosus. Spine length and texture also varied in hybrids and some were without spines. Hybrid 16Ci was short compared to all others and had succulent leaves and stems, which easily separated from the plant body. These hybridizations resulted in morphologically distinct types with acquisition of physical traits intermediate to the type specimens which may drive evolution of these species.

Italian Ryegrass (Lolium perenne ssp. multiflorum) and Corn (Zea mays) Competition

Italian ryegrass is an annual/biennial grass that is typically used as a pasture crop or a cover crop along roadsides, rights-of-way, and industrial areas. Glyphosate-resistant (GR) Italian ryegrass populations have been documented around the world, mostly in orchard and vineyard situations. The first evidence of evolved GR Italian ryegrass in row/agronomic crops was reported from Washington County, Mississippi in 2005. GR Italian ryegrass populations can jeopardize preplant burndown options in reduced-tillage crop production systems, thereby, delaying planting operations. The effects of competition of Italian ryegrass on crop growth and yield are poorly understood. A field study was conducted in the 2012 growing season and repeated in the 2013 growing season. GR and susceptible (GS) Italian ryegrass populations were established in the greenhouse and transplanted in prepared corn row beds in the fall of 2011 and 2012 at 0, 1, 2, 3, and 4 plants·meter> of crop row. Italian ryegrass plants overwintered and developed over the following spring-summer. Glyphosate was applied at 1.26 kg·ae/ha (1.5× of labeled rate) in the spring to burndown the Italian ryegrass plants and corn was planted into the ryegrass residue 2 - 3 wk later. Current corn production practices were followed. Corn density (early and late season), height (early season), and yield and Italian ryegrass biomass (early-mid season) measurements were recorded during both years. Corn height was greater in 2012 than that in 2013 at comparable stages of the growing season, due to a cooler and wetter early season in 2013 than that in 2012. Averaged across weed densities, corn density (both early and late season) and yield were higher in the GS than those in the GR population, but Italian ryegrass biomass was similar for both populations. Averaged across Italian ryegrass populations, corn density (both early and late season), and yield were inversely proportional to Italian ryegrass density. In summary, Italian ryegrass significantly reduced corn density and yield and reduction was greater with the GR than that with the GS population. Studies are underway to study inter population competition in Italian ryegrass and investigate allelopathic effects of Italian ryegrass on selected crops.

Hyperspectral Imaging for Differentiating Glyphosate-Resistant and Glyphosate-Susceptible Italian Ryegrass

Glyphosate is widely used in row crop weed control programs of glyphosate-resistant (GR) crops. With the accumulation of glyphosate use, several weeds have evolved resistance to glyphosate. In order to control GR weeds for profitable crop production, it is critical to first identify them in crop fields. Conventional method for identifying GR weeds is destructive, tedious and labor-intensive. This study developed hyperspectral imaging for rapid sensing of Italian ryegrass (Lolium perenne ssp. multiflorum) plants to determine if each plant is GR or glyphosate-susceptible (GS). In image analysis, a set of sensitive spectral bands was determined using a forward selection algorithm by optimizing the area under the receiver operating characteristic between GR and GS plants. Then, the dimensionality of selected bands was reduced using linear discriminant analysis. At the end the maximum likelihood classification was conducted for plant sample differentiation of GR Italian ryegrass from GS ones. The results indicated that the overall classification accuracy is between 75% and 80%. Although the accuracy is lower than the classification of Palmer amaranth (Amaranthus palmeri S. Wats.) in our previous study, this study provides a rapid, non-destructive approach to differentiate between GR and GS Italian ryegrass for improved site-specific weed management.

Influence of Water Quality, Formulation, Adjuvant, Rainfastness, and Nozzle Type on Efficacy of Fomesafen on Palmer Amaranth (<i>Amaranthus palmeri</i>) Control

Protoporphyrinogen oxidase (PPO) inhibitors are one of the few remaining postemergence herbicide options for controlling Palmer amaranth in soybean growing areas of Mississippi, USA. Most Palmer amaranth populations in Mississippi are resistant to both glyphosate and acetolactate synthase inhibitors. Resistance to PPO inhibiting herbicides in Palmer amaranth has very recently been reported in Arkansas, Tennessee, and isolated pockets of Mississippi. A significant proportion of reports of PPO inhibitor failures in Mississippi are not considered to be resistance-related at this time. Therefore, the objective of this research was to evaluate factors affecting the efficacy of fomesafen on Palmer amaranth including: quality of spray carrier (water), formulations, adjuvant, rainfastness, and nozzle type. All water samples and formulation combinations provided >95% control of Palmer amaranth 3 WAT. Some combinations of water samples and formulations did not result in complete control of the treated plants, with one or two individuals surviving 3 WAT. Formulation 1 provided 99% control compared to 95% from formulation 2. Irrespective of combinations of herbicide, adjuvant and height, control of Palmer amaranth was ≥91%. Formulation 1 provided 94% control compared to 88% from formulation 2. The adjuvant x height interaction was significant, owing to a 10% reduction in control of larger plants (86%) compared to smaller plants (96%) in presence of COC. COC provided better control (93%) than NIS (88%). Simulated rainfall applied ≥60 min after herbicide application did not adversely affect efficacy on Palmer amaranth when formulation 1 was applied in combination with NIS, with control ranging from 94% to 100%. Formulation 1 with COC provided ≥93% control at all rainfall application times, except 30 min after herbicide treatment, which resulted in 79% control. Formulation 2 provided better control with COC (79% to 100%) than NIS (71% to 90%), in general, across the rainfall treatments applied at various times following herbicide application. All nozzle and weed height combinations resulted in 89% or better control of Palmer amaranth. In summary, water quality, formulation, adjuvant, rainfastness, or nozzle type did not affect the activity of fomesafen under optimal application conditions in the greenhouse.

Late-Season Grass Weed Management with In-Crop and Post-Harvest Herbicides in Twin-Row Glyphosate-Resistant Soybean

Emergence of grasses late in the season has become a problem in glyphosate-resistant (GR) soybean production in the southern US. A 3-yr field study was conducted from 2011 to 2013 at Stoneville, MS to determine efficacy of post-harvest and pyroxasulfone-based in-crop herbicides on late-season grasses and yield in twin-row glyphosate-resistant soybean. Experiments were conducted in a split-plot arrangement of treatments in a randomized complete block design with fall herbicides (with and without pendimethalin at 1.12 kg ai ha-1 and paraquatat 0.84 kg ai ha-1) as main plots and in-crop herbicides as subplots with four replications. The six in-crop herbicide programs were: glyphosate applied early postemergence (EPOST) at 0.84 kg·aeha-1 followed by (fb) glyphosate late postemergence (LPOST) at 0.84 kg·ha-1 with and without pyroxasulfone preemergence (PRE) applied at 0.18 kg ai ha-1, pyroxasulfone PRE fb glyphosate at 0.84 kg·ha-1 LPOST or glyphosate at 0.84 kg·ha-1 + S-metolachlor at 1.68 kg ai ha-1 EPOST, pyroxasulfone PRE fb S-meto- lachlor at 1.12 kg·ha-1 + fomesafen at 0.27 kg ai ha-1 EPOST fb clethodim at 0.14 kg ai ha-1, and a no-herbicide control. Browntop millet, Digitaria spp., and junglerice densities at 2 weeks after LPOST, grass weed dry biomass at harvest, and soybean yield were similar regardless of post- harvest herbicides in all three years. At 2 weeks after LPOST, browntop millet, Digitaria spp. and junglerice densities were greatly reduced in all five in-crop herbicide treatments compared with no herbicide plot in all three years. Grass weed dry biomass in no-herbicide plots was 3346, 6136, and 6916 kg·ha-1 in 2011, 2012, and 2013, respectively and the five herbicide treatments reduced grass weed dry biomass by at least 87%, 84%, and 99% in 2011, 2012, and 2013, respectively. Soybean yield was higher with all five in-crop herbicide treatments compared to no herbicide control in all three years. These results indicate that browntop millet, Digitaria spp., and junglerice infestations can be reduced with pyroxasulfone-based in-crop herbicide programs in twin-row GR soybean.

Glyphosate Resistance in Giant Ragweed (<i>Ambrosia trifida</i>L.) from Mississippi Is Partly Due to Reduced Translocation

A giant ragweed population from a glyphosate-resistant (GR) soybean field in Mississippi, USA was suspected to be resistant to glyphosate. Greenhouse and laboratory studies were conducted to confirm and quantify the magnitude of glyphosate resistance in a resistant biotype selected from this population and to elucidate possible physiological and molecular mechanisms of glyphosate resistance. Glyphosate dose response studies indicated that ED50 (effective dose required to reduce plant growth by 50%) values for glyphosate-resistant (GR-MS) and glyphosate-susceptible (GS-MS) biotypes, based on percent injury, were 0.52 and 0.34 kg ae/ha glyphosate, respectively, indicating a 1.5-fold level of resistance in GR-MS. The absorption pattern of 14C-glyphosate in the two giant ragweed biotypes was similar throughout the measured time course of 168 h after treatment (HAT). The amount of 14C-glyphosate that translocated out of treated leaves of the GR-MS and GS-MS plants was similar up to 24 HAT. However, the GS-MS biotype translocated more (71% and 76% of absorbed at 48 and 96 HAT, respectively) 14C-glyphosate than the GR-MS biotype (44% and 66% of absorbed at 48 and 96 HAT, respectively) out of the treated leaf. No target site mutation was identified at the Pro106 location of the EPSPS gene of the GR-MS biotype. The mechanism of resistance to glyphosate in giant ragweed from Mississippi, at least, is due to reduced glyphosate translocation.

Morpho-Physiological Characterization of Glyphosate-Resistant and -Susceptible Horseweed (Conyza canadensis) Biotypes of US Midsouth

Horseweed is traditionally considered a non-cropland weed. However, populations resistant to glyphosate have eventually become established in no-till agronomic cropping systems. Growth chamber and greenhouse experiments were conducted to compare selected biological and physiological parameters of glyphosate-resistant (GR) and -susceptible (GS) horseweed biotypes from Mississippi with a broader goal of fitness characterization in these biotypes. Vegetative growth parameters (number of leaves, rosette diameter and area, shoot and root fresh weights) were recorded weekly from 5 to 11 wk after emergence and reproductive attributes [days to bolting (production of a flowering stalk) and flowering] and senescence were measured for both GR and GS biotypes under high (24°C/20°C) and low (18°C/12°C) temperature regimes, both with a 13-h light period. Physiological traits such as net photosynthesis, phenolic content, and cell membrane thermostability, all in the presence and absence of glyphosate, and leaf content of divalent cations such as Ca2+ and Mg2+ were assayed in the two biotypes under the high temperature regime. All horseweed vegetative growth parameters except root fresh weight were higher in the high temperature regime compared to that in low temperature regime in both biotypes. Number of leaves, rosette diameter and area, shoot and root fresh weight were 40 vs. 35, 9.3 vs. 8.7 cm, 51 vs. 43 cm2, 3.7 vs. 3.2 g, and 3.5 vs. 4.2 g under high and low temperature conditions, respectively, when averaged across biotypes and weekly measurements. All growth parameters listed above were higher for the GR biotype compared to the GS biotype. Number of leaves, rosette diameter and area, shoot and root fresh weight were 38 vs. 37, 9.1 vs. 8.9 cm, 50.2 vs. 44 cm2, 3.9 vs. 3.1 g, and 4.3 vs. 3.5 g for GR and GS biotypes, respectively, averaged across the temperature treatments and weekly measurements. Reproductive developmental data of these biotypes indicated that the GS biotype bolted earlier than the GR biotype. The GS biotype had more phenolic content and exhibited higher cell membrane thermostability, but less net photosynthetic rate compared to the GR biotype. At 48 h after treatment with glyphosate, there was no change in phenolic content of both GR and GS biotypes. However, glyphosate reduced cell membrane thermostability and net photosynthetic rate more in the GS biotype than that in the GR biotype. Chemical analysis of GR and GS leaf tissue did not reveal any differences in levels of divalent cations such as Ca2+ and Mg2+. Further studies are needed to determine if some of the differences between the two biotypes observed above relate to fitness variation in a natural environment.