Galen Brunk

The importance of analytical techniques in allelopathy studies with the reported allelochemical catechin as an example

Allelopathy can be challenging to demonstrate. Developing rigorous analytical techniques to detect and quantify compound(s) of interest from soil or liquid media lays the foundation for designing ecologically relevant experiments that incorporate candidate allelochemicals. In this paper, fundamental components of analytical techniques, including method development, validation, and appropriate controls are discussed. Research on the candidate allelochemical from spotted knapweed, catechin, is used as an example to demonstrate the importance of including these components both during data collection and in subsequent publications. This example shows how contrasting results between research groups can be difficult to interpret when information on controls and method validation are not included in publications. Recent research suggests that catechin is not likely driving spotted knapweed’s invasion, and thus future research on this system should focus on alternate candidate toxins from spotted knapweed. By employing appropriate analytical techniques, such as those outlined here, a strong foundation can be laid for ecologically oriented experiments that examine the role of allelochemicals in structuring communities.

Aminopyralid and Clopyralid Absorption and Translocation in Canada Thistle (Cirsium arvense)

Abstract Aminopyralid is a new auxinic herbicide that provides Canada thistle control at lower use rates than clopyralid. Studies were conducted to determine if differences in absorption, translocation, or metabolism account for aminopyralids greater biological activity. Radiolabeled aminopyralid and clopyralid were applied to individual leaves of rosette-stage Canada thistle plants. Nonionic surfactant was used for the absorption studies because it provided higher aminopyralid absorption than methylated seed oil or crop oil concentrate. Clopyralid was absorbed very rapidly, reaching 72% 24 h after treatment (HAT) and remaining near or above 80% during a 192-h time course. During the same time period, aminopyralid absorption increased from 34 to 60%. Clopyralid translocation out of the treated leaf was significantly higher than aminopyralid, 39% compared with 17%, respectively, 192 HAT. More of applied clopyralid translocated to aboveground tissue 192 HAT (27%) than to roots (12%), whereas aminopyralid translocation was similar in aboveground tissue (10%) and roots (7%) 192 HAT. Neither aminopyralid nor clopyralid was metabolized 192 HAT. Although aminopyralid is effective at lower use rates than clopyralid, clopyralid absorption and translocation were higher in Canada thistle. These results suggest that aminopyralids chemical structure may provide for greater biological activity at the target site than clopyralid. Nomenclature: Aminopyralid; clopyralid, Canada thistle, Cirsium arvense (L.) Scop. CIRAR

Vapor Movement of Synthetic Auxin Herbicides: Aminocyclopyrachlor, Aminocyclopyrachlor-Methyl Ester, Dicamba, and Aminopyralid

Abstract Aminocyclopyrachlor, a newly discovered synthetic auxin herbicide, and its methyl ester, appear to control a number of perennial broadleaf weeds. The potential volatility of this new herbicide and its methyl ester were determined under laboratory conditions and were also compared to dicamba and aminopyralid with the use of enclosed chamber and open-air plant bioassays. Bioassays consisting of visual estimates of epinastic responses and kidney bean and soybean leaf-width measurements were developed to measure vapor release from glass and plastic. Vapor release of aminocyclopyrachlor from glass surfaces was undetectable under laboratory conditions, and no phytotoxic responses were observed when plants were exposed to vapors emanating from various surfaces. Results were similar to those of aminopyralid, indicating the risk of plant injury from vapor movement of aminocyclopyrachlor and aminopyralid was very low. When combined with 1% methylated seed oil, vapor release of aminocyclopyrachlor-methyl ester reached 86% 192 h after application to glass surfaces. Phytotoxic responses of plants exposed to vapors emanating from various surfaces treated with aminocyclopyrachlor-methyl ester were similar to responses to dicamba under enclosed incubation conditions, but were less in outdoor, open-air environments. Studies are needed to understand better the risk of injury to nontarget plants due to vapor movement of aminocyclopyrachlor-methyl ester under field applications. Nomenclature: Aminocyclopyrachlor aminopyralid kidney bean, Phaseolus vulgaris, ‘Green potted bush’ soybean, Glycine max (L.) Merr. ‘Pioneer 94B53’

Herbivory and novel weapons: no evidence for enhanced competitive ability or allelopathy induction of Centaurea diffusa by biological controls

Biological control of weeds by arthropod herbivores is thought to work by reducing the competitive ability of the weed relative to the surrounding vegetation. However, the assumption that herbivory reduces plant competitive ability has not been tested in most biological control systems, and counter to expectation, recent research on the impact of biological control agents on invasive Centaurea species suggests that this genus may respond to herbivory by increased competitive ability through enhanced plant re-growth and/or by inducing increased production of phytotoxic allelochemicals. We examined the impact of two biological control agents of the invasive plant diffuse knapweed (C. diffusa) to see if feeding by either of these insects would enhance the plant’s competitive ability or allelochemical output. Sub-lethal herbivory by either of the biological control agents significantly reduced knapweed performance when the plant was grown in competition with either of two native species. Competition with knapweed significantly reduced the performance of both native species (Artemisia frigida and Bouteloua gracilis), and herbivory by one of the biocontrol agents resulted in a small but significant increase in both native species’ performance. Diffuse knapweed’s putative allelochemical 8-hydroxyquinoline was not detected in experimental or field collected soils from knapweed-infested sites. In contrast to other studies on the impacts of biological control on other Centaurea species, these data support the premise that biological control agents may reduce invading plant competitive ability. We find no evidence for diffuse knapweed allelopathy mediated by 8-hydroxyquinoline or enhanced allelopathy in response to herbivory by biological control agents.

Aminocyclopyrachlor Absorption, Translocation and Metabolism in Field Bindweed (Convolvulus arvensis)

Abstract 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. Nomenclature: Field bindweed, Convolvulus arvensis L.; aminocyclopyrachlor.

Role of Soil Sorption and Microbial Degradation on Dissipation of Mesotrione in Plant-Available Soil Water

Mesotrione is a carotenoid biosynthesis-inhibiting herbicide labeled for pre-emergence and postemergence weed control in corn production. Understanding the factors that influence the dissipation of mesotrione in soil and in the plant-available water (PAW) is important for the environmental fate assessment and optimal weed management practices. The present research investigated the role of soil properties and microbial activities on the interrelated sorption and degradation processes of mesotrione in four soils by direct measurements of PAW. We found that mesotrione bound to the soils time dependently, with approximately 14 d to reach equilibrium. The 24-h batch-slurry equilibrium experiments provided the sorption partition coefficient ranging from 0.26 to 3.53 L kg(-1), depending on soil organic carbon and pH. The dissipation of mesotrione in the soil-bound phase was primarily attributed to desorption to the PAW. Degradation in the PAW was rapid and primarily dependent on microbial actions, with half-degradation time (DT(50)) <3 d in all four soils tested. The rapid degradation in the PAW became rate limited by sorption as more available molecules were depleted in the soil pore water, resulting in a more slowed overall process for the total soil-water system (DT(50) <26 d). The dissipation of mesotrione in the PAW was due to microbial metabolism and time-dependent sorption to the soils. A coupled kinetics model calibrated with the data from the laboratory centrifugation technique provided an effective approach to investigate the interrelated processes of sorption and degradation in realistic soil moisture conditions.

Comparison of the Interactions of Aminopyralid vs. Clopyralid with Soil

Abstract Laboratory studies were conducted to compare the soil adsorption of aminopyralid and clopyralid with the use of batch-slurry and centrifugation assays. The calculated soil binding constants for both herbicides varied between the two techniques, but the centrifugation assay had a lower coefficient of variation compared to the batch-slurry assay. These results indicate that a centrifugation assay is a more accurate procedure for measuring the interaction of aminopyralid and clopyralid with soils. Aminopyralid adsorbed more tightly than clopyralid to six of the eight soils tested. Adsorption Kd values ranged from 0.083 to 0.364 for clopyralid and 0.106 to 0.697 for aminopyralid. Pearson correlation analysis indicated that binding of both herbicides was highly correlated to soil organic matter and texture but not to soil pH. On average, soil thin-layer chromatography indicated that aminopyralid was less mobile (Rf  =  0.82) than clopyralid (Rf  =  0.91), although both were mobile. These results suggest that aminopyralid will have a lower leaching potential than clopyralid. Lower potential aminopyralid soil leaching, coupled with low use rates, suggests it may be the herbicide of choice in areas where potential for leaching could be a concern. Nomenclature: Aminopyralid; clopyralid.

Typhula blight development in Poa annua and Poa pratensis as influenced by persistence of the fungicides chlorothalonil and fludioxonil under snow cover

Abstract Typhula blight is a major problem on golf courses where snow cover persists for long periods. The disease is primarily managed by applying a mixture of fungicides in the autumn prior to winter snow cover. The fungicides chlorothalonil and fludioxonil are often included in these mixtures even though they do not completely suppress Typhula blight when applied alone. We studied whether minimal control was due to a lack of persistence of these fungicides under snow, or whether certain isolates of Typhula ishikariensis and T. incarnata exhibited tolerance to these fungicides. The persistence of chlorothalonil and fludioxonil residues in the turf was determined during the winters of 2005–06, 2006–07 and 2007–08. Concentrations of chlorothalonil and fludioxonil in the verdure decreased or remained nearly the same at most sampling dates, indicating these fungicides did not dissipate rapidly under snow. Despite this, only marginal control of Typhula blight was observed in the fungicide-treated plots. The in vitro sensitivity of T. ishikariensis and T. incarnata isolates to chlorothalonil and fludioxonil was tested. Growth of most isolates (70%) on agar amended with 1 µg mL−1 chlorothalonil was inhibited by more than 50% relative to growth on non-amended agar. However, almost all isolates exhibited some growth at concentrations as high as 500 µg mL−1. A high proportion of isolates (85%) were inhibited by greater than 80% at 1 µg mL−1 fludioxonil. Therefore, at least some growth of these isolates at high fungicide concentrations may explain why chlorothalonil and fludioxonil are not completely effective in suppressing Typhula blight.