Vince M. Davis

Review: Confirmation of Resistance to Herbicides and Evaluation of Resistance Levels

Abstract 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.

Influence of Weed Management Practices and Crop Rotation on Glyphosate-Resistant Horseweed (Conyza canadensis) Population Dynamics and Crop Yield-Years III and IV

Abstract Horseweed is an increasingly common and problematic weed in no-till soybean production in the eastern cornbelt due to the frequent occurrence of biotypes resistant to glyphosate. The objective of this study was to determine the influence of crop rotation, winter wheat cover crops (WWCC), residual non-glyphosate herbicides, and preplant application timing on the population dynamics of glyphosate-resistant (GR) horseweed and crop yield. A field study was conducted from 2003 to 2007 in a no-till field located at a site that contained a moderate infestation of GR horseweed (approximately 1 plant m−2). The experiment was a split-plot design with crop rotation (soybean–corn or soybean–soybean) as main plots and management systems as subplots. Management systems were evaluated by quantifying in-field horseweed plant density, seedbank density, and crop yield. Horseweed densities were collected at the time of postemergence applications, 1 mo after postemergence (MAP) applications, and at the time of crop harvest or 4 MAP. Viable seedbank densities were also evaluated from soil samples collected in the fall following seed rain. Soybean–corn crop rotation reduced in-field and seedbank horseweed densities vs. continuous soybean in the third and fourth yr of this experiment. Preplant herbicides applied in the spring were more effective at reducing horseweed plant densities than when applied in the previous fall. Spring-applied, residual herbicide systems were the most effective at reducing season-long in-field horseweed densities and protecting crop yields since the growth habit of horseweed in this region is primarily as a summer annual. Management systems also influenced the GR and glyphosate-susceptible (GS) biotype population structure after 4 yr of management. The most dramatic shift was from the initial GR : GS ratio of 3 : 1 to a ratio of 1 : 6 after 4 yr of residual preplant herbicide use followed by non-glyphosate postemergence herbicides. Nomenclature: Glyphosate; horseweed, Conyza canadensis (L.) Cronq. ERICA.

A Field Survey to Determine Distribution and Frequency of Glyphosate-Resistant Horseweed (Conyza Canadensis) in Indiana

In-field surveys, which directly estimate weed population densities, typically utilize either random or nonrandom field selection methods. We used both methods to characterize the distribution and frequency of glyphosate-resistant (GR) horseweed populations and other late-season soybean weed escapes and to develop a database for tracking weed shifts, control failures, and the presence of other herbicide-resistant biotypes over time in Indiana. In-field surveys were conducted in a total of 978 Indiana soybean fields during September and October of 2003, 2004, and 2005. Information from fields with horseweed was obtained from 158 sites (19%) sampled through a systematic random site selection method and 128 fields through a nonrandom site selection method. When present, horseweed seed was collected and germinated in the greenhouse; rosettes 5 to 10 cm wide were sprayed with 1.72 kg ae/ha of glyphosate. Populations with less than 60% control at 28 d after treatment were determined to be glyphosate resistant. A selected subset of glyphosate-resistant populations was confirmed resistant by subsequent glyphosate dose response experiments. All populations in the subset with less than 60% control at the 1.72 kg ae/ha rate of glyphosate demonstrated 4- to 110-fold levels of resistance (R : S ratios). Glyphosate-resistant populations were found in all regions of Indiana; however, the highest frequencies were in the southeastern (SE) region with 38% of fields sampled and only 1, 2, and 2% of fields sampled in the northwestern (NW), northeastern (NE), and southwestern (SW) regions, respectively. Information gathered in this survey can assist in the development of applied research, as well as reactive glyphosate-resistant horseweed management education in the SE region of the state. Moreover, detecting resistance at low frequencies can direct proactive resistance education to farmers and practitioners in the other regions of the state as a means of providing an early warning system to address glyphosate resistance in weeds. Nomenclature: Glyphosate, horseweed, Conyza canadensis (L.) Cronq. ERICA; soybean, Glycine max L. Merr

Cross-resistance of horseweed (Conyza canadensis) populations with three different ALS mutations

BACKGROUND Horseweed is a weed commonly found in agronomic crops, waste areas and roadsides. Resistance to ALS-inhibiting herbicides in horseweed was first reported in 1993 in a population from Israel. Resistance to ALS-inhibiting herbicides in horseweed is now widespread, but, as of now, the resistance mechanism has not been reported. RESULTS Two of three populations evaluated (P116 and P13) were found to be uniform for resistance (>98% of individuals survived 8.8 g AI ha(-1) of cloransulam), whereas a third population, P525, contained about 85% resistant individuals. Cross-resistance to cloransulam, chlorimuron, imazethapyr and bispyribac was observed in the P116 population. P525 and P13 were both sensitive to imazethapyr but resistant to chlorimuron, imazethapyr and bispyribac. Enzyme activity assays indicated that resistance in P13 was due to an altered target site. Southern blot analysis indicated that the ALS target site is encoded by a single copy gene. Overlapping ALS gene regions were amplified and sequenced from each population. Amino acid substitutions of Ser for Pro at position 197 (P197S) was detected from P13, Ala for Pro (P197A) was identified from P525 and substitution of Glu for Asp (D376E) at position 376 was found in P116. Molecular markers were developed to differentiate between wild-type and resistant codons at positions 197 and 376 of horseweed ALS. CONCLUSION Resistance to ALS-inhibiting herbicides in horseweed is conferred by target-site mutations that have also been identified in other weed species. Identification of the mutations within horseweed ALS gene sequence enables molecular assays for rapid detection and resistance diagnosis.

Growth and seed production of horseweed (Conyza canadensis) populations resistant to glyphosate, ALS-inhibiting, and multiple (glyphosate+ALS-inhibiting) herbicides.

Abstract Horseweed populations with mixtures of biotypes resistant to glyphosate and acetolactate synthase (ALS)–inhibiting herbicides as well as biotypes with multiple resistance to glyphosate + ALS-inhibiting herbicides have been documented in Indiana and Ohio. These biotypes are particularly problematic because ALS-inhibiting herbicides are commonly tank mixed with glyphosate to improve postemergence horseweed control in soybean. The objective of this research was to characterize the growth and seed production of horseweed populations with resistance to glyphosate or ALS-inhibiting herbicides, and multiple resistance to glyphosate + ALS-inhibiting herbicides. A four-herbicide by four-horseweed population factorial field experiment was conducted in the southeastern region of Indiana in 2007 and repeated in 2008. Four horseweed populations were collected from Indiana or Ohio and confirmed resistant to glyphosate, ALS inhibitors, both, or neither in greenhouse experiments. The four herbicide treatments were untreated, 0.84 kg ae ha−1 glyphosate, 35 g ai ha−1 cloransulam, and 0.84 kg ae ha−1 glyphosate + 35 g ai ha−1 cloransulam. Untreated plants from horseweed populations that were resistant to glyphosate, ALS-inhibiting, or multiple glyphosate + ALS-inhibiting herbicides produced similar amounts of biomass and seed compared to populations that were susceptible to those herbicides or combination of herbicides. Furthermore, aboveground shoot mass and seed production did not differ between treated and untreated plants. Nomenclature: Cloransulam; glyphosate; horseweed, Conyza canadensis L. ERICA; soybean, Glycine max L. Merr.

Glyphosate-resistant Horseweed (Conyza Canadensis) Emergence, Survival, and Fecundity in No-till Soybean

Abstract Horseweed has generally been considered a winter annual weed species, but efforts to control horseweed as a winter annual weed in no-till soybean production with glyphosate have routinely failed in southeast Indiana. The objective of this study was to determine emergence timing, plant survival, and fecundity of a glyphosate-resistant (GR) horseweed biotype in the presence or absence of other winter annual weeds or soybean. A field study was conducted from October 2003 to October 2004 and repeated from October 2004 to October 2005 in fields following no-till soybean production. Horseweed emergence was not observed in the fall of 2003. Winter survival of plants that emerged in the fall of 2004 was 20% by late April 2005 and was inversely related to fall rosette size. Horseweed population densities were the highest in mid-May of both years, and over 90% of the plants observed at this time emerged in the spring. Plant survival from mid-May to mid-October was 3% and 21% in 2004 and 2005, respectively. Horseweed with flower heads above the soybean canopy by early August had greater late-season survival and produced more seed than plants growing below the canopy. Horseweed with flower heads above the soybean canopy produced an average of 27,200 and 58,320 seeds plant−1 in 2004 and 2005, respectively. Our research indicates that this southeast Indiana horseweed biotype behaves primarily as a summer annual weed and produces significant amounts of seed when uncontrolled in no-till soybean production. Nomenclature: Glyphosate; horseweed, Conyza canadensis (L.) Cronq. ERICA; soybean, Glycine max (L.) Merr

Influence of Weed Management Practices and Crop Rotation on Glyphosate-Resistant Horseweed Population Dynamics and Crop Yield

Abstract Horseweed is an increasingly problematic weed in soybean because of the frequent occurrence of glyphosate-resistant (GR) biotypes. The objective of this study was to determine the influence of crop rotation, winter wheat cover crops (WWCC), residual nonglyphosate herbicides, and preplant herbicide application timing on the population dynamics of GR horseweed and crop yield. A field study was conducted at a site with a moderate infestation of GR horseweed (approximately 1 plant m−2) with crop rotation (soybean–corn or soybean–soybean) as main plots and management systems as subplots. Management systems were evaluated by quantifying horseweed plant density, seedbank density, and crop yield. Crop rotation did not influence in-field horseweed or seedbank densities at any data census timing. Preplant herbicides applied in the spring were more effective at reducing horseweed plant densities than when applied in the previous fall. Spring-applied, residual herbicide systems were the most effective at reducing season long horseweed densities and protecting crop yield because horseweed in this region behaves primarily as a summer annual weed. Horseweed seedbank densities declined rapidly in the soil by an average of 76% for all systems over the first 10 mo before new seed rain. Despite rapid decline in total seedbank density, seed for GR biotypes remained in the seedbank for at least 2 yr. Therefore, to reduce the presence of GR horseweed biotypes in a local no-till weed flora, integrated weed management (IWM) systems should be developed to reduce total horseweed populations based on the knowledge that seed for GR biotypes are as persistent in the seed bank as glyphosate-sensitive (GS) biotypes. Nomenclature: Glyphosate; horseweed, Conyza canadensis L. ERICA; corn, Zea mays L; soybean, Glycine max (L.) Merr; winter wheat, Triticum aestivum L.

Control of Horseweed (Conyza canadensis) with Growth Regulator Herbicides

Abstract The growth regulator herbicides 2,4-D and dicamba are used to control glyphosate-resistant horseweed before crops are planted. With the impending release of 2,4-D–resistant and dicamba-resistant crops, use of these growth regulator herbicides postemergence will likely increase. The objective of this study was to determine the effectiveness of various growth regulators on Indiana horseweed populations. A greenhouse dose–response study was conducted to evaluate the effectiveness of 2,4-D ester, diglycolamine salt of dicamba, and dimethylamine salt of dicamba on control of four populations of horseweed in the greenhouse. Population 66 expressed twofold levels of tolerance to 2,4-D ester and diglycolamine salt of dicamba. Population 43 expressed an enhanced level of tolerance to diglycolamine salt of dicamba but not to the other herbicides. Diglycolamine salt of dicamba provided the best overall control of populations 3 and 34. Additionally, a field study was conducted to evaluate standard use rates of 2,4-D amine, 2,4-D ester, diglycolamine salt of dicamba, and dimethylamine salt of dicamba on control of various sized glyphosate-resistant horseweed plants. Control of plants 30 cm or less in height was 90% or greater for all four herbicides. On plants greater than 30 cm tall, diglycolamine salt of dicamba provided 97% control while 2,4-D amine provided 81% control. Diglycolamine salt of dicamba provided the highest level of control of glyphosate-resistant horseweed, followed by dimethylamine salt of dicamba, 2,4-D ester and 2,4-D amine, respectively. This research demonstrates that horseweed populations respond differently to the various salts of 2,4-D and dicamba, and it will be important to determine the appropriate use rates of each salt to control glyphosate-resistant horseweed. Nomenclature: 2,4-D; dicamba; glyphosate; horseweed, Conyza canadensis (L.) Cronq. ERICA.

Field Presence of Glyphosate-Resistant Horseweed (Conyza canadensis), Common Lambsquarters (Chenopodium album), and Giant Ragweed (Ambrosia trifida) Biotypes with Elevated Tolerance to Glyphosate

Late-season field surveys conducted in Indiana from 2003 to 2005 showed that common lambsquarters and giant ragweed plants were present in 11 and 22%, respectively, of randomly sampled soybean fields that also contained glyphosate-resistant horseweed. In the fall of 2005 and 2006, seed from 13 common lambsquarters and 22 giant ragweed populations were collected from previously surveyed fields that had confirmed glyphosate-sensitive or -resistant horseweed. The objective of this study was to determine whether the presence of glyphosate-resistant horseweed was correlated with the presence of common lambsquarters and giant ragweed biotypes with elevated tolerance to glyphosate. Through a series of greenhouse screens, 57% of common lambsquarters and 31% of giant ragweed populations collected from fields that had glyphosate-resistant horseweed expressed elevated levels of glyphosate tolerance. However, elevated tolerance to glyphosate was expressed by 33% of giant ragweed and 100% of common lambsquarters populations collected in fields that had glyphosate-sensitive horseweed. Therefore, under the parameters of this experiment and through different types of analyses, we concluded there was not a strong correlation between the late-season presence of glyphosate-resistant horseweed and common lambsquarters and giant ragweed populations with elevated glyphosate tolerance in the same field. A number of the weed populations expressed significant stunting from exposure to glyphosate, but were able to resume growth. Thus, researchers should evaluate plant regrowth in addition to biomass suppression when making assessments of glyphosate resistance in weed populations through greenhouse and field screening. Nomenclature: Glyphosate; common lambsquarters, Chenopodium album L. CHEAL; giant ragweed, Ambrosia trifida L. AMBTR; horseweed, Conyza canadensis (L.) Cronq. ERICA; soybean, Glycine max (L.) Merr

Response and Survival of Rosette-Stage Horseweed (Conyza canadensis) after Exposure to 2,4-D

Abstract 2,4-D is often used as a preplant burndown herbicide to help control horseweed and other broadleaf weeds before planting in no-till corn and soybean production. Isolated instances of poor horseweed control have occurred in production fields. The objective of this research was to evaluate the response of various horseweed populations to 2,4-D. In the first study, 478 horseweed populations from Indiana were subjected to 280 g ae ha−1 of 2,4-D amine in the greenhouse. This rate of 2,4-D caused visible injury and prevented all biotypes from forming new leaves for 28 days. There were specific populations where all plants sprayed were alive at 28 days after treatment (DAT), and approximately 10% of all populations had a least one plant that survived 280 g ae ha−1 2,4-D, resumed growth, and produced seed. In a dose-response study, we observed populations with three-fold more tolerance to 2,4-D. The most tolerant population had a GR90 of 513 g ae ha−1 and the most susceptible population had a GR90 of 121 g ae ha−1 based on dry weights. Growth suppression with 2,4-D was not affected by rosette size for rosettes between 0.5 and 10 cm in width. Nomenclature: 2,4-D; horseweed, Conyza canadensis (L.) Cronq. ERICA; corn, Zea mays L.; soybean, Glycine max (L.) Merr.