As the acreage of glyphosate-resistant (GR) cropping systems increases, so does the occurrence of herbicide-resistant weeds in the landscape. Biotypes of GR Conyza canadensis were first observed in Delaware, USA, in 2000. Since the first documentation of GR C. canadensis, there have been many instances of independent evolution of GR C. canadensis biotypes. The ecology of GR C. canadensis and any potential fitness consequences of GR trait are still unknown. If there is no fitness cost or a fitness increase associated with the GR trait, GR C. canadensis may increase in prevalence in the landscape even in the absence of glyphosate application. With the consideration that fitness is a difficult parameter to measure, other variables may be used as surrogate measures of fitness, such as reproduction, growth rate, phenology, survivorship, etc. This research seeks to determine if differences exist in surrogate measures of fitness - patterns of growth, reproductive allocation, and competitive ability - and if so, how differences may apply to future population changes, for glyphosate-resistant and -susceptible C. canadensis. The first two experiments presented test two populations of GR plants and two populations of GS plants, first in a greenhouse study of shading effects and intraspecific competition (Chapter 2), and second in a field survivorship study within two habitat types (Chapter 3). The third study tests for the prevalence of GR C. canadensis in field margins of GR cropping systems and seeks correlations in the occurrence of resistance and factors related to geography and field management (Chapter 4). The greenhouse study (Experiment 1, Chapter 2) of surrogate measures of fitness (growth, reproduction) in response to intraspecific competitive ability and shading showed that, while all populations had the same response to shading, there were differences between populations in growth and reproduction. While all measures were greater for individual plants with no competition, plants competing with individuals from one of the GR populations (R2) often had measures that were not different from the no competition treatment or were greater than the other competition treatments (R1, S1, and S2). The field survivorship study compared growth, survival, and reproduction of the previously studied four populations of C. canadensis is two habitats, a old-field in the second year of succession (ruderal) and a soybean field planted with a GR cultivar (agrestal) (Experiment 2, Chapter 3). One of the GR populations (R1) had the highest survivorship of the four populations in the ruderal habitat, while there was no difference in survivorship of the four populations in the agrestal habitat. While there was little difference in growth between ruderal populations, in the agrestal habitat, one GS population (S1) consistently had the smallest diameter, height, and leaf number. One of the agrestal GR populations (R2) was larger than the others, though not always different from R1. R1 was the first population to senesce regardless of habitat. Both agrestal GR populations produced more capitulae (seed heads) than the GS populations, and R1 also showed high reproductive success in the ruderal habitat. A test for the occurrence of GR C. canadensis in field margins of GR cropping systems (Experiment 3, Chapter 4) utilized a discriminating spray test on seed collected from the margins of 17 agricultural fields in 2008 from Illinois, Indiana, and Nebraska. The relationships between the occurrence of GR C. canadensis in 2008 field margins and 1) geography, 2) field management practices, 3) herbicide diversity, 4) weed community diversity, 5) field-interior changes in C. canadensis populations (λ), and 6) knowledge or suspicion of difficult-to-control C. canadensis populations, were tested. Geography was the clearest relationship in these data and was related to geography, with greater, more variable occurrence of GR C. canadensis in 2008 field margins in the southern and eastern sites than the northern and western sites. Management practices in field-interiors from year 2006 to 2008 had little relationship to occurrence of GR C. canadensis in 2008 field margins, except for the increased use of tillage and the increased use of non-glyphosate herbicide modes of action in 2007 field-interiors of fields with the highest occurrence of GR C. canadensis in the field margins in 2008. Additionally, the greatest occurrence of GR C. canadensis was associated with high diversity in the weed community in 2007 field-interiors. These results suggest that GR C. canadensis persisted in 2008 field margins even though there was a perceivable increase in management intensity the previous year. In conclusion, although there were no differences in fitness clearly associated with the GR trait in C. canadensis, GR populations may possess equal or greater vigor in growth, reproduction, and competition as GS populations. Based on these results, GR C. canadensis may persist in agricultural field-margins or other ruderal, unmanaged habitats and act as a seed source for future field-interior infestations. Depending on the characteristics of the GR biotype, GR C. canadensis may increase in frequency in the landscape.

The introduction of glyphosate-resistant (GR) crops in the late 1990s changed the way producers used herbicides to control weeds. Since the introduction of GR crops producers have relied on glyphosate alone for weed control instead of utilizing multiple modes of action for weed control. This over-reliance resulted in several weed species developing resistance to glyphosate. This has resulted in organizations from the public and private sector questioning the sustainability of GR cropping systems. Researchers from Illinois, Indiana, Iowa, Mississippi, Nebraska, and North Carolina established 156 onarm trials to determine the sustainability of GR cropping systems. The objectives of this study were: to determine the economics of a university weed resistance best management practice (BMP) versus a producers’ normal production practice; to evaluate when a producer that is risk neutral (profit maximizing) or risk averse should adopt a weed resistance BMP; and to compare the influences of using a university weed resistance BMP to a producer’s normal production practice on the 27 most common weed species in Mississippi. In all instances, the university weed resistance BMP utilized multiple modes of action in conjunction with glyphosate. A university weed resistance BMP can provide the same level of control on 27 of the most common weeds in Mississippi that a producer has become accustomed to with a glyphosate alone system, while delaying or controlling GR weeds. A university weed resistance BMP resulted in an increase in weed control cost, but similar yields and economic returns when compared to a producer’s normal production practice. Rotating a GR crop with a different GR crop resulted in higher economic returns when compared to a continuous GR cropping system or a GR crop followed by a non-GR crop rotation. Producers are often reluctant to adopt a weed resistance BMP because of the perceived increased cost for weed control. A risk neutral or risk averse producer should adopt a weed resistance BMP and feel confident that their decision will provide weed control equivalent to a glyphosate alone weed control program before resistance developed, delay or control GR weeds and be economically sound.

A survey was conducted by phone to nearly 1,200 growers in six states (Illinois, Indiana, Iowa, Mississippi, Nebraska, and North Carolina) in 2005. The survey measured producers’ cropping history, perception of glyphosate-resistant (GR) weeds, past and present weed pressure, tillage practices, and herbicide use as affected by the adoption of GR crops. The objectives of this study were to determine the effect of GR crop use on producers’ tillage practices; changes in herbicide use patterns after adoption of a GR crop; effect of grower awareness of GR weeds on sources of information growers’ use; and growers’ perceptions on resistance management based on knowledge of GR weeds in their farming operation. The adoption of GR cropping systems contributed to large increases in the percentage of growers using no-till and reduced-till systems. Tillage intensity declined more in continuous GR cotton and GR soybean (45 and 23%, respectively) than in rotations that included GR corn or non-GR crops. Tillage intensity declined more in the states of Mississippi and North Carolina than in the other states, with 33% of the growers in these states shifting to more conservative tillage practices after the adoption of a GR crop. This was in part due to the lower amount of conservation tillage adoption in these states prior to GR crop availability. With respect to herbicide use patterns, frequently used herbicides for fall applications were 2,4-D and glyphosate; these herbicides were often used for preplant, burndown weed control in the spring. As expected, crop rotations using GR crops had a high percentage of respondents that made one to three POST applications of glyphosate per year. Overall, glyphosate use has continued to increase, with concomitant decreases in utilization of other herbicides. Concerning grower awareness of GR weeds and perceptions of resistance management in 2005, the majority of the growers (88%) were aware of a weed’s potential to develop resistance to glyphosate, while 44% were aware of state-specific, documented cases of glyphosate weed resistance. Growers that have had experience with GR weeds were more knowledgeable about resistance management practices that could be used to mitigate them.

Doctor of Philosophy / Department of Agronomy / Phillip W. Stahlman / Kochia [Kochia scoparia (L.) Schrad.] is a weed of great economic importance in the Great Plains and western United States and Canada. This weed is prone to evolving resistance to herbicides. Glyphosate is the most widely used herbicide in glyphosate-resistant crops and chemical fallow, and is extremely valuable to crop production. Anecdotal reports of kochia control failure with glyphosate in western Kansas arose during the mid-2000’s. The objectives of this research were to (1) confirm and characterize glyphosate resistance in kochia and measure its impact in western Kansas, (2) gather information on grower weed management practices before and since glyphosate resistance in kochia was confirmed, and (3) determine if altered absorption and translocation of glyphosate contributes to glyphosate resistance in kochia. Dose-response studies on greenhouse and outdoor grown plants, and shikimate accumulation assays confirmed one kochia population collected in 2007 and eight populations collected in 2010 tolerated three- to eleven-times more glyphosate compared to a known glyphosate-susceptible (GS) population. Furthermore, 40 kochia populations collected in 2012 showed varied response, from slightly elevated tolerance to resistance to 0.84 kg ae ha-1 glyphosate. Further analysis suggested these populations were at different stages of resistance evolution. An online survey revealed that growers increased glyphosate use rate and application frequency, but decreased exclusive use of glyphosate and diversified weed management practices during post- compared to pre-glyphosate confirmation periods. Most survey respondents reported presence of glyphosate-resistant (GR) kochia in at least in few fields, and half reported GR kochia in a majority of fields. Thus, together with the resistance confirmation studies, it is estimated that at least one-third of western Kansas kochia populations have evolved resistance to glyphosate. Nominal differences in absorption and translocation of 14C-glyphosate observed between GS and GR kochia populations likely do not contribute to differential response of these populations to glyphosate. Glyphosate-resistant kochia has become widespread in western Kansas in a short period of time. Use of weed resistance best management practices (BMP) is imperative to sustain the utility of glyphosate in the region.

Master of Science / Agronomy / Mithila Jugulam / Common waterhemp (Amaranthus rudis) is a problematic weed species of cropping systems throughout the Midwestern states, including Kansas. Recently, waterhemp populations from Kansas were found to have evolved resistance to the widely used herbicide glyphosate as a result of amplification of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), the enzyme target of glyphosate. The objectives of this research were to 1) perform glyphosate dose-response study and determine the relationship between relative EPSPS genomic copies and EPSPS gene expression in glyphosate-resistant waterhemp, and 2) characterize the genomic configuration and distribution of EPSPS copies using florescence in situ hybridization (FISH) in three glyphosate-resistant waterhemp populations. Waterhemp populations from eastern Kansas were screened with 868 g ae haˉ¹ (field used rate) of glyphosate, and genomic DNA and total RNA was isolated from the survivors to determine the EPSPS genomic copies and EPSPS gene expression relative to the acetolactate synthase (ALS) gene using qPCR. Furthermore, waterhemp specific EPSPS probes were synthesized to perform florescence in situ hybridization (FISH) on these glyphosate-resistant plants. Results of these experiments indicate a positive correlation between level of glyphosate resistance, EPSPS copies, and their expression. As expected, a negative correlation was found between shikimate accumulation and EPSPS copies. Sequencing of the EPSPS gene showed no presence of the proline 106 mutation, which is known to be associated with glyphosate resistance suggesting that an insensitive EPSPS enzyme was not involved in the mechanism of glyphosate resistance. FISH analysis of resistant plants illustrated presence of amplified EPSPS copies on two homologous chromosomes, likely near the centromeric region. . This is the first report demonstrating a positive relationship between EPSPS copies and expressions, as well as chromosome configuration of EPSPS copies in glyphosate- resistant waterhemp from Kansas.

Giant ragweed is an extremely competitive weed and poor control in soybean could lead to significant yield losses for Ontario producers. In 2008, a giant ragweed biotype near Windsor, ON was not controlled with glyphosate and further testing confirmed it as the first glyphosate-resistant (GR) weed in Canada. Giant ragweed seed was collected from 102 locations in Essex (70), Kent (21), Lambton (10) and Waterloo (1) counties to document the occurrence and distribution of GR giant ragweed in Ontario. Giant ragweed seedlings were sprayed with glyphosate at 1800 g a.e. ha-1, and evaluated 1, 7, 14 and 28 days after application (DAA). Results from the survey concluded that there are 47 additional locations in southwestern Ontario with GR giant ragweed. The majority of the sites were found in Essex county, but there was one location in both Chatham-Kent and Lambton counties. Field trials were established at six sites with GR giant ragweed during the 2010 and 2011 growing seasons. The objectives were to determine the level of giant ragweed control with increasing doses of glyphosate, and glyphosate tank mixes applied either preplant or postemergence. Control of giant ragweed increased with higher doses of glyphosate, but only at doses that are not economical for producers. The most effective glyphosate tank mixes were 2, 4-D ester, saflufenacil, linuron, and cloransulam-methyl providing up to 98, 94, 99 and 97% control 4 weeks after application (WAA), respectively. Glyphosate plus dicamba in dicamba-tolerant soybean provided up to 100% giant ragweed control, 4 WAA at the three confined field trial locations. / Monsanto Canada Inc.; Canadian Agricultural Adaptation Program; Grain Farmers of Ontario

The expression of glyphosate resistance can vary within single field populations of common waterhemp and Palmer amaranth. This variability in expression can translate into control ranging from 20 to 80%, which could be the difference in a minor versus a major failure in weed management. Certain application parameters that have been previously associated with glyphosate efficacy, such as glyphosate application time of day and plant stress may exacerbate this variability and lead to failed control of plants on the lower end of the resistance spectrum. Greenhouse research was conducted in 2011 to determine the influence of glyphosate application time of day on the expression of resistance in common waterhemp and Palmer amaranth. Control of both glyphosate-susceptible (GS) and -resistant (GR) weed species showed similar trends in response to glyphosate with respect time of application. Decreased sensitivity of all Amaranthus biotypes was greatest at 9:00 pm and may be attributed to an observed shift in leaf orientation from horizontal to vertical at the time of glyphosate application in response to low-light conditions. The altered leaf orientation most likely reduced herbicide spray coverage. The magnitude of resistance, the difference in the sensitivity of the resistant versus susceptible biotypes, was unaffected by glyphosate application time of day; however, these results indicate that even in resistant populations glyphosate applied at suboptimal times of day such as the evening can cause a further increase in weed escapes from glyphosate. Greenhouse and field experiments were conducted in 2011 and 2012 to determine the influence the soil nutrient amendments on glyphosate sensitivity and growth rate and of GS and GR common waterhemp and Palmer amaranth. In both the GR and GS biotypes of common waterhemp the sensitivity to glyphosate was increased as fertilizer was introduced. However, only the sensitivity of the susceptible biotype of Palmer amaranth was increased with the addition of fertilizer. The lack of response in the GR Palmer amaranth population to fertilizer can be associated with the fact that due to carrier volume limitations enough glyphosate could not be applied to achieve 50% control. The magnitude of resistance was decreased numerically with the addition of fertilizer in both weed species; however, only in common waterhemp was the magnitude of resistance significantly different with the use of high rates fertilizer. The use of fertilizer also had an influence on the growth rate and dormancy of axillary buds. Lateral branching (broken dormancy in axillary buds) was increased in both common waterhemp and Palmer amaranth with the addition of fertilizer. Converting dormant buds to active meristems favors glyphosate translocation and could be responsible for increased glyphosate efficacy. In the field, glyphosate efficacy in GR common waterhemp and Palmer amaranth was also increased with addition of fertilizer; however, this effect was variable. Optimizing the efficacy of glyphosate when applied to even mixed populations of GS and GR Palmer amaranth and common waterhemp can reduce surviving weeds that can produce seed and perpetuate the frequency of glyphosate resistance in the field. Furthermore, greater efficacy of glyphosate may translate into relatively less significant failures in glyphosate applications allowing for successful rescue herbicide treatments and minimal impact on crop yield compared with a complete glyphosate failure with dramatic implications on reduced crop yield and increased weed seed production.

Master of Science / Department of Agronomy / Randall S. Currie / Mithila Jugulam / Italian ryegrass (Lolium perenne L. ssp. multiflorum (Lam.) Husnot), one of the problem weeds of the US, evolved resistance to multiple herbicides including glyphosate due to selection in Arkansas (AR). Glyphosate is a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibitor and amplification of EPSPS gene, the molecular target of glyphosate confers resistance to this herbicide in several weed species, including Italian ryegrass from AR. The objective of this study was to determine the expression of EPSPS gene and protein as well as distribution of EPSPS copies on the genome of glyphosate-resistant Italian ryegrass (ARR) using a known susceptible Italian ryegrass (ARS) from AR. EPSPS gene copies and expression of ARR and ARS were determined using quantitative PCR with appropriate endogenous controls. EPSPS protein expression was determined using Western blot analysis. Fluorescence in situ hybridization (FISH) was performed on somatic metaphase chromosomes to determine the location of EPSPS copies. Based on qPCR analysis, ARR plants showed a wide range of 12 to 118 EPSPS copies compared to a single copy in ARS. EPSPS gene expression correlated with the gene copy number in both ARR and ARS. Individuals with high EPSPS copies showed high protein expression in Western blot analysis. FISH analysis showed presence of brighter EPSPS signals, distributed randomly throughout the genome of ARR individuals compared to a faint signal in ARS plants. Random distribution of EPSPS copies was previously reported in glyphosate-resistant Palmer amaranth. Overall, the results of this study will help understand the origin and mechanism of EPSPS gene amplification in Italian ryegrass.

From 2019 to 2020, greenhouse and field research was conducted at North Dakota State University to investigate the canopy temperature response of waterhemp (Amaranthus rudis), kochia (Kochia scoparia), common ragweed (Ambrosia artemisiifolia), horseweed (Conyza canadensis), Palmer amaranth (Amaranthus palmeri), and red root pigweed (Amaranthus retroflexus) after glyphosate application to identify glyphosate resistance. In these experiments, thermal images were captured of randomized glyphosate resistant populations and glyphosate susceptible populations of each weed species. The weed canopies' thermal values were extracted and submitted to statistical testing and various classifiers in an attempt to discriminate between resistant and susceptible populations. Glyphosate resistant horseweed, when collected within greenhouse conditions, was the only biotype reliably classified using significantly cooler temperature signatures than its susceptible counterpart. For field conditions, image based machine learning classifiers using thermal data were outperformed by classifiers made using additional multispectral data, suggesting thermal is not a reliable predictor of glyphosate resistance.

Made available in DSpace on 2016-12-08T16:44:47Z (GMT). No. of bitstreams: 1 PGPV14MA144.pdf: 861442 bytes, checksum: c0bd112f70401acc3674e49bcc979cf3 (MD5) Previous issue date: 2014-07-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of the present work is to analyze the response of plants of ryegrass (Lolium multiflorum) and wavy-leaved fleabane (Conyza bonariensis) to increasing doses of glyphosate to detect herbicide susceptible and resistant biotypes and to evaluate the physiological quality of seeds of these biotypes to verify the relationship between the resistance and the physiological seed quality of these weeds. Ryegrass and wavy-leaved fleabane plants were exposed to increasing doses of glyphosate, ranging from 0 up to 1440 g ae ha-1, and the fresh weight were weighted 21 days after application, for dose-response tests. Seeds were subjected to tests of physiological quality (germination, cold test and accelerated aging). The experimental design was completely randomized, using ten replicates for ryegrass and six replicates for wavy-leaved fleabane in the dose-response tests, and four replicates for tests of physiological seed quality. All studied biotypes of ryegrass and wavy-leaved fleabane showed differential response due to increasing doses of glyphosate. Ryegrass resistant biotypes showed resistance factors (RF) of 3.0 (Passo Fundo) and 8.3 (Vacaria), while the susceptible ones showed FR of 1.5 (Ponte Serrada) and 1.0 (Lages). Wavy-leaved horseweed resistant biotypes showed FR 2.0 (Papanduva) and 15.5 (Campos Novos), and the susceptible biotype FR 1.0 (Lages). In the tests of physiological seed quality, ryegrass biotype of Lages showed the best performance for germination (64%) and accelerated aging (86%) tests compared to the other biotypes. For the cold test, the biotype of Vacaria showed the best result with 86% of germination, while the biotype of Passo Fundo showed the worst results in all tests. Biotypes of wavy-leaved fleabane did not differ in germination. For Lages and Papanduva wavy-leaved fleabane, itwas found the biotypes responded positively to cold stress (66 and 61%, respectively). The biotype of Campos Novos, with high resistance to the herbicide, did not withstand the stress exposure, presenting similar to germination (34%) compared to the cold test (28%) result. The seeds of the wavy-leaved fleabane biotypes responded negatively to accelerated aging, in that the biotype of Lages showed the best result (12%), while biotypes of Papanduva and Campos Novos showed 4.0 and 2.0%, respectively. Biotypes of ryegrass with differential response to glyphosate present distinct percentage of germination and vigor of seeds, although it is not directly dependent on herbicide resistance. Susceptible biotypes of wavy-leaved fleabane and the biotype with low degree of resistance to glyphosate are more vigorous than the resistant biotype. The physiological seed quality of ryegrass and wavy-leaved fleabane is not directly related to the resistance to glyphosate, and the influence of environment is probably more significant than the resistance / Objetivou-se com o presente trabalho analisar a resposta de plantas de azevém (Lolium multiflorum) e buva (Conyza bonariensis) a doses crescentes do glyphosate para detecção de biótipos suscetíveis e resistentes ao herbicida e avaliar a qualidade fisiológica de sementes desses biótipos para verificar a relação entre a resistência e a qualidade fisiológica de sementes dessas plantas daninhas. Plantas de azevém e buva foram expostas a doses crescentes de glyphosate, variando de 0 a 1.440 g e.a. ha-1, com avaliação da massa fresca após 21 dias da aplicação. As sementes foram submetidas a testes de qualidade fisiológica (germinação, teste de frio e envelhecimento acelerado). O delineamento experimental utilizado foi o inteiramente casualizado, com dez repetições para o azevém e seis repetições para a buva no teste de dose-resposta e com quatro repetições para os testes de qualidade fisiológica de sementes. Todos os biótipos estudados de azevém e buva apresentaram resposta diferencial ao aumento de dose do herbicida glyphosate. Os biótipos de azevém resistentes apresentaram fatores de resistência (FR) igual a 3,0 (Passo Fundo) e 8,3 (Vacaria) enquanto os susceptíveis 1,5 (Ponte Serrada) e 1,0 (Lages). Os biótipos de buva resistentes apresentaram FR de 2,0 (Papanduva) e 15,5 (Campos Novos) e o biótipo susceptível FR de 1,0 (Lages). Nos testes de qualidade fisiológica o biótipo de azevém de Lages apresentou melhor desempenho para os testes de germinação (64%) e envelhecimento acelerado (86%) em relação aos demais biótipos. Para o teste de frio, o biótipo de Vacaria apresentou o melhor resultado com 86%. Enquanto que o biótipo de Passo Fundo apresentou os piores resultados em todos os testes realizados. Os biótipos de buva não se diferenciaram no teste de germinação. Para os biótipos Lages e Papanduva foi verificado que o vigor desses biótipos responderam positivamente ao estresse pelo frio (66 e 61% respectivamente). O biótipo de Campos Novos, com elevada resistência ao herbicida, não suportou a exposição ao estresse, apresentando resultado semelhante ao teste de germinação (34%) em relação ao teste de frio (28%). As sementes dos biótipos de buva responderam negativamente ao teste de envelhecimento acelerado, em que o biótipo de Lages apresentou o melhor resultado com 12%, enquanto os biótipos de Papanduva e Campos Novos apresentaram 4,0 e 2,0% respectivamente. Os biótipos de azevém com resposta diferencial ao glyphosate apresentam porcentagem de germinação e vigor de sementes distintos, porém não diretamente dependentes da resistência ao herbicida. Os biótipos de buva suscetível e com baixo grau de resistência ao herbicida glyphosate são mais vigorosos que o biótipo resistente. A qualidade fisiológica de sementes de azevém e buva não está diretamente relacionada com a resistência ao herbicida glyphosate, sendo a influência do ambiente, provavelmente, mais significativa do que a resistência