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<p>PPO inhibitors are a valuable group of herbicides that provide soil-residual and foliar control of glyphosate resistant <em>Amaranthus</em> species. The ΔG210 mutation in the <em>PPX2</em> gene confers PPO-inhibitor resistance and has been present in the Midwest for more than a decade. Until recently, PPO-inhibitor resistance in waterhemp was attributable to just the ΔG210 mutation in the <em>PPX2</em> gene, but recently, several new PPO-resistant biotypes have been discovered in waterhemp and Palmer amaranth. A possible explanation is a change in PPO-inhibitor use patterns and commonly used active ingredients.</p> <p>A direct comparison of the ΔG210 mutation and a new mutation, R128G, was conducted in the greenhouse. Results indicate that the R128G mutation in waterhemp is not substantially better than the ΔG210 mutation with respect to conferring resistance to PPO inhibitors applied preemergence. Furthermore, there is no evidence that the utility of PPO inhibitors applied preemergence will diminish any further as a result of the R128G mutation increasing in frequency. A set of field trials was conducted to investigate how a new PPO inhibitor, trifludimoxazin, will select for resistant biotypes in the field. Overall, a greater number of resistant plants survived the foliar herbicide applications than emerged through soil applications. Trifludimoxazin did not increase the frequency of PPO-resistant individuals when applied to soil, but when applied to foliage, increased the frequency of PPO-resistant individuals by 2.5 to 2.6-fold, similar to other PPO inhibitors applied to foliage. In other experiments, investigations of waterhemp populations with unique resistance phenotypes were conducted. In populations that were more resistant than others, yet had the same ΔG210 mutation, there was no evidence of herbicide metabolism contributing to the greater resistance phenotype. In resistant populations that had no target site mutations, target site expression experiments and lipid peroxidation experiments were inconclusive. However, there was some evidence of increased target-site expression or increased antioxidant capacity as being causal mechanisms, although no mechanisms have been fully ruled out.</p>

Repeated use of acetolactate synthase (ALS)-inhibiting herbicides in recent years has resulted in the selection of 89 weed species resistant to these herbicides. One management strategy that can eliminate or slow the development of resistance is applying mixtures of herbicides with different modes of action. This research involved the characterization of ALS-inhibiting herbicide-resistant smooth pigweed (<i>Amaranthus hybridus</i> L.), as well as investigations on weed management programs in corn (<i>Zea mays</i> L.) utilizing mesotrione, a triketone, in mixtures with other herbicides. ALS-inhibiting herbicide-resistant smooth pigweed biotypes were collected from fields in Virginia, Delaware, Maryland, and Pennsylvania to evaluate response to ALS-inhibiting herbicides and to determine the molecular mechanisms of resistance. Sequencing of the ALS genes from these biotypes revealed two amino acid substitutions known to confer resistance, Ala₁₂₂ to Thr and Ser₆₅₃ to Asn, and one that has not been previously reported in plants, Asp₃₇₆ to Glu. The smooth pigweed biotype with an Asp₃₇₆ substitution displayed resistance to four classes of ALS-inhibiting herbicides that included imidazolinone (IMI), sulfonylurea (SU), pyrimidinylthiobenzoate (PTB), and triazolopyrimidine sulfonanilide (TP) chemistries. Transformation of this smooth pigweed ALS gene into <i>Arabidopsis thaliana</i> confirmed that the Asp₃₇₆ substitution is responsible for the resistance. Other biotypes that had a substitution at Ala₁₂₂ exhibited resistance to an IMI herbicide, little to no resistance to SU herbicides, and increased sensitivity to a PTB and a TP herbicide, whereas, biotypes that had a substitution at Ser₆₅₃ exhibited high-level resistance to an IMI herbicide and lower resistance to PTB and SU herbicides. Experiments were also conducted to investigate the effectiveness of mesotrione in preemergence (PRE) and postemergence (POST) corn weed management programs in Virginia. Mesotrione applied PRE in mixtures with <i>S</i>-metolachlor and atrazine controlled common lambsquarters (<i>Chenopodium album</i> L.), smooth pigweed, common ragweed (<i>Ambrosia artemisiifolia</i> L.), and morningglory (<i>Ipomoea</i> spp.) species when a timely rainfall followed application. POST applications of mesotrione controlled common lambsquarters and smooth pigweed, but common ragweed and morningglory species were not always controlled. Common ragweed and morningglory species were controlled by mesotrione in a mixture with atrazine POST. Large crabgrass [<i>Digitaria sanguinalis</i> (L.) Scop.] and giant foxtail (<i>Setaria faberi</i> Herrm.) control was generally better when the ALS-inhibiting herbicides nicosulfuron plus rimsulfuron or rimsulfuron plus thifensulfuron plus atrazine were applied in a mixture with mesotrione. Mixtures of mesotrione with other POST herbicides in a total POST program produced corn yields comparable to standard PRE followed by POST weed management programs. / Ph. D.

Sustainable weed control in managed agricultural systems requires the judicious use of multiple weed control tactics and prevents over-reliance on any one tactic. In this context, sustainable weed management plays a critical role in the mitigation of one of agriculture's most pressing problems- herbicide resistance. Research conducted in Virginia sought to explore the effects of integrating multiple weed management tactics in corn and cool-season turfgrass. Additionally, research was conducted to confirm nicosulfuron and glyphosate herbicide resistance in Virginia johnsongrass and elucidate the molecular mechanisms conferring those resistances. Rye and hairy vetch cover crop residues, combined with reduced rates of preemergence herbicide and postemergence glyphosate applications, were shown to provide sufficient weed control and corn yield. Cover crop type or residue level did not augment weed control in corn production systems, but the use of glyphosate was essential for late-season weed control. Rye and vetch biculture as a cover crop increased corn yield compared to rye cover crop alone. In cool-season turfgrass, the addition of reduced preemergence herbicide rates to corn gluten meal, an organic herbicide product, reduced crabgrass 25%. Moreover, control was dependent on herbicide choice. Herbicides applied at half of recommended labeled rates or less did not control crabgrass at a commercially-acceptable level, regardless of corn gluten meal addition. In field experiments, Virginia johnsongrass expressed resistance to nicosulfuron and glyphosate. Glyphosate at 0.88 kg ae ha-1 controlled johnsongrass 65%. Nicosulfuron at 0.14 kg ai ha-1 controlled the same population 10%. Greenhouse experiments confirmed differential sensitivity of putative herbicide-resistant johnsongrass seedlings to nicosulfuron and glyphosate when compared to a susceptible population. Herbicide resistance was not conferred via target-site mutation. Five ALS-gene site mutations were confirmed absent in Virginia johnsongrass, while three others were located in coding regions that could not be elucidated in johnsongrass. Further investigations showed glyphosate resistance was not conferred via reduction in herbicide absorption or translocation. The susceptible johnsongrass caused an increase in a polar metabolite at Rf = 0.17 with concomitant reduction in glyphosate over time. Although the mechanism is not clear, these data suggests that glyphosate resistance in johnsongrass may be associated with differential metabolism. / Ph. D.

Using data envelopment analysis with principal components, we calculate an adoption-intensity index for herbicide-resistance best management practices (BMPs). Empirical results for over 1,100 farmers in twenty-two U.S. states suggest that many farmers could improve their herbicide resistance BMP adoption. Two-limit truncated regression results show that higher yields and a greater proportion of acres planted with Roundup Ready (R) seeds motivate weed BMP adoption. While soybean and corn farmers have lower adoption intensity than cotton farmers, farmer educational attainment and greater concern for herbicide effectiveness and for human and environmental safety are found to help increase the adoption of weed BMPs.

Made available in DSpace on 2017-07-21T19:59:47Z (GMT). No. of bitstreams: 1 Stela Becher.pdf: 1500518 bytes, checksum: a9d6338bf319b6f8a5407fc7f6e00bdb (MD5) Previous issue date: 2009-09-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Mesotrione, 2-(4-Mesyl-2-nitrobenzoyl)-1,3-cyclohexanedione, is a trycetone herbicide chemical class, with high leaching potential, and competitively inhibit 4-hydroxyphenylpyruvate dioxygenase (HPPD), that converts tyrosine to plastoquinone and α-tocopherol, and was found in plants, fungus, and bacteria, including Acinetobacter. The aim of this work was to characterize pathways of Brazilian strains of Acinetobacter facing mesotrione. Acinetobacter calcoaceticus and Acinetobacter sp. were selected in up to 15.00 mM (resistance) and 0.03 mM (tolerance) concentrations of mesotrione and tested at 35 mM buffered solution in spectrophotometry until 60 hours, showing to be incapable to degrade mesotrione. Computer bacterial cells measures (ImageJ 1.37v) indicate starvation process, probably due to non use of mesotrione as carbon source, and this characteristic could be consider a marker for herbicide degradation. Mesotrione herbicide activity is major in N, and enhanced by S, and has a strong electron withdrawing. For this reason, we studied antioxidative stress enzymes catalase, superoxide dismutase and glutathione reductase, by SDS-PAGE, non-denaturatig PAGE and spectrophotometry. Acinetobacter calcoaceticus showed a higher response to oxidative stress than Acinetobacter sp., with more soluble proteins and lower number of electrophoresis bands isoforms (12 against 23), because its enzymes were selected for this herbicide and show better adaptation than Acinetobacter sp.. A microbial strategy of herbicide resistance/tolerance and specific oxidative stress raised by mesotrione herbicide could give additional non-degradating options to microorganisms, not necessarily leading to fast herbicide degradation, so determining the herbicide fate in soils and groundwater. / Mesotrione, 2 - (4-mesil-2-nitrobenzoil) -1,3-cicloexanodionas, é um herbicida da classe química tricetona, com alto potencial de lixiviação, inibe competitivamente a 4-hidroxifenilpiruvato dioxigenase (HPPD), que converte a tirosina em plastoquinona e α-tocoferol, e foi encontrado em plantas, fungos e bactérias, incluindo Acinetobacter. O objetivo deste trabalho foi caracterizar modos de adaptação de linhagens brasileiras de Acinetobacter ao mesotrione. Acinetobacter calcoaceticus e Acinetobacter sp. foram selecionadas em até 15.00 mM (resistência) e 0,03 mM (tolerância) de mesotrione e avaliadas para degradação em experimentos de até 60 horas a 35 mM do herbicida em espectrofotometria, mostrando-se incapazes de degradação. Medidas das células bacterianas em computador (ImageJ 1.37V) indicam processo de inanição, provavelmente devido à não utilização de mesotrione como fonte de carbono, e essa característica pode ser considerada como um marcador para a degradação do herbicida. A forte atividade oxidativa do herbicida mesotrione está vinculada ao radical N da molécula, e reforçada pelo radical S. Por este motivo, foi estudado o estresse oxidativo para as enzimas catalase, superóxido dismutase e glutationa redutase, por SDS-PAGE não denaturante, PAGE e espectrofotometria. Acinetobacter calcoaceticus mostrou uma maior resposta ao estresse oxidativo do que Acinetobacter sp., apresentando mais proteínas solúveis e menor número de isoformas de bandas em eletroforese (12 contra 23), provavelmente devido a suas enzimas terem sido selecionadas para este herbicida e mostraram melhor adaptação que Acinetobacter sp.. Estratégias de resistência microbiana ao herbicida e padrões específicos de enzimas anti-oxidativas poderiam dar opções não degradativas aos microorganismos, levando á manutenção de maiores quantidades herbicidas em solos e águas subterrâneas, e por mais tempo.

Progress in tree biotechnology initially trailed behind agricultural crops due to their long life cycle, difficult tissue culture and regeneration protocols, and their abundance in natural forests. However, rapid global deforestation rates, together with an increased world demand for pulp, paper and timber products, have prompted scientific and commercial focus to improve genetic timber stocks. South Africa, a tree-poor country (where indigenous forests are protected), has relied almost solely on exotic plantations to meet its demand for timber. A pioneer study investigating the feasibility of using direct (biolistic) and indirect (Agrobacterium-mediated) methods for gene transfer was undertaken in Pinus patula Schiede et Deppe, a Mexican softwood and a forerunner for saw timber, pulpwood and paper in the South African forest industries. The aim of the transformation methods was to impart herbicide resistance to the trees. This was achieved via the introduction of a bar-GUS pAHC25 cassette under the control of the ubiquitin promoter. To provide target material for transformation, two in vitro micropropagation pathways were used: somatic embryogenesis and organogenesis. Both embryonal suspensor masses (ESM) and somatic embryos at various stages of development were initially used as target explants for the biolistic study using an established in vitro protocol. A stepwise selection was implemented in order to allow transformed (particularly bombarded) cultures the opportunity to regenerate under selection pressure using MSG3 maintenance medium supplemented with BASTA® herbicide at 1 mg l ¯¹ followed by 3 mg l¯¹ active ingredient at the next subculture. Biolistic transgene delivery was more efficient when sorbitol was included in the pre-bombardment medium enabling use of higher vacuum and shooting pressures, without lowering the regeneration potential of ESM significantly. Bombarded material from two genotypes (Lines 2 and 3) was regenerated to produce mature somatic embryos using an optimized regeneration regimen. The indirect study with Agrobacterium tumefaciens (LBA4404), transformed with the pAHC25 vector via triparental mating or heat shock, used a variety of target tissues including: mature somatic embryos, ESM and mature zygotic embryos (MZE's) - a novel in vitro system for P. patula. The Agrobacterium-mediated method resulted in optimized decontamination conditions using a combination of liquid MSG3 (or sterile dH₂O for mature embryos) supplemented with 500 mg l ¯¹ cefotaxime, with rotation, and sterile 65 mm Whatman No. 3 filter paper stacks, which avoided excess filtering and stress to transformation material. Further efforts to aid regeneration during the indirect study included L-proline post-transformation, though no mature somatic embryos were regenerated at the conclusion of the Agrobacterium-mediated study. Recovery of transformed ESM in both studies was best during the active growth phase 4-6 d after subculture. Regeneration with good somatic embryo potential was an exigent aspect in both transformation studies. Expression of positive histochemical GUS activity in all transformed material was confirmed by polymerase chain reaction (PCR) analysis indicating that Pinus patula tissue was amenable to transformation. A new bar PCR regime was implemented in P. patula. In the biolistic study, a higher transformation efficiency of bar amplicons (53%) than GUS amplicons (45%) was observed, reflecting their non-linked status on the pAHC25 transformation vector. This is the first report of biolistic transformation of P. patula that will allow for the production of transgenic ESM. The production of transgenic P. patula holds great promise for commercial development in the South African forestry industry. The application of transgenic trees in the timber industry is numerous but the aims most relevant to P. patula include wood modification and disease resistance to pathogens like pitch canker fungus. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

Thesis (M.S.)--University of Tennessee, Knoxville, 2009. / Title from title page screen (viewed on Oct. 22, 2009). Thesis advisor: Lawrence E. Steckel. Vita. Includes bibliographical references.

Ce travail a eu pour but premier d’étudier le risque d’évolution de la résistance aux herbicides inhibiteurs de l’acétolactate synthase (ALS) chez l’ambroisie à feuilles d’armoise (Ambrosia artemisiifolia L.) à travers quatre points : (i) la pression de sélection (étude de l’efficacité d’une gamme d'herbicides inhibiteurs de l’ALS), (ii) la capacité de réponse adaptative de l’adventice (détermination de la variation de la sensibilité aux inhibiteurs de l’ALS entre plantes et mise en place d’un programme de sélection récurrente), (iii) une étude de terrain (recherche de résistance aux inhibiteurs de l’ALS au champ en France), (iv) l’étude des mécanismes de résistance (liée à la cible – RLC – et non liée à la cible – RNLC – par une approche de transcriptomique). Le second objectif fut d’étudier la connectivité des populations d’A. artemisiifolia dans des paysages agricoles à l’aide de marqueurs microsatellites développés lors de ce travail afin de déterminer les facteurs qui pourraient faciliter la dispersion de cette espèce et de la résistance à l’échelle du paysage agricole.En ce qui concerne la résistance aux herbicides :-La réponse de d’A. artemisiifolia aux herbicides inhibiteurs de l’ALS est très variable entre substances.-Des plantes ayant survécu à la dose maximale autorisée et à des doses supérieures de metsulfuron ont été sélectionnées pour débuter un programme de sélection récurrente. Après deux cycles de sélection, on observe une intensification de la résistance au metsulfuron et une émergence de la résistance à l’imazamox et au tribénuron.-Trois cas de résistance à l’imazamox ont été identifiés au champ dont deux cas de pure RNLC et un cas de coexistence RLC – RNLC.-Un transcriptome d’A. artemisiifolia a été généré grâce à la technique de séquençage PacBio pour rechercher des gènes impliqués dans les mécanismes de RNLC (approche RNAseq). 62 gènes candidats ont été identifiés dont des transporteurs ABC, des cytochromes P450 ainsi que des glutathione-S-transférases connus pour être impliqués dans la dégradation des herbicides.Pour l’étude de la connectivité des populations agricoles :-26 marqueurs microsatellites ont été développés et ont révélé une forte variabilité génétique. La structuration génétique a été étudiée à grande échelle pour des populations d’A. artemisiifolia d’Europe (aire d’invasion) et d’Amérique du Nord (aire d’origine).-À une échelle plus fine (paysage agricole), la structure génétique des populations reste influencée par les événements de colonisation. Les événements de migration qui ont été identifiés entre zones de présence de l’ambroisie suggèrent des flux de gènes (pollen/semences) et une connectivité modérés à l’échelle d’un territoire agricole. Dans les environnements agricoles, la dispersion des allèles de résistance aux herbicides pourrait se faire facilement de proche en proche via les flux de pollen, et également à plus longue distance via des dispersions de graines. Les activités anthropiques jouent un rôle majeur dans la dispersion des semences (machineries agricoles, lots de semences contaminés…).-L’analyse du système de reproduction a confirmé que cette espèce est allogame ce qui entraîne des flux de gènes intra- et inter-populations importants.Les connaissances acquises au cours de ce travail pourront aider à développer des stratégies de contrôle mieux adaptées, pour lutter efficacement contre A. artemisiifolia afin de limiter son expansion, telles que :-Des stratégies de désherbage diversifiées : combinaison de lutte mécanique (dont faux semis) et chimique (diversification des modes d’action herbicides).-Un allongement et une diversification des rotations de cultures en favorisant des cultures d’hiver et/ou des cultures couvrantes et compétitrices.Ces connaissances pourront aussi être utilisées dans la lutte contre une autre espèce adventice du genre Ambrosia, Ambrosia trifida.Mots-clés (6) : Ambrosia artemisiifolia L., ambroisies, résistance aux herbicides, / The first aim of this work was to investigate the risk for the evolution of resistance to acetolactate synthase inhibitor (ALS) herbicides in the common ragweed (Ambrosia artemisiifolia L.) through four points: (i) the selection pressure (effectiveness of a range of ALS inhibitor herbicides), (ii) the adaptive response of Ambrosia artemisiifolia (recurrent selection experiment), (iii) a resistance monitoring in fields in France, and (iv) the investigation of the mechanisms underlying herbicide resistance (target-site (TSR) and non-target-site resistance (NTSR) using transcriptomic analyses). The second aim was to study the connectivity of A. artemisiifolia populations in agricultural landscapes using microsatellite markers developed during this work, to determine factors that could facilitate the spread of this invasive weed species and the spread of herbicide resistance.In regards to herbicide resistance:-The sensitivity of A. artemisiifolia to ALS-inhibiting herbicides is variable between active ingredients.-Plants that survived the French maximum authorized field rate and higher rates of metsulfuron were selected to implement a recurrent breeding program. After two selection cycles, the resistance level to metsulfuron increased and resistance to imazamox and tribenuron emerged.-Three cases of imazamox resistance were identified in the field, including two cases of pure NTSR and one case of TSR - NTSR coexistence.-A transcriptome for A. artemisiifolia, AMBELbase, was generated using the PacBio sequencing technology to search for genes involved in NTSR mechanisms (RNAseq approach). 62 candidate contigs were identified including ABC transporters, cytochromes P450 and glutathione S-transferases known to be involved in the degradation of herbicides.In regards to population connectivity:-26 microsatellite markers were developed and revealed high genetic variability. Genetic structuring has been studied on a large scale for populations of A. artemisiifolia from Europe (invasion range) and North America (native range).-On a finer scale (agricultural landscape), the genetic structure of populations was influenced by colonization events. Migration events detected among the areas colonized by A. artemisiifolia suggested moderate pollen/seed flows and connectivity at the farmland scale. In agricultural environments, herbicide resistant alleles could be easily spread among neighbouring populations via pollen flow, and also at longer distances via seed dispersal. Human-related activities play a major role in the dispersal of seeds (agricultural machinery, contaminated seed lots, etc.).-The mating system analysis confirmed that A. artemisiifolia is an obligate outcrossing species which leads to important intra- and inter-population gene flow.The knowledge acquired during this work may help to foster the development of better management strategies to effectively control A. artemisiifolia to limit its spread, such as:-Diversified weed control strategies: combination of mechanical (including false-seed) and chemical weeding (diversification of herbicide modes of action).-Longer diversified crop rotations including more winter crops and/or cover and competitive crops to break the life cycle of A. artemisiifolia.These knowledge may also be used to better control of another weed species of the genus Ambrosia, Ambrosia trifida L.

Weed control is an integral part of a successful overall production strategy in row- cropping systems and has the potential to reduce or eliminate yield losses that negatively affect profitability. Timely and correctly selected herbicide applications are the major keys for effective weed control in a majority of instances. However, there are negative factors that contribute to ineffectiveness and weed escape issues that currently lack viable options for management. Sparsely populated late-season weeds that emerge after lay-by herbicide applications and weeds that have become tolerant and resistant to traditional herbicide chemistries are of greatest concern. Historically, these weeds would have been pulled or chopped by hand or removed by cultivation, but with current production strategies built around conservation tillage and herbicide management practices, blanket disturbance of the soil through plowing is not a viable option. There is an immediate need for site-specific weed management to address these weed escapes while minimizing soil disturbance that reduces residual herbicide efficacy and lessens moisture losses that negatively effects the growing crop