Thesis (PhD)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: Please refer to fulltext for abstract / AFRIKAANSE OPSOMMING: Sien asb volteks vir opsomming

Doctor of Philosophy / Department of Agronomy / Tesfaye T. Tesso / The lack of effective post-emergence weed control options is often highlighted as one of the major factors behind dwindling acreage under sorghum (Sorghum bicolor (L.) Moench) in the United States. The discovery of herbicide resistance sources in wild sorghum population and subsequent efforts to incorporate them into cultivated sorghum was received with much optimism to change weed management practices in sorghum. As the development of the technology advances, especially of the Acetolactate synthase (ALS) resistance, concerns over the temporary interveinal chlorosis and reduced seedling vigor in some of the resistant families became heightened. This thesis research is designed to shed light on the genetic basis of seedling chlorosis and assess its impacts on yield potential. The study has three parts; the first part is focused on identifying the genetic causes and plant mechanisms associated with the chlorotic phenotype. ALS herbicide resistant sister-lines expressing normal and chlorotic phenotypes were analyzed via RNA sequencing at four time points during seedling growth. The study identified several variants of genes coding chloroplast precursors and those that cause epigenetic modifications. Once confirmed, genetic markers can be developed to track these gene variants in the breeding population and eliminate segregates genetically prone to chlorosis/yellowing. The second part of the study focuses on assessing the effect of ALS resistance associated chlorosis on agronomic and nutritional parameters of sorghum inbred lines. A set of ALS resistant lines expressing different levels of the chlorotic phenotype were evaluated in replicated field trials and laboratory methods. Results showed that interveinal chlorosis delays flowering but does not have negative effect on yield and nutritional parameters with and without herbicide treatment. The last part addresses whether there is any yield drag that may be associated with herbicide resistance traits and foliar interveinal chlorosis. For this, we synthesized a large set (182) of hybrids from ALS resistant, ACCase resistant and regular (susceptible) seed and pollinator parents. The hybrids were then evaluated in three sets at multiple locations during the 2014 and 2015 crop seasons along with commercial checks. The results revealed that resistance to both herbicides do not cause any drag to grain yield. The traits also do not have any negative impact on grain and nutritional quality of resistant hybrids.

<p>Control of weeds that have survived a postemergence (POST) herbicide often need to be controlled in order to prevent seed production and interference with crops. The most efficacious herbicides and timings used for respray applications has not been determined in many problematic weed species. Previous research has demonstrated that weeds clipped to simulate a failed herbicide application responded differently to herbicide applications to regrowth based on herbicide used and weed species. Other research is conflicting as to the optimum timing of an herbicide respray application with various herbicides. Gaining a better understanding of how to maximize respray herbicide performance will help growers and land managers to preserve crop yield and prevent weed seed production in the event of POST contact herbicide failure. The objectives of this research were to determine the optimum respray herbicide and timing combinations for control of four problematic weed species in the midwestern United States that have survived an application of either glufosinate or fomesafen: waterhemp<i> </i>[<i>Amaranthus tuberculatus</i> (Moq.) J. D. Sauer], Palmer amaranth (<i>Amaranthus palmeri</i> S. Watts), giant ragweed (<i>Ambrosia trifida</i> L.), and horseweed (<i>Erigeron canadensis</i> L). Through a series of field and greenhouse experiments we determined that respray herbicide, respray application timing, initial herbicide, and level of injury from the initial application influence efficacy of the respray herbicide in a species-specific manner. Waterhemp regrowth following a failed glufosinate application was controlled most effectively by applying glufosinate or fomesafen 7 to 11 days after initial treatment. When following fomesafen, applications of 2,4-D 3-7 days after initial treatment or glufosinate 7 to11 days after initial treatment were most effective. Control of Palmer amaranth regrowth following either initial herbicide is best achieved with respray applications of glufosinate, fomesafen, or 2,4-D applied no later than 7 days after initial treatment. The best strategy to control giant ragweed regrowth following a failed fomesafen applications is to apply 2,4-D, dicamba, fomesafen, or glufosinate at any timing between 3 and 11 days after initial treatment. Efficacy of the respray glufosinate application was maximized when applied 11 days after the initial application rather than 3 days after initial application. Horseweed regrowth was best controlled by 2,4-D, dicamba, or glufosinate applied at any timing between 3 and 11 days after the initial application. Where injury from the initial herbicide application is high, there were fewer differences among herbicide treatments and treatment timings. A greenhouse bioassay revealed that as waterhemp injury from an initial glufosinate application increases, control with a respray herbicide also increases. Therefore, complete control of weed regrowth is achieved more easily with increasing injury from the initial application. This research suggests that timing of herbicide respray applications is more urgent than previously thought, so scouting must be done within days of a contact herbicide application to ensure adequate control. </p>

The objective of this Ph.D. research was to identify new and novel mechanisms of wild radish (Raphanus raphanistrum L.) resistance to photosystem II (PSII) inhibitors, auxinics, and acetohydroxyacid synthase (AHAS) inhibitors. PSIIinhibitor resistance was demonstrated to be target-site based, and conferred by a Ser264 to Gly substitution of the D1 protein. Auxinic resistance was associated with reduced herbicide translocation to the meristematic regions of resistant wild radish plants. Two new resistance mutations of wild radish AHAS were discovered, including one encoding the globally rare Asp376 to Glu substitution, and another encoding an Ala122 to Tyr substitution, which has never been identified or assessed for resistance in plants previously. Characterization of the frequency and distribution of AHAS resistance mutations in wild radish from the WA wheatbelt revealed that Glu376 was widespread, and that some mutations of AHAS are more common than others. Computer simulation was used to examine the molecular basis of resistance-endowing AHAS target-site mutations. Furthermore, through the computer-aided analysis, residues were identified with the potential to confer resistance upon substitution, but which have not previously been assessed for this possibility. Results from this Ph.D. research demonstrate that diverse, unrelated mechanisms of resistance to PSII inhibitors, auxinics, and AHAS inhibitors have evolved in wild radish of the WA wheatbelt, and that these mechanisms have accumulated in some populations.

Over the last decades the amount of weeds in our fields have noticeably decreased. The easily annihilated and susceptible species were almost eradicated by means of herbicides and agrotechnical operations. Their place was taken by more resistant species, which are a huge problem for the farmers. For this purpose I decided to make an experiment under the working circumstances. I observed the occurrence of the weeds in certain localities. Especially I concentrated on the silky bent grass (Apera spica-venti), on its occurrence, regulation, extension and susceptibility or resistance to certain herbicides. The effect of the tested herbicide combinations was statistically significant for silky bent grass as well as for the other weeds (P 0.05). The coach grass was the only exception, where the effect of these preparations was not proved, as well as Vicia cracca, Lamium purpurem, Galium aparine, Geranium pusillum (P > 0.05). However the coach grass appeared among these inconclusively responsive weeds more frequently. (P >0.05). As the results of experiment showed, the most effective combinations for elimination of the silky bent grass (Apera spica-venti) were Sumimax + Glean, Cugar Forte + Glean + Dural a Cugar Forte + Logran. All these combinations were proved to be 100% effective against the silky bent grass (Apera spica-venti). The other combinations were not 100% effective. These comprised Rapsan + Command+ Grounded with 89 % effectivity, Sumimax + Logran 91 % effectivity a Butisan+ Clomate+ Grounded 67 % effectivity. The results show that the silky bent grass (Apera spica-venti) is more susceptible to some herbicide combinations and in the fields it can be successfully reduced. On the contrary the silky bent grass (Apera spica-venti) is resilient to some other agents. From this point of view the right choice of the herbicides is crucial in next year.

Au cours de la dernière décennie, l’usage intensif des herbicides en agriculture a provoqué plusieurs crises sanitaires et environnementales. La recherche de nouveaux herbicides bio-inspirés est donc devenue une urgence, en particulier afin de réduire les risques de pollution. Les porphyrines, naturellement présentes dans les végétaux, sont des molécules photosensibles. En présence d’oxygène, leur photo-activation conduit à la production d’Espèces Réactives de l’Oxygène capables d’induire la mort cellulaire. Ce principe utilisé en thérapie photodynamique peut être transféré aux plantes, par exemple à l’aide de porphyrines chargées. Nous avons étudié les propriétés physicochimiques (absorption UV-Visible, émission de fluorescence, photo-stabilité et production d’EROs) ainsi que les effets sur des cellules de tabac TBY-2 d’une série de porphyrines chargées. Pour étudier les mécanismes d’action des porphyrines en tant qu’herbicides, ces molécules doivent être tracées et localisées dans la plante. Dans ce but, nous avons synthétisé des porphyrines liées de manière covalente à un marqueur fluorescent par plusieurs bras espaceurs ; ces derniers ont été choisis grâce à une étude en modélisation moléculaire de leur flexibilité conformationnelle. Les propriétés photo-physiques de ces nouvelles dyades ont été étudiées expérimentalement et théoriquement. / Over the past decade, intensive use of herbicide in agriculture has caused several sanitary and environmental problems. Finding new bio-inspired herbicides preventing pollution has appeared crucial. Naturally present in plants, porphyrins are photosensitive. In the presence of oxygen, their photo-activation leads to production of Reactive Oxygen Species, which induce cell death. Already used in Photodynamic Therapy, this effect can be used to plant. In that purpose, a series of charged porphyrins (commercial and synthesized) were selected, and their physicochemical properties (e.g. UV-Vis absorption, fluorescence emission, photostability, ROS production) as well as their effects on TBY-2 (Tobacco Bright Yellow) cells were evaluated. Second, localizing molecules in plants is mandatory to understand mechanisms of the herbicide action. In this context, porphyrins were covalently grafted to a fluorescent marker, by a series of spacers that were chosen according to a preliminary molecular modeling evaluation of their conformational flexibility. The new dyads obtained were thoroughly studied both theoretically and experimentally for their photophysical properties.

Quizalofop and florpyrauxifen-benzyl are both new herbicides for rice in the midsouthern U.S. Quizalofop is only effective for control of monocot weed species; therefore, mixtures of florpyrauxifen-benzyl with quizalofop could be beneficial in acetyl CoA carboxylase (ACCase)-resistant rice. Field experiments were conducted at the Delta Research and Extension Center in Stoneville, MS, in 2017 and 2018 to evaluate control of monocot and dicot weed species with sequential applications of quizalofop including auxinic herbicides in the first or second treatment. Other field experiments in 2017 and 2018 evaluated sequential applications of different rates of quizalofop with florpyrauxifen-benzyl included in treatments immediately prior to flooding. A final field experiment in 2017 and 2018 evaluated growth and yield of six ACCase-resistant rice cultivars and advanced lines following POST applications of florpyrauxifen-benzyl.

Doctor of Philosophy / Department of Agronomy / Kassim Al-Khatib / Mitchell R. Tuinstra / Sorghum acreage is declining throughout the United States because management options and yield have not maintained pace with maize improvements. The most extreme difference has been the absence of herbicide technology development for sorghum over the past twenty years. The objectives of this study were to evaluate the level of resistance, type of inheritance, and causal mutation of wild sorghums that are resistant to either acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides or acetohydroxyacid synthase (AHAS)-inhibiting herbicides. ACCase-inhibiting herbicides used in this study were aryloxyphenoxypropionate (APP) family members fluazifop-P and quizalofop-P along with cyclohexanedione (CHD) family members clethodim and sethoxydim. The level of resistance was very high for APP herbicides but low to nonexistent to CHD herbicides. With genetic resistance to APP herbicides, the resistance factors, the ratio of resistance to susceptible, were greater than 54 to 64 for homozygous individuals and greater than 9 to 20 for heterozygous individuals. Resistance to CHD herbicides was very low with resistance factors ranging from one to about five. Genetic segregation studies indicate a single gene is the cause of resistance to APP herbicides. Sequencing identified a single mutation that results in cysteine replacing tryptophan (Trp-2027-Cys). Trp-2027-Cys has previously been reported to provide resistance to APP but not CHD herbicides. The other wild sorghum evaluated in this study was resistant to AHAS-inhibiting herbicides including imidazolinone (IM) family member, imazapyr, and sulfonylurea (SU) family member, nicosulfuron. Resistance factors in this genotype were very high, greater than 770 for the IM herbicide and greater than 500 for the SU herbicide, for both herbicide chemical families. Genetic segregation studies demonstrate that resistance was controlled by one major locus and two modifier loci. DNA sequencing of the AHAS gene identified two mutations, Val-560-Ile and Trp-574-Leu. Val-560-Ile is of unknown importance, but valine and isoleucine are similar and residue 560 is not conserved. Trp-574 is a conserved residue and Leu-574 is a known mutation that provides strong cross resistance to IM and SU herbicides. The results of these studies suggest that these sources of APP, SU, and IM resistance may provide useful herbicide resistance traits for use in sorghum.

Herbicides have become a key component in modern agricultural production. Meanwhile, there is a concern that some herbicides persist past the growing season of the treated crop, and negatively influence the production of the subsequently planted crops. Amongst various herbicides used in western Canada, acetohydroxyacid synthase (AHAS)-inhibiting herbicides warrant special attention given their residual properties and acute plant toxicity at low concentrations in soil. Soil residual AHAS inhibitors have the potential to influence both leguminous host plants and their bacterial symbiotic partners; consequently, the use of an AHAS inhibitor in a given year can negatively influence the inoculation success and grain yield of legumes cropped in the following year. <p>The present thesis project focused on one of the AHAS inhibiting herbicides (flucarbazone) and studied its potential for carryover injury and negative influence on the success of inoculation in field pea. A series of growth chamber and field experiments were conducted to test the following null hypothesis: the presence of residual flucarbazone in soil does not affect nodulation of field pea by inoculum rhizobia. <p>A growth chamber experiment clearly demonstrated the susceptibility of field pea to the presence of flucarbazone in soil where the lowest concentration of flucarbazone amendment (5 ìg kg1) significantly reduced the crop growth. In contrast, a field study failed to reveal any negative effects of flucarbazone use on crop growth and N2 fixation. <p>It was concluded that if the weather and soil conditions favour decomposition of flucarbazone as described in the present study, flucarbazone applied at the recommended field rate will not persist into the following season at high enough concentrations to negatively influence field pea growth, grain yields, and inoculation success. To ensure safety of rotational crops, it is important to strictly adhere to the herbicide application guidelines. Additionally, producers are cautioned to be particularly aware of the environmental and soil conditions that may reduce the rate of herbicide degradation.

Herbicides have become a key component in modern agricultural production. Meanwhile, there is a concern that some herbicides persist past the growing season of the treated crop, and negatively influence the production of the subsequently planted crops. Amongst various herbicides used in western Canada, acetohydroxyacid synthase (AHAS)-inhibiting herbicides warrant special attention given their residual properties and acute plant toxicity at low concentrations in soil. Soil residual AHAS inhibitors have the potential to influence both leguminous host plants and their bacterial symbiotic partners; consequently, the use of an AHAS inhibitor in a given year can negatively influence the inoculation success and grain yield of legumes cropped in the following year. <p>The present thesis project focused on one of the AHAS inhibiting herbicides (flucarbazone) and studied its potential for carryover injury and negative influence on the success of inoculation in field pea. A series of growth chamber and field experiments were conducted to test the following null hypothesis: the presence of residual flucarbazone in soil does not affect nodulation of field pea by inoculum rhizobia. <p>A growth chamber experiment clearly demonstrated the susceptibility of field pea to the presence of flucarbazone in soil where the lowest concentration of flucarbazone amendment (5 ìg kg1) significantly reduced the crop growth. In contrast, a field study failed to reveal any negative effects of flucarbazone use on crop growth and N2 fixation. <p>It was concluded that if the weather and soil conditions favour decomposition of flucarbazone as described in the present study, flucarbazone applied at the recommended field rate will not persist into the following season at high enough concentrations to negatively influence field pea growth, grain yields, and inoculation success. To ensure safety of rotational crops, it is important to strictly adhere to the herbicide application guidelines. Additionally, producers are cautioned to be particularly aware of the environmental and soil conditions that may reduce the rate of herbicide degradation.