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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
11

Phylogeny, identification and pathogenicity of the Botryosphaeriaceae associated with collar and root rot of the biofuel plant Jatropha curcas in Brazil, with a description of new species. / Phylogeny, identification and pathogenicity of the Botryosphaeriaceae associated with collar and root rot of the biofuel plant Jatropha curcas in Brazil, with a description of new species

Machado, Alexandre Reis 19 July 2012 (has links)
Made available in DSpace on 2015-03-26T13:37:52Z (GMT). No. of bitstreams: 1 texto completo.pdf: 6115511 bytes, checksum: f9a6de95b1f160207c633c70ff99d635 (MD5) Previous issue date: 2012-07-19 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / A partir da iniciativa do Governo Federal em introduzir o biodiesel na matriz energética brasileira, surgiu a necessidade de se pesquisar plantas oleaginosas potenciais para produção de matéria-prima para este biocombustível. O pinhão manso (Jatropha curcas) tem se destacado por ser uma planta perene, de fácil manejo, além de produzir sementes com alto teor de óleo. A grande expansão das áreas de cultivo tem sido acompanhada pelo surgimento de diversas enfermidades, do qual pouco se conhece sobre os reais agentes etiológicos. Atualmente, em diversas áreas do Brasil, tem-se relatado a ocorrência de uma nova doença que não apenas reduz a produtividade, como tem causado a morte das plantas. Esta doença está associada a uma podridão das raízes e do colo das plantas. Alguns patógenos já foram relatados para essa doença, sendo mais frequente a ocorrência de fungos da família Botryosphaeriaceae, grupo conhecido pela dificuldade de separação das espécies utilizando características morfológicas. Assim, o objetivo deste trabalho foi identificar os possíveis agentes etiológicos, investigar a diversidade de Botryosphaeriaceae associado a essa doença, utilizando características morfológicas aliadas às ferramentas moleculares, bem como provar a potogenicidade dos isolados. Foram realizadas coletas nos estados de Minas Gerais e Espírito Santo. Em adição a estes, foram obtidas amostras dos estados do Piauí e São Paulo. Isolados monospóricos foram obtidos e armazenados. Estes tiveram o DNA extraído e as regiões ITS e TEF1-α sequenciadas. A partir dos resultados das análises filogenéticas, foram separados dois a três isolados de cada espécie para a caracterização morfológica. Nove espécies de Botryosphaeriaceae foram identificadas, sendo Lasiodiplodia theobromae, L. parva, L. pseudotheobromae, L. iraniensis, Neoscytalidium dimidiatum, Macrophomina phaseolina e três a serem propostas como novas espécies (Lasiodiplodia sp.1, Lasiodiplodia sp.2 e Macrophomina sp.1). Todas as espécies distinguiram morfologicamente e filogeneticamente, com exceção de Macrophomina sp.1 que não esporulou em meio de cultura. Até o momento, apenas Lasiodiplodia theobromae, Lasiodiplodia sp.1 e Neoscytalidium dimidiatum tiveram a patogenicidade comprovada. Pelos testes de Blotter foram encontrados associados às sementes: Lasiodiplodia theobromae, Macrophomina phaseolina e Macrophomina sp.1. Espécies de Botryosphaeriaceae ocorrem em uma ampla gama de hospedeiros e ambientes, e são frequentemente referidos como endofíticos, patógenos latentes ou oportunistas, devido a manifestação da doença estar diretamente associada à ocorrência de estresse do hospedeiro. A expansão das áreas de pinhão manso no mundo tem contribuído para o surgimento de várias doenças que até o momento não tinham seus agentes etiológicos conhecidos. Este estudo fornece informações novas para futuros estudos de manejo da doença, programas de quarentena e especialmente para o desenvolvimento de variedades resistentes à podridão do colo e raiz do pinhão-manso. / The introduction of biodiesel in the Brazilian energy matrix by initiative of the Federal Government, encouraged the search for potential oleaginous crops to supply raw material for biofuel production. The physic nut (Jatropha curcas) has been highlighted since it is a perennial plant, easy to manage, and produces seeds with a high oil content. The expansion of areas of Jatropha in the world has contributed to the emergence of various diseases. Currently in Brazil, the occurrence of a new disease that not only reduces the productivity, since it causes death of the plants. This disease is associated with a collar and root rot of plants. A number of pathogens have been associates with this disease, the occurrence of Botryosphaeriaceae fungi being the most frequent, which is a group known to be difficult to delimit species based on morphological characters. Thus, the purposes of this work was to investigate the diversity of Botryosphaeriaceae associated with collar and root rot of J. curcas, with the aid of morphology and molecular tools and to asses the pathogenicity of the species involved. Samples were colleted in states of Minas Gerais and the Espirito Santo. In addition, samples were obtained from Piauí and São Paulo states. Single spore cultures were obtained and stored. These had their DNA extracted and the ITS and TEF1-α regions were sequenced. From the results of the phylogenetic analyses, two to three isolates of each species were separated for the morphological characterization and pathogenicity tests. With the purpose of investigating the association of the pathogens in seeds, Blotter tests were performed. Nine Botryosphaeriaceae species were identified: Lasiodiplodia theobromae, L. parva, L. pseudotheobromae, L. iraniensis, Neoscytalidium dimidiatum, Macrophomina phaseolina and three to be proposed as new species (Lasiodiplodia sp.1, Lasiodiplodia sp.2 and Macrophomina sp.1). All species were distinguished morphologically and phylogenetically, except Macrophomina sp.1 that failed sporulate in culture.Currently, only Lasiodiplodia theobromae, Lasiodiplodia sp.1 and Neoscytalidium dimidiatum have proven to be pathogenic. Lasiodiplodia theobromae, Macrophomina phaseolina and Macrophomina sp.1 were associated with seeds. Botryosphaeriaceae species occur in a wide range of hosts and environments, and are often referred to as endophytes and latent or opportunistic pathogens, because manifestation of the disease is directly linked with host stress. The great expansion of Jatropha areas in the world have contributed to the emergence of several diseases, which so far, the etiological agent have remained unknown. This study will provide new information for future studies of disease management, quarantine programs and especially, the development of resistant varieties for collar and root rot of J. curcas.
12

Asymptomatic infections of Euphorbia lathyris by Macrophomina phaseolina.

Himmel, Phyllis Terry January 1988 (has links)
In November of 1984 and 1985, Euphorbia lathyris was planted into a field naturally infested with Macrophomina phaseolina located at the Campbell Avenue Farm in Tucson, Arizona. Plants without foliar symptoms and rhizosphere soil were sampled regularly from emergence until the following May or June. Soil rhizosphere populations ranged from 0.7-3.0 cfu/g soil in 1985 to 8.0-24.1 cfu/g soil in 1986, and did not change significantly over either growing season (P > 0.05). Both the incidence of disease and the number of infection sites per cm of root increased significantly (P < 0.05) over each growing season and were not related to rhizosphere soil populations of M. phaseolina (P > 0.05). The distribution of infection sites along the tap root over both growing seasons remained the same in that most were located in the top 0-7 cm of tap root. Infected E. lathyris without apparent symptoms were subjected to low-water and high-temperature stress treatments in growth chambers. Root infection was not found to be dependent upon any stress. Lesion development was significantly dependent upon the imposition of any stress treatment, and further root colonization was significantly dependent upon low-water stress (P < 0.05). M. phaseolina was consistently recovered from asymptomatic roots. A consistently lower leaf water potential was measured on infected E. lathyris than from non-infected controls when no stress treatment was applied. Polyclonal antisera made against hyphae and microsclerotia of M. phaseolina was not successful in detecting this pathogen in E. lathyris by I-ELISA. Antisera applied to fresh thin sections of infected plant tissue was effective in staining hyphae of M. phaseolina when used with a second antibody conjugated to fluorescence isothiocyanate or to an enzyme (to which a substrate was added to "stain" hyphae).
13

Interferência de patógenos nos resultados dos testes de vigor em sementes de feijoeiro /

Frigeri, Thaís. January 2007 (has links)
Resumo: O objetivo dessa pesquisa foi verificar a influência de Macrophomina phaseolina, Colletotrichum dematium f. truncata e Colletotrichum lindemuthianum na qualidade fisiológica de sementes de feijoeiro, em especial nos resultados do teste de condutividade elétrica. Foram utilizadas sementes das cultivares Carioca e FT Nobre. As sementes foram infectadas com os fungos em meio de cultura BDA sem e com restrição hídrica (acrescido de manitol a -1,0 MPa). Nos tratamentos testemunhas foram utilizados os mesmos meio de cultura, porém, sem a presença dos fungos. Para cada tratamento as sementes foram sobrepostas nos meios de cultura por 16 horas para M. phaseolina e por 48 horas para o caso de C. dematium f. truncata e C. lindemuthianum. Após secagem natural foram realizados testes de sanidade, germinação em areia e de vigor, como, índice de velocidade de emergência, peso da matéria seca da plântula, teste de frio, teste de envelhecimento acelerado e condutividade elétrica. Também foram realizadas análises na água de embebição das sementes no teste de condutividade elétrica, determinando-se as concentrações de potássio, cálcio e magnésio, assim como nos meios de cultura utilizados com e sem a sobreposição de sementes, para verificação da hipótese de consumo de nutrientes das sementes pelos fungos...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The aim of this research was to verify the influence of Macrophomina phaseolina, Colletotrichum dematium f. truncata and Colletotrichum lindemuthianum in the physiological quality of bean seeds, special in the results of electrical conductivity test. There were used seeds from Carioca and FT Nobre cultivars. The seeds were artificially inoculated with fungi M. phaseolina, C. dematium f. truncata and C. lindemuthianum in BDA medium culture with and without hydric restriction (it was added -1,0 MPa manitol). In the control treatment there was used the same medium culture without the presence of the fungi. For each treatment the seeds were placed on the medium culture for 16 hours for M. phaseolina and 48 hours for C. dematium f. truncata and C. lindemuthianum. After a nature dry, seeds were evaluated, by the blotter test, sand germination and vigor tests: speed of emergence index, seedlings dry weigh, cold, accelerated aging and electrical conductivity tests. Analyses from the imbibition water of the seeds in the electrical conductivity test, were also done a measuring of concentrations of potassium, calcium and magnesium, as well as in the medium culture used with and without the seeds, to verify the hypothesis of seeds nutrients consumption by the fungi...(Complete abstract, click electronic address below) / Orientadora: Rita de Cássia Panizzi / Coorientador: Nelson Moreira de Carvalho / Banca: Margarete Camargo / Banca: Juliana Altafin Galli / Mestre
14

Dissecting the molecular responses of Sorghum bicolor to Macrophomina phaseolina infection

Bandara, Y.M. Ananda Yapa January 1900 (has links)
Doctor of Philosophy / Department of Plant Pathology / Christopher R. Little / Charcoal rot, caused by the necrotrophic fungus, Macrophomina phaseolina (Tassi) Goid., is an important disease in sorghum (Sorghum bicolor (L.) Moench). The molecular interactions between sorghum and M. phaseolina are poorly understood. In this study, a large-scale RNA-Seq experiment and four follow-up functional experiments were conducted to understand the molecular basis of charcoal rot resistance and/or susceptibility in sorghum. In the first experiment, stalk mRNA was extracted from charcoal-rot-resistant (SC599) and susceptible (Tx7000) genotypes and subjected to RNA sequencing. Upon M. phaseolina inoculation, 8560 genes were differentially expressed between the two genotypes, out of which 2053 were components of 200 known metabolic pathways. Many of these pathways were significantly up-regulated in the susceptible genotype and are thought to contribute to enhanced pathogen nutrition and virulence, impeded host basal immunity, and reactive oxygen (ROS) and nitrogen species (RNS)-mediated host cell death. The paradoxical hormonal regulation observed in pathogen-inoculated Tx7000 was characterized by strongly upregulated salicylic acid and down-regulated jasmonic acid pathways. These findings provided useful insights into induced host susceptibility in response to this necrotrophic fungus at the whole-genome scale. The second experiment was conducted to investigate the dynamics of host oxidative stress under pathogen infection. Results showed M. phaseolina’s ability to significantly increase the ROS and RNS content of two charcoal-rot-susceptible genotypes, Tx7000 and BTx3042. Over-accumulation of nitric oxide (NO) in stalk tissues in the pathogen-inoculated susceptible genotypes was confirmed using a NO-specific fluorescent probe and confocal microscopy. Significantly increased malondialdehyde content confirmed the enhanced oxidative stress experienced by the susceptible genotypes after pathogen inoculation. These findings suggested the contribution of oxidative stress-associated induced cell death on charcoal rot susceptibility under infection. In the third functional experiment, the behavior of the sorghum antioxidant system after pathogen inoculation was investigated. M. phaseolina significantly increased the glutathione s-transferase (GST), glutathione peroxidase (GPX), glutathione reductase (GR), and peroxidase activities of the susceptible genotypes (Tx7000, BTx3042) but not in the resistant genotypes (SC599, SC35). Increased activities of these enzymes in susceptible genotypes may contribute to reduced oxidative stress thus lowering charcoal rot susceptibility. The fourth functional experiment was designed to quantify induced host-derived cell wall degrading enzymes (CWDEs) using crude enzyme mixtures from stalks. A gel diffusion assay revealed significantly increased pectinesterase activity in pathogen-inoculated Tx7000 and BTx3042 while significantly increased polygalacturonase activity was determined by absorbance. Fluorimetric determination of cell extracts revealed significantly increased cellulose degrading enzyme activity in M. phaseolina-inoculated Tx7000 and BTx3042. These findings revealed the pathogen’s ability to promote charcoal rot susceptibility in grain sorghum through induced host CWDEs. The last functional study was designed to profile the stalk tissue lipidome of Tx7000 and SC599 after M. phaseolina inoculation using automated direct infusion electrospray ionization-triple quadrupole mass spectrometry (ESI-MS/MS). M. phaseolina significantly decreased the phytosterol, phosphatidylserine, and ox-lipid contents in Tx7000 while significantly increasing stigmasterol:sitosterol ratio. Except for ox-lipid content, none of the above was significantly affected in resistant SC599. Results suggested the lethal impacts of M. phaseolina inoculation on plastid- and cell- membrane integrity and the lipid-based signaling capacity of Tx7000. Findings shed light on the host lipid classes that contribute to induced charcoal rot susceptibility in grain sorghum.
15

The potential of eliminating the grain sink for enhancing biofuel traits in sweet sorghum hybrids

Jebril, Jebril January 1900 (has links)
Doctor of Philosophy / Department of Agronomy / Tesfaye Tesso / Sweet Sorghum [Sorghum bicolor (L.) Moench] is a type of cultivated sorghum grown primarily for its sugar-rich stalks. Because of its high fermentable sugar content, the crop is widely recognized as an alternative feedstock source for bio-fuel production. The extent to which stalk sugar accumulation occurs may be determined by several factors including the sink size. Grain is the most important sink in sorghum and other grain crops. Three experiments were conducted in this study to determine the extent to which the grain sink can reduce sugar accumulation in the stalks, to test and validate a genetic system that allows development of sterile sweet sorghum hybrids, and to assess the potential of sugar-rich hybrids to overcome stalk rot diseases. The first experiment, based on 22 sweet sorghum genotypes, was undertaken to study the effect of eliminating the grain sink (removing the head prior to anthesis) on stalk juice yield, sugar accumulation, and biomass. The data showed that the grain sink had a significant effect on all traits measured. Elimination of the grain sink significantly increased oBrix % (17.8%), dry biomass (27.8%), juice yield (23.9%), and total sugar yield (43.5%). The second experiment was aimed at validating the role of A3 genetic male sterility system for producing sterile sweet sorghum hybrids. Ten sweet sorghum pollinator lines of variable sugar content were selected among the entries included in the previous experiment. The lines were crossed to four A1 and A3 cytoplasmic male sterile (CMS) lines using a Design II mating scheme. The A3 females did not have effective restorers so that the hybrids were expected to be sterile. The parental lines and corresponding hybrids were evaluated for biomass production, oBrix, juice and sugar yield using a randomized complete block design. All A3 hybrids were sterile and did not produce seed when heads were covered prior to pollination. The effect of grain sink represented by the A1 vs. A3 CMS were highly significant for Brix%, biomass, juice, and sugar yield. Comparison of parents vs. crosses component was highly significant, indicating marked heterosis effect for the traits. Both general (GCA) and specific (SCA) combining ability effects were also significant for all traits, indicating the role of both additive and dominance genetic effects in the inheritance of the characters. Earlier studies have shown positive relationships between stalk sugar concentration and stalk rot disease resistance in sorghum. Thus, the objective of the third experiment was to study the effects of the CMS mediated differential accumulation of stalk sugar on severity of charcoal rot disease caused by Macrophomina phaseolina. The experiment provided an opportunity to test the effect of variable stalk sugar in the same genetic backgrounds. The data indicated that hybrids produced from A3 cytoplasm were more resistant to charcoal rot (7.1cm lesion length) compared to those produced from the A1 hybrids (9.5 cm lesion length). The enhanced resistance of hybrids with higher sugar yield could have significant agronomic advantage in sugar based bio-fuel feedstock production.
16

Differentiation of Fungal Phytopathogens by FT-IR and MALDI-TOF MS

Atkinson, Curtis Muldrow 14 December 2013 (has links)
The use of matrix assisted laser desorption ionization time-oflight mass spectrometry, Fourier transform infrared spectroscopy, and other analytical means of identifying and differentiating microorganisms hold much promise. These analytical tools have been extensively assessed for their ability to differentiate bacteria and fungi. Most of this research has been coordinated in medically relevant microorganisms, but the technology can work just as well with agriculturally important microorganisms. In this thesis, these technologies were reviewed and then subsequently studied for their ability to differentiate Aspergillus species (that devastate corn and other crops yearly with aflatoxin contamination), as well as Macrophomina phaseolina and Thielaviopsis basicola which limit yields on soybean and other crops yearly. With the use of these technologies, harmful plant pathogens could be identified and subsequently treated to improve crop yields and also help to protect our nation and state’s food supply.
17

The Strawberry Rhizosphere Microbiome: Role on Plant Health and Nutrition

Boyd, Eric Michael 01 June 2020 (has links) (PDF)
Microbial-root associations are important to help plants cope with abiotic and biotic stressors. Managing these interactions offers an opportunity for improving the efficiency and sustainability of agricultural production. By characterizing the bacterial and archaeal community (via 16S rRNA sequencing) associated with the bulk and rhizosphere soil of sixteen strawberry cultivars in two controlled field studies, we explored the relationships between the soil microbiome and plant resistance to two soilborne fungal pathogens of strawberry (Verticillium dahliae and Macrophomina phaseolina). Overall, the plants had a distinctive rhizosphere microbiome relative to the bulk soil, with higher abundances of known beneficial bacteria such as Pseudomonads and Rhizobium. Plant genotype, biomass, leaf nutrient content and mortality were influenced differently by the rhizosphere microbiome in each of the two trials. In the V. dahliae trial, the rhizosphere microbiome was associated with plant biomass and leaf nutrient content and only indirectly to the disease resistance. In the M. phaseolina trial, the rhizosphere microbiome was associated to plant biomass, but not nutrient content; furthermore, resistant cultivars had larger abundances of Pseudomonas and Arthrobacter in their rhizosphere relative to susceptible cultivars. The mechanisms involved in these beneficial plant-microbial interactions and their plasticity in different environments should be studied further for the design of low-input disease management strategies.
18

Effects of Nitrogen Management and Cultivar on Strawberry Production Under Disease Pressure

Garcia-Brucher, Kamille A 01 December 2021 (has links) (PDF)
Effects of nitrogen management and cultivar on strawberry production under disease pressure Kamille Garcia-Brucher California strawberry growers face increasing regulatory pressures to manage nitrogen (N) applications in their production system. Standard practice in the California strawberry industry is to apply a synthetic pre-plant controlled release fertilizer (CRF) to ensure the crop has sufficient N during winter establishment. Some research from the UC Cooperative Extension suggests this practice is not efficient at delivering N to the crop since most of the N is released from CRF before strawberry crop N uptake is significant. Another concern for California strawberry growers is loss of their crop to a myriad of soilborne pathogens. Compost is commonly applied as a soil amendment in California strawberry fields as it offers both agronomic and environmental benefits including the potential for disease suppression. In light of legislation restricting N in some California cropping systems, Ag Order 4.0, and incentives programs established to promote soil conservation practices, compost may be a viable substitute for synthetic pre-plant CRF N. In this study, we investigated the effects of pre-plant fertilizer and strawberry cultivar on fruit yield, disease incidence, soil and plant N dynamics and soil carbon (C) at the Cal Poly Strawberry Center, San Luis Obispo, CA in a field infested with Macrophomina phaseolina. Pre-plant fertilizer treatments included 100 lb N/ac Cal Poly certified organic compost, 100 lb N/ac synthetic CRF and a control treatment (0 lb N/ac). Strawberry cultivars included three UC varieties, ‘Monterey,’ ‘Albion,’ and ‘San Andreas,’ and one Driscoll’s proprietary cultivar. Fruit yield and plant mortality data were collected throughout the growing season. Soil C was measured from soil samples collected in the root zone (6 in) while soil nitrate was measured from pore water samples collected in and below the root zone (6 and 12 in, respectively). Strawberry crop N uptake was determined using destructive plant samples while fruit N concentration was determined from subsamples of harvested fruit taken in April, May, June, and July each year. Although compost application did not significantly affect C sequestration and did not reduce disease incidence, there was no significant difference in total yield between compost and CRF treatments suggesting that compost can substitute for synthetic CRF without negatively affecting yield. There was significantly less plant mortality in control treatments compared with compost and CRF treatments suggesting excessive pre-plant N impacts disease incidence by M. phaseolina but more research is needed to better understand the mechanisms of infection by this soilborne pathogen. Total yield in this experiment was lower compared with statewide averages and crop N concentration was lower compared with the literature which is likely a result of disease pressure. Fruit N concentrations for the cultivars in this study were lower than the conversion coefficient defined by the Ag Order which means growers are removing less N through harvest allowing them more room in their N budget. Based on our results, compost may be substituted for synthetic CRF without negatively affecting yield and perhaps even make desirable soil improvements in this production system. And in fields with significant levels of M. phaseolina in the soil, N applications should be considered as it was seen to impact disease incidence. Keywords: compost, controlled release fertilizer, M. phaseolina, nitrogen uptake, Ag Order 4.0
19

Plant Compound Pest Control in California Strawberry (Fragaria × ananassa) Production

Weissman, Eli Mahanes 01 February 2017 (has links) (PDF)
Allelopathy occurs when one organism releases a compound into the environment that affects the functioning of another organism. Scientists have long suspected that alleopathic plant compounds could offer novel, softer chemistries to the ongoing battle of controlling pests in agricultural fields. Strawberry growers rely on toxic fumigants to kill soilborne fungal pests, weeds, nematodes, and insects. Increased regulations have reduced the use of fumigants (including methyl bromide), and strawberry growers need new sustainable pest control solutions. We selected four putative allelochemicals with known fungicidal and herbicidal activity (ferulic acid, gallic acid, juglone, and p-Coumaric acid). We assessed the pesticidal activity of these plant compounds both in agar and in soil on two emerging soilborne fungal pathogens (Macrophomina phaseolina and Fusarium oxysporum f.sp. fragariae), and four annual weeds commonly found in strawberry production fields (Malva parviflora, Melilotus officinalis, Poa annua, and Senecio vulgaris). We also assayed lettuce (Lactuca sativa ‘Inferno’), which served as a positive control plant species due to its sensitivity to phytotoxic compounds. Fitted sigmoidal dose-response curves predicted EC50 and EC75 values for each combination of plant compound and pest. All plant compounds inhibited the in vitro radial mycelial growth of the two soilborne fungal pathogens in a dose-dependent manner. Fusarium oxysporum f.sp. fragariae was more sensitive to the plant compounds than Macrophomina phaseolina. Average EC50 values for the radial mycelial growth of two F. oxysporum f.sp. fragariae isolates were 75.1 parts per million by weight (ppmw) juglone, 469 ppmw p-Coumaric acid, and 687 ppmw ferulic acid. Average EC50 values for the radial mycelial growth of two M. phaseolina isolates were 196 ppmw juglone, 2869 ppmw p-Coumaric acid, and 5716 ppmw ferulic acid. The three compounds we assayed in vitro also reduced M. phaseolina colony forming unit counts in soil and the EC50 values were 476 ppmw ferulic acid, 612 ppmw juglone, and 827 ppmw p-Coumaric acid. Metconazole, the conventional fungicide control, did not inhibit M. phaseolina colony forming unit counts in soil at its label high rate. The plant compounds required similar or lower rates to inhibit colony forming units that grew from M. phaseolina overwintering structures (microsclerotia) in soil as to inhibit radial mycelial growth in vitro. Based on the EC50 value in soil assays, ferulic acid was the least expensive plant compound to apply on a per acre basis to inhibit M. phaseolina ($74,226). In F.oxysporum f.sp. fragariae soil assays, the compounds induced hormesis at lower rates and may be germination stimulant candidates. Metconazole and the high rates of every compound effectively or completely inhibited F. oxysporum f.sp. fragariae colony forming units in soil. The plant compounds were more herbicidal than fungicidal in vitro. When combining the in vitro seedling length results for L. sativa, M. parviflora, P. annua, and S. vulgaris the EC50 values differed significantly (p < .0001) and were: 47 ppmw juglone, 120 ppmw p-Coumaric acid, 189 ppmw ferulic acid, and 297 ppmw gallic acid. At least one rate of ferulic acid, juglone, and p-Coumaric acid inhibited the germination of all plant species, while gallic acid only inhibited the germination of P. annua at 1000 ppmw (p < .05). In soil, visible microbial contamination in individual wells of 24-well plates and seed dormancy made it difficult to fit curves to weed seedling length data. The soil assay L. sativa seedling length EC50 values 11 days after initial treatment were slightly higher than in vitro, although plant compounds were in the same order of phytotoxicity: 129 ppmw juglone, 616 ppmw p-Coumaric acid, 644 ppmw ferulic acid, and 1584 ppmw gallic acid. Based on the EC50 value in soil assays, the least expensive compound to inhibit L. sativa seedling length on a per acre basis was gallic acid ($21,676). Germination 26 days after initial soil treatment generally declined in a dose-dependent manner for each compound. There was a direct relationship between plant compound rate and seedling damage in soil with the higher rates of all compounds, except p-Coumaric acid, inducing damage comparable to a conventional herbicide (pendimethalin or oxyfluorfen). Contaminated treatments appeared to be due to an interaction between plant compounds and microorganisms because herbicide and water controls had almost no microbial growth 11 days after initial treatment. Further, there was a significant positive linear relationship between level of contamination in phenolic acid-treated wells (ferulic acid, gallic acid, and p-Coumaric acid, p < .0001) and the in soil rate. This relationship was slightly negative in juglone soil treatments (p = .0167), which may be due to its greater antimicrobial activity than the phenolic acids. We propose that herbicidal effects in soil were due to the joint effect of the plant compounds themselves, and the microbial growth in wells. Microbial growth was either antagonistic or additive to the inhibitory action of the plant compounds. The plant compounds we assayed were inhibitory of emerging fungal pathogens in strawberry production and common annual strawberry field weeds. Evidence presented in this thesis correlates well with past research that not only found plant compounds to be herbicidal and fungicidal, but also described their modes-of-action (such as the production of reactive oxygen species that causes necrotic lesions on roots, and inhibition of glycolytic enzyme activity that prevents germination), and implicate plant compounds as carbon sources for a variety of microorganisms. Compound prices are currently exorbitant, but may decline as demand increases. Whether or not they provide effective pest control may depend on soil texture, organic matter, microbial diversity, and other edaphic factors.
20

Identification and evaluation of mycotoxins produced by Macrophomina phaseolina

Khambhati, Vivek Hemant 06 August 2021 (has links)
The fungus Macrophomina phaseolina (Tassi) Goidanich (Mp) is the causal agent of charcoal rot in soybean and infects over 500 plant species worldwide. Mp produces various mycotoxins and is suspected of utilizing a toxin-mediated process to penetrate host tissue. Identification and evaluation of secondary metabolites produced by Mp will further elucidate the pathogenesis mechanisms used by the fungus. Mp cultures isolated from soybean were evaluated for phytotoxicity in a hydroponic soybean bioassay and chemically analyzed by LC-MS/MS. All Mp cultures at two dilutions induced phytotoxicity symptoms including chlorosis, necrosis, wilting, stunting, and death. Analysis identified 13 unreported secondary metabolites including mellein, a compound with various biological activities. The phytotoxicity of mellein was evaluated against soybean seedlings in hydroponic culture, and symptoms of wilting and stunting were observed at levels above 40 MUg/L. Observations suggest that mellein does not directly contribute to the phytotoxic effects of Mp cultures.

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