[No abstract submitted] / Science, Faculty of / Botany, Department of / Graduate

Stemphylium blight is a defoliating fungal disease caused by <i>Stemphylium botryosum</i>. It has become more prevalent in Saskatchewan. Although not much is known about the biology of the fungus, increasing lentil (Lens culinaris) yield losses of up 62% have been reported in Bangladesh and India. The infection of lentil by <i>S. botryosum</i> was investigated under a range of temperatures (5 to 30°C), wetness periods (0 to 48 h) and wetness periods interrupted by dry periods of 6 to 24 h. The experiments involved testing the impact of environmental conditions on germination of conidia on glass slides and stemphylium blight infection on lentil (cv. CDC Milestone). Generalised linear models and non-parametric tests were used to determine the effects of these factors on conidial germination and disease development. Infection levels increased with increasing temperature and wetness duration. A latent period of 48 h was observed at 25°C and 30°C under continuous wetness. The duration of the latent period increased with decreasing temperatures and decreasing wetness duration. <i>S. botryosum</i> required warm temperatures (above 25°C) and a minimum wetness period of 8 h for optimal disease development. Low levels of infection were observed within the first 2 h of incubation at 10°C and increased with longer wetting periods up to 48 h and temperatures up to 30°C. The pathogen could maintain infectivity during interrupted wetness periods despite its requirement for prolonged wetness periods. Infection levels were not significantly affected by interrupting dry periods of 6 to 24 h although long dry periods (24 h) combined with higher temperatures (30°C) resulted in a decrease in stemphylium blight severity. Germination studies on glass slides supported these findings. Response surface models were developed that provided a good fit for the response of conidial germination to temperature and wetness duration. The coefficients of determination for the regression of observed against predicted effects ranged from 0.88 to 0.97. The general additive model could also be used to predict stemphylium blight severity responses to temperature and wetness duration (scaled deviance = 1.04). However, that model tended to overestimate infection levels especially at lower temperatures. The coefficients of determination for the observed against predicted effects at 5 to 30ºC ranged from 0.77 to 0.92 for the general additive model.

Stemphylium blight of lentil caused by <i>Stemphylium botryosum</i> Wallr., is a serious problem in Bangladesh, northeast India and Nepal causing more than 60 % yield losses under epidemic conditions. The pathogen started to appear on lentil in Saskatchewan in recent years and is widely distributed throughout western Canada but it is not well understood. An investigation of inheritance of resistance to stemphylium blight was done in the lentil cross Barimasur-4 × CDC Milestone. In order to develop a reliable indoor screening technique for this inheritance study, a suitable isolate of <i>Stemphylium botryosum</i>, a suitable culture medium for inoculum production and an appropriate plant age for indoor inoculation were identified. The maximum differential of disease severity was observed when lentil genotypes were inoculated at 14 days after planting (DAP). At 14 DAP, lentil plants rapidly defoliated but were capable of regrowth which caused variability in scoring for disease reaction. Inoculation at 42 DAP, close to the flowering stage, was found to be better for consistently scoring disease reaction. V8P medium was most suitable for inducing conidia production. Based on ability to sporulate, the isolate SB-19 from Saskatchewan was identified as suitable for conducting genetic studies of resistance to stemphylium blight. It was compared to isolate SB-BAN from Bangladesh for aggressiveness on two lentil cultivars. The SB-BAN isolate was found to be more aggressive. A preliminary screening of local and exotic germplasm done with the two isolates revealed considerable variability for disease resistance. Resistance to <i>S. botryosum</i> appeared to be quantitatively inherited in the cross Barimasur-4 × CDC Milestone according to both field and indoor screenings. The results of this study also confirmed that Precoz, one of the parents of Barimasur-4, was resistant to <i>S. botryosum</i>.

Stemphylium blight is a defoliating fungal disease caused by <i>Stemphylium botryosum</i>. It has become more prevalent in Saskatchewan. Although not much is known about the biology of the fungus, increasing lentil (Lens culinaris) yield losses of up 62% have been reported in Bangladesh and India. The infection of lentil by <i>S. botryosum</i> was investigated under a range of temperatures (5 to 30°C), wetness periods (0 to 48 h) and wetness periods interrupted by dry periods of 6 to 24 h. The experiments involved testing the impact of environmental conditions on germination of conidia on glass slides and stemphylium blight infection on lentil (cv. CDC Milestone). Generalised linear models and non-parametric tests were used to determine the effects of these factors on conidial germination and disease development. Infection levels increased with increasing temperature and wetness duration. A latent period of 48 h was observed at 25°C and 30°C under continuous wetness. The duration of the latent period increased with decreasing temperatures and decreasing wetness duration. <i>S. botryosum</i> required warm temperatures (above 25°C) and a minimum wetness period of 8 h for optimal disease development. Low levels of infection were observed within the first 2 h of incubation at 10°C and increased with longer wetting periods up to 48 h and temperatures up to 30°C. The pathogen could maintain infectivity during interrupted wetness periods despite its requirement for prolonged wetness periods. Infection levels were not significantly affected by interrupting dry periods of 6 to 24 h although long dry periods (24 h) combined with higher temperatures (30°C) resulted in a decrease in stemphylium blight severity. Germination studies on glass slides supported these findings. Response surface models were developed that provided a good fit for the response of conidial germination to temperature and wetness duration. The coefficients of determination for the regression of observed against predicted effects ranged from 0.88 to 0.97. The general additive model could also be used to predict stemphylium blight severity responses to temperature and wetness duration (scaled deviance = 1.04). However, that model tended to overestimate infection levels especially at lower temperatures. The coefficients of determination for the observed against predicted effects at 5 to 30ºC ranged from 0.77 to 0.92 for the general additive model.

Stemphylium blight of lentil caused by <i>Stemphylium botryosum</i> Wallr., is a serious problem in Bangladesh, northeast India and Nepal causing more than 60 % yield losses under epidemic conditions. The pathogen started to appear on lentil in Saskatchewan in recent years and is widely distributed throughout western Canada but it is not well understood. An investigation of inheritance of resistance to stemphylium blight was done in the lentil cross Barimasur-4 × CDC Milestone. In order to develop a reliable indoor screening technique for this inheritance study, a suitable isolate of <i>Stemphylium botryosum</i>, a suitable culture medium for inoculum production and an appropriate plant age for indoor inoculation were identified. The maximum differential of disease severity was observed when lentil genotypes were inoculated at 14 days after planting (DAP). At 14 DAP, lentil plants rapidly defoliated but were capable of regrowth which caused variability in scoring for disease reaction. Inoculation at 42 DAP, close to the flowering stage, was found to be better for consistently scoring disease reaction. V8P medium was most suitable for inducing conidia production. Based on ability to sporulate, the isolate SB-19 from Saskatchewan was identified as suitable for conducting genetic studies of resistance to stemphylium blight. It was compared to isolate SB-BAN from Bangladesh for aggressiveness on two lentil cultivars. The SB-BAN isolate was found to be more aggressive. A preliminary screening of local and exotic germplasm done with the two isolates revealed considerable variability for disease resistance. Resistance to <i>S. botryosum</i> appeared to be quantitatively inherited in the cross Barimasur-4 × CDC Milestone according to both field and indoor screenings. The results of this study also confirmed that Precoz, one of the parents of Barimasur-4, was resistant to <i>S. botryosum</i>.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-04-27T18:09:30Z No. of bitstreams: 1 2015 - Jessica Coelho Melo.pdf: 1651769 bytes, checksum: 37df9227b9a25c24297ba5364064be27 (MD5) / Made available in DSpace on 2018-04-27T18:09:30Z (GMT). No. of bitstreams: 1 2015 - Jessica Coelho Melo.pdf: 1651769 bytes, checksum: 37df9227b9a25c24297ba5364064be27 (MD5) Previous issue date: 2015-08-21 / CNPq / The gray leaf spot caused by Stemphylium solani, has occasioned losses in the tomato crop (Solanum lycopersicum L.) in several regions of Brazil. Although there are resistant cultivars, the control has been made, essentially, with application of fungicides. These, despite the practicality and efficiency can lead to the accumulation of polluting substances in the air and in the fruits. This study aimed to evaluate: 1) the chemical control efficiency of the disease with commercial fungicides registered for crop and with alternative products such as syrups and plant extracts; 2) the influence of these treatments on the development and production of tomato; 3) the contribution of metals in the system and its contents in leaves and fruits; 4) to compare the contents of metals in the fresh mass of the fruits with the limits allowed by the legislation. Three trials were carried out, one in the greenhouse and two in the field, and different products were compared: mancozebe, tebuconazole, copper oxychloride, vi?osa mixture, bordeaux mixture, lime sulfur, garlic extract and water. Were evaluated the progress of the disease, the production and the supply of metals in the soil, plants and fruits (Cd, Co, Cr, Cu, Fe, Mn, Pb, Ni and Zn). The disease occurred on average at low intensity and did not affect the fruit production, which did not allow an adequate evaluation of the efficiency of the treatments. Among all the products tested, bordeaux and vi?osa mixtures and mancozebe were the most efficient in controlling the disease, however, without affecting the accumulation of fresh and dry mass of the plants and productivity. Bordeaux and vi?osa mixtures were the products that contributed the most to the incorporation of metals to the agrosystem followed by mancozebe and copper oxychloride. The concentration of metals in the fruits was below the maximum limit allowed by the MERCOSUL Technical Regulation, however, it deserves attention in view of fact that the Cd values found in the fruits are very close to the allowed limit, especially in the treatments with vi?osa mixture, oxychloride copper and bordeaux mixture. The continuous use of fungicides and vi?osa and bordeaux mixtures can contribute to the accumulation of heavy metals in the agrosystem. New studies addressing this topic should be made. / A mancha-de-estenf?lio, causada por Stemphylium solani, tem ocasionado perdas na cultura do tomateiro (Solanum lycopersicum L.) em v?rias regi?es do Brasil. Apesar de existirem cultivares resistentes, o controle tem sido feito, essencialmente, com aplica??o de fungicidas. Estes, apesar da praticidade e efici?ncia podem levar ao ac?mulo de subst?ncias poluentes no ar e nos frutos. O presente trabalho teve como objetivos avaliar: 1) a efici?ncia do controle qu?mico da doen?a com fungicidas comerciais registrados para a cultura e com produtos alternativos como caldas e extratos de plantas; 2) a influ?ncia destes tratamentos no desenvolvimento e produ??o do tomateiro; 3) o aporte de metais no sistema e os seus teores nas folhas e frutos; 4) comparar os teores de metais na massa fresca dos frutos com os limites permitidos pela legisla??o. Realizaram-se tr?s ensaios, um em casa-de-vegeta??o e dois no campo e compararam-se diferentes produtos: mancozebe, tebuconazole, oxicloreto de cobre, calda vi?osa, calda bordalesa, calda sulfoc?lcica, extrato de alho e ?gua. Avaliaram-se o progresso da doen?a, a produ??o e o aporte de metais no solo, plantas e frutos (Cd, Co, Cr, Cu, Fe, Mn, Pb, Ni e Zn). A doen?a ocorreu em m?dia a baixa intensidade e n?o afetou a produ??o de frutos, o que n?o permitiu uma adequada avalia??o da efici?ncia dos tratamentos. Dentre todos os produtos testados, as caldas bordalesa e vi?osa e o mancozebe foram os mais eficientes no controle da doen?a, por?m, sem afetar o ac?mulo de massa fresca e seca das plantas e a produtividade. As caldas bordalesa e vi?osa foram os produtos que mais contribu?ram para a incorpora??o de metais ao agrossistema seguido do mancozebe e do oxicloreto de cobre. A concentra??o de metais nos frutos esteve abaixo do limite m?ximo permitido pelo Regulamento T?cnico MERCOSUL, no entanto, merece aten??o tendo em vista o fato dos valores de Cd encontrados nos frutos estarem muio pr?ximo do limite permitido, especialmente nos tratamentos com calda vi?osa, oxicloreto de cobre e calda bordalesa. O uso cont?nuo de fungicidas e das caldas vi?osa e bordalesa podem contribuir para o ac?mulo de metais pesados no agrossistema. Novos estudos abordando este tema devem ser feitos.

<p>As part of development of a human pathogen suppressing in plantae model system within <i>Spinacia oleracea</i>, a known pathogen transmitting produce plants, the secretome of the <i>Stemphylium</i> strain PNW2016-02, an endophytic fungal isolate of spinach plants, was studied. This strain was previously isolated in Purdue University Northwest laboratories in the Biological Sciences Department. As the secondary metabolite secretions of PNW2016-02 were shown to have antibiotic properties against a broad range of bacteria, we sought to improve our understanding of these properties and other characteristics of PNW2016-02 by sequencing and annotating the genome of this strain. Chemical compound characterization was achieved using HPLC-MS, which provided the ability to identify and quantify chemical compounds contained within the PNW2016-02 secretome. Through multi-gene phylogenetic analysis and comparative genomics, we found that PNW2016-02 clusters with a clade representing <i>Stemphylium vesicarium</i>; however, genome annotation also uncovered several genes unique to PNW2016-02. To assist in more fully understanding the secondary metabolites PNW2016-02, a semi-automated bioinformatic pipeline was developed in the R statistical environment in order to reconstruct and characterize the metabolic pathways, especially those pertinent to antibiotic production. Analysis using the bioinformatic pipeline revealed the presence of a number of antimicrobial metabolites, such as flavonoids, and suggests these compounds are produced and transformed in pathways, such as the phenylpropanoid synthesis pathway. These findings also suggest an important vital role of the shikimate pathway for antimicrobial metabolite production within PNW2016-02. Overall, the findings presented here have implications for understanding the antimicrobial strategies employed by <i>Stemphylium</i> PNW2016-02 and its potential for use in the above-mentioned model system.</p>

L'estemfiliosi de la perera és una malaltia fúngica ocasionada pel fong Deuteromicet Stemphylium vesicarium. Els símptomes característics d'aquesta malaltia són lesions necròtiques en fulles i fruits, provocant el depreciament econòmic d'aquests últims. Si les condicions climàtiques són favorables aquesta malaltia pot arribar a provocar la pèrdua total de la producció. La malaltia es localitza bàsicament en la conca mediterrània, sent les zones fructícoles de Girona a Catalunya i Emilia-Romagna a Itàlia els llocs on les pèrdues de producció són més grans. Actualment l'únic mètode de control és la cobertura permanent dels òrgans de la planta amb fungicides. Per obtenir aquesta protecció és necessari realitzar aplicacions preventives amb una cadència fixa, setmanal o quinzenal, de diferents fungicides en els arbres. Aquesta cadència d'aplicació comporta que al llarg del període vegetatiu de la perera es realitzin entre 20 i 25 aplicacions de fungicida, per cadències setmanals, o entre 10 i 13 aplicacions si la cadència és quinzenal. Aquest elevat nombre de tractaments comporta uns efectes toxicològics a nivell mediambiental i per a la salut humana, un augment del cost econòmic de la producció i en alguns casos afavorir l'aparició de resistències del patogen a determinats fungicides. Molts d' aquests tractaments es realitzen sense que les condicions ambientals siguin favorables a l'inici d'infeccions del fong, i són per tant innecessàries. Amb l'objectiu d'obtenir una eina per determinar el risc d'infeccions en funció de les condicions climàtiques i realitzar d'aquesta manera els tractaments amb fungicides només quan són necessaris, es va elaborar el Sistema pel Tractament Racionalitzat de l'Estemfiliosi de la Perera (STREP). El model consisteix en relacionar quantitativament amb la severitat de la malaltia els paràmetres climàtics de durada del període d'humectació i temperatura mitjana durant aquests períodes. En aquest treball s'ha determinat l'eficàcia en el control de la malaltia per diferents fungicides aplicats de manera preventiva amb una cadència fixa en diferents finques comercials i durant varis anys. S'ha constatat que no hi ha cap producte totalment eficaç en el control de la malaltia, sent l'eficàcia obtinguda mitjana. Aquesta eficàcia de control esta en funció del nivell de pressió de la malaltia, obtenint eficàcies inferiors quan el nivell en els testimonis no tractats és elevat. En assajos en condicions controlades s'ha determinat que els fungicides analitzats aplicats de manera curativa o postinfecciosa, quan ja s'han iniciat les infeccions, presenten una eficàcia més baixa que quan s'apliquen preventivament. El tiram és el producte amb una eficàcia més elevada en la major part de les condicions experimentals. S' ha analitzat durant 5 anys, 1992-1996, en diferents plantacions de perera de Girona, la dinàmica dels paràmetres ambientals de durada deis períodes amb humectació, la temperatura mitjana de l'aire, la temperatura mitjana durant els períodes d'humectació, la durada dels períodes amb humitat relativa superior al 90% i la pluviometria. S 'ha constatat que la humectació deguda a la rosada és la principal causa d'humectació i que és un procés lent i continu. Les durades més freqüents d'humectació han estat períodes inferiors a 12 hores i les temperatures més freqüents durant aquests períodes han estat entre 11 i 21°C. S'ha determinat també que existeixen períodes amb humitat relativa elevada però sense formació de pel·lícula d'aigua i s'ha observat que ocasionalment existeixen períodes breus d'interrupció de la humectació en que poden existir humanitats relatives elevades o baixes.La capacitat predictiva del model STREP ha estat avaluada mitjançant 42 assajos en condicions naturals i controlades, determinant que en més del 95 % dels casos avaluats les prediccions de malaltia s'han correspost amb les observades, i que només en dos casos el model ha infraestimat el nivell de risc d'infecció. Demostrant així que el model STREP és fiable en les seves prediccions. S'ha estudiat l'efecte de la humitat relativa durant els períodes interromputs d'humectació i les durades d'aquests períodes, en les infeccions produïdes pel fong. S'ha constatat que el model STREP hauria d'incorporar com a períodes separats els períodes d'humectació interromputs amb relatives elevades (>98%) s'haurien de considerar com a períodes continus. Per una altra banda s'ha comprovat que els períodes amb humitats relatives elevades sense formació d'humectació en condicions naturals no són suficients per provocar l'inici d'infeccions de S. Vesicarium.S'ha utilitzat el model STREP com a eina per guiar els tractaments fungicides en diferents parcel·les de finques comercials durant dos anys. D'aquests assajos es pot concloure que utilitzant valors de risc acumulat SA:0.4 com a llindar per a realitzar els tractaments es controla la malaltia amb una eficàcia semblant als tractaments setmanals, però amb una reducció del nombre d'aplicacions necessàries entre un 25 i un 50%. Utilitzar un llindar SA:0.6 en finques amb varietats sensibles i una pressió d'inòcul del patogen elevada presenta bons resultats però inferiors als obtinguts amb tractaments setmanals, no obstant l'estalvi de tractaments ha estat molt important (50-70%).De tots aquests resultats es conclou que el model STREP és adequat per ésser utilitzat en programes pilot a escala comercial en el control racional de l'estemfiliosi de la perera. En funció d'aquests resultats es considerarà la seva implementació en Estacions d'Avisos Fitosanitaris. / Brown spot of pear, caused by Stemphylium vesicarium, is an important disease in fruit-growing areas of Europe, mainly in Girona (Catalonia) and Emilia-Romagna (Italy). Infections and necrosis occur on leaves and fruits. If the environmental conditions are favourable for the disease, the loss of production will be very high. The control of brown spot of pear is based on protecting sprays of fungicide applied, al 7-to 15-day intervals depending on the type of fungicide. The high number of fungicide applications needed to maintain acceptable levels of disease can produce toxicological effects on ecosystems, humans and increased costs of pear production. We have determined the efficacy of disease control by different fungicides, sprayed with a preventive strategy in 3 trials during 2 years. None of the fungicides showed high efficacy of disease control, and the efficacy was related to the disease level. Experiments under controlled-conditions showed that treatments done with post-¬infection strategies have a low efficacy. Thiram was the fungicide with better efficacy for both preventive and after-infection strategies. During five years,1992-1996, we studied the dynamics of environmental conditions of wetness period, temperature, high relative humidity period (>90%) and rainfall. The most frequent wetness periods were shorter than 12h. The temperature during these periods was between 11-21°C. There were periods with high relative humidity but without wetness formation, and there were also wetness periods interrupted by dry periods. The relative humidity during these dry periods was high or low. The research has been focused to eliminate unnecessary sprays, and an infection model for S. vesicarium (STREP) was previously developed, which quantifies the effect of wetness duration and temperature on the severity of infection by S. vesicarium. The model was evaluated in 42 field trials and was validated in relation lo its capacity to predict the intensity of disease. Most part of disease level predictions (95%) were coincident with disease levels observed, showing that STREP model was a reasonable good predictor. We studied the effect of relative humidity during interrupted wetness periods, and the effect on disease progression of periods with high relative humidity but without wetness on disease levels. Interrupted periods with low relative humidity can be considered as two separated periods, but if the relative humidity is high (>98 %) it can be considered as a continuous period. Periods shorter than 12-18 h with high relative humidity but without wetness were insufficient to start infections. The STREP model was evaluated during two years and was used for scheduling treatments with fungicides, and showed that the efficacy with SA:0.4 threshold was similar to weekly sprays, but saving 25-50 % of fungicide sprays. The utilisation of a threshold SA:0.6 showed a smaller efficacy than weekly sprays but it saved applications about 50-70 %. From these results it can be accepted say that the STREP model can be used in pilot trials as a forecaster for scheduling sprays in commercial orchards for rational control of brown spot of pear. If the results are acceptable, the model will be integrated on Grower's Warning Stations.