The cowpea, Vigna unguiculata (L.) Walp., is known as feijão-de-corda and feijão-verde , among others, is one of the main cultures exploited by small producers of the Northeast region of Brazil. Among the phytopathogens that affects its productivity, Sclerotium rolfsii Sacc. is noteworthy, causing the stem rot in several cultures around the world. The objective of this work was to evaluate the alternative control of S. rolfsii Sacc. in saplings of V. unguiculata (L.) Walp. trough the biocontrol of antagonists, organic residues incorporation to the soil, utilization of essential oils, plant extracts and mineral nutrition. The work was developed in the Laboratory de Phytopathology and in the vegetation house of CECA/UFAL. The pathogen was obtained trough the isolations of cowpea with symptoms of the disease and, afterwards, it was cultivated in sterilized rice. For in vitro control, the antagonists isolates were matched with the pathogen in PDA medium, for the purpose of evaluating the reduction of growth and the hyperparasitism. For the biofumigations of the soil, the organic materials poultry litter, mussel, sugar cane bagasse, bean residue, cassava scuff were dehydrated in stove at 55ºC for 96h, grinded in the concentrations of 10% and 20% (v/v) and incorporated to the substrate, infested for 20 days and compared to the group treated with methyl thiophanate and to the control. After thirty days, the seedlings were evaluated about the incidence and the suppression of the disease. In the in vivo control the seeds were microbiolized with antagonists (C110, C21, ENF24, R14 and Trichoderma harzianum), the fungicide and saline solution for the control. The substrate was infested with the pathogen, two days after the sow, and after 30 days were evaluated. For the natural substances, 21 days old seedlings were pulverized with cassava flour wastewater extract (40%), eucalyptus oil (1%), peppermint (1%), Ecolife® (2%), methyl thiophanate (0,7 g/L) and water for the control and after two days, the substrate was infested with the pathogen. Six days after the inoculation, a new pulverization was done. The mineral fertilization was done in the sow trough Sarruge solution and doses of calcium silicate and sodium, 50, 100, 500 and 1000mg/L-1 and water for the control. The substrate was infested two days after that and a second fertilization was done 10 days after the sow. After 30 days the evaluations took place. The antagonists R14, C16, ENF 24 and T. harzianum inhibited the pathogen with RC from 42 up to 57%. Trichoderma has the hyperparasitic capacity. The incorporation of organic material was not efficient in the control of the disease. The in vivo antagonists reduced the incidence of the disease, but it was effective in the suppression. The oils and plant extracts were not efficient in reduced the incidence. The mineral fertilizations was not able to suppress the disease. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O feijão-caupi, Vigna unguiculata (L.) Walp., é conhecido como feijão-de-corda e feijão-verde sendo uma das principais culturas exploradas pelos pequenos produtores no Nordeste brasileiro. Dentre os fitopatógenos que afetam sua produtividade, destaca-se Sclerotium rolfsii Sacc. que causa a podridão de colo em diversos cultivos do mundo. O objetivo do trabalho foi avaliar o controle alternativo de S. rolfsii Sacc. em mudas de V. unguiculata (L.) Walp. através do uso de antagonistas, incorporação de resíduos orgânicos ao solo, utilização de óleos essenciais, extratos vegetais e ecolife® e nutrição mineral. O trabalho foi desenvolvido no Laboratório de Fitopatologia e em casa de vegetação do CECA/UFAL. O patógeno foi obtido pelo isolamento de folhas de feijão-caupi com sintomas da doença e depois cultivados em arroz esterilizado. Para controle in vitro , os isolados de antagonistas foram pareados com o patógeno em meio de BDA, para avaliar a redução de crescimento micelial (RC) e o hiperparasitismo. Para a biofumigação do solo, as matérias orgânicas cama de frango, marisco, bagaço de cana, resíduo de feijão, raspa de mandioca foram desidratadas em estufa 55ºC por 96h, moídas e incorporadas ao substrato infestado, em concentrações de 10% e 20% (v/v) por 20 dias e comparadas ao tiofanato metílico e a testemunha. Após 30 dias, as plantas foram avaliadas quanto à incidência da doença e desenvolvimento da planta. No controle in vivo as sementes foram microbiolizadas com os antagonistas (C110, C21, ENF24, R14 e Trichoderma harzianum), o fungicida e água salina para testemunha. O substrato foi infestado com o patógeno, dois dias após o semeio foram avaliadas com 30 dias. Para as substâncias naturais, plantas de feijão-caupi, com 21 dias de idade foram pulverizadas com extrato de manipueira (40%), óleo de eucalipto (1%), hortelã pimenta (1%), Ecolife® (2%), tiofanato metílico (0,7 g/L) e água para a testemunha. Após dois dias, o substrato foi infestado com o patógeno. Ao completar seis dias da infestação, uma nova pulverização foi realizada. A adubação mineral foi realizada no semeio através de solução de Sarruge e doses de silicato de cálcio e sódio, 50, 100, 500 e 1000mg/L-1 e água para testemunha. O substrato foi infestado dois dias depois e uma segunda adubação foi feita 10 dias após o semeio. Os antagonistas R14, C16, ENF 24 e T. harzianum inibiram o patógeno com RC de 42 a 57%. Trichoderma teve capacidade hiperparasitária. A incorporação da matéria orgânica não foi eficiente no controle da doença. Os antagonistas in vivo reduziram a incidência da doença e contribuiu para o desenvolvimento das plantas. Os óleos e extratos vegetais não foram eficientes em reduzir a incidência da doença. A adubação mineral não foi capaz de suprimir a doença.

Made available in DSpace on 2016-12-08T16:44:50Z (GMT). No. of bitstreams: 1 PGPV15MA169.pdf: 682086 bytes, checksum: e66cb9ecfd276d4b0dca8a7c5e02653d (MD5) Previous issue date: 2015-02-27 / The state of Santa Catarina is a major producer and consumer of corn kernels. Much of this cereal is used by agribusinesses in the production of animal protein. The stalk rot (PC) and ear can harm the quality and quantity of grain produced. The objective of this study was to quantify damage caused by PC in different altitudes in the state of Santa Catarina, identify the pathogens involved, assess the quality of grains and relate the presence of disease in the stalk to damaged kernels. Samples were taken in nine municipalities in the 2012/13 crop (Quilombo, Descanso, Petrolândia, Cunha Pora, Ituporanga, Papanduva, Campos Novos, Fraiburgo and Vargeão) and six in the 2013/14 crop (Quilombo, Palmitos, Bela Vista do Toldo, Fraiburgo and Caçador). The areas were divided according to the altitude of origin: low areas (0-400 meters), medium (400-800 m) and high (above 800 m). The work was conducted in partnership with the corn seed company Pioneer, using its commercial assays for data collection. We evaluated four commercial hybrids: 30F53YH, 30R50YH, AG8025 (PRO and PRO2) and DKB240 (PRO and PRO2). The experimental design was completely randomized factorial design with four replications. Cases and identification of causative agents were quantified at harvest, which was separated ears of patients from healthy plants. The spikes were threshed separately, the beans were weighed and dried to determine the damage caused by PC and analysis of the percentage of burnt grain (GA). The PC incidence ranged from 8.7% to 63.5%, average 26.35% in the 2012/13 crop, and 0.6 to 13.8%, averaging 5.52% in the 2013 harvest / 14. The damage caused by stalk rot ranged from 171.2 to 1248.02 kg ha-1 (average of 453.42 kg ha-1) in the first harvest and 17.1 to 338.24 kg ha-1 (mean 138.59 kg ha-1) in the second. Higher incidences and PC damage was detected in the areas of medium and high altitude. The hybrid with higher incidence of PC was DKB240 (in PRO versions and PRO2). The disease more present in Santa Catarina areas was fusarium. There were significant linear correlation between incidence and damage by 60% of the assessed areas. In the season 2012/13 the Fraiburgo had a higher incidence of GA for the four hybrids, average of 14.9%, while Campos Novos the lowest, 0.5%. In the harvest 2013/14 was not a trend, each hybrid responded differently to the site. The relationship between PC and GA for the four hybrids was low / O Estado de Santa Catarina é um importante produtor e consumidor de grãos de milho. Grande parte desse cereal é utilizado pelas agroindústrias na produção de proteína animal. As podridões do colmo (PC) e de espiga podem prejudicar a qualidade e quantidade de grãos produzidos. O objetivo deste trabalho foi quantificar danos causados por PC em altitudes diferenciadas no estado de Santa Catarina, identificar os patógenos envolvidos, avaliar a qualidade dos grãos produzidos e relacionar a presença de doença no colmo com grãos ardidos. Foram realizadas coletas em nove municípios na safra 2012/13 (Quilombo, Descanso, Petrolândia, Cunha Porã, Ituporanga, Papanduva, Campos Novos, Fraiburgo e Vargeão) e seis na safra 2013/14 (Quilombo, Palmitos, Bela Vista do Toldo, Fraiburgo e Caçador). As áreas foram divididas de acordo com a altitude de origem: áreas baixas (0 a 400 metros), médias (400 a 800 m) e altas (acima de 800 m). O trabalho foi conduzido em parceria com a empresa de sementes de milho Pioneer, utilizando seus ensaios comerciais para coleta de dados. Foram avaliados quatro híbridos comerciais: 30F53YH, 30R50YH, AG8025 (PRO e PRO2) e DKB240 (PRO e PRO2). O delineamento experimental foi inteiramente casualizado, fatorial, com quatro repetições. A incidência de PC e identificação de agentes causais foram quantificadas na colheita, onde separou-se espigas de plantas doentes de sadias. As espigas foram trilhadas separadamente, os grãos foram pesados e secados para determinação do dano causado por PC e análise da porcentagem de grão ardido (GA). As incidências de PC variaram de 8,7% a 63,5%, média 26,35%, na safra 2012/13, e de 0,6 a 13,8%, média de 5,52%, na safra 2013/14. O dano causado por podridão de colmo variou de 171,2 a 1.248,02 Kg ha-1 (média de 453,42 Kg ha-1) na primeira safra e de 17,1 a 338,24 Kg ha-1 (média de 138,59 Kg ha-1) na segunda safra. Maiores incidências e danos de PC foram detectados nas áreas de média e alta altitude. O híbrido com maior incidência de PC foi DKB240 (nas versões PRO e PRO2). A doença mais presente nas áreas catarinenses foi fusariose. Existiu correlação linear positiva entre incidência e dano em 60% das áreas avaliadas. Na safra 2012/13 o município de Fraiburgo apresentou maior incidência de GA para os quatro híbridos avaliados, média de 14,9%, enquanto Campos Novos a menor, 0,5%. Na safra 2013/14 cada híbrido respondeu de forma diferenciada ao local. A relação entre PC e GA para os quatro híbridos avaliados foi baixa

The damping-off and stem rot disease syndrome is harmful to many cultivated crops. Damping-off and stem rot caused by Sclerotium rolfsii Sacc. on cowpea results in yield losses with serious socio-economic implications. The objectives of the present study were to investigate the occurrence of the diseases in Benin, study etiology and factors influencing the diseases, and develop strategies for the control of the diseases in the field. Results showed that the diseases are distributed countrywide. Sclerotium rolfsii was the main causal agent but minor pathogens, namely Pythium sp., Rhizoctonia solani Kühn and Phoma pomorum Thüm were also recorded. In the Ouémé valley, the diseases were favoured by soil moisture and S. rolfsii initial inoculum that were higher closer to the river. Sclerotium rolfsii isolates collected in the valley showed genetic diversity in terms of pathogenicity, mycelial compatibility groups and ITS rDNA sequences. A paper-based screening method was found to be a rapid laboratory method for screening for resistance in cowpea cultivars. Furtheremore, Moringa oleifera L. leaf extracts, Trichoderma Kd 63 and Trichoderma IITA 508 significantly reduced the disease incidence. The best disease control was recorded in the field when M. oleifera seed treatment was integrated with a soil sprinkle of Trichoderma. The present work provides information on damping-off and stem rot of cowpea in Benin and control strategies for ecologically sustainable cowpea production. / Thesis (DPhil (Microbiology and Plant Pathology))--University of Pretoria, 2006. / Microbiology and Plant Pathology / unrestricted

<p>  </p> <p><em>Sclerotinia sclerotiorum </em>is a soilborne pathogen of soybean that causes Sclerotinia stem rot, alternatively called white mold. Sclerotinia stem rot can cause significant yield losses under cool and wet environmental conditions. Two biofungicides, <em>Coniothyrium minitans </em>and <em>Bacillus amyloliquefaciens, </em>are currently available and labeled to limit or suppress <em>S. sclerotiorum</em> in soybean. These biofungicides can be applied in place of synthetic foliar fungicides to provide an alternative mode of action for the control of Sclerotinia stem rot. However, limited information is available regarding the efficacy of <em>C. minitans </em>and <em>B. amyloliquefaciens </em>as biocontrol agents of <em>S. sclerotiorum </em>in soybean and the sensitivity of the biofungicides biological activity on <em>S. sclerotiorum </em>to pesticides commonly used in soybean production systems. This research aims to provide management recommendations for <em>S. sclerotiorum </em>in soybean using <em>C. minitans </em>and <em>B. amyloliquefaciens </em>and to develop guidelines for how to incorporate the biofungicides into an established soybean pest management program. To assess the effectiveness of <em>C. minitans </em>and <em>B. amyloliquefaciens </em>as biocontrol agents of <em>S. sclerotiorum </em>dual culture, amended media, and soil plate assays were conducted along with experiments in the growth chamber and field. The presence of a distinct inhibition zone surrounding the <em>B. amyloliquefaciens </em>colony in the dual culture assay and the absence of mycelial growth on the media plates amended with <em>B. amyloliquefaciens </em>confirmed that the bacteria can control the mycelial growth of <em>S. sclerotiorum </em>through antibiosis. The absence of an inhibition zone surrounding the <em>C. minitans </em>isolate in the dual culture assay along with the degradation of sclerotia following treatment with <em>C. minitans </em>in the soil plate assay indicates an inability to limit the mycelial growth of <em>S. sclerotiorum </em>and confirms that the primary mode of action is mycoparasitism. In the growth chamber, <em>B. amyloliquefaciens</em> at 14.03 L/ha applied using the dip method significantly reduced Sclerotinia stem rot lesion length when compared to the non-treated control and resulted in the lowest lesion area under the disease progress curve (lAUDPC). When <em>B. amyloliquefaciens </em>and <em>C. minitans </em>were applied in the field, no differences were observed between treatments for soybean moisture, test weight, or yield. To evaluate the sensitivity of <em>B. amyloliquefaciens </em>and <em>C. minitans</em> biological activity on <em>S. sclerotiorum </em>to pesticides commonly used in soybean production systems a poison plate assay as well as soil plate, growth chamber, and field experiments were conducted. In the poison plate assay <em>C. minitans </em>was most sensitive to the preemergence herbicide flumioxazin and the synthetic fungicides boscalid and fluazinam, while <em>B. amyloliquefaciens </em>was sensitive only to the synthetic fungicide fluazinam. In the soil plate assay the mycoparasitic activity of <em>C. minitans </em>on sclerotia of <em>S. sclerotiorum </em>was sensitive to flumioxazin, metribuzin, glyphosate, picoxystrobin, and boscalid. In the controlled environment experiments, none of the pesticides tested decreased the efficacy of <em>B. amyloliquefaciens</em>. There were no significant interactions between <em>C. minitans </em>and <em>B. amyloliquefaciens </em>with preemergence herbicides, postemergence herbicides, and synthetic fungicides for soybean moisture, test weight, and yield. This research demonstrates that <em>B. amyloliquefaciens </em>and <em>C. minitans </em>are effective biocontrol agents of <em>S. sclerotiorum </em>in soybean. However, antagonistic relationships exist between the biofungicides and certain preemergence, postemergence, and synthetic fungicides used in soybean production systems.</p>

Sclerotinia sclerotiorum is a pathogenic fungus that infects around 400 species of host    plants. Stem rot disease caused by this fungus is economically disastrous for Brassica napus cultivators in Sweden. Due to the lack of disease resistant cultivars, disease management has been solely dependent on fungicide application. The current disease  prediction models are not scientifically accurate and take into account factors such as   weather, previous disease incidence, and conomic effects which often result in unnecessary and excessive use of fungicides by cultivators. Real-Time Polymerase Chain Reaction has proven to be the fastest, most accurate and reliable technique for detecting plant pathogens as it gives an idea about disease severity by measuring pathogen concentration in environmental samples. Reproducible and able qPCR assays have the potential of being the main principle on which more scientifically accurate plant disease prediction and management models an be developed. The aim of this study was to validate a previously established qPCR assay to detect S. sclerotiorum. An absolute quantification experiment     was performed by using plasmid DNA cloned with a target gene as template. Further,   three different qPCR machines  were compared  to make a plausible conclusion regarding    their sensitivity and efficiency in detecting minuscule amounts of DNA from the   environment. While a solid conclusion could not be reached regarding the sensitivity of    each of these machines, this study pointed out some basic trends about each machine    that may help researchers in selecting the most efficient qPCR system when working with detection of plant pathogens.