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Predacious fungi that capture and kill nematodes live, amongst other places, in the soil where it is assumed that they participate in the natural control of nematodes, including the plant parasitic species that cause tremendous losses to farmers. There are predacious fungi in the soil which prey on amoebae, rotifers, rhizopods and other small animals but they were not dealt with in this study. [...]

Resumo: O interesse pelo controle biológico de nematóides vem aumentando ano após ano, estimulado pelas crescentes restrições ao uso de agrotóxicos. Entre os agentes do controle biológico de nematóides, os fungos predadores vêm se destacando, dada a facilidade de crescimento em meios artificiais e substratos diversos e a comprovada eficácia. No presente estudo, confirmou-se que espécies de Arthrobotrys e de Monacrosporium estão entre os fungos nematófagos mais comuns nos solos do Brasil. Esses fungos crescem em subprodutos da agroindústria, potencializando a formulação desses agentes a baixo custo e pode tornar-se o recurso mais vantajoso para o manejo de nematóides em cultivos de hortifrutigranjeiros em volta de centros urbanos. Bagaço de cana misturado com farelo de arroz, na proporção de 2:1, é um substrato adequado à formulação de fungos nematófagos. Para o cultivo de alface, uma única aplicação, antes do plantio, é suficiente para o manejo dos nematóides. A adição de formulações dos fungos ao substrato para a produção de mudas pode facilitar a utilização desse recurso, e formulações contendo mais de uma espécie com habilidades predatórias diferentes, incluindo parasitos de ovos, podem ser mais eficazes que formulações de uma única espécie. A especificidade dos fungos dificulta o comércio de formulações desses agentes, dado o risco de comprometimento à credibilidade dos produtos, se formulações não-específicas para os nematóides que ocorrem em certas áreas, forem utilizadas. Por conseguinte, as formulações desses agentes são mais adequadas para a distribuição assistida, já que os nematóides que ocorrem nos locais onde serão empregadas, podem ser previamente identificados. / Abstract: The interest in the biological control of nematodes is increasing year after year, stimulated by the increasing restrictions to the use of chemical defensives. Among the agents for the biological control of nematodes, the predators fungi have become the group of higher interest since they grow easily on different kinds of material and they also present known efficacy. In the present study, it was demonstrated that species of Arthrobotrys and Monacrosporium are among the most common nematophagous fungi in Brazilian soils. These fungi grow on by-products of the agroindustry, making formulations of these agents of low cost and can become the most advantageous resource for the nematode control in vegetable production areas around urban centers in Brazil. Sugar cane bagasse mixed with rice bran in the ratio of 2:1 is an adequate substratum for the formulation of nematophagous fungi. For the lettuce culture, only one application, before the planting date, is enough for the control of nematodes. The addition of fungi to the substratum for the production of seedlings in nurseries can facilitate the use of this resource. Formulations containing more than one species with different predatory abilities, including egg parasites, can be more efficient than formulations with only one species. The specificity of the fungi makes difficult the commerce of formulations of these agents, since the risk of unsuccessful use can put down the credibility on the products, if non-specific formulations for the nematodes that occur in certain areas are used. Therefore, the formulations of these agents are more suitable for attended distribution, since the nematodes that occur in places where the formulation will be used can be previously identified. / Orientador: Jaime Maia dos Santos / Coorientador: José Carlos Barbosa / Banca: Clélia Aparecida Iunes Lapera / Banca: Marineide Mendonça Aguillera / Banca: Carlos Amadeu Leite de Oliveira / Banca: Júlio César Galli / Doutor

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A dissertation submitted to the Faculty of Science, University of the Witwatersrand, Johannesburg, in fulfilment of requirements for the degree of Master of Science. Johannesburg, 2016. / The entomopathogenic nematodes (EPNs) have emerged as an important group of insect pests. The EPNs which comprise the Steinernema genus share symbiotic associations with Xenorhabdus bacteria. This research project focused on isolating and characterizing a novel and indigenous EPN isolate with its associated bacteria. The biological control potential of the nematode was investigated in the areas of host infectivity, infective juvenile recovery and progeny yield. These processes were investigated at three different factors. These included time, population size and temperature. The infectious abilities of the symbiotic bacteria were also evaluated without the contributions of the nematode partner at different bacterial doses, time intervals and temperature regimens. The genome of the bacteria was thereafter acquired through whole-genome sequencing and annotation techniques to elucidate the virulence mechanisms and genes involved in temperature adaptation. The species isolated in this investigation was novel. The species shared an 85 % maximum identity to and taxonomically grouped with the species Steinernema khoisanae. The two species shared a common ancestor but the extended branch length of the species under investigation substantiated its novelty. The EPNs infected hosts at different time intervals, population densities and temperature regimens. However, the EPNs performed these processes to different extents. Host mortality significantly increased with time. The EPNs also infected insect hosts at the two experimental temperatures. However, host mortality was higher at the temperature regimen of 20° C and lower at 30° C. Host infections were not significantly different at two tested population densities of 500 and 1000 infective juveniles. The levels of interaction between temperature and time and temperature population density were not statistically significant. The subsequent biological process of recovery was evaluated. The EPNs recovered at both population densities and temperature regimens. The infective juvenile recoveries were statistically insignificant at both population densities and temperature regimens. Since recovery was based on the mere presence of progeny infective juveniles, the percentages were high which contributed to the statistical insignificant findings. This also contributed to the non-significant interaction between population density and temperature. The last biological process investigated was the progeny yield of infective juveniles. The yields were significantly different between both population densities of infective juveniles and temperature regimens. Higher yields were obtained at the temperature regimen of 20° C and 25° C. Lower yields were obtained at 30° C. The unexpected finding was higher progeny yields obtained from the lower population densities of infective juveniles. This contributed to the significant interaction present between population density and temperature. The bacteria were thereafter molecularly characterized. The symbiotic bacteria shared a 99 % sequence similarity to the species Xenorhabdus sp. strain GDc328. It was interesting to observe the infectious abilities of the bacteria without contributions from the EPNs. This study was measured at different bacterial doses, time intervals and temperature regimens. Host mortality was achieved without contributions from the EPN. Host mortality significantly increased with bacterial dose and time. Host mortality was also significantly different between each temperature regimen. Higher mortalities were observed at 30° C and lower mortalities were observed at 20° C. The differences in the performance between the EPN-bacterial partnership and the bacteria alone were attributed to the manner in which adaptation occurred. Since the EPN-bacteria existed as a bi-partite entity, the partners evolved as a bi-partite complex. The bacteria were removed from the symbiosis and cultured individually. External factors may have re-shaped the performance of the bacteria at the different temperature regimens. To further understand the genetic mechanisms of temperature adaptation, host infectivity and symbiosis, the draft whole genome sequence of the bacteria was then acquired. The genome of the bacteria comprised several genes which encoded the flagella system of the bacteria. Also pairs of co-localized toxin-antitoxin genes were discovered. Temperature acclimatization was performed through different cold and heat shock proteins and lastly several molecular chaperones. The studies showed that the species Steinernema spp. and its associated symbiotic bacteria Xenorhabdus sp. strain GDc328 were good bio-pesticide candidates for application against endemic insect pests. / LG2017

Submitted by Michele Mologni (mologni@unoeste.br) on 2019-01-24T16:37:31Z No. of bitstreams: 1 Lorrayne Guimarães Bavaresco.pdf: 1579894 bytes, checksum: be828e9da52619462a559db48496ff9a (MD5) / Made available in DSpace on 2019-01-24T16:37:31Z (GMT). No. of bitstreams: 1 Lorrayne Guimarães Bavaresco.pdf: 1579894 bytes, checksum: be828e9da52619462a559db48496ff9a (MD5) Previous issue date: 2018-08-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Among the forms of biological control, the use of rhizobacteria has shown to be promising in the suppression and protection of plants against the attack of phytonematoids. The objective of this work was to evaluate the effect of Bacillus subtilis on the reproduction of Meloidogyne spp., associated to changes in rhizosphere colonization and growth, nutrition and enzymatic activity of tomato cultivars tolerant and susceptible to root-knot nematodes. The methods of application consisted of the inoculation of Bacillus subtilis (AP-3) in pre-planting and post-planting of the tomato. Two experiments were conducted in a plant growth chamber. In the first experiment the tomato plants were cultivated on a sterile substrate, inoculating 5000 eggs of Meloidogyne incognita. In the second, tomato cultivation was carried out in sandy soil naturally infested with root-knot nematodes. The plant growth, nematode reproduction, soil and rhizosphere colonization by bacteria of the genus Bacillus spp., and biochemical changes were evaluated at 40 days after transplanting of the tomato. In both modes of application, the Bacillus subtilis was efficient in increasing root mass and in reducing the reproduction of the root-knot nematode in susceptible tomato cultivated in naturally infested soil. In sterile substrate, inoculation of B. subtilis in post-planting promoted reduction of the number of active forms of the nematode and both applications promoted the decrease of eggs of Meloidogyne incognita in the roots of the tomato. The application of B. subtilis, as a sterile substrate, induced changes in the nutritional uptake of tomato plants, increasing calcium levels and reducing leaf potassium. The application of B. subtilis contributed to the reduction of the peroxidase activity in leaves of the tomato susceptible to root-knot nematodes, in sterile condition. The rhizobacteria, after planting, allowed the increase in the proline content in the leaves of the tomato, cultivated in soil naturally infested with root-knot nematodes. Inoculations with B. subtilis promoted an increase in the number of Bacillus spp. in the sterile substratum and in the rhizosphere of the tomato, being confirmed the persistence of these bacteria at 40 DAP. / Dentre as formas de controle biológico, a utilização de rizobactérias tem se mostrado promissora na supressão e proteção de plantas ao ataque de fitonematoides. O presente trabalho teve como objetivo avaliar o efeito da aplicação de Bacillus subtilis na reprodução de Meloidogyne spp., associado as alterações na colonização da rizosfera e no crescimento, nutrição e atividade enzimática de cultivares de tomateiro tolerante e suscetível aos nematoides-das-galhas. Os modos de aplicação estudados consistiram na inoculação do Bacillus subtilis (AP-3) em pré-plantio e pós-plantio do tomateiro. Foram conduzidos dois experimentos em câmara de crescimento de plantas. No primeiro experimento as plantas de tomateiro foram cultivadas em substrato esterilizado, com inoculação de 5000 ovos de Meloidogyne incognita. No segundo, o cultivo do tomateiro foi realizado em solo arenoso naturalmente infestado por nematoides-das-galhas. Aos 40 dias após o transplantio das mudas, foram avaliados o crescimento das plantas, a reprodução do nematoide, a colonização do solo e da rizosfera por bactérias do gênero Bacillus spp. e as alterações bioquímicas do tomateiro. Em ambos os modos de aplicação, o Bacillus subtilis foi eficiente em aumentar a massa radicular e em reduzir a reprodução do nematoide-das-galhas em tomateiro suscetível, cultivado em solo naturalmente infestado. Em substrato estéril, a inoculação de B. subtilis em pós-plantio promoveu redução do número de formas ativas do nematoide e ambas as aplicações promoveram a diminuição de ovos de Meloidogyne incognita nas raízes do tomateiro. A aplicação de B. subtilis, na condição de substrato estéril, induziu mudanças na absorção nutricional do tomateiro, aumentando os teores de cálcio e reduzindo o potássio foliar. A aplicação de B. subtilis contribuiu para redução da atividade peroxidase nas folhas do tomateiro suscetível a nematoides-das-galhas, em condição estéril. A rizobactéria, em pós-plantio, possibilitou o aumento no teor de prolina nas folhas do tomateiro, cultivado em solo naturalmente infestado com nematoides-das-galhas. As inoculações com B. subtilis promoveram aumento do número de Bacillus spp. no substrato estéril e na rizosfera do tomateiro, sendo comprovada a persistência destas bactérias aos 40 DAP.

Thesis (M.Agric. (Horiticulture)) --Unversity of Limpopo, 2013 / The ground leaching technology (GLT) system, using crude extracts of wild cucumber (Cucumis myriocarpus) and wild watermelon (Cucumis africanus) fruits, had been widely researched and developed in management of the root-knot (Meloidogyne species) nematodes in tomato (Solanum lycopersicon) production. In the GLT system, experiments were harvested at 56 days after inoculation with nematodes, which was approximately three generations of Meloidogyne species. Also, studies in GLT systems demonstrated that effective micro-organisms (EM) were not essential in the release of chemicals from crude extracts for nematode suppression, with suggestions that the system exclusively relied upon irrigation or rainwater for leaching out chemicals. However, the system had hardly been tested on other nematode species with longer life cycles and crops. The objective of this study was to investigate the influence of crude extracts of C. myriocarpus (cucurbitacin A-containing phytonematicide) and C. africanus (cucurbitacin B-containing phytonematicide) with and without EM on suppression of population densities of the citrus nematode (Tylenchulus semipenetrans) on rough lemon (Citrus jambhiri) over three generations of the nematode. Two studies, one on C. myriocarpus and the other on C. africanus, with and without EM, were conducted separately using the GLT system under greenhouse conditions with three generations of T. semipenetrans as the standard of application interval of the materials. Citrus seedlings were transplanted in 7-L plastic pots contain 6.5-L pasteurised river sand and Hygromix (3:1 v/v) and inoculated with approximately 25 000 second-stage juveniles (J2s) in 2 × 2 × 2 factorial experiment, where treatments were arranged in a randomised complete block design (RCBD), with six replications. At 150 days after treatment, nematode and plant variables were collected and subjected to factorial analysis of variance. Under C. myriocarpus (Cm), EM × Cm interaction was not significant for nematodes (juveniles + eggs) in roots and juveniles in soil, while under C. africanus (Ca), EM × Ca interaction was highly significant for nematodes, but not for juveniles. Crude extracts of C. myriocarpus and C. africanus fruits contributed 21-36% and 38-59% to total treatment variation in nematodes, respectively. Relative to untreated control, crude extracts of C. myriocarpus fruit reduced nematodes by 22% in roots, but increased juveniles in soil by 93%. Similarly, C. africanus fruit reduced nematodes in roots by 80%, but increased juveniles in soil by 178%. The increase of juveniles in the soil was explained on the basis of opposing forces on nematode population densities under crude extracts of Cucumis and untreated control, along with the inherent nature of cyclic population growth in plant-parasitic nematodes. In plant variables, certain significant (P ≤ 0.05) interactions consistently occurred under both Cucumis species. However, effects of the interactions were not consistent under the two Cucumis species. In most of the variables, the non-significant effects of EM × Cm interactions supported the view that the GLT systems were independent of microbial activities, while significant (P ≤ 0.05) EM × Ca interactions suggested that the systems under C. africanus fruit could be viewed as being dependent upon microbial degradation activities. Growth of rough lemon rootstock was, to a certain extent, suppressed by application of crude extracts from Cucumis fruits, suggesting that the material were phytotoxic to this citrus rootstock. Under low nematode population densities, T. semipenetrans infection supported the view that nematode numbers below the damage threshold levels have stimulatory effects on growth of plants as observed in plant height under conditions of this study. In conclusion, the approximately three nematode-generation-application interval of 150 days for crude extracts of fruits in Cucumis species was rather too long for the efficacies of the materials on suppression of the population densities of T. semipenetrans in rough lemon seedlings. Consequently, shorter application intervals, as demonstrated for Meloidogyne species would be appropriate, although caution has to be taken to ensure that phytotoxicity to the rootstock was avoided.

Made available in DSpace on 2014-06-11T19:32:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-12-15Bitstream added on 2014-06-13T20:03:01Z : No. of bitstreams: 1 soares_plm_dr_jabo.pdf: 8552234 bytes, checksum: 46b7a86535ee8fa96f0e072e9d4b1dee (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O interesse pelo controle biológico de nematóides vem aumentando ano após ano, estimulado pelas crescentes restrições ao uso de agrotóxicos. Entre os agentes do controle biológico de nematóides, os fungos predadores vêm se destacando, dada a facilidade de crescimento em meios artificiais e substratos diversos e a comprovada eficácia. No presente estudo, confirmou-se que espécies de Arthrobotrys e de Monacrosporium estão entre os fungos nematófagos mais comuns nos solos do Brasil. Esses fungos crescem em subprodutos da agroindústria, potencializando a formulação desses agentes a baixo custo e pode tornar-se o recurso mais vantajoso para o manejo de nematóides em cultivos de hortifrutigranjeiros em volta de centros urbanos. Bagaço de cana misturado com farelo de arroz, na proporção de 2:1, é um substrato adequado à formulação de fungos nematófagos. Para o cultivo de alface, uma única aplicação, antes do plantio, é suficiente para o manejo dos nematóides. A adição de formulações dos fungos ao substrato para a produção de mudas pode facilitar a utilização desse recurso, e formulações contendo mais de uma espécie com habilidades predatórias diferentes, incluindo parasitos de ovos, podem ser mais eficazes que formulações de uma única espécie. A especificidade dos fungos dificulta o comércio de formulações desses agentes, dado o risco de comprometimento à credibilidade dos produtos, se formulações não-específicas para os nematóides que ocorrem em certas áreas, forem utilizadas. Por conseguinte, as formulações desses agentes são mais adequadas para a distribuição assistida, já que os nematóides que ocorrem nos locais onde serão empregadas, podem ser previamente identificados. / The interest in the biological control of nematodes is increasing year after year, stimulated by the increasing restrictions to the use of chemical defensives. Among the agents for the biological control of nematodes, the predators fungi have become the group of higher interest since they grow easily on different kinds of material and they also present known efficacy. In the present study, it was demonstrated that species of Arthrobotrys and Monacrosporium are among the most common nematophagous fungi in Brazilian soils. These fungi grow on by-products of the agroindustry, making formulations of these agents of low cost and can become the most advantageous resource for the nematode control in vegetable production areas around urban centers in Brazil. Sugar cane bagasse mixed with rice bran in the ratio of 2:1 is an adequate substratum for the formulation of nematophagous fungi. For the lettuce culture, only one application, before the planting date, is enough for the control of nematodes. The addition of fungi to the substratum for the production of seedlings in nurseries can facilitate the use of this resource. Formulations containing more than one species with different predatory abilities, including egg parasites, can be more efficient than formulations with only one species. The specificity of the fungi makes difficult the commerce of formulations of these agents, since the risk of unsuccessful use can put down the credibility on the products, if non-specific formulations for the nematodes that occur in certain areas are used. Therefore, the formulations of these agents are more suitable for attended distribution, since the nematodes that occur in places where the formulation will be used can be previously identified.