Lantana camara L. (Verbenaceae) remains a highly invasive and ecologically damaging weed in South Africa, despite some 50 years of biological control efforts. Lack of success has been ascribed to varietal differences, climate and predation of agents but these have not been tested. In this study, the effects of ant predation were tested on populations of three biological control agents for L. camara. Colonies of two species, Crematogaster sp. 1 and 2 were investigated. Crematogaster sp. 1 colonies were offered no choice between immature stages of the agents Hypena laceratalis Walker (Lepidoptera: Noctuidae), Falconia intermedia Distant (Hemiptera: Miridae) or Teleonemia scrupulosa Stål (Hemiptera: Tingidae) on lantana shoots. Density-dependent predation on F. intermedia and T. scrupulosa nymphs on lantana shoots was tested using Crematogaster sp. 2 colonies. In choice experiments Crematogaster sp. 2 colonies were offered F. intermedia or T. scrupulosa nymphs on potted lantana plants. Preliminary food trials confirmed that colonies foraged for protein, thereby validating results of no-choice experiments. Crematogaster sp.1 foragers removed 50% of F. intermedia nymphs, followed by 45% of H. laceratalis larvae and only 9% of T. scrupulosa nymphs. Foragers recruited most actively to H. laceratalis larvae and significantly more H. laceratalis biomass was removed than either F. intermedia or T. scrupulosa. A trade-off existed in prey size selection because larger larvae provided considerably more biomass but required forager cooperation and a longer time to subdue than did smaller prey. This increases both forager energy expense and mortality risk by other predators. This study showed that all Crematogaster sp. 1 colonies removed small (≤10mm) H. laceratalis larvae more frequently than larvae larger than 10mm. Thus, of these biological control agents, predators probably prefer small H. laceratalis larvae. Significantly more F. intermedia than T. scrupulosa nymphs were removed by Crematogaster sp. 1, while Crematogaster sp. 2 colonies removed comparable numbers of both agent species. Falconia intermedia nymphs' fast movement triggered a predatory response by these ant species. In contrast, the relatively immobile behaviour of T. scrupulosa nymphs was identified as a highly effective predator avoidance strategy. Since T. scrupulosa nymphs are unable to escape predators by moving, they appear to depend on the presence of alternative prey attracting predator attention. At high agent and/or forager density, T. scrupulosa nymphs attempted escape, but foragers identified them as prey once they moved and caught them. Predation on F. intermedia was also density dependent in that at high nymph and/or forager densities, escape routes were congested and nymphs were more easily caught. Survival of F. intermedia and T. scrupulosa nymphs in particular was low on ant-accessed shrubs in choice experiments and high on ant-excluded shrubs. It is likely that ants significantly depress F. intermedia populations in the field since besides predation, ant foragers probably interrupt F. intermedia feeding and ovipositioning. The combination of parasitism and predation on early instar larvae may explain why H. laceratalis occurs across lantana's range in South Africa but populations remain low. It is unlikely that T. scrupulosa nymphs are habitually preyed on by ant species unless they attract attention by being mobile. Although biological control of L. camara is influenced by climate and physiological defence mechanisms, this study has shown that predation by two ant species severely impacts leaf-feeding agents for L. camara. Thus, it is recommended that future selection of additional agents to control lantana should exclude leaf-feeding .

Pereskia aculeata Miller (Cactaceae) is an environmental weed that is damaging to natural ecosystems in South Africa. The plant is native to Central and South America and was first recorded in South Africa in a botanical garden in 1858. In this thesis, research into the biological control of P. aculeata was conducted with the intention of improving the control of the weed. A pre-release study of the relationship between P. aculeata density and native plant biodiversity indicated that P. aculeata has a negative impact on native biodiversity. The native plant biodiversity associated with different P. aculeata densities was used to determine threshold values and goals for the control of the weed. A threshold value of 50% P. aculeata density was calculated, indicating that P. aculeata density must be maintained below 50% in order to conserve native plant biodiversity. The ultimate goal of the control programme should be to maintain P. aculeata densities below 30%. At these densities there was no significant difference in native plant biodiversity from if the weed were absent from the ecosystem. The biological control agent, Phenrica guérini Bechyne (Chrysomelidae), has been released in South Africa but the potential of the agent to impact P. aculeata is not known and no post release evaluation has been conducted. Impact assessment studies indicate that P. guérini does not impact P. aculeata, even at high densities, but the results of greenhouse experiments should be interpreted with caution because of problems with extrapolation into the field. Although observations in the field suggest that P. guérini has reduced P. aculeata densities at one site, it is clear that new biological control agents are needed to reduce the weed to acceptable levels. Identifying the origin of the South African P. aculeata population was believed to be important to the biological control programme due to the disjunct native distribution and intraspecific variation of the species. Natural enemies associated with plant genotypes in different parts of the native distribution may have developed specialised relationships with certain intraspecific variants of the plant, resulting in differences in agent efficacy on certain host plant genotypes. A molecular study indicated that the closest relatives to the South African weed population found in the native distribution were in Rio de Janeiro Province, Brazil. A bioassay experiment in which fitness related traits of the biological control agent, P. guérini, were measured on various P. aculeata genotypes was conducted to determine the importance of host plant intraspecific variation. There was little variation in fitness traits between genotypes and no evidence of intraspecific host plant specialization. Although intraspecific variation had no effect on agent efficacy in the case of P. guérini, it is possible that other natural enemies may be more specialized. Genotype matching is expected to be more important when natural enemies likely to be specialised to individual genotypes are considered for biological control. Potential biological control agents were prioritized from data collected on surveys in the native distribution. The most promising of these, based on the presence of feeding, incidence, predicted host range, climatic matching, genotype matching and mode of damage, are two species of Curculionidae, the current biological control agent P. guérini and the stem boring moth, Maracayia chiorisalis Walker (Crambidae). The two curculionid species and M. chlorisalis should be considered priorities for host specificity studies. Releases of P. guérini and any new biological control agents should be made at sites where the pre-release study was conducted so that post-release evaluation data can be compared with the pre-release data and the impact of biological control can be evaluated. Retrospective analyses of biological control programmes provide important ways of improving aspects of biological control programmes, such as methods of agent selection. The evaluation of success in biological control programmes is essential for retrospective analyses because factors that have lead to successes or failures can be analysed. Retrospective analyses of biological control programmes, such as this thesis, may improve weed management, thereby contributing to the conservation of natural resources.

Cydia pomonella (Linnaeus) (Family: Tortricidae), the codling moth, is considered one of the most significant pests of apples and pears worldwide, causing up to 80% crop loss in orchards if no control measures are applied. Cydia pomonella is oligophagous feeding on a number of alternate hosts including quince, walnuts, apricots, peaches, plums and nectarines. Historically the control of this pest has been achieved with the use of various chemical control strategies which have maintained pest levels below the economic threshold at a relatively low cost to the grower. However, there are serious concerns surrounding the use of chemical insecticides including the development of resistance in insect populations, the banning of various insecticides, regulations for lowering of the maximum residue level and employee and consumer safety. For this reason, alternate measures of control are slowly being adopted by growers such as mating disruption, cultural methods and the use of baculovirus biopesticides as part of integrated pest management programmes. The reluctance of growers to accept baculovirus or other biological control products in the past has been due to questionable product quality and inconsistencies in their field performance. Moreover, the development and application of biological control products is more costly than the use of chemical alternatives. Baculoviruses are arthropod specific viruses that are highly virulent to a number of lepidopteran species. Due to the virulence and host specificity of baculoviruses, Cydia pomonella granulovirus has been extensively and successfully used as part of integrated pest management systems for the control of C. pomonella in Europe and around the world, including South Africa. Commercial formulations have been typically based on the Mexican strain of CpGV. However due to long-term multiple applications of CpGV and the reliance on CpGV in organic farming practices in Europe, resistance to the CpGV-M strain has developed in a number of field populations of C. pomonella. This study aimed to identify and characterize novel isolates of CpGV in South Africa and compare their virulence with the commercial standard CpGV-M. Secondly, since C. pomonella is difficult to culture on a large scale, an alternate method of CpGV production was investigated in order to determine if CpGV could be produced more efficiently and at a reduced cost without negatively impacting the quality of the product. Several isolates of CpGV were recovered either from field collected larvae or from a laboratory-reared C. pomonella colony. Characterisation of DNA profiles using a variety of restriction enzymes revealed that only a single isolate, CpGV-SA, was genetically different from the Mexican strain of the virus used in the commercially available CpGV based products in South Africa. In dose-response bioassays using CpGV-SA, LC₅₀ and LC₉₀ values for neonate C. pomonella larvae were 3.18 x 10³ OBs/ml and 7.33 x 10⁴ respectively. A comparison of these values with those of CpGV-M indicated no significant difference in the virulence of the two isolates under laboratory conditions. This is a first report of a genetically distinct CpGV isolate in South Africa. The biological activity and novelty of CpGV-SA makes this isolate a potentially important tool for CpGV resistance management in South Africa. In order to justify production of CpGV in an alternative host, studies on the comparative biological performance of C. pomonella and T. leucotreta based on oviposition, time to hatch, larval developmental times and rearing efficiency as well as production costs were performed. Thaumatotibia leucotreta was found to be more fecund and to have significantly shorter egg and larval developmental times. In addition, larval production per unit of artificial diet was significantly higher than for C. pomonella. This resulted in T. leucotreta being more cost effective to produce with implications for reduced insectary space, sanitation practices as well as the labour component of production. Virus yield data generated by inoculation both C. pomonella and T. leucotreta with nine concentrations of CpGV resulted in comparable virus yields, justifying the continuation of the research into production of CpGV in T. leucotreta. It was important to determine the LC and LT values required for mass production of CpGV in late instar T. leucotreta larvae. Dose- and time-response bioassays with CpGV-M were conducted on artificial diet to determine these values. Fourth instar LC₅₀ and LC₉₀ values were 5.96 x 10³ OBs/ml and 1.64 x 10⁵ OBs/ml respectively. LT50 and LT90 values were 81.10 hours and 88.58 hours respectively. Fifth instar LC₅₀ and LC₉₀ values were 6.88 x 10⁴ OBs/ml and 9.78 x 10⁶ OBs/ml respectively. LT₅₀ and LT₉₀ values were 111.56 hours and 137.57 hours respectively. Virus produced in fourth instar T. leucotreta larvae was bioassayed against C. pomonella neonate larvae and compared to CpGV-M to establish if production in the heterologous host negatively affected the virulence of the isolate. No significant difference in virulence was observed between virus produced in T. leucotreta and that produced in C. pomonella. The data generated in the bioassays was used in CpGV mass production trials to evaluate production. All production methods tested produced acceptable virus yields. To examine the quality of the virus product, genomic DNA was extracted from larval cadavers and subjected to REN analysis with HindIII. The resulting DNA profiles indicated that the virus product was contaminated with the homologous virus, CrleGV. Based on the above results, the use of T. leucotreta as an alternate host for the in vivo production of CpGV on a commercial basis is not at this stage viable and requires further investigation before this production methodology can be reliable used to produce CpGV. However, this study has shown that CpGV can be produced in a homologous host, T. leucotreta and significant strides have been made towards developing a set of quality control standards that are essential for further development of successful production methodology. Finally a novel isolate of CpGV has been identified with comparable virulence to the CpGV-M. This is an important finding as it has broad reaching implications for resistance management of CpGV products in South Africa.

Identifier les leviers permettant de stimuler la régulation naturelle des bioagresseurs tout en préservant la biodiversité est indispensable pour concevoir des paysages agricoles fonctionnels. A partir d’une méta-analyse et d’une étude empirique reposant sur 42 parcelles viticoles localisées dans Nouvelle Aquitaine (France), nous avons cherché à évaluer l’impact du déploiement de l’agriculture biologique à de large échelles spatiales sur 1) les communautés d’ennemis naturels, 2) les services de régulation naturelle, 3) les taux d’infestation par les bioagresseurs. Dans ce travail, nous avons montré que la proportion d’agriculture biologique est un facteur structurant plus les communautés d’ennemis naturels que la proportion d’habitats semi-naturels dans le paysage. De plus, nous avons montré que les communautés de bioagresseurs rencontrées dans les vignes ne sont pas influencées par la proportion d’agriculture biologique alors qu’elles répondent plutôt négativement à la proportion d’habitats semi-naturels. Par ailleurs, nous avons montré que l’agriculture biologique, à l’échelle globale et indépendamment du type de culture considérée est un système de culture stimulant la régulation naturelle des bioagresseurs. En viticulture, elle permet de réduire l’utilisation des produits phytosanitaires, comparé à l’agriculture conventionnelle. Enfin, nos analyses ont révélé qu’au-delà de la différence de systèmes de culture, un certain nombre de facteurs locaux (e.g., âge des parcelles, fréquence de traitements, productivité) permettent d’expliquer la structure des communautés d’ennemis naturels et des services de régulation naturelle des bioagresseurs. Tout en produisant des connaissances sur les processus permettant d’expliquer les assemblages des communautés d’ennemis naturels et les niveaux de services de régulation rendus, notre travail suggère des pistes pour l’aménagement des paysages viticoles permettant de concilier préservation de la biodiversité et maximisation des régulations naturelles. / Identifying landscape context and farming systems that enhance natural pest control while maintaining biodiversity is crucial to design functional agricultural landscapes. Using a meta-analysis and an empiric study based on 42 vineyards in Nouvelle Aquitaine (France), we investigated the effect of the deployment of organic farming at a landscape scale on 1) natural enemy communities, 2) natural pest control and 3) pest infestation levels. Here, we showed that the proportion of organic farming structured more natural enemy communities than the proportion of semi-natural habitats. On the opposite, pest and pathogen infestations were never influenced by the proportion of organic farming while they were negatively influenced by the proportion of semi-natural habitats. Furthermore, at a global scale and for every crop types, organic farming, per se, enhances natural pest control. In viticulture, it is less dependent of synthetic agrochemicals than conventional farming. Moreover, local factors such as the treatment frequency index, the field age and the crop productivity had important effects on natural enemy communities and natural pest control. Finally, we yielded knowledge on processes that impact natural enemy assembly and natural pest control in agrosystems. For vineyards-dominated landscapes, our work suggests some tracks for landscape planning that support biodiversity conservation and natural pest control.

Submitted by Deyse Queiroz (deysequeirozz@hotmail.com) on 2018-05-22T15:00:44Z No. of bitstreams: 1 JESSÉ MALVEIRA GOMES - DISSERTAÇÃO PPGHT - 2017.pdf: 1522466 bytes, checksum: cc3a2a2cb3b9b312f031097d4500a8df (MD5) / Made available in DSpace on 2018-05-22T15:00:44Z (GMT). No. of bitstreams: 1 JESSÉ MALVEIRA GOMES - DISSERTAÇÃO PPGHT - 2017.pdf: 1522466 bytes, checksum: cc3a2a2cb3b9b312f031097d4500a8df (MD5) Previous issue date: 2017-03 / O milho é uma das culturas mais produzida e importante no mundo, sendo utilizada na alimentação humana e animal. O Brasil é um dos maiores produtores desse grão, sendo o terceiro maior produtor do mundo. Apesar de uma produção elevada, a produtividade é comprometida pela ação de insetos pragas, sendo a Spodoptera frugiperda Smith a de maior atuação nessa cultura. Em meio a esse desafio, o uso constante de inseticidas é utilizado com intuito de minimizar as perdas nas lavouras, porém, ocasionando grandes impactos ambientais, bem como a contaminação de trabalhadores rurais. Uma das estratégias viáveis e promissoras na supressão de pragas tem sido o manejo integrado de pragas (MIP), que tem como alicerce o uso isolado ou consorciado de técnicas de controles de pragas, sendo o controle biológico uma das ferramentas mais sustentáveis do ponto de vista ecológico. Desta forma, o presente trabalho teve como objetivo avaliar a toxicidade da cepa bacteriana 358.1 isolada do petróleo e o sobrenadante da cultura de células sobre o controle de lagartas de 2º e 3º instares de S. frugiperda, comparando as taxas de mortalidade e efeitos sobre seu desenvolvimento e com inseticidas químicos comerciais. O experimento consistiu de duas etapas, o primeiro ensaio foi realizado com lagartas de 3º instar com três diluições diferentes da cepa 358.1, sobrenadante da cultura de células, solução salina como controle negativo e o inseticida fenpropatrina como controle positivo. No segundo ensaio utilizou-se lagartas de 3º e 2º instares, a cultura celular da cepa 358.1 em sua concentração total (4 x 108 UFC/mL), seu sobrenadante da cultura, como controle negativo água destilada e testemunha positiva o produto comercial flubendiamida. Os experimentos com a cepa 358.1, sobrenadante da cultura de células e os produtos químicos testados não resultaram em uma taxa de mortalidade de S. frugiperda significante, porém, a cepa 358.1 e seu sobrenadante da cultura resultaram no aumento da duração da fase larval e em distúrbios morfofisiológicos do inseto, ocasionando, inclusive, no surgimento de adultos com asas atrofiadas, sendo desta maneira, promissores para à realização de novos testes. / Corn is one of the most produced and important crops in the world, utilized for human and animal feeding. Brazil is one of the top producers of this grain, being the third biggest producer worldwide. Despite the great production, the productivity is compromised due to the activity of insect pest. Because of this challenge, the continuous use of insecticides is done aiming to minimize lost in the crops. However, it causes major environmental impact and it is also contaminating rural workers. One of the viable and promising strategies to the pest suppression has been the pest integrated management (PIM), which is based on the use of isolated or consortium of pest controlling techniques, the biological control being one of the most sustainable tools in an ecological perspective. Therefore, the present study had the goal of evaluating the toxicity of the 358.1 bacterium strain isolated from the oil and supernatant of the cell culture under control of caterpillars from 2nd and 3rd instars of S. frugiperda, comparing the death rates and effects on their development, and also comparing with commercial chemical insecticides. The experiment consisted in two stages; the first assay was done with 3rd instar caterpillar with three different dilutions from the 358.1 strain, supernatant of the cell culture, saline solution as negative control, and fenpropatrina insecticide as positive control. On the second assay, 3rd and 2nd instars caterpillars were used, the cell culture from the 358.1 strain on its total concentration (4 x 108 UFC/mL), supernatant of the cell culture, distilled water as negative control and flubendiamida, a commercialized product, as the positive control. The experiments with the 358.1 strain, supernatant of the cell culture and the tested chemical products didn’t show a significant death rate for S. frugiperda, however, the 358.1 strain and supernatant of the cell culture resulted in an increase in the larval phase duration and on morphophysiological disturbs on the insect, resulting yet in the emergence of adults with atrophied wings, thus being promising to the realization of new tests.

A viticultura é uma atividade de grande importância econômica, destacando-se a sustentabilidade da pequena propriedade e o desenvolvimento territorial associados às atividades ligadas ao turismo. As plantas cultivadas se tornam vulneráveis a patógenos e insetos-praga e a videira apresenta diversas espécies consideradas pragas que reduzem sua produção e rentabilidade, entre estas, Spodoptera frugiperda, causando danos em diferentes partes da planta. Fungos entomopatogênicos podem oferecer uma alternativa aos pesticidas convencionais para o controle de pragas, pois produzem enzimas que degradam o exoesqueleto do inseto como quitinases e proteases facilitando o modo de infecção. Neste trabalho foi avaliado o potencial inseticida do fungo Nomurea rileyi, linhagem UCS03, contra S. frugiperda e o perfil eletroforético por SDS-PAGE das proteínas secretadas por N. rileyi em presença da cutícula do inseto em diferentes intervalos de tempo em gel unidimensional. O fungo N. rileyi apresentou virulência contra S. frugiperda, determinando um CL50 de 2 x 109conídios/mL com a linhagem UCS03 demonstrando atividade bionseticida. Na avaliação do perfil proteico do secretoma de N. rileyi em presença da cutícula do inseto, em diferentes tempos de cultivo, foi possível verificar um perfil altamente diferenciado. A maior concentração de proteína foi encontrada no 14° dia de incubação (0,3507 mg/mL) reduzindo a quantidade de proteínas após este período. Na análise por SDS-PAGE foi possível verificar diferentes proteínas de diferentes massas moleculares, nos intervalos de tempo considerados, sendo muitas inferiores a 75 kDa. Estas proteínas com diferentes massas moleculares podem estar envolvidas no metabolismo do fungo. Desta forma, estes resultados podem contribuir para a compreensão do processo de infecção de N. rileyi em S. frugiperda, oferecendo potencial para o desenvolvimento de novas pesquisas e aplicações destas em processos biotecnológicos. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2017-03-14T12:21:21Z No. of bitstreams: 1 Dissertacao Ana Carolina Ruiz.pdf: 656327 bytes, checksum: 52c5a7db081b0ec94339972595956f52 (MD5) / Made available in DSpace on 2017-03-14T12:21:21Z (GMT). No. of bitstreams: 1 Dissertacao Ana Carolina Ruiz.pdf: 656327 bytes, checksum: 52c5a7db081b0ec94339972595956f52 (MD5) Previous issue date: 2017-03-14 / Viticulture is an activity of great economic importance with emphasis on the sustainability of small property and territorial development associated with tourism related activities. Cultivated plants become vulnerable to pathogens and insect pests and the vine has several species considered as pests that reduce their production and profitability, among them, Spodoptera frugiperda, with causes damage in different parts of the plant. Entomopathogenic fungi can provide an alternative to conventional pesticides for controlling pests, they produce enzymes that degrade the insect exoskeleton, such aschitinases and proteases facilitating the infection. In this work, the insecticide potential of the fungus Nomurea rileyi and S. frugiperda was evaluated, as well as the proteins secreted by N. rileyi in the presence of insect cuticle at different time intervals in one-dimensional gel. The fungus N. rileyi presented virulence against S. frugiperda, determining a CL50 of 2 x 109 con / mL with UCS03 strain demonstrating bionseticida activity. In assessing the protein profile of secretome N. rileyi in the presence of insect cuticle, at different times of cultivation, a highly differentiated profile was verified. The highest concentration of protein was found at day 14 of incubation (0.3507 mg / ml) reducing the amount of protein after this period. In the one-dimensional gel analysis was verified different molecular weights of proteins, in the time interval considered being many less than 75kDa. These proteins with different molecular weights may be involved in fungal metabolism. Thus, these results can contribute to the understanding of the infection process of N. rileyi in S. frugiperda, offering potential for the development of new researches and applications in biotechnological processes.

O Brasil é o maior produtor de citros, sendo um dos seus principais produtos de exportação da cadeia agroindustrial. Um dos fatores que afetam a qualidade e a produção do produto final obtido é a Mancha Marrom de Alternaria, causado pelo fungo Alternaria alternata. A doença provoca desfolha, seca de ramos e queda de frutos em tangerinas e seus híbridos. Para controle dessa doença são utilizados fungicidas, porém, a mancha marrom é uma doença de difícil controle podendo ser necessárias de 7 a 10 aplicações do fungicida, aparecimento de resistência dos fungos, intoxicação nas pessoas e contaminação do meio ambiente devido ao emprego de fungicidas. Métodos alternativos como o controle biológico com aplicações de micro-organismos antagonistas e a nutrição das plantas são opções de menor impacto ao meio ambiente. Sendo assim, o objetivo deste trabalho foi avaliar o potencial de Trichoderma spp., Bacillus subtilis e fertilizante organomineral, no controle da Mancha Marrom de Alternaria em plantas cítricas de tangerina Ponkan. O experimento foi realizado com delineamento casualizado, constituído de oito tratamentos com quatro repetições. Os experimentos foram avaliados quanto a incidência da doença. Os dados mostraram que o melhor controle foi o tratamento químico, porém os tratamentos com Trichoderma spp., Trichoderma spp. + fertilizante e Trichoderma spp. + Bacillus subtilis apresentaram ótimas taxas de controle, podendo inclusive substituir o tratamento químico no controle desta doença. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2017-06-13T14:28:19Z No. of bitstreams: 1 Dissertacao Bianca Luzardo Porto.pdf: 1070166 bytes, checksum: 4e890666fce3c2b0d8ce25ef2c71241f (MD5) / Made available in DSpace on 2017-06-13T14:28:19Z (GMT). No. of bitstreams: 1 Dissertacao Bianca Luzardo Porto.pdf: 1070166 bytes, checksum: 4e890666fce3c2b0d8ce25ef2c71241f (MD5) Previous issue date: 2017-06-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES. / Brazil is the largest citrus producer, being one of its main exports of agro-industrial chain. One of the factors that affect the quality and production of the final product is the Brown Spot Alternaria, caused by the fungus Alternaria alternata. The disease causes leaf drop, dry branches and fruit drop in tangerines and their hybrids. To control this disease fungicides are used, however, the brown spot is a difficult to control disease, may be required from 7 to 10 applications of fungicide, appearance of resistance of fungi, intoxication in people and environment contamination owing the application of fungicides. Alternative methods such as biological control with applications antagonistic microorganisms and plant nutrition are options of smaller impact to the environment. Therefore, the objective of this work was to evaluate the potential of Trichoderma spp., Bacillus subtilis and fertilizer, in control of Alternaria Brown Spot in citrus plants Ponkan tangerine. The experiment was carried out in a randomized design, consisting of eight treatments with four replicates. The experiments were evaluated for the incidence of the disease. The data showed that the best control was the chemical treatment, but the treatments with Trichoderma spp., Trichoderma spp. + Fertilizer and Trichoderma spp. + Bacillus subtilis presented excellent control rates, and may even replace chemical treatment in the control of this disease.

Mealybugs (Planococcus citri) and thrips (Scirtothrips aurantii) are common and extremely damaging citrus crop pests which have proven difficult to control via conventional methods, such as chemical pesticides and insect growth regulators. The objective of this study was to determine the efficacy of entomopathogenic fungi against these pests in laboratory bioassays. Isolates of Metarhizium anisopliae and Beauveria bassiana from citrus orchards in the Eastern Cape, South Africa were maintained on Sabouraud Dextrose 4% Agar supplemented with Dodine, chloramphenicol and rifampicin at 25°C. Infectivity of the fungal isolates was initially assessed using 5th instar false codling moth, Thaumatotibia leucotreta, larvae. Mealybug bioassays were performed in 24 well plates using 1 x 107 ml-1 conidial suspensions and kept at 26°C for 5 days with a photoperiod of 12 L:12 D. A Beauveria commercial product and an un-inoculated control were also screened for comparison. Isolates GAR 17 B3 (B. bassiana) and FCM AR 23 B3 (M. anisopliae) both resulted in 67.5% mealybug crawler mortality and GB AR 23 13 3 (B. bassiana) resulted in 64% crawler mortality. These 3 isolates were further tested in dose-dependent assays. Probit analyses were conducted on the dose-dependent assays data using PROBAN to determine LC₅₀ values. For both the mealybug adult and crawlers FCM AR 23 B3 required the lowest concentration to achieve LC₅₀ at 4.96 x 10⁶ conidia ml-1 and 5.29 x 10⁵ conidia ml-1, respectively. Bioassays on adult thrips were conducted in munger cells with leaf buds inoculated with the conidial suspensions. Isolate GAR 17 B3 had the highest mortality rate at 70% on thrips while FCM AR 23 B3 resulted in 60% mortality. Identification of the isolates, FCM AR 23 B3, GAR 17 B3 and GB AR 23 13 3, were confirmed to be correct using both microscopic and molecularly techniques. ITS sequences were compared to other sequences from GenBank and confirmed phylogenetically using MEGA6. Mealybug infection was investigated using scanning electron microscopy, mycosis was confirmed but the infection process could not be followed due to the extensive waxy cuticle. These results indicate that there is potential for the isolates FCM AR 23 B3 and GAR 17 B3 to be developed as biological control agents for the control of citrus mealybug and thrips. Further research would be required to determine their ability to perform under field conditions.

A viticultura é uma atividade de grande importância econômica, destacando-se a sustentabilidade da pequena propriedade e o desenvolvimento territorial associados às atividades ligadas ao turismo. As plantas cultivadas se tornam vulneráveis a patógenos e insetos-praga e a videira apresenta diversas espécies consideradas pragas que reduzem sua produção e rentabilidade, entre estas, Spodoptera frugiperda, causando danos em diferentes partes da planta. Fungos entomopatogênicos podem oferecer uma alternativa aos pesticidas convencionais para o controle de pragas, pois produzem enzimas que degradam o exoesqueleto do inseto como quitinases e proteases facilitando o modo de infecção. Neste trabalho foi avaliado o potencial inseticida do fungo Nomurea rileyi, linhagem UCS03, contra S. frugiperda e o perfil eletroforético por SDS-PAGE das proteínas secretadas por N. rileyi em presença da cutícula do inseto em diferentes intervalos de tempo em gel unidimensional. O fungo N. rileyi apresentou virulência contra S. frugiperda, determinando um CL50 de 2 x 109conídios/mL com a linhagem UCS03 demonstrando atividade bionseticida. Na avaliação do perfil proteico do secretoma de N. rileyi em presença da cutícula do inseto, em diferentes tempos de cultivo, foi possível verificar um perfil altamente diferenciado. A maior concentração de proteína foi encontrada no 14° dia de incubação (0,3507 mg/mL) reduzindo a quantidade de proteínas após este período. Na análise por SDS-PAGE foi possível verificar diferentes proteínas de diferentes massas moleculares, nos intervalos de tempo considerados, sendo muitas inferiores a 75 kDa. Estas proteínas com diferentes massas moleculares podem estar envolvidas no metabolismo do fungo. Desta forma, estes resultados podem contribuir para a compreensão do processo de infecção de N. rileyi em S. frugiperda, oferecendo potencial para o desenvolvimento de novas pesquisas e aplicações destas em processos biotecnológicos. / Viticulture is an activity of great economic importance with emphasis on the sustainability of small property and territorial development associated with tourism related activities. Cultivated plants become vulnerable to pathogens and insect pests and the vine has several species considered as pests that reduce their production and profitability, among them, Spodoptera frugiperda, with causes damage in different parts of the plant. Entomopathogenic fungi can provide an alternative to conventional pesticides for controlling pests, they produce enzymes that degrade the insect exoskeleton, such aschitinases and proteases facilitating the infection. In this work, the insecticide potential of the fungus Nomurea rileyi and S. frugiperda was evaluated, as well as the proteins secreted by N. rileyi in the presence of insect cuticle at different time intervals in one-dimensional gel. The fungus N. rileyi presented virulence against S. frugiperda, determining a CL50 of 2 x 109 con / mL with UCS03 strain demonstrating bionseticida activity. In assessing the protein profile of secretome N. rileyi in the presence of insect cuticle, at different times of cultivation, a highly differentiated profile was verified. The highest concentration of protein was found at day 14 of incubation (0.3507 mg / ml) reducing the amount of protein after this period. In the one-dimensional gel analysis was verified different molecular weights of proteins, in the time interval considered being many less than 75kDa. These proteins with different molecular weights may be involved in fungal metabolism. Thus, these results can contribute to the understanding of the infection process of N. rileyi in S. frugiperda, offering potential for the development of new researches and applications in biotechnological processes.

O Brasil é o maior produtor de citros, sendo um dos seus principais produtos de exportação da cadeia agroindustrial. Um dos fatores que afetam a qualidade e a produção do produto final obtido é a Mancha Marrom de Alternaria, causado pelo fungo Alternaria alternata. A doença provoca desfolha, seca de ramos e queda de frutos em tangerinas e seus híbridos. Para controle dessa doença são utilizados fungicidas, porém, a mancha marrom é uma doença de difícil controle podendo ser necessárias de 7 a 10 aplicações do fungicida, aparecimento de resistência dos fungos, intoxicação nas pessoas e contaminação do meio ambiente devido ao emprego de fungicidas. Métodos alternativos como o controle biológico com aplicações de micro-organismos antagonistas e a nutrição das plantas são opções de menor impacto ao meio ambiente. Sendo assim, o objetivo deste trabalho foi avaliar o potencial de Trichoderma spp., Bacillus subtilis e fertilizante organomineral, no controle da Mancha Marrom de Alternaria em plantas cítricas de tangerina Ponkan. O experimento foi realizado com delineamento casualizado, constituído de oito tratamentos com quatro repetições. Os experimentos foram avaliados quanto a incidência da doença. Os dados mostraram que o melhor controle foi o tratamento químico, porém os tratamentos com Trichoderma spp., Trichoderma spp. + fertilizante e Trichoderma spp. + Bacillus subtilis apresentaram ótimas taxas de controle, podendo inclusive substituir o tratamento químico no controle desta doença. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES. / Brazil is the largest citrus producer, being one of its main exports of agro-industrial chain. One of the factors that affect the quality and production of the final product is the Brown Spot Alternaria, caused by the fungus Alternaria alternata. The disease causes leaf drop, dry branches and fruit drop in tangerines and their hybrids. To control this disease fungicides are used, however, the brown spot is a difficult to control disease, may be required from 7 to 10 applications of fungicide, appearance of resistance of fungi, intoxication in people and environment contamination owing the application of fungicides. Alternative methods such as biological control with applications antagonistic microorganisms and plant nutrition are options of smaller impact to the environment. Therefore, the objective of this work was to evaluate the potential of Trichoderma spp., Bacillus subtilis and fertilizer, in control of Alternaria Brown Spot in citrus plants Ponkan tangerine. The experiment was carried out in a randomized design, consisting of eight treatments with four replicates. The experiments were evaluated for the incidence of the disease. The data showed that the best control was the chemical treatment, but the treatments with Trichoderma spp., Trichoderma spp. + Fertilizer and Trichoderma spp. + Bacillus subtilis presented excellent control rates, and may even replace chemical treatment in the control of this disease.