Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Planococcus citri (Risso) (Hemiptera: Pseudococcidae), the citrus mealybug, is a highly destructive pest of citrus in South Africa. The ability of mealybugs to develop resistance to chemicals, as well as their cryptic nature and protective wax coverings, all individually and combined, impair the ability of insecticides to control them. Furthermore, chemicals deplete natural enemy populations that usually establish control over mealybug populations well before harvest in March to May. The inability of chemicals to control P. citri successfully and the growing public awareness of the detrimental environmental effects and health risks that are associated with pesticides are pressuring citrus growers to find alternative methods to chemical control. Entomopathogenic nematodes of the Rhabditida order, belonging either to the families Heterorhabditidae or Steinernematidae, have proven to be valuable biocontrol agents of a variety of insect pest species. These nematodes are, however, not yet commercially available in South Africa. Various laboratory bioassays were conducted to determine the potential of entomopathogenic nematodes to control P. citri. Adult female P. citri were screened for susceptibility to six indigenous nematode species. Planococcus citri was found to be most susceptible to Steinernema yirgalemense and Heterorhabditis zealandica, causing 97% and 91% mortality, respectively. Both H. zealandica and S. yirgalemense completed their life cycles inside adult female P. citri. Under optimal conditions, H. zealandica and S. yirgalemense, at an application rate of 170 infective juveniles (IJs) / insect (LD90), were able to control P. citri efficiently with a discriminating dosage of 11(LD50). The water activity (aw) bioassay indicated that S. yirgalemense was two times more tolerant to lower levels of free water than H. zealandica, with aw50 = 0.96 and aw90 = 0.99, compared to H. zealandica with aw50 = 0.98 and aw90 = 1.01. Furthermore, S. yirgalemense proved able to locate and infect P. citri at a faster rate than H. zealandica. Nematode activity was not significantly affected when exposed to 15°C, 20°C and 25°C. Results of the exposure trial also showed that the first 2−4 h post-application was the most decisive time for establishing successful infection of mealybugs. Further experiments were conducted to determine the ability of two polymer products, Zeba® and Xanthan gum, and a surfactant, Nu-Film-P®, to improve nematode application suspensions. Despite not being as effective as Xanthan gum, Zeba® still retarded sedimentation significantly. The addition of 0.02% Xanthan gum did not improve the ability of nematodes to control P. citri at 60% and 80% relative humidity containing H. zealandica and 0.03% Zeba® increased mortality by 14% at 60% RH and by 22% at 80% RH. The same polymer formulation was tested with S. yirgalemense, with mortality being found to have increased by 21% at 60% RH and by 27% at 80% RH. The combined addition of Nu-Film-P® and Zeba® was able to retard sedimentation significantly, increasing the average number of nematodes deposited on 2-cm2 leaf discs. To illustrate those factors that should be taken into consideration before applying nematodes in an integrated pest management (IPM) programme for citrus, the compatibility of H. zealandica and S. yirgalemense with biological control agents and agrochemicals to which they are most likely to be exposed was determined. Bioassays showed the coccinellid beetle, Cryptolaemus montrouzieri, to be susceptible to both nematode species. Beetle larvae proved to be highly susceptible, with H. zealandica obtaining 80%, and S. yirgalemense obtaining 92% control. Adult beetles were found to be twice as susceptible to S. yirgalemense, with 64% mortality recorded, as to H. zealandica, with 30% mortality recorded. Tolerance of H. zealandica and S. yirgalemense infective juveniles to the affect of aqueous solutions of an insecticide (Cyperphos 500 E.C.®), two biopesticides (CryptogranTM and HelicovirTM), and two adjuvants (Nu-Film-P® and Zeba®) on the infectivity and survival of H. zealandica and S. yirgalemense was evaluated. Heterorhabditis zealandica proved to be highly compatible with all products tested. Significant increase in mortality of S. yirgalemense was recorded after 12 h exposure to CryptogranTM, HelicovirTM and Cyperphos 500 E.C.®, and after 6 h exposure to Nu-Film-P®. However, there was no decrease in the ability of S. yirgalemense to infect the insect hosts. Towards selecting the appropriate nematode species for field studies, the ability of S. yirgalemense and H. zealandica to control P. citri was first evaluated under less harsh simulated glasshouse conditions. The ability of both the above-mentioned nematode species to control P. citri, with and without the addition of 0.03% Zeba® and 0.06% Nu-Film-P®, was evaluated in a growth chamber at 75 ± 8% relative humidity and 22°C. All treatments resulted in significantly higher mortality of adult female P. citri than did the control. The addition of 0.03% Zeba® and 0.06% Nu-Film-P® to an H. zealandica and an S. yirgalemense suspension increased resultant mortality from 26% to 30%, and from 34% to 45%, respectively. This increase in mortality was, however, not significant. The ability of this formulation to prolong the ability of S. yirgalemense to infect P. citri and to prolong nematode survival was also evaluated under the same conditions and showed the formulation to improve both infectivity and survival for up to 2–3 h post-application. In the semi-field trial, S. yirgalemense was unable to obtain significant control of P. citri without the addition of 0.03% Zeba® to nematode application suspensions. However, with the addition of 0.03% Zeba® S. yirgalemnse was able to obtain up to 53% control. The study established that the polymer product Zeba® improves the ability of S. yirgalemense to infect P. citri by retarding desiccation and by buffering nematodes from suboptimal environmental conditions. / AFRIKAANSE OPSOMMING: Planococcus citri (Risso) (Hemiptera: Pseudococcidae), die sitrus witluis, is ʼn baie skadelike pes van sitrus in Suid Afrika. Die vermoë van witluise om weerstand teen chemiese middels te ontwikkel, hul kriptiese lewenswyse en die beskermende waslaag wat hul liggame omhul inhibeer gesamentlik en individueel die vermoë van insektisiede om witluis bevolkings te beheer. Chemiese middels verminder ook die natuurlike vyande wat gewoonlik witluis bevolkings beheer voor die sitrus oestydperk in Maart tot Mei. Die onvermoë van chemiese middels om P. citri suksesvol te beheer en verhoogde bewustheid van die publiek rond om die vernietigende omgewings impak en gesondheidsrisiko’s verbonde aan chemiese insek beheer, noodsaak sitrus produsente om alternatiewe beheermetodes te ontwikkel. Entomopatogeniese nematodes, van die orde Rhabditida wat aan die families Heterorhabditidae of Steinernematidae behoort, is bekend as effektiewe biologiese beheeragente van ʼn verskeidenheid insek pes spesies. Hierdie nematodes is egter tans nie kommersieel beskikbaar in Suid- Afrika nie. Om ten einde die vermoë van plaaslike nematode spesies te bepaal om P. citri te beheer, is verskeie biotoetse in die laboratorium uitgevoer. ʼn Vinnige siftings proses is uitgevoer om vas te stel watter plaaslike nematode spesies die hoogste persentasie mortaliteit van P. citri wyfies veroorsaak. Daar is bevind dat P. citri die mees vatbaarste is vir Steinernema yirgalemense en Heterorhabditis zealandica wat 97% en 91% mortaliteit respektiewelik veroorsaak het. Die ontwikkeling van beide H. zealandica en S. yirgalemense na die infektering van volwasse P. citri wyfies is gevolg en daar is bevind dat beide nematode spesies hul lewensiklusse kon voltooi in insek kadawers. Onder optimale toestande was H. zealandica en S. yirgalemense in staat om P. citri effektief te beheer, as hul teen ʼn konsentrasie van 170 infektiewe larwes (JIs) per insek (LD90) toegedien word, met ʼn diskriminerende toedienings dosis van 11 (LD50). ʼn Water aktiwiteit biotoets het gewys dat S. yirgalemense twee keer so verdraagsaam is teenoor laer vlakke van vrye water as wat H. zealandica is met aw50 = 0.96 en aw90 = 0.99, in vergelyking met H. zealandica met aw50 = 0.98 en aw90 = 1.01. Resultate van ʼn blootstellings toets het ook gewys dat S. yirgalemense die vermoë het om P. citri vinniger op te spoor en te infekteer as H. zealandica. Die blootstellings toets was uitgevoer teen 15°C, 20°C en 25°C en daar is bevind dat die aktiwiteit van H. zealandica nie beduidend verhoog het met ʼn verhoging in temperatuur nie. Resultate vir die blootstellings toets het ook gewys dat die eerste 2 tot 4 uur na toediening van nematodes die mees kritieke tyd is om suksesvolle infektering van witluise te bevestig. Verdere eksperimente is uitgevoer om te bepaal of die byvoeging van twee polimeer produkte nl. Zeba® en Xanthan gum en ʼn benatter Nu-Film-P®, nematode toedienings suspensies kan verbeter. Deur 0.02% Xanthan gum by nematode toedienings suspensies te voeg is die vermoë van nematodes om P. citri by 60% en 80% relatiewe humiditeit (RH) te beheer nie beduidend verhoog nie, terwyl die byvoeging van 0.03% Zeba® wel ʼn beduidende verhoging in mortaliteit veroorsaak het. ʼn Suspensie van H. zealandica en 0.03% Zeba® het beheer met 14% by 60% RH en met 22% by 80% RH verhoog. Die invloed van dieselfde polimeer formulasie was ook getoets op S. yirgalemense en mortaliteit het verhoog met 21% by 60% RH en met 27% by 80% RH. Ten spyte daarvan dat “Xanthan gum” nie so effektief was om afsakking van nematodes te verhoed nie, het die byvoeginging van 0.03% Zeba® by toedienings suspensies steeds ʼn beduidende invloed gehad. Die gekombineerde byvoeging Nu-Film-P® en Zeba® was in staat om die gemiddelde aantal nematodes gedeponeer op 2-cm2 blaar skyfies te verhoog. Om te wys watter faktore in ag geneem moet word voor EPNs as deel van ʼn geïntegreerde pes beheer program toegedien word, is die verenigbaarheid van H. zealandica en S. yirgalemense vir biologiese beheer agente en landbouchemikalieë, waaraan hulle heel waarskynlik blootgestel gaan word, bepaal. Biotoetse het gewys dat die liewenheersbesie, Cryptolaemus montrouzieri, vatbaar is vir beide nematode spesies. Die larwale fase was hoogs vatbaar met mortaliteit van 80% en 92% verkry deur die toedeining van H. zealandica en S. yirgalemense respektiewelik. Daar is ook bepaal dat volwasse besies twee maal so vatbaar is vir S. yirgalemense wat 64% mortaliteit veroorsaak het, as vir H. zealandica met 30% mortaliteit aangeteken. Verdraagsaamheid van IJs van H. zealandica en S. yirgalemense met oplossings van ʼn insektisied (Cyperphos 500 E.C.®), twee bio-plaagdoders (CryptogranTM en HelicovirTM), en twee byvoegmiddels (Nu-Film-P® en Zeba®), vir infektiwiteit en oorlewing is bepaal. Resultate het gewys dat H. zealandica hoogs verenigbaar is met alle produkte wat getoets is. ʼn Beduidende verhoging in mortaliteit van S. yirgalemense is aangeteken na 12 ure se blootstelling aan CryptogranTM, HelicovirTM en Cyperphos 500 E.C.®, en na 6 ure se blootstelling aan Nu-Film-P®. Resultate het egter ook gewys dat die vermoë van S. yirgalemense om om gashere te infekteer nie deur beinvloed word nie. Om ten einde te bepaal watter een van H. zealandica of S. yirgalemense die beter isolaat sou wees om te gebruik in veldproewe, is hul vermoë om P. citri te beheer eers evalueer onder minder ongunstige gesimuleerde glashuis toestande. Die vermoë van beide nematode spesies om P. citri te beheer is bepaal in ʼn groeikamer by 75 ± 8% (RH) met en sonder die byvoeging van 0.03% Zeba® en 0.06% Nu-Film-P®. Mortaliteit verkry deur alle behandelings was beduidend hoër as die kontrole. Deur 0.03% Zeba® en 0.06% Nu-Film-P® by toedienings suspensies van H. zealandica en S. yirgalemense te voeg is die gevolglike mortaliteit van P. citri verhoog van 26% na 30% en van 34% na 45%, respektiewelik. Hierdie verhoging in mortaliteit was egter nie beduidend nie. Die vermoë van die bo-genoemde formulasie om oorlewing van S. yirgalemense en infeksie tydperk van P. citri te verleng was bepaal onder dieselfde toestande en daar is gevind dat die formulasie beide die infektiwiteit en oorlewing van nematodes met 2 tot 3 uur na toediening verleng het. Tydens die semi-veldproef was S. yirgalemense nie in staat om beduidend hoër mortaliteit van P. citri te verkry, in vergelyking met die kontrole, sonder die byvoeging van 0.03% Zeba® by toedienings suspensies nie. Met die byvoeging van Zeba® is daar tot 53% beheer verkry. Die studie het onweerlegbaar gewys dat die polimeer produk Zeba® die vermoë van S. yirgalemense om P. citri te infekteer verbeter deur die uitdroging van nematodes te vertraag en deur hul te beskerm teen ongunstige omgewings toestande.

Thesis (MSc (Conservation Ecology and Entomology))--University of Stellenbosch, 2011. / Please refer to full text to view abstract.

No description available.

Yellow toadflax, Linaria vulgaris (L.) Mill. (Plantaginaceae), is a non-native invasive plant. Rhinusa pilosa Germar (Coleoptera: Curculionidae) is a proposed biocontrol agent. Gall development by R. pilosa was described using histological methods and compared between plant populations from native and introduced ranges. Key stages of oviposition were isolated histologically to determine their importance in gall induction. Rhinusa pilosa galled and developed on four geographically distinct Canadian populations in a pre-release quarantine study. Low agent densities only negatively affected one population. High densities of R. pilosa reduced potential reproductive output and plant biomass. Conducting detailed investigations into the biology, impact, and development of R. pilosa on populations from invasive and native ranges may help predict the efficacy of R. pilosa in the field if approved for release and.goes beyond current pre-release testing requirements. / ix, 168 leaves : ill. (chiefly col.) ; 29 cm

Gibberella zeae (anamorph: Fusarium graminearum) is the causal agent of fusarium head blight (FHB) and maize ear rot, two major diseases of wheat and corn in Eastern Canada. / In Quebec, Microsphaeropsis sp., an antagonist of Venturia inaequalis, the causal agent of apple scab, was isolated from the apple leaf litter. This fungus, well adapted to Quebec climate, can reduce the initial inoculum of V. inaequalis. FHB and Gibberella ear rot are similar to apple scab because the major inoculum source comes from melanized structures produced on crop residues. Consequently, we evaluated the potential of Microsphaeropsis sp. (isolate P130A) to inhibit ascospore production of G. zeae when applied to crop residues as post harvest or pre-planting applications. Under in vitro conditions, the antagonist significantly reduced ascospore production on wheat and corn residues, when applied prior to (82% and 92% respectively) or at the same time as the pathogen (36% and 58% respectively). Under field conditions, the antagonist had no effect on the pattern of perithecia maturation but significantly reduced the number of ascospores produced on two sampling dates, May 1998 and July 1999. (Abstract shortened by UMI.)

In Quebec, the swede midge (Contarinia nasturtii), a newly invasive pest, was found for the first time in 2003. During a two year study (2004-2005) I studied the seasonal development of the swede midge, the presence of natural enemies and tested entomopathogenic nematodes for its control. Results showed that the overwintering generation of the swede midge emerged during the second half of June. Based on adult captures and abundance of larvae in the field, there were three to four overlapping generations. Late transplants were more heavily attacked by the swede midge and broccoli and cauliflower were the most susceptible crops. No parasitoids were found in the experimental sites during either year of the study and two polyphagous coccinellid predators (Harmonia axyridis and Coccinella septempunctata) were field-collected for further evaluation under laboratory conditions. Although these two coccinellid species fed on swede midge larvae in plastic containers, further experiments demonstrated that H. axyridis was not able to prey on swede midge larvae on infested broccoli plants. The susceptibility of swede midge larvae to three species of entomopathogenic nematodes (Steinernema feltiae, S. carpocapsae, and Heterorhabditis bacteriophora) was also evaluated in laboratory assays. Heterorhabditis bacteriophora was the only species that caused significant mortality to swede midge larvae and, at a concentration of 1000 IJs/larva, caused 90-100% mortality in loam, sandy loam, clay and muck soil. This is the first study describing the seasonal development of the swede midge in Quebec. In addition, the potential use of entomopathogenic nematodes as biological control agents against the swede midge is demonstrated.

Water hyacinth, Eichhornia crassipes Mart. Solms-Laubach (Pontederiaceae), a freefloating aquatic macrophyte of Neotropical origin, was introduced into South Africa as an ornamental aquarium plant in the early 1900’s. By the 1970’s it had reached pest proportions in dams and rivers around the country. Due to the sustainability, cost efficiency and low environmental risk associated with biological control, this has been a widely used method in an attempt to reduce infestations to below the threshold where they cause economic and ecological damage. To date, five arthropod and one pathogen biocontrol agents have been introduced for the control of water hyacinth but their impact has been variable. It is believed that their efficacy is hampered by the presence of highly eutrophic systems in South Africa in which plant growth is prolific and the negative effects of herbivory are therefore mitigated. It is for these reasons that new, potentially more damaging biocontrol agents are being considered for release. The water hyacinth grasshopper, Cornops aquaticum Brüner (Orthoptera: Acrididae), which is native to South America and Mexico, was brought into quarantine in Pretoria, South Africa in 1995. Although the grasshopper was identified as one of the most damaging insects associated with water hyacinth in its native range, it has not been considered as a biocontrol agent for water hyacinth anywhere else in the world. After extensive host-range testing which revealed it to be safe for release, a release permit for this candidate agent was issued in 2007. However, host specificity testing is no longer considered to be the only important component of pre-release screening of candidate biocontrol agents. Investigating biological and ecological aspects of the plant-herbivore system that will assist in determination of potential establishment, efficacy and the ability to build up good populations in the recipient environment are some of the important factors. This thesis is a pre-release evaluation of C. aquaticum to determine whether it is sufficiently damaging to water hyacinth to warrant its release. It investigated interactions between the grasshopper and water hyacinth under a range of nutrient conditions found in South African water bodies as well as the impact of the grasshopper on the competitive performance of water hyacinth. Both plant growth rates and the response of water hyacinth to herbivory by the grasshopper were influenced by nutrient availability to the plants. The ability of water hyacinth to compensate for loss of tissue through herbivory was greater under eutrophic nutrient conditions. However, a negative linear relationship was found between grasshopper biomass and water hyacinth performance parameters such as biomass accumulation and leaf production, even under eutrophic conditions. Water hyacinth’s compensatory ability in terms of its potential to mitigate to detrimental effects of insect feeding was dependent on the amount of damage caused by herbivory by the grasshopper. Plant biomass and the competitive ability of water hyacinth in relation to another freefloating aquatic weed species were reduced by C. aquaticum under eutrophic nutrient conditions, in a short space of time. It was also found that grasshopper feeding and characteristics related to their population dynamics such as fecundity and survival were significantly influenced by water nutrient availability and that environmental nutrient availability will influence the control potential of this species should it be released in South Africa. Cornops aquaticum shows promise as a biocontrol agent for water hyacinth but additional factors that were not investigated in this study such as compatibility with the South African climate and the current water hyacinth biocontrol agents need to be combined with these data to make a decision on its release. Possible management options for this species if it is to be introduced into South Africa are discussed.

Potential vegetative and reproductive increases in yield, as well as the biological efficacy against Fusarium verticillioides and F. proliferatum causing ear and stem rot in maize crops of commercially-formulated micro-organism formulation T-Gro (Trichoderma harzianum isolate DB103 WP) combined with Spartacus (Beauveria bassiana isolate DB 105 WP), T-Gro combined with Armenius (Bacillus subtilis isolate DB 109 WP), T-Gro combined with Maximus (Bacillus subtilis isolate DB 108 WP), T-Gro combined with Shelter (Bacillus subtilis isolate DB 101), T-Gro combined with Bismarck (Microbacterium maritypicum isolate DB 107 WP), as well as individual treatments of T-Gro, Armenius, Bismarck, Maximus and Shelter, were investigated when applied to maize seed and soil under field conditions. All the micro-organism treatments were compared with Thiram 750WP (750g/kg thiram WP) and an untreated control. The micro-organism treatments showed an increase in vegetative as well as reproductive yields when compared to the reference product Thiram 750 WP and the untreated control. There were no observations of adverse effects on the germination of maize seed in all the treatments that were applied. The three isolates B. subtilis, T. harzianum, and M. maritypicum, showed a significant reduction in vascular tissue discolouration of the main and ear stems, indicating a potential to be used in the reduction and control of diseases caused by Fusarium spp. Results also showed poor to very good increases of stem and foliage biomass as well as cob yield per plant produced by the micro-organism treatments when compared to the untreated control. The highest cob yield per plant that differed significantly from the untreated control was produced by T-Gro and Shelter. No phytotoxicity of any kind was observed with the application of the micro-organism formulations and they could therefore be deemed suitable to be used for the treatment of maize seed. The micro-organism formulations containing fungal and bacterial biological control agents have the potential to be used in commercial maize production to increase vegetative and reproductive yields and reduce the severity of ear and stem rot in maize. / Agriculture Animal Health and Human Ecology / M.Sc. (Agriculture)

LIMA, Jaqueline Rabelo de. Controle da antracnose pós-colheita do mamão com leveduras killer. 2013. 135 f. : Tese (doutorado) - Universidade Federal do Ceará, Pró-Reitoria de Pesquisa e Pós-Graduação, Programa de Pós-Graduação em Biotecnologia, Renorbio, Fortaleza-CE, 2013. / Submitted by demia Maia (demiamlm@gmail.com) on 2016-05-23T14:38:07Z No. of bitstreams: 1 2013_tese_jrlima.pdf: 3900571 bytes, checksum: 4b74f4790b64e178ecb28e6fbdaeb326 (MD5) / Approved for entry into archive by demia Maia (demiamlm@gmail.com) on 2016-05-23T14:44:00Z (GMT) No. of bitstreams: 1 2013_tese_jrlima.pdf: 3900571 bytes, checksum: 4b74f4790b64e178ecb28e6fbdaeb326 (MD5) / Made available in DSpace on 2016-05-23T14:44:00Z (GMT). No. of bitstreams: 1 2013_tese_jrlima.pdf: 3900571 bytes, checksum: 4b74f4790b64e178ecb28e6fbdaeb326 (MD5) Previous issue date: 2013 / This study aimed to isolate yeast able to produce and excrete the toxin from killer tropical fruits, to act in the biological control of plant pathogens in postharvest. A total of 580 yeasts strains, isolated from Ceara State of Brasil, were evaluated for their ability to produce killer toxin. Of these strains, 29 tested positive for the killer phenotype and were further evaluated for their ability to control Colletotrichum gloeosporioides germination in vitro. All yeast strains that expressed the killer phenotype were characterized by sequencing the D1/D2 regions of the large subunit of the rRNA gene. Five yeast strains provided a significant reduction in mycelial growth and conidial germination of C. gloeosporioides in vitro, especially Meyerozyma guilliermondii, which was able to reduce the fungal mycelial growth on solid medium (PDA) by 60% and block 100% of conidia germination in liquid media (PDB). Filtering and autoclaving the liquid cultures had no effect on the growth of the pathogen. These results indicate the potential use of antagonist yeasts isolated from tropical fruits in the control of anthracnose caused by C. gloeosporioides in papaya. Further elucidation of main mechanisms involved on anthracnose control by these yeasts could be helpful for the development of biocontrol techniques related to the management of this disease in tropical fruits. The efficiency of two killer yeast strains, with better results in vitro were tested in vivo against C. gloeosporioides, Wickerhamomyces anomalus (strain 422) and Meyerozyma guilliermondii (strain 443), as biocontrol agents against C. gloeosporioides, a postharvest anthracnose agent of papaya and other tropical fruits, was assessed. These strains were previously selected through in vitro assays, but in the present study, their in vivo action was assessed. In addition, the influence of phytopathogen inoculation time on the fruit in combination with the use of the biocontrol agent was also assessed. Through the use of scanning electron microscopy (SEM), we assessed mycoparasitism as an antagonistic mechanism of action. In addition, two hydrolytic enzymes, chitinase and β-1, 3 glucanase, were assayed. Our results indicated that W. anomalus (strain 422) and M. guilliermondii (strain 443) reduced the disease occurrence by 24.62% and 20.68%, respectively, for up to 6 days after inoculation, when applied 3 hours before the phytopathogen and incubated in a wet chamber (95% relative humidity) at 28°C. The time of yeast inoculation had a significant effect on its antagonistic action. Application of the yeasts 12 or 24 hours before the phytopathogen inoculation resulted in 13.75% and 30% of disease reductions for W. anomalus (strain 422) and 31.35% and 41.17% reductions for M. guilliermondii (strain 443), respectively. Electron micrographs confirmed mycoparasitism by clearly indicating the interaction of the yeasts with C. gloeosporioides hyphae, causing, in some cases, a loss of turgor and yeast penetration of walls with marked concavity formation on hypha cell walls. The efficiency of two killer yeasts, Wickerhamomyces anomalus (strain 422) and Meyerozyma guilliermondii (strain 443), which showed better results in tests in vtro and in vivo, associated with five different application vehicles was assessed for the protection of papayas postharvest. In this study, after 90 days of incubation at 4°C, W. anomalus (strain 422) and M. guilliermondii (strain 443) were viable with all application vehicles tested. Fruits treated with different formulations (yeasts + application vehicles) had a decreased incidence of disease (by at least 30%) compared with untreated fruits. The treatment of W. anomalus (strain 422) + 2% starch lowered disease occurrence by 48.3%. The most efficient treatments using M. guilliermondii (strain 443) were those with 2% gelatin or 2% liquid carnauba wax, both of which reduced anthracnose by 50% in postharvest papayas. Electron micrographs of the surface tissues of the treated fruits showed that all application vehicles provided excellent adhesion of yeast to the surface. The formulations based on starch (2%), gelatin (2%) and carnauba wax (2%) were the most efficient at controlling fungal diseases in postharvest papayas / Este trabalho objetivou isolar leveduras capazes de produzir e excretar a toxina killer a partir de frutos tropicais, para atuarem no controle biológico de fitopatogenos em pós-colheita. Inicialmente, foram isoladas 580 leveduras a partir de 87 amostras de frutos tropicais (mamão, caju, sapoti, murici, manga e acerola), dentre as quais 29 exibiram o fenótipo killer. Todas as cepas killer foram identificadas pelo sequenciamento da região D1/D2 do 28S rRNA, em que ficou demonstrada a presença de Candida aaseri, Wickerhamomyces anomalus, Pichia kluyveri, Meyerozyma guilliermondii, Kodamaea ohmeri. Cinco leveduras foram capazes de inibir em 100% a germinação de conídios em meio líquido e reduzir o crescimento micelial, em meio sólido de Colletotrichum gloeosporioides in vitro, com destaque para M. guilliermondii (cepa 443) que foi capaz de reduzir o crescimento micelial do fitopatogeno em 60% em meio sólido. As duas leveduras com melhores resultados in vitro foram testadas in vivo, contra C. gloeosporioides, agente da antracnose em pós-colheita de mamão e outros frutos tropicais, W. anomalus (cepa 422) e M. guilliermondii (cepa 443). Também foi investigada a ocorrência de micoparasitismo como mecanismo de ação do antagonista por meio de microscopia eletrônica de varredura – MEV e detecção das enzimas hidrolíticas, quitinase e β-1-3 glucanase. Os resultados demonstraram que, quando aplicada simultaneamente ao fitopatógeno e incubadas em câmara úmida (95% U.R.) a 28 °C, as leveduras W. anomalus (cepa 422) e M. guilliermondii (cepa 443) foram capazes de reduzir a ocorrência da doença em 24,62%, 7,0% e 20,68%, respectivamente, até 06 dias após a inoculação. Verificou-se que o tempo de inoculação da levedura teve significativa influência sobre sua ação antagonista; a aplicação dos agentes com 24 ou 12 horas de antecedência em relação ao fitopatógeno resultou em redução de 30% e 13,75% para W. anomalus (cepa 422), em 40% e 35% para W. anomalus (cepa 440) e em, 41,17 e 31,35% para M. guilliermondii (cepa 443) respectivamente. A ocorrência de micoparasitismo foi confirmada através de eletromicrografias que evidenciaram a união das leveduras às hifas do C. gloeosporioides, provocando, em alguns casos, perda de turgidez e até ruptura dessas; tudo isso associado à produção de β 1-3-glucanase. As leveduras killer, W. anomalus (cepa 422) e M. guilliermondii (cepa 443) que apresentaram melhores resultados nos testes in vivo foram testadas associação com cinco diferentes veículos de aplicação na proteção em pós-colheita de mamão. Após 90 dias de incubação a 4 °C, essas leveduras mantiveram-se viáveis em todos os veículos de aplicação testados e durante todo o período de incubação, frutos tratados com formulações (leveduras + veículos de aplicação) apresentaram incidência de doença, pelo menos 30% menores, quando comparadas aos frutos não tratados. Para W. anomalus (cepa 422), o tratamento que utilizou amido (2%) reduziu em 48,3% a ocorrência de doenças, já para M. guilliermondii, (cepa 443), os tratamentos mais eficientes no controle da doença foram os que utilizaram gelatina e cera líquida de carnaúba (2%) como veículos de aplicação, ambos foram capazes de reduzir em 50% a ocorrência de doenças em pós-colheita de mamões. Eletromicrograficas revelaram que todos os veículos de aplicação foram eficientes em permitir a fixação das leveduras na superfície do fruto. Leveduras killer podem atuar no biocontrole em pós-colheita de mamão e estes microrganismos atuam através de uma variedade de mecanismos de ação, o que potencializa seu efeito protetor e amplia sua eficiência de ação.

Made available in DSpace on 2014-06-11T19:25:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-16Bitstream added on 2014-06-13T20:32:40Z : No. of bitstreams: 1 fraga_df_me_jabo.pdf: 1036947 bytes, checksum: 765bfbd32f9b458cda31ae6bb29934a4 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O conhecimento do comportamento de infestação dos insetos em sistemas agrícolas facilita as amostragens, economiza tempo sem perder a confiabilidade para a tomada de decisões no sistema de manejo de pragas. Assim o objetivo foi estudar a distribuição vertical e temporal de ovos de Alabama argillacea e de Heliothis virescens, e o parasitismo de ovos por Trichogramma pretiosum em cultivares de algodoeiro. O delineamento experimental foi em blocos ao acaso, com cinco tratamentos (cultivares DeltaOPAL, FMX-933, FMT-701, FMX-910 e NuOPAL), com oito repetições. As avaliações foram semanais, a partir da emergência das plantas. Para a distribuição vertical, as plantas foram avaliadas nas partes superior, média e inferior, anotando-se o número de ovos. Os ovos de A. argillacea e de H. virescens foram parasitados por Trichogramma pretiosum. Não houve preferência para oviposição por A. argillacea e por H. virescens durante os estágios fenológicos das plantas, tal como o grau de parasitismo por T. pretiosum também não foi influenciado. Quanto à distribuição vertical dos ovos, o terço superior e o médio das plantas foram os mais preferidos para oviposição por A. argillacea nas cultivares NuOPAL, DeltaOPAL e FMX-910, enquanto que H. virescens preferiu ovipositar no terço superior das plantas. T. pretiosum preferiu ovipositar em ovos de A. argillacea presentes no terço superior e médio das plantas, sendo que ovos de H. virescens presentes no terço superior das plantas foram mais parasitados por T. pretiosum / The knowledge of a pest infestation behavior on agricultural areas help the sampling and reduce time without lose confidence on decisions in integrated pest management. This work aimed to study the vertical and temporal distribution of Alabama argillacea and Heliothis virescens eggs, as well as parasitism of its eggs by Trichogramma pretiosum in cotton cultivars. The experimental design was randomized blocks with five treatments with five treatments (cultivars DeltaOPAL, FMX-933, FMT-701 and FMX-910 and NuOPAL) and with eight replicates. Evaluations were performed weekly since plant emergence. For the vertical distribution, plants were divided into three parts, upper, middle and bottom. The number of eggs present in plants was recorded. A. argillacea and H. virescens eggs were parasitized by Trichogramma pretiosum. There was no preference for oviposition by A. argillacea and H. virescens during the plants phenological stages, as well as T. pretiosum parasitism. Concerning the vertical distribution of eggs, the upper and middle parts of the plants were the most preferred for oviposition by A. argillacea on cultivars NuOPAL, DeltaOPAL and FMX-910, and H. virescens preferred to oviposit in the upper part of the plants. T. pretiosum preferred to oviposit in A. argillacea eggs on the upper and middle parts of the plants, and in H. virescens on the upper part of the plants