Thesis (PhD)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Apple stem grooving virus (ASGV) is globally associated with latent infection in commercial apple trees. Little is known about this plant-­‐virus interaction. This study made use of next-­‐generation sequencing to investigate the effect of virus-­‐infection on the expression of the different small RNA (sRNA) species namely, miRNAs, nat-­‐siRNAs, phasiRNAs, rasiRNAs, tRNA-­‐derived sRNAs and vsiRNAs. Broad and narrow size-­‐range datasets were generated using sRNA libraries prepared from total and size-­‐selected RNA, respectively. Through bioinformatic data analyses, 130 genomic loci were predicted to give rise to miRNAs, 85 of which were novel MIR genes. Targets were predicted for the majority of miRNAs, a few of which could be validated with a publicly available degradome dataset. Cis-­‐ and trans-­‐natural antisense transcripts (NATs) were identified, of which only the latter were highly enriched for sRNAs in their overlapping regions. Transcript as well as genomic regions were also identified that can give rise to phasiRNAs. For 25 of these loci an in-­‐phase miRNA target site was identified, half of which could be validated with the degradome dataset. Nearly all apple repeat sequences in Repbase were associated with sRNA synthesis. sRNAs derived from both ends of mature tRNAs were identified. These sRNAs corresponded to tRFs and tRNA-­‐halves. Reads associated with tRNA-­‐halves were prominent in the broad range datasets. sRNAs, originating from the central regions of tRNAs, were also observed. Analysis of the vsiRNAs suggested the presence of two ASGV genetic variants in two of the samples, while the third sample was infected with only one variant. Comparison of the vsiRNA profiles generated from the two datasets highlighted the influence of library preparation on the interpretation of results. Differential expression analysis of the identified apple sRNA species showed no variation between healthy and infected plants, except for the tRNA-­‐derived sRNAs, which did show altered expression levels. Taken together, the various sRNA species characterised in this study significantly extended the existing knowledge of apple sRNAs and provide a broad platform for future functional studies in apple. This study also presents the first and most comprehensive report on sRNAs involved in ASGV infection in apple. / AFRIKAANSE OPSOMMING: Appel gleufstam virus (ASGV) word wêreldwyd geassosieer met latente infeksie in kommersiële appelbome. Min inligting oor hierdie plant-­‐virus interaksie is beskikbaar. Hierdie studie het van volgende-­‐generasie volgordebepaling gebruik gemaak om die effek van virusinfeksie op die uitdrukking van verskillende klein RNA (sRNA) spesies, nl. miRNAs, nat-­‐siRNAs, phasiRNAs, rasiRNAs, tRNA-­‐afkomstige sRNAs en vsiRNAs, te ondersoek. Datastelle met breë en smal grootte-­‐verspreiding is gegenereer m.b.v. sRNA biblioteke wat onderskeidelik voorberei is vanaf totale RNA en RNA van ‘n bepaalde grootte. Deur middel van bioinformatiese data-­‐ontleding is 130 genomiese loci voorspel wat aanleiding kan gee tot miRNAs, waarvan 85 nuwe MIR gene is. Teikens is voorspel vir die meerderheid van die miRNAs en 'n aantal daarvan kon bevestig word m.b.v. 'n publiek-­‐beskikbare degradoom datastel. Cis-­‐ en trans-­‐natuurlike antisense transkripte (NATs) is geïdentifiseer, waarvan slegs die laasgenoemde verryk was vir sRNAs in hul oorvleuelende areas. Transkrip sowel as genomiese areas, wat aanleiding kan gee tot phasiRNAs, is ook geïdentifiseer. Vir 25 van hierdie loci is 'n in-­‐fase miRNA teiken geïdentifiseer, waarvan die helfte bevestig kon word met die degradoom datastel. Byna al die appel herhalende volgordes in Repbase was geassosieer met sRNA sintese. sRNAs afkomstig van beide kante van volwasse tRNAs is geïdentifiseer. Hierdie sRNAs het ooreengestem met tRFs en tRNA-­‐helftes. Volgordes geassosieer met tRNA-­‐helftes was prominent in die breë grootte-­‐verspreiding datastelle. sRNAs, afkomstig van die sentrale dele van tRNAs, is ook waargeneem. Ontleding van die vsiRNAs het die teenwoordigheid van twee ASGV genetiese variante in twee van die monsters aangetoon, terwyl die derde monster met slegs een variant geïnfekteer was. Die vergelyking van vsiRNA profiele, gegenereer vanaf die twee datasteltipes, beklemtoon die invloed van biblioteek voorbereiding op die interpretasie van resultate. Ontleding van die differensiële uitdrukking van die geïdentifiseerde appel sRNA spesies het geen verskil tussen gesonde en geïnfekteerde plante getoon nie, behalwe vir die tRNA-­‐afkomstige sRNAs, wat wel verandering die vlak van uitdrukking getoon het. Die verskillende sRNA spesies wat in hierdie studie geïdentifiseer is, het die bestaande kennis van appel sRNAs aansienlik uitgebrei en bied 'n breë platform vir toekomstige funksionele studies in appel. Hierdie studie bied ook die eerste, en mees omvattende verslag oor sRNAs betrokke in ASGV infeksie in appel.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The codling moth, Cydia pomonella (Lepidoptera: Tortricidae), is the key pest of apples and pears worldwide. The withdrawal of certain fundamental chemicals from codling moth management spray programmes, due to concerns about human, environmental and ecosystem health, has resulted in the search for softer, more environmentally friendly, and safer control measures. Entomopathogenic nematodes (EPNs), naturally occur in the soil, and actively search for hosts. The interest in using EPNs from the families Heterorhabditidae and Steinernematidae as a control measure was sparked in 1953, when an EPN was discovered in an insect. The aim is to incorporate EPNs in an integrated pest management (IPM) programme, to ensure minimal residue and eventually residue-free pome fruit production in South Africa. In order to ensure EPN success, both the environmental and technical factors influencing their efficacy, were investigated in this study. The biocontrol potential of three imported EPN isolates, being Steinernema feltiae and two isolates of Heterorhabditis bacteriophora (Hb1, Hb2), as well as a local isolate, Steinernema yirgalemense, were evaluated for the control of the codling moth under local conditions. All concentrations of S. yirgalemense, applied by immersion in a suspension of nematodes, gave > 98% control. The two formulated isolates of H. bacteriophora, Hb1-f and Hb2-f, gave < 30% control. When using the same nematode isolates, produced in vivo, S. yirgalemense still resulted in a higher codling moth control of > 90%, compared to 54% and 31% control of the H. bacteriophora Hb1 and Hb2 isolates, respectively. In follow up field trials, S. feltiae resulted in ≥ 80% control, and was thus more effective than both S. yirgalemense and the H. bacteriophora (Hb1) isolates, with 66% and 24%, and 24% and 9% control, for two separate trials, respectively. To validate the data obtained from the field trials, subsequent laboratory bioassays were conducted evaluating temperature regimes, following the same cycle as under natural conditions, with a constant humidity of 100%. Steinernema feltiae proved to be most effective, causing > 90% mortality, followed by S. yirgalemense, with 78% mortality. The two H. bacteriophora isolates (Hb1, Hb2) under the above-mentioned laboratory conditions, resulted in 73% and 59% control, respectively. Humidity thus seems to be the most important factor affecting EPN efficacy during above-ground applications. Steinernema feltiae proved to be a better candidate than S. yirgalemense for the control of the codling moth. The efficacy of different EPN isolates in controlling diapausing codling moth larvae at different temperatures was also evaluated, under local conditions, using spray application. Steinernema feltiae and two isolates of H. bacteriophora Hb1 and Hb2, including two local isolates, S. yirgalemense and Steinernema jeffreyense, were evaluated. The use of S. jeffreyense resulted in the most effective control, with 67% mortality, followed by H. bacteriophora (Hb1) with 42%, and then by S. yirgalemense with 41%. Laboratory bioassays simulating field conditions revealed that S. feltiae was most virulent to codling moth larvae, with 67% mortality by infection, followed by S. yirgalemense with 58%, the H. bacteriophora strain Hb1 with 48%, and the Hb2 strain with 24%. A comparison of the infection and penetration rate of two isolates of H. bacteriophora (Hb1, Hb2), S. feltiae and S. yirgalemense, which was carried out in multiwell plates at 14°C and 25°C, respectively, confirms the dramatic effect of temperature on EPN efficacy. At 14°C, all treatments with EPN species resulted in slower codling moth mortality than they did at 25°C, as after 48 h, < 15% mortality was recorded for all species, whereas at the warmer temperature, > 98% mortality was recorded for all species. After the exposure of washed, cool-treated larvae to 25°C for 24 h, the application of both S. feltiae and S. yirgalemense resulted in 100% mortality, whereas the application of the two H. bacteriophora isolates, Hb1 and Hb2, resulted in 68% and 54% control, respectively, over the same time period. At 14°C, S. feltiae had the highest average penetration rate of 20 IJs/insect, followed by S. yirgalemense with 14 IJs/insect, whereas S. yirgalemense had the highest penetration rate at 25°C, with 39 IJs/insect, followed by S. feltiae, with 9 IJs/insect. The two H. bacteriophora isolates had higher average penetration rates at the higher temperature. This study has highlighted the biocontrol potential of S. jeffreyense, as well as showing that S. feltiae is a cold-active nematode, whereas the other three EPN isolates prefer warmer temperatures. Stacked wooden fruit bins are regarded as preferred overwintering sites for codling moth diapausing larvae. Control strategies against the codling moth in South Africa have been hampered by the reinfestation of orchards by nearby stacked infested fruit bins or by the movement of bins between orchards. Worldwide, wooden fruit bins are systematically being replaced with plastic bins, which, in South Africa, will only be phased out over a few years. The objective of this study was to evaluate the potential of H. bacteriophora, S. feltiae, and S. yirgalemense, to disinfest miniature wooden fruit bins under controlled conditions in the laboratory. After dipping minibins in a suspension of 25 IJs/ml of all three EPN species, under optimum conditions of temperature and humidity, the highest percentage of control was obtained using S. feltiae (75%) followed by S. yirgalemense (57%), and then by H. bacteriophora (Hb1) (27%). The addition of adjuvants significantly increased (p < 0.001) S. feltiae infectivity to > 95%, whereas it did not result in a significant increase in H. bacteriophora or S. yirgalemense infectivity. The results indicated that H. bacteriophora would not be a suitable candidate to use for the control of the codling moth larvae in wooden fruit bins. The current preferred candidate for control would be S. feltiae, whose efficacy could be increased by means of the addition of an adjuvant. During winter, when the whole codling moth population are larvae and in diapause, no control measures are applied in orchards. This study has shown that EPNs can be sprayed in orchards to lower the codling moth cohort emerging after winter, as well as be included in an IPM programme. EPNs can act as a second line of defence, through supplementary control, and ensure effective control of the codling moth larvae which survived chemical spray applications, to safeguard against resistant codling moth populations in the next season. / AFRIKAANSE OPSOMMING: Kodlingmot, Cydia pomonella (Lepidoptera: Tortricidae), is ‘n belangrike plaag van appels en pere wêreldwyd. Die onttrekking van sekere fundamentele chemikalieë vanuit die kodlingmot beheerprogram weens die kommer oor menslike, omgewings en ekosisteemgesondheid, het gelei tot die soektog na sagter, meer omgewingsvriendelike en veiliger beheermaatreëls. Entomopatogeniese nematodes (EPNs) kom natuurlik in die grond voor en soek aktief na gashere. Die belangstelling in die gebruik van EPNs van die families Heterorhabditidae en Steinernematidae as 'n beheermaatreël is te danke aan die ontdekking van 'n EPN in ‘n insek in 1953. Die doel is om EPNs in 'n geïntegreerde plaagbeheerprogram (GPB) te inkorporeer om sodoende minimale residue te verseker en uiteindelik residu vrye produksie van kernvrugte in Suid-Afrika. Ten einde die sukses van EPNs te verseker, is beide die omgewings- en tegniese faktore wat hul doeltreffendheid beïnvloed in die studie ondersoek. Die biologiese beheer potensiaal van drie ingevoerde EPN isolate, Steinernema feltiae en twee Heterorhabditis bacteriophora (Hb1, Hb2) isolate, sowel as 'n plaaslike isolaat, Steinernema yirgalemense, is vir die beheer van kodlingmot onder plaaslike toestande geëvalueer. Alle konsentrasies van S. yirgalemense, wat deur indompeling in ‘n suspensie van nematodes toegedien is, het > 98% beheer tot gevolg gehad. Die twee geformuleerde isolate van H. bacteriophora, Hb1-f en Hb2-f, het < 30% beheer gegee. Met die gebruik van dieselfde nematode isolate, wat in vivo geproduseer is, het S. yirgalemense nog steeds 'n hoër kodlingmot beheer van > 90% opgelewer, in vergelyking met die 54% en 31% beheer van die H. bacteriophora Hb1 en Hb2 isolate, onderskeidelik. Steinernema feltiae het in opvolg veldproewe ≥ 80% beheer tot gevolg gehad en was dus meer effektief as beide S. yirgalemense en die H. bacteriophora (Hb1) isolate, met 66% en 24% en 24% en 9% beheer onderskeidelik in twee afsonderlike veldproewe. Om die resultate van die veldproewe te bevestig, is daaropvolgende laboratorium biotoetse uitgevoer en temperatuur regimes is geëvalueer deur die selfde siklus as onder natuurlike toestande te volg, met 'n konstante humiditeit van 100%. Die studie het bewys dat S. feltiae die mees doeltreffende isolate was met > 90% mortaliteit, S. yirgalemense het gevolg met 78% mortaliteit. Die twee H. bacteriophora isolate (Hb1, Hb2) het onderskeidelik onder bogenoemde laboratorium toestande 73% en 59% beheer tot gevolg gehad. Humiditeit blyk dus die belangrikste faktor te wees wat EPN se doeltreffendheid tydens bogrondse toediening affekteer. Die studie het bewys dat S. feltiae 'n beter kandidaat as S. yirgalemense vir die beheer van kodlingmot is. Die doeltreffendheid van verskillende EPN isolate vir die beheer van diapause kodlingmot larwes sowel as EPN se aktiwiteit by verskillende temperature is ook onder plaaslike toestande, deur bogrondse bespuitings, geëvalueer. Steinernema feltiae en twee isolate van H. bacteriophora (Hb1, Hb2), S. yirgalemense en ‘n ander plaaslike isolaat, Steinernema jeffreyense, is geëvalueer. Die gebruik van S. jeffreyense, het tot die mees effektiewe beheer gelei, met 67% mortaliteit, gevolg deur H. bacteriophora (Hb1) met 42%, en dan S. yirgalemense met 41%. Laboratorium biotoetse wat veldtoestande simuleer, het bewys dat S. feltiae die mees doeltreffend teen kodlingmot larwes is, met 67% mortaliteit tydens infeksie, gevolg deur S. yirgalemense met 58%, die H. bacteriophora Hb1 isolaat met 48%, en die Hb2 isolaat met 24%. 'n Vergelyking van die infeksie- en penetrasie tempo van twee isolate van H. bacteriophora (Hb1, Hb2), S. feltiae en S. yirgalemense wat in 12-put plate teen 14°C en 25°C uitgevoer is, het die dramatiese effek van temperatuur op EPN doeltreffendheid bevestig. By 14°C het alle EPN spesies behandelings stadiger kodlingmot mortaliteit as by 25°C na 48h tot gevolg gehad. ‘n Mortaliteit van < 15% is vir alle spesies aangeteken terwyl by die warmer temperature is > 98% mortaliteit vir alle spesies aangeteken. Na die blootstelling van afgespoelde, koel behandelde larwes aan 25°C vir 24 uur, het die toediening van beide S. feltiae en S. yirgalemense, 100% mortaliteit van larwes tot gevolg gehad terwyl die toediening van die twee H. bacteriophora isolate, Hb1 en Hb2, onderskeidelik 68% en 54% beheer tot gevolg gehad, oor dieselfde tydperk. By 14°C, het S. feltiae die hoogste gemiddelde penetrasie tempo van 20 ILs/ larwe, gevolg deur S. yirgalemense met 14 ILs/ larwe tot gevolg gehad, terwyl S. yirgalemense die hoogste penetrasie tempo getoon het by 25°C met 39 ILs/ insek, gevolg deur S. feltiae met 9 ILs/ insek. Die twee H. bacteriophora isolate (Hb1 en Hb2) het ook hoër gemiddelde penetrasie tempo by die hoër temperatuur getoon. Hierdie studie het die biobeheer potensiaal van S. jeffreyense beklemtoon, asook weereens bevestig dat S. feltiae ‘n koue-aktiewe nematode is, terwyl die ander drie EPN isolate warmer temperature verkies. Hout vrugtekratstapels, word beskou as ‘n ideale oorwintering skuiling vir kodlingmot diapause larwes. In Suid-Afrika word beheerstrategieë teen kodlingmot in die wiele gery deur die herbesmetting van boorde deur nabygeleë besmette hout vrugtekratte of deur die beweging van kratte tussen boorde. Hout vrugtekratte word wêreldwyd stelselmatig vervang met plastiek kratte. Dit sal egter eers oor ‘n aantal jare in Suid-Afrika uitgefaseer word. Die doel van hierdie studie was om die potensiaal van H. bacteriophora, S. feltiae, en S. yirgalemense te evalueer deur miniatuur hout vrugtekratte onder gekontroleerde toestande in die laboratorium te disinfekteer. Na die onderdompeling van die mini vrugtekratte in 'n nematode suspensie van 25 ILs/ml van al drie EPN spesies, onder optimale toestande van temperatuur en humiditeit, is die hoogste persentasie van beheer met die gebruik van S. feltiae (74,85% ± 3.64%) verkry. Die byvoeging van toevoegings middels het S. feltiae se vermoë om te infekteer betekenisvol (p <0,001) tot > 95% verhoog, maar dit het nie tot 'n betekenisvolle toename in die infektiwiteit van H. bacteriophora of S. yirgalemense gelei nie. Die resultate dui daarop dat H. bacteriophora nie 'n geskikte kandidaat is om te gebruik vir die beheer van kodlingmot larwes in besmette hout kratte nie. Die voorkeurkandidaat tans vir beheer is S. feltiae, waarvan die doeltreffendheid verhoog kan word deur middel van die byvoeging van 'n bymiddel. Gedurende die winter wanneer die hele kodlingmot populasie as larwes in diapause is, word geen beheer in boorde toegepas nie. Hierdie studie het getoon dat EPNs in boorde gespuit kan word om sodoende die opkomende kodlingmot populasie na die winter te verlaag en kan ook ingesluit word in 'n GPB program. Die EPNs kan as 'n tweede verdedigingslinie optree en doeltreffende beheer van kodlingmot larwes verseker wat chemiese bespuitings oorleef het, en sodoende beskerming teen weerstandige kodlingmot populasies in die volgende seisoen bied.

O trabalho objetiva apresentar as características da certificação de maçãs e analisar o comportamento do consumidor do município de Santa Maria, localizado na região central do Rio Grande do Sul, Brasil. A análise compreende a discussão dos aspectos relacionados à valoração da certificação e das características desse produto agroalimentar no momento da compra. Como procedimentos metodológicos, adotou-se a revisão bibliográfica para compreender os processos de certificação e regulamentação dos mercados agroalimentares e para discutir aspectos relacionados ao comportamento do consumidor. Além disso, usaram-se dados secundários para compreender a realidade produtiva da maçã no Brasil e no mundo. Para discutir a relação entre os consumidores e o mercado de maçãs, construiu-se um formulário com questões abertas e fechadas sobre o perfil do consumidor, os aspectos considerados no momento da compra e seu nível de conhecimento acerca da certificação. Os formulários foram aplicados para uma amostra significativa para o município de Santa Maria, em quatro espaços supermercadistas. Para as análises, adotaram-se procedimentos estatísticos e descritivos, como testes paramétricos, não paramétricos e a estimação de um modelo de regressão logística para avaliar como o público consumidor valora a certificação. Os resultados apontam que o público estudado é heterogêneo. Constatou-se que o consumidor não se detém por informações sobre segurança transmitidas e apresentadas nas frutas através dos selos de certificação. Poucos consumidores afirmaram saber o que, de fato, é a certificação. Uma parcela maior afirmou que, além de não saber o significado, não considera a certificação da fruta relevante no momento da compra, valorizando mais outros aspectos e fatores, tais como aparência, tamanho e o preço da fruta. Concluise que o nível de conhecimento acerca das informações sobre segurança, inspeção e controle e certificação tendem a não ser tão importantes no momento da compra. Por final, concluiu-se que se a produção de maçãs fosse totalmente destinada ao mercado santa-mariense, de acordo com a demanda representada pelos consumidores, o fato da oferta dessa fruta possuir algum selo de certificação seria pouco relevante. / This paper aims to present the characteristics of the certification apples and analyze consumer behavior in the municipality of Santa Maria, located in the central region of Rio Grande do Sul, Brazil. The analysis includes the discussion of issues related to the valuation of certification and characteristics of agro-food product at the purchase time. As methodological procedures in the study, we adopted the literature review to understand the processes of certification and regulation of agrifood markets and to discuss issues related to consumer behavior. In addition, they used secondary data to understand the reality of apple production in Brazil and worldwide. To discuss about the relationship between consumers and the market for apples, built up a form with closed questions about the consumer profile, the aspects considered at the time of purchase and their level of knowledge about certification. The forms were applied to a significant sample for the municipality of Santa Maria in four supermarket spaces. For analysis, were adopted descriptive and statistical procedures such as parametric tests, as nonparametric tests, and the estimation of a logistic regression model to assess how the consuming public values the certification. The results show that the studied public is heterogeneous. It was found that the consumer do not value safety information transmitted and presented in fruits through the certification seals. Some few consumers know what certification is. A larger portion said that in addition to not knowing the meaning, does not consider the certification something relevant at the time of purchase, giving more importance to other aspects and factors, such as appearance, size and fruit price. It is concluded that the level of knowledge about information security, inspection and control and certification tend to not be as important at the time of purchase. By the end, it is concluded if that the production of apples was fully intended to Santa Maria market, according to the demand represented by consumers, the fact that the supply of this fruit have a certification label would not be relevant.

Made available in DSpace on 2014-06-11T19:31:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-02Bitstream added on 2014-06-13T20:21:44Z : No. of bitstreams: 1 celli_mg_dr_sjrp.pdf: 756707 bytes, checksum: 4e5cc119fa93aadaf2ac40ff2235e038 (MD5) / A patulina, toxina com potencial carcinogênico, mutagênico e teratogênico produzida por Penicillium spp., Aspergillus spp. e Byssoclamys spp., pode ser encontrada em maçãs, sucos comerciais e outros produtos não fermentados constituindo-se num problema neste setor agroindustrial. Em bebidas fermentadas, porém, não é detectável mesmo que a matéria-prima esteja visivelmente contaminada. O presente trabalho visou quantificar a patulina tanto no tecido deteriorado de maçãs in natura quanto na parte sadia ao redor da lesão, monitorar a degradação no processo fermentativo típico utilizando Saccharomyces cerevisiae e degradar a toxina presente em produtos derivados de maçã pela ação do composto bioativo produzido pela levedura. A micotoxina foi quantificada por CLAE em sistema isocrático de fase reversa e detector UV a 275 nm. O grau de recuperação alcançado foi de 86,24% e os limites de detecção e de quantificação foram 4,3 µg/L e 8,6 µg/L, respectivamente. Foram analisados 35 frutos de maçã in natura cultivar Fuji, tendo sido constatada a presença de patulina em 32 amostras, em concentrações que variaram de 1,01 a 120,4 mg/Kg de tecido na porção deteriorada e de 0,02 a 5,02 mg/Kg de tecido, na sadia. Para avaliação da cinética de degradação de patulina, suco de maçã contendo 5,0 µg de patulina/mL foi inoculado com 0,25g de células de levedura seca ativa/L, sendo observada uma maior redução no teor de toxina de 81,6% nas primeiras 48 horas de fermentação. Este tempo foi utilizado como parâmetro para obtenção do extrato bruto contendo o composto bioativo capaz de degradar a toxina. Na cinética de degradação de patulina pelo extrato bruto estéril inoculado com 3,0 µg de patulina/mL, foi obtida a maior taxa de degradação (50%) da toxina nas primeiras 48 horas, seguido de uma degradação mais lenta de apenas 5,7% nas 48 horas... / Patulin, a toxin with potential carcinogenic, mutagenic and teratogenic effects produced by Penicillium sp., Aspergillus sp. and Byssoclamys sp., can be found in apples, juices and other commercial non-fermented products making up a problem in the agribusiness sector. However, patulin is not detectable in fermented beverages even if the raw material is visibly contaminated. This paper aimed to quantify patulin in both the tissue deteriorated fresh apples and at the sound around the lesion, to monitor the degradation in typical fermentation process using Saccharomyces cerevisiae and degrade the toxin present in products derived from apple by the action of the bioactive compound produced by yeast. The mycotoxin was quantified by HPLC in isocratic system of reversed phase and UV detection at 275 nm. The degree of recovery achieved was 86.24% and the limits of detection and quantification were 4.3 µg/L and 8.6 µg/L, respectively. Thirty five fresh apple fruits Fuji were analyzed and it was, found the presence of patulin in 32 samples in concentrations that ranged from 1.01 to 120,4 mg/Kg of tissue in the damaged portion and 0.02 to 5, 02 mg/Kg of tissue in sound. To evaluate the kinetics of degradation of patulina, apple juice with 5.0 µg patulin/mL was inoculated with 0.25 g of cells active dry yeast/L, being observed a greater reduction in the toxin content of 81.6% in the first 48 hours of fermentation. This time was used as a parameter to obtain the crude extracts containing bioactive compound capable of degrading the toxin. The kinetics of patulin degradation by the crude extract inoculated with 3.0 µg patulin/mL showed the highest rate of the toxin degradation (50%) in the first 48 hours, followed by only 5.7% degradation after the remaining 48 hours of analysis. The kinetics of degradation of patulin by the extract dialysate inoculated with 3.0 µg patulin/mL showed... (Complete abstract click electronic access below)

Competitive exclusion has been the mechanism hypothesized to account for the biological control of fire blight disease of pear and apple by the bacterium Pseudomonas fluorescens A506 (A506). Recent laboratory assays demonstrated, however, that A506 produces an antibiotic that is toxic to the fire blight pathogen, Erwinia amylovora, when cultured on media amended with iron (Fe����� or Fe�����). This study investigated this iron-dependent antibiosis by A506 by: 1) examining bioavailability of iron to A506 on blossom surfaces, 2) mutagenizing A506 to disrupt genes involved in antibiotic production, and 3) evaluating suppression of fire blight by A506 when co-treated with an iron chelate (FeEDDHA). Bioavailability of iron on blossoms was investigated with an iron biosensor [iron-regulated promoter (pvd) fused to an ice nucleation reporter gene (inaZ)] in A506. A506 (pvd-inaZ) expressed high ice nucleation activity (INA) on blossoms indicating a low-iron environment unlikely to induce antibiosis by A506. Spraying blossoms with FeEDDHA at concentrations ���0.1 mM significantly suppressed INA by A506 (pvd-inaZ). Transposon mutagenesis was used to generate and select mutants of A506 exhibiting altered antibiotic production profiles. One antibiotic-deficient mutant, A506 Ant���, was recovered; this mutant showed reduced epiphytic fitness on blossoms of apple and pear trees compared to the parent stain, A506. Another mutant, A506 Ant���, lost the characteristic fluorescent phenotype and exhibited iron-independent antibiotic production in defined culture media. A506 Ant��� established high populations on blossoms of apple and pear trees, similar to populations attained by A506, and reduced incidence of fire blight between 20 to 40%, levels comparable to A506 in orchard trials. In orchard trials, A506 was co-treated with FeEDDHA and fire blight suppression was evaluated. Bacterial strains established high populations on blossoms when co-treated with 0.1 mM FeEDDHA or in water. Significantly enhanced suppression of fire blight incidence by antibiotic producing strains of A506 amended with 0.1 mM FeEDDHA was observed in 2 of 5 trials, providing some evidence that iron-induced antibiosis can be a contributing mechanism in disease control. Lack of disease control by the antibiotic deficient strain, A506 GacS, and by 0.1 mM FeEDDHA alone added support to this hypothesis. / Graduation date: 2004

The biology and behavior of the codling moth, Cydia pomonella (L.), was studied to determine factors responsible for differences in susceptibility to moth damage among apple and pear cultivars. In release experiments, codling moth adults were intercepted in significantly greater numbers in host (apple, pear) than in non-host (maple) trees, suggesting a directed response rather than random movement to host trees. There was no correlation between adult preference for certain hosts and the susceptibility of the hosts to codling moth attack. Adults were guided by cues from the tree canopy, foliage and fruit. Cues from host fruit were less important at the beginning of the season. Similar adult behavior patterns were observed in different coding moth strains. However, moths reared on apples were guided more by olfactory cues from fruit than sterilized moths reared on artificial diet. The percentage of moths mating under caged conditions was higher in apple and pear than in maple trees. In host plants, stimuli from fruit were not essential for mating. The egg distribution in the field varied through the season depending on the host cultivar. However, mean distance of eggs to fruit, as well as larval travel speed, was not different among host cultivars. Selection of an oviposition site by the adult female was affected by several factors, including visual, chemical, and tactile stimuli of host fruit, as well as anatomical (pubescence) and chemical (oviposition stimulants) properties of leaves. There were no differences in natural mortality and developmental rates of the egg stage on host cultivars. However, there were significant differences in first instar larval mortality among hosts over most of the season. Mortality was much higher on 'Anjou' than on 'Bartlett' and 'Red Delicious', except at the end of the season. Differential first instar mortality was due to the different rate of success in entering the fruit. The ability to penetrate fruit was correlated with infestation levels observed in the field and related both to neonate behavior and anatomical characteristics of host fruits. Larval food source did not affect larval and pupal development rates, adult fecundity, or egg viability. / Graduation date: 2001

Intra- and interplant movement and dispersal of the predator mites Neoseiulus californicus (McGregor) and Neoseiulus fallacis (Garman) were studied on both lima bean (Phaseolus lunatus L.) and apple (Malus pumila Miller) branch plant systems that were seeded with excess numbers of the spider mite Tetranychus urticae Koch. Individuals of either predator were randomly selected from colonies of well-fed, mixed-age adult females and moved to test plants. When tested separately in each plant system, median leaf positions of N. californicus from the point of release were greater than those of N. fallacis at 1-8 d. A fan placed in front of a common source unit (1x3m) containing T. urticae and near equal densities of both predaceous mites, provided continuous air (wind) to 3 isolated receiver units located 2.5, 5, and 7.5 m downwind. Receiver units consisted of continuous bean foliage with excess T. urticae, but no predators. Wind speeds at the source and each receiver unit averaged 2.2, 0.9, 0.4, and 0.03 m/s respectively. As predators eliminated prey, N. fallacis dispersed earlier at higher prey densities and further downwind than N. californicus, but cumulative densities of each predator in all receiver units were alike after 20 d. Results of both plant movement studies supported the hypothesis that N. californicus has dispersal traits more like those of a generalist predator of spider mites than does N. fallacis. Spatial patterns of dispersal and biological control of pest mites are discussed in relation to predation types within the Phytoseiidae. / Graduation date: 1997

A two year experiment demonstrated electrical energy savings through evaporator fan cycling in part of a full-sized refrigerated controlled atmosphere apple storage facility in Yakima, Washington. Room cooling demand and evaporator fan operation was controlled by the bulk fruit temperature. Refrigerant temperature was regulated by a computer controlled ramping sequence and a weighted average of three thermistors rather than a room thermostat. Evaporator fans remained off 60-65% of the time during periods of fan cycling operation. Seasonal average fan energy savings approached 50-55% when the product remained in storage for a typical length of time. Room environment conditions, product mass loss, and fruit quality were not compromised. Bulk fruit temperature was exceptionally stable as compared to fruit stored in non-fan cycled rooms controlled by traditional means. Less fan motor heat input in the fruit storage space also results in compressor energy savings. Total electrical energy savings were projected to an 18 room fruit storage facility implementing computer controlled evaporator fan cycling. Projected electrical energy savings achievable through evaporator fan cycling for the state of Washington were made based on reported apple storage capacity. / Graduation date: 1993

Computer decision aids can help integrate and apply diverse sources of information and expertise to problems of integrated pest management (IPM) in agriculture and forestry. AppleMgr combines a rule-based expert system with databases and spreadsheets in a prototype decision aid intended to be expanded and modified for use by extension workers in the Northwest U.S. The program requires an IBMcompatible microcomputer with hard disk. AppleMgr concentrates on the two most important insect pests on apple in the Northwest--codling moth, Cydia, pomonella (L.), and San Jose scale, Quadraspidiotus perniciosus Comstock, and on phytophagous mites, whose control largely depends on predators. The primary goal of AppleMgr is to demonstrate an improved process of decision making in apple IPM. AppleMgr is composed of modules for diagnosis of pest injury, identification of pest and natural enemy specimens, and management. The first two modules arrive at conclusions through backward-chaining inference from user observations. The management module uses backward chaining supplemented with external calculation programs to find the net benefit of a pesticide application. A method is included to predict yield and fruit size from crop samples. Cullage from codling moth and San Jose scale, mite effect on fruit size, probability of biological mite control and pesticide efficacy are predicted from researchers' data and estimates. Selected relative beneficial and adverse side effects of pesticides are presented in spreadsheets. An analysis of packing house records for apple crops from eight orchards at three yields using 1987 and 1988 prices and packing charges showed that net crop value varied by up to $8000 per acre. The variability in crop value and the importance of adverse side effects of pesticides suggest that the commonly-used "action thresholds" for treatment are seriously inadequate. AppleMgr may point the way toward more dynamic and realistic methods of IPM decision making. / Graduation date: 1990

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