Thesis (MSc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Plants are constantly exposed to biotic and abiotic stress inducing factors that threaten their existence. Biotic factors such as pathogens are the cause of huge yield losses to crop plants worldwide with fungal pathogens debatably constituting the worst damage. Fungal pathogens such as Botrytis cinerea, which has a wide host range, release cell wall degrading enzymes called endopolygalacturonases (ePGs) during plant infection. These ePGs break down the pectin component of the cell wall, thus providing an entry route, as well as nutrients for the fungus. Plants have evolved mechanisms to counteract and suppress the action of the ePGs. This is achieved through the action of cell wall associated proteins called polygalacturonaseinhibiting proteins, PGIPs. PGIPs directly inhibit ePGs and their inhibitory action also prolongs the existence of longer chain oligogalacturonide residues which are believed to elicit a cascade of defence responses. In grapevine, a PGIP encoding gene, VvPGIP1, was previously isolated and characterised. VvPGIP1, as well as nine non-vinifera grapevine PGIPs have been expressed in tobacco and shown to be potent antifungal proteins that caused the transgenic tobacco to have strong resistance phenotypes against Botrytis in whole plant infection assays. Following on the tobacco study, two of the non-vinifera PGIPs were expressed in cultivars of the susceptible Vitis vinifera. Characterisation of the putative transgenic population showed that transgene integration was successful, the transgenes were being expressed and there were at least 29 transgenic lines with independent integration events. The transgenic lines were confirmed to have active PGIPs (transgene-derived) in their leaves. Crude protein extracts from 22 lines exhibited 100% inhibition against crude B. cinerea PGs (BcPGs). The plant lines with positive transgene integration, expression, independent integration events and exhibiting 100% transgene-derived PGIP activity were further selected for whole plant and detached leaf antifungal assays where they were challenged with B. cinerea. The whole plant infection assay showed that expression of the non-vinifera PGIPs in V. vinifera promotes susceptibility to B. cinerea, not resistance. This surprising result could perhaps be explained by a quicker and stronger recognition between the pathogen and the host and the stronger activation of defence responses in the host. A more active hypersensitive response in the host would benefit Botrytis being a necrotroph. The type of lesions and the onset and speed of lesion development observed on the transgenics lines versus the wild type support this possibility. Knowledge gaps with regards to the efficiency of the ePG inhibition by the nonvinifera PGIPs during infection of grapevine tissue; the potential changes that might be caused by expressing PGIPs in a grapevine host with a native PGIP with high homology to the transgenes (including potential gene silencing) and the potential impact on defence signalling and defence responses all provides further avenues of study to elucidate this very interesting phenotype further. Overall, this study provides a comprehensively characterised population of transgenic plants that provides useful resources for in vivo analysis of PGIP function in defence, where the host plant harbours a native copy of the PGIP encoding gene. / AFRIKAANSE OPSOMMING: Plante word voortdurend blootgestel aan biotiese en abiotiese faktore, wat stres veroorsaak en hul bestaan bedreig. Biotiese faktore, soos patogene, veroorsaak groot verliese in wêreldwye gewasopbrengste, met swampatogene wat moontlik die grootste skade veroorsaak. Swampatogene, soos Botrytis cinerea, wat ‘n wye reeks gasheerplante kan infekteer, stel selwand-afbrekende ensieme tydens plantinfeksie vry, wat as endo-poligalakturonases (ePG’s). bekend staan. Hierdie ePG’s breek die pektienkomponent van die selwand af, wat gevolglik as ‘n ingangspunt dien,asook voedingstowwe vir die swam verskaf. Plante het meganismes ontwikkel om die aktiwiteit van hierdie ePG’s te bekamp en te onderdruk. Die aktiwiteit van die selwand-geassosieërde proteïene, genaamd poligalakturonase-inhiberende proteïene (PGIP’s), speel hier ‘n rol. PGIP’s inhibeer ePG’s direk en hul inhiberende aktiwiteit verleng ook die bestaan van langketting oligogalakturoniedresidu’s, wat blykbaar ‘n kaskade van weerstandsreaksies kan inisieer. ‘n PGIP-koderende geen, VvPGIP1, is voorheen uit wingerd geïsoleer en gekarakteriseer. VvPGIP1, asook nege nie-vinifera wingerd-PGIP’s is voorheen in tabak uitgedruk en bevestig as proteïene met sterk anti-swamaktiwiteit, soos bevestig deur die bevinding dat die transgeniese tabak ‘n weerstandsfenotipe teen Botrytis in heelplant-infeksietoetse het. Ná die tabakstudie is twee van die nie-vinifera PGIP’s uitgedruk in vatbare V. vinifera-kultivars. Karakterisering van die vermeende transgeniese bevolking het getoon dat die transgeen-integrasie suksesvol was, dat die transgeen uitgedruk word en dat daar ten minste 29 transgeniese lyne met onafhanklike integrasie gebeurtenisse geskep is. Daar is verder bevestig dat die transgeniese lyne aktiewe PGIP’s (transgeen-afkomstig) in hul blare het. Ongesuiwerde proteïenekstrakte van 22 lyne het 100% inhibisie teen ‘n mengsel van ongesuiwerde B. cinerea PGs (BcPGs) getoon. Die plantlyne met positiewe transgeenintegrasie en -uitdrukking, asook onafhanklike integrasiegebeure en wat 100% transgeen-afkomstige PGIP-aktiwiteit getoon het, is verder aan heel-plant en verwyderde blaarswaminfeksies met B cinerea onderwerp. Die heelplantinfeksietoetse het getoon dat uitdrukking van nie-vinifera PGIP’s in V. vinifera ‘n toename, in plaas van ‘n afname, in vatbaarheid teen B. cinerea veroorsaak. Hierdie verbasende resultaat kan moontlik toegeskryf word aan ‘n vinniger en sterker herkenningsreaksie tussen patogeen en gasheer en die moontlike sterker stimulering van weerstandsreaksies in die gasheer. ‘n Meer aktiewe hipersensitiewe reaksie in die gasheer sal tot die voordeel van Botrytis, wat ‘n nektrotroof is, wees. Die tipe letsel, asook die aanvang en spoed van letselontwikkeling wat waargeneem is in transgeniese lyne teenoor die wilde-tipe ondersteun hierdie moontlikheid. Gapings in kennis ten opsigte van die doeltreffendheid van die ePG-inhibisie deur die nievinifera PGIP’s tydens infeksie van wingerdweefsel, die moontlike veranderinge (insluitend ‘n moontlike geenuitdowingseffek) wat veroorsaak kan word deur die uitdrukking van PGIP-gene in ‘n kultivar met ‘n inheemse en baie homoloë PGIP-geen, kon ‘n invloed op weerstandseine en weerstandsreaksies gehad het. Hierdie aspekte lewer verdere studiemoontlikhede om hierdie interessante fenotipe verder te verklaar.Algeheel lewer hierdie studie ‘n breedvoeriggekarakteriseerde bevolking trangeniese plante, wat dien as nuttige hulpbronne vir in vivoanalise van PGIP se funksie in siekteweerstandbiedendheid, veral waar die gasheerplant ‘n inheemse kopie van die PGIP-koderende geen huisves.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The global wine industry has shifted to a more adopted ‘consumer-preference’ production. Modern wine consumers are more knowledgeable and cultivated in their understanding of wine quality, value and style. The quality of red wines mainly depends on grape composition, the wine making process and the ability of tasters to recognise sensory attributes. The harvest date/stage has an influence on the grape composition, thus making the decision on when to harvest an important factor in the production of quality wines or different wine styles. The traditional indicators used in the wine industry to determine time of harvest are more related to the perception of taste and mouthfeel and give little indication of the style of wine in terms of aromatic profile. A new physiological indicator using berry sugar accumulation for the purpose of sequential harvest is proposed to assist the winemaker in producing wines with possible different sensory profiles. This indicator can be used in association with the classical indicators of ripening to affect the diversity of wine styles from a single vineyard or a group of vineyards. The wines could thus have different potential aroma profiles, depending on when the grapes were harvested. The main aim of this study was to assess the effect of performing sequential harvest using a physiological indicator on red wine’s sensory composition. This was done to study the possible relation between harvest time (e.g. fruit composition evolution) and the wine styles/sensory attributes across the different harvest times, thereby possibly increasing the diversity of wine styles. A theoretical berry sugar loading concept was compiled and displays a phase of rapid sugar loading starting at véraison followed by a plateau phase. Depending on whether grapes were harvested in the beginning, mid or end of the plateau phase of fruit sugar accumulation the wines could have different potential aroma profiles. Three main stages: fresh fruit (FF), neutral (N) or pre ripe and mature fruit (MF) has been previously proposed using the sugar loading concept and in terms of harvesting dates. Cabernet Sauvignon and Merlot grapes form Eikendal Vineyards, Stellenbosch were used to make wines according to sequential harvest. Four harvest stages were considered, pre fresh fruit (Pre FF), fresh fruit (FF), mature fruit (MF) and over ripe (OR). The wines were tasted and analysed using two different sensory techniques. In both Merlot and Cabernet Sauvignon wines, the PreFF and OR stages could be more easily discriminated than the two harvest stages in-between, FF and MF. The results suggested that the wines made from the FF and MF stages could not be distinguished from each other in general when the attribute citation frequency method or sorting tasks were performed. However, a trend could be observed for both Cabernet Sauvignon and Merlot wines in terms of aroma attributes with attributes changing from green to ripe fruit during ripening using expert tasters. Relevant research should be engaged to refine sequential harvest in order to obtain more diverse wine styles from a single site or a group of vineyards. / AFRIKAANSE OPSOMMING: Die wêreldwye wynbedryf het ’n verskuiwing ondergaan na ’n verbruikersvoorkeurbenadering in produksie. Wynverbruikers is deesdae beter ingelig en meer ontwikkeld ten opsigte van hulle kennis van wyngehalte, wynstyl, asook die waarde van wyn. Die gehalte van rooiwyn hang hoofsaaklik af van die druifsamestelling, die wynmaakproses en die vermoë van proewers om sensoriese eienskappe te herken. Aangesien die oesdatum/-fase ’n invloed het op druifsamestelling, is die besluit oor wanneer daar geoes moet word ’n belangrike faktor in die vervaardiging van gehaltewyne of verskillende wynstyle. Die tradisionele aanwysers wat in die wynbedryf gebruik word om oestyd te bepaal, hou verband met die waarneming van smaak en mondgevoel en gee weinig aanduiding van die wynstyl op grond van die aromatiese profiel. ’n Nuwe fisiologiese aanwyser wat gebruik maak van suikerakkumulasie in die druiwekorrel in opeenvolgende oeste, het ten doel om die wynmaker te help om wyne met verskeie moontlike sensoriese profiele te vervaardig. Hierdie aanwyser kan saam met die klassieke aanwysers van rypwording gebruik word om ’n verskeidenheid wynstyle uit een wingerd of wingerdgroep te vervaardig. Die wyne kan dus potensieel oor verskillende aromatiese profiele beskik, afhangend van wanneer die druiwe geoes is. Die hoofdoel van die studie was om die invloed van opeenvolgende oes te toets deur ’n fisiologiese aanwyser op rooiwyn se sensoriese samestelling toe te pas. Dit word gedoen deur die moontlike verhouding tussen die oestyd (bv. ontwikkeling van vrugsamestelling) en die wynstyle of wyn se sensoriese kenmerke op verskillende oestye te bestudeer ten einde die verskeidenheid wynstyle potensieel te vermeerder. ’n Teoretiese konsep van druifsuikeropname is saamgestel wat dui op ’n fase van vinnige suikeropname wat by véraison begin, gevolg deur ’n plato-fase. Wyn kan oor verskillende moontlike aromatiese profiele beskik, afhangend daarvan of die druiwe aan die begin, middel of einde van die plato-fase van suikeropname geoes is. Drie hooffases is al voorheen voorgestel deur gebruik te maak van die konsep van suikeropname volgens oesdatum, te wete vars vrugte (VV) (“fresh fruit”, FF), neutraal (N) (“neutral”, N) of voor ryp (“pre ripe”), en ryp vrugte (RF) (“mature fruit”, MF). Cabernet Sauvignon- en Merlot-druiwe van Eikendal, Stellenbosch, se wingerde is gebruik om wyn volgens opeenvolgende oes te maak. Vier oesfases is oorweeg, te wete voor vars vrugte (VVV) (“pre fresh fruit”, Pre FF), vars vrugte (VV) (“fresh fruit”, FF), ryp vrugte (RV) (“mature fruit”, MF), en oorryp (OR) (“over ripe”, OR). Die wyn is geproe en geanaliseer deur gebruik te maak van twee verskille sensoriese tegnieke. In die geval van beide die Merlot- en Cabernet Sauvignon-wyn kon die VVV- en OR-fases makliker onderskei word as die twee tussenin-fases, VV en RV. Resultate dui daarop dat wyn wat van die VV- en RV-fases gemaak is, oor die algemeen nie van mekaar onderskei kan word wanneer die frekwensie van kenmerkaanhaling-metode en sorteringstaak uitgevoer word nie. ’n Tendens kon egter waargeneem word vir Cabernet Sauvignon- én Merlot-wyn ten opsigte van aromatiese kenmerke, deurdat kenmerke gedurende rypwording van groen na ryp vrugte verander het indien ekspertproewers gebruik is. Verdere navorsing moet gedoen word om opeenvolgende oes te verfyn ten einde ’n wyer verskeidenheid wynstyle van ’n enkele area of wingerdgroep te verkry.

The vine mealybug, Planococcus ficus (Signoret) (Homoptera: Pseudococcidae) is a pest with significant economic impact on the grape growing industry in South Africa and other parts of the world. With the isolation and synthesizing of the vine mealybug sex pheromone in 2001, new control options for the integrated management of the vine mealybug have been created. The status of sex pheromone monitoring as a tool in the integrated management of the vine mealybug has been evaluated from different perspectives. A significant quantitative difference in male vine mealybug trap catch numbers has been observed between wine and table grape vineyards and results indicated that there were differences in the susceptibility of grape cultivars to vine mealybug. Currently, the delta trap design is the accepted trap design for vine mealybug monitoring. No studies have yet been conducted to determine the optimum trap parameters like size or design. Population pressure may have an influence on the qualitative efficiency of various trap designs. The basis for degree-day forecasting models has been established adequately. However, refinements need to be done and the incorporation of factors such as humidity and regionality also need to be considered. Daily maximum temperatures fluctuating around the upper developmental threshold temperature for prolonged periods of time seemed to suppress population numbers. Different vineyard management practices exist for wine and table grape production. While an action threshold of 65 vine mealybug males per trap per two-week period seems an acceptable threshold for table grape production, it may not be appropriate for wine grape (or raisin grape) production. Using sex pheromone traps for population monitoring is a valid technique in the arsenal of management tactics against the vine mealybug. However, refinements and validation of research results must be done further to build credibility into the monitoring system. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2006.

La uva de mesa es la fruta más exportada en Chile. Sobre ella recaen una serie de condiciones que debe cumplir para poder ser exportada, como por ejemplo, que posea un alto calibre de bayas (gran tamaño) y apirenia (ausencia de semillas) en sus frutos. Sin embargo, el desarrollo simultáneo de ambas características es incompatible de forma natural, principalmente por motivos hormonales. Esta memoria se desarrolló en el marco del proyecto Genoma II en uva de mesa, que busca el desarrollo de marcadores genéticos que permitan la identificación temprana de los caracteres antes mencionados. El presente trabajo tuvo como principal objetivo el estudio de la relación entre estos caracteres (apirenia y tamaño de baya) y los perfiles de expresión de los genes de la proteína inhibidora de pectin metilesterasa (PMEI) y de Spindly (SPY). Ambos genes fueron identificados previamente en una búsqueda realizada utilizando el método de mapeo por QTL (Quantitative Trait Locus) y estarían estrechamente ligados con la condición de apirenia estenoespermocárpica y la expresión del carácter tamaño de baya en Vitis vinifera. En este Trabajo de Memoria se diseñaron partidores para aislar por PCR fragmentos de tres secuencias génicas de PMEI (1, 2 y 4) y tres secuencias génicas distintas de SPY (A, B y C) en el genoma de Vitis vinifera, los que se caracterizaron parcialmente en su secuencia nucleotídica y aminoacídica. Se identificó, caracterizó y analizó el patrón de expresión de estos genes, mediante el uso de la técnica de PCR cuantitativo en tiempo real. En este estudio se utilizaron cuatro individuos segregantes con fenotipos contrastantes, en tamaño de baya y presencia/ausencia de semilla, en siete diferentes estadíos de desarrollo. Dichos individuos provienen de la recombinación de dos parentales apirenos estenoespermocárpicos (Ruby Seedless y Thompson Seedless). Adicionalmente, se analizó la posible participación de estos productos protéicos en vías metabólicas, encontrándose que SPYpodría estar participando en la vía de las giberelinas, actuando sobre represores e inhibidores de GA, y PMEI participaría en la vía de la degradación de pectinas inhibiendo el accionar de PME. Al comparar los perfiles de expresión entre individuos con fenotipos contrastantes de baya y semilla, se observó que a medida que las bayas se van desarrollando, ocurren cambios de los niveles de expresión en la mayoría de los genes analizados. Se detectó que nuevos genes identificados en este estudio como PMEI-1 y SPY-C,estarían relacionados con el desarrollo de la baya, mientras que SPY-B estaría asociado a los cambios relacionados con el tamaño de la semilla. En cambio, los dos genes candidatos obtenidos de los estudios por QTL (PMEI-2 y SPY-A), no presentaron cambios en su perfil de expresión que se puedan asociar al fenotipo tamaño de baya o semilla. Este resultado, presenta una inconsistencia con respecto a lo indicado por el método de mapeo por QTL y por ende, se recomienda realizar un nuevo mapeo por QTL con una mayor resolución aumentando el número de marcadores moleculares. Los resultados de esta investigación generaron información genética y biológica de ambos genes, lo que permitirá en el futuro realizar una aproximación hacia el conocimiento de los caracteres en estudio. Sin embargo, se sugiere un nuevo análisis con todas las isoformas génicas encontradas en este estudio, en el que se diferencien tejidos (de baya, de piel y de semilla), y se utilicen a lo menos tres o más segregantes por fenotipo, con el propósito de realizar ensayos con una mayor certeza y precisión a nivel estadístico. Este análisis permitiría obtener datos confiables que relacionen un gen con un fenotipo determinado y no con un individuo en particular.

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La vigne (Vitis vinifera) est sensible à de nombreuses maladies, nécessitant l'utilisation massive de produits phytosanitaires. Des méthodes alternatives doivent être développées. Chez la plante modèle Arabidopsis thaliana, les gènes AtNPR1, AtNDR1 et AtEDS1 interviennent dans la voie de signalisation contrôlée par l'acide salicylique (SA) en réponse aux agents pathogènes biotrophes. Nous avons identifié par une approche gène candidat chez V. vinifera, de potentiels orthologues de ces trois gènes (VvNPR1.1, VvNPR1.2, VvNHL1 et VvEDS1).Nos travaux ont montré que les protéines VvNPR1 fusionnées à la GFP sont localisées dans le noyau. De plus, la surexpression transitoire des VvNPR1 stimule l'expression de gènes de défense en système hétérologue et homologue. La surexpression de VvNPR1.1 chez le mutant npr1 restaure les différents phénotypes du mutant, contrairement à la surexpression de VvNPR1.2. VvNPR1.1 est semble être l'orthologue d'AtNPR1. La surexpression de VvNHL 1 chez le mutant ndr1 ne rétablit pas la résistance à Pseudomonas syringae. Cependant, ces plantes sont plus sensibles à Botrytis cinerea, probablement par l'accentuation d'un processus de mort cellulaire. L'expression de VvNHL 1 est réprimée en réponse à B. cinerea, constituant un mécanisme de défense contre les nécrotrophes. VvNHL1 semble donc promouvoir la mort cellulaire. L'expression de VvEDS1 est stimulée en réponse à des agents pathogènes et à un traitement par le SA. L'étude de sa fonction est en cours par surexpression chez des mutants eds1. La compréhension des voies de signalisation des réponses de défense chez la vigne devrait permettre l'obtention de vignes plus résistantes aux agents pathogènes. / Vitis vinifera is susceptible to many pathogens, thus requiring a massive use of phytochemicals. Alternative methods have to be developed. In the madel plant Arabidopsis thaliana, the AtNPR1, AtNDR1 and AtEDS1 genes are components of the salicylic acid (SA) signalling pathway involved in resistance to biotrophic pathogens. Using a candidate gene approach, we have identified putative orthologs of these genes in the grapevine genome (VvNPR1.1, VvNPR1.2, VvNHL 1 and VvEDS1).Our work shows thal VvNPR1-GFP fusion proteins are localized in the nucleus. Moreover, transient overexpression of VvNPR1 genes induces defence gene expression, in bath heterologous and homologous systems. Overexpression of VvNPR1.1 in npr1 mutants restores the different phenotypes of the mutants, while VvNPR1.2 overexpression does not. These results strongly suggest that VvNPR1.1 is the AtNPR1 ortholog. Overexpression of VvNHL 1 in ndr1 mutants does not restore resistance to Pseudomonas syringae. However, resistance to Botrytis cinerea in these plants is weakened, probably because of stimulation of cell death. Furthermore, VvNHL1 repression following B. cinerea infection in V. vinifera could be considered as a defence mechanism against necrotrophic pathogens and suggests thal VvNHL1 is involved in promotion of cell death. VvEDS1 expression is induced by pathogens infection and SA treatment. VvEDS1 functional characterization is in progress using eds1 Arabidopsis mutants overexpressing VvEDS1. Knowledge of defence signalling pathways in V. vinifera will contribute to obtain pathogen resistant grapevines.

A ocorrência de doenças em uvas de mesa gera perdas desde o cultivo até a comercialização, o que afeta diretamente o potencial de exportação do Brasil. A prevenção na realização de injúrias nos frutos é essencial para garantir a eficácia dos tratamentos aplicados tanto em pré- quanto em pós-colheita. Visando o emprego de produtos alternativos que auxiliem no manejo sustentável da cultura, o fosfito de potássio encontra-se como ferramenta útil. O efeito do fosfito no controle de doenças causadas por oomicetos já foi relatado, porém, como este ingrediente ativo atua sobre microrganismos do reino Fungi ainda é desconhecido. Neste contexto, este trabalho teve como objetivo avaliar o fosfito de potássio no controle das doenças podridão mole, mofo cinzento e podridão da uva madura, causadas por Rhizopus stolonifer, Botrytis cinerea e Colletotrichum gloeosporioides, respectivamente, em bagas de uva ´Itália´ e seus efeitos na qualidade físico-química dos frutos. Também objetivou-se estudar os mecanismos de ação envolvidos na inibição do desenvolvimento do micélio de R. stolonifer na presença do produto e na indução de respostas de defesa em bagas de uva. Foram realizados ensaios in vitro para avaliação do efeito do fosfito sobre o crescimento micelial, esporulação e germinação dos esporos para R. stolonifer, B. cinerea e C. gloeosporioides. Experimentos utilizando-se de bagas foram conduzidos, visando avaliar o efeito no uso do fosfito como fungicida produtor ou curativo para os três patógenos. Cachos foram utilizados para se determinar os efeitos da aspersão do fosfito sobre a qualidade físico-química dos frutos. Para avaliação dos mecanismos de ação do fosfito sobre o micélio de R. stolonifer, foram determinados os efeitos sobre a permeabilidade da membrana plasmática, a síntese de parede celular e a síntese de proteínas. A atividade de guaiacol peroxidase e de compostos fenólicos totais foi determinada em bagas de uva tratadas ou não com fosfito. Houve inibição do crescimento micelial, da esporulação e da germinação dos esporos dos patógenos desenvolvidos em diferentes concentrações de fosfito de potássio. O tratamento protetor e curativo das bagas evidenciou reduções significativas na área abaixo da curva de progresso das doenças estudadas. Não houve efeito do produto sobre a qualidade físico-química dos cachos de uva. Quanto aos mecanismos de ação, foram observados efeitos do fosfito sobre a permeabilidade de membrana, devido à elevada perda de eletrólitos do micélio. O fosfito proporcionou elevação na síntese de proteínas totais no micélio do patógeno e redução na atividade de ?-1,3- glucanase e quitinase, que são enzimas relacionadas à síntese de parede. A atividade de guaiacol peroxidase e a concentração de compostos fenólicos totais não aumentou com a aplicação do fosfito nas bagas. Desta forma, pode-se concluir que o fosfito de potássio controlou a podridão mole, o mofo cinzento e a podridão da uva madura e não teve efeito sobre a qualidade físico-química dos frutos. Ainda, o fosfito atuou sobre a permeabilidade de membrana, síntese de proteínas e de parede celular do micélio de R. stolonifer e não proporcionou indução da atividade de guaiacol peroxidase e compostos fenólicos totais em bagas de uva. / The occurrence of table grape diseases causes losses from the field to the commercialization, which affects the Brazil potential to export. The prevention of injuries in fruits is essential to ensure the pre- and postharvest treatment efficiency. In order to use alternative products to support the crop sustainable management, phosphite may be a useful tool. The phosphite effect on control of diseases caused by oomycetes has been reported, however, how the active ingredient acts on microorganisms from the Fungi kingdom is unknown. Thus, the objective was to evaluate potassium phosphite on the control of soft rot, grey mold and ripe rot, caused by Rhizopus stolonifer, Botrytis cinerea e Colletotrichum gloeosporioides, respectively, in ´Itália´ grape berries and its effect on physico-chemical properties of the fruits. Moreover, the objective was to study the mode of action involved on the inhibition of R. stolonifer grown in the presence of phosphite and on the induction of defense responses in grape berries. In vitro assays were carried out to evaluate the effect of phosphite on mycelial growth, sporulation and spore germination in R. stolonifer, B. cinerea e C. gloeosporioides. Single berries were used in order to evaluate the effect of phosphite as a protective or curative fungicide against the three pathogens. Grape clusters were used to determine the effects of phosphite spraying on physico-chemical properties. To evaluate the mode of action of phosphite on R. stolonifer mycelium the plasma membrane permeability, the protein synthesis and the cell wall synthesis were monitored. The guaiacol peroxidase activity and phenolic compound concentration were evaluated in grape berries treated or not with phosphite. There were inhibition on mycelial growth, sporulation and spore germination on the three pathogens in different potassium phosphite concentrations. The protective and curative treatment in grape berries showed significant reductions on the area under disease progress curve of the studied diseases. There was no effect on physico-chemical properties of the grapes. Regarding the mode of action, the phosphite affected membrane permeability what was seen as high electrolyte leakage from the mycelium. Phosphite caused high protein synthesis in the mycelium and reduction on ?-1,3-glucanase and chitinase activities, which are enzymes related to cell wall synthesis. The guaiacol peroxidase activity and phenolic compound concentration did not increase due the phosphite application. Thus, it may be concluded that potassium phosphite controlled soft rot, grey mold and ripe rot in grape berries and did not affect the physico-chemical properties of the fruits. Besides that, phosphite still acted on membrane permeability, protein synthesis and cell wall synthesis on the R. stolonifer mycelium and did not changes in guaiacol peroxidase activity and phenolic compound concentration in the grape berries.

Xylella fastidiosa é uma bactéria fitopatogênica transmitida por insetos vetores conhecidos como cigarrinhas (Hemiptera: Cicadellidae, Subfamília Cicadellinae). Uma estirpe desta bactéria causa o Mal de Pierce em videira nos EUA e México. No Brasil, esta bactéria ainda não foi detectada colonizando videira, embora esta cultura tenha importância em algumas regiões Nesta pesquisa foi feito um levantamento faunístico de cigarrinhas da família Cicadellinae por meio de armadilhas adesivas amarelas em vinhedos dos Estados do Rio Grande do Sul e Pernambuco, com o objetivo de identificar potenciais vetoras de X. fastidiosa e sua flutuação populacional. Para a obtenção das cigarrinhas foram realizadas coletas com cartões adesivos amarelos em quatro parreirais comerciais de Vitis vinifera para cada Estado. Em cada parreiral foram instalados 20 cartões, distribuídos em 10 pontos espaçados de 40 x 40 m, com duas alturas de amostragem (45 cm do solo e 45 cm acima da lâmina foliar de videira). Os cartões adesivos foram trocados quinzenalmente no período de setembro/2004-setembro/2006 e junho/2005-junho/2007 no Rio Grande do Sul e Pernambuco, respectivamente. Baseando-se em análise faunística, determinaram-se as espécies de cigarrinhas mais abundantes, constantes, freqüentes e dominantes, as quais foram avaliadas quanto à flutuação populacional. No Rio Grande do Sul, 34 espécies de cicadelíneos e 6 de cercopídeos foram encontradas. Porém, a maioria (98,4%) dos 3,893 espécimes coletados foram cicadelídeos, distribuídos nas subfamílias Cicadellinae (n = 2.344; 23 espécies), Gyponinae (n = 1.327; 9 espécies), Deltocephalinae (n = 147; 1 espécie) e Coelidinae (n = 13; 1 espécie). Os insetos da subfamília Cicadellinae foram divididos nas tribos Cicadellini (n = 1.606; 12 espécies) e Proconiini (n = 738; 11 espécies). Dentre os cicadelíneos, 5 espécies de Cicadellini (Bucephalogonia xanthophis, Dilobopterus dispar, Macugonalia cavifrons e a morfo-espécie Cicadellini sp. 1) e 5 de Proconiini (Molomea consolida, Oncometopia facialis, Oncometopia fusca e Tapajosa rubromarginata) prevalecem nos vinhedos do Rio Grande do Sul, de acordo com as análises faunísticas. Nos vinhedos de Pernambuco, verificou-se uma menor diversidade de espécies de cigarrinhas em relação aos do Rio Grande do Sul. Um total de 4.106 cicadelídeos foram coletados, pertencentes a duas subfamílias: Cicadellinae (n = 4.094; 4 espécies) e Gyponinae (n = 12; 2 espécies). A espécie mais abundante foi a H. spottii com 3.965 indivíduos encontrados. Esta espécie utiliza a videira como hospedeiro de oviposição e desenvolvimento. Os períodos de maior ocorrência de cigarrinhas nos vinhedos são de outubro a agosto na Serra Gaúcha, e de janeiro a junho em Pernambuco. Neste estudo também foi testada a possibilidade de transmissão por cigarrinhas de uma estirpe de X. fastidiosa de citros, causadora da Clorose variegada dos citros (CVC), para videira e ameixeira. Não houve transmissão para videira, indicando que a estirpe de X. fastidiosa de citros pode não ser capaz de estabelecer infecção sistêmica em videira, após a inoculação por inseto vetor. No entanto, a diversidade e abundância de cigarrinhas potenciais vetoras nos Estados de Pernambuco e Rio Grande do Sul indicam um grande risco para disseminação do Mal de Pierce em videira caso uma estirpe de X. fastidiosa patogênica a esta cultura seja introduzida ou evolua a partir de estirpes existentes no Brasil. / Xylella fastidiosa is plant-pathogenic bacterium transmitted by leafhoppers (Hemiptera: Cicadellidae) in the subfamily Cicadellinae, commonly known as sharpshooters. In the United States and Mexico, a particular strain of this bacterium causes Pierce\'s disease (PD) in grapevines. PD has not been reported in Brazil, although grape is a major crop in some regions of this country. In this study, a 2-year survey of Cicadellidae leafhoppers was carried out by yellow sticky traps in vineyards of the States of Rio Grande do Sul and Pernambuco, in order to identify potential vectors of X. fastidiosa as well as their seasonal patterns of occurrence in the crop. The survey was conducted in four commercial plantings of Vitis vinifera L. per State, by using 20 traps distributed in 10 sampling points and 2 heights (45 cm above soil and 45 cm above the crop canopy) per vineyard. The cards were changed fortnightly during the periods of September/2004-September/2006 and June/2005-June/2007 in the States of Rio Grande do Sul and Pernambuco, respectively. Faunistic analyses of the trapping data from each vineyard were run to determine the most abundant, constant, frequent and dominant sharpshooter species, for which the population fluctuation was studied. In Rio Grande do Sul, 34 leafhopper and 6 spittlebugs (Hemiptera: Cercopidae) species were trapped, but most (98.4%) of the 3,893 specimens collected were leafhoppers, which were distributed in the subfamilies Cicadellinae (n = 2,344; 23 species), Gyponinae (n = 1,327; 9 species), Deltocephalinae (n = 147; 1 species) and Coelidinae (n = 13; 1 species). The sharpshooter (Cicadellinae) specimens were divided in the tribes Cicadellini (n = 1,606; 12 species) and Proconiini (n = 738; 11 species). Among the sharpshooters, 5 species of Cicadellini (Bucephalogonia xanthophis, Dilobopterus dispar, Macugonalia cavifrons and the morpho-species Cicadellini sp. 1) and 5 of Proconiini (Molomea consolida, Oncometopia facialis, Oncometopia fusca and Tapajosa rubromarginata) are prevalent in vineyards of Rio Grande do Sul based on the faunistic indices. In the vineyards of Pernambuco State, a different species composition and a lower diversity of sharpshooters were found. A total of 4,106 leafhopper specimens were trapped, distributed in two subfamilies: Cicadellinae (n = 4,106; 4 species) and Gyponinae (n = 12; 2 species). H. spottii was the most abundant sharpshooter, with 3,965 specimens. The periods of higher sharpshooter populations in the vineyards are from October to August in Rio Grande do Sul, and from January to June in Pernambuco. The possibility of transmission of a Citrus variegated chlorosis (CVC) strain of X. fastidiosa from citrus to grape was tested by using the sharpshooter B. xanthophis as a vector. No transmission to the test plants was recorded, suggesting that the CVC strain may not establish systemic infections in grape after vector inoculation. However, the diversity and abundance of native sharpshooters found in Rio Grande do Sul and Pernambuco indicate a high risk of PD spread in vineyards if a pathogenic strain of X. fastidiosa to grapes evolves or is introduced in Brazil.

Existe uma crescente demanda por alternativas de controle de doenças para se reduzir o uso do controle químico, como por exemplo, o estudo da indução de resistência em plantas, através de fitotoxinas como a taxtomina A e outras substâncias. O vírus do mosaico do pepino (Cucumber mosaic virus - CMV) e o vírus do mosaico amarelo da abobrinha (Zucchini yellow mosaic virus - ZYMV) têm causado grandes prejuízos em culturas importantes, assim como o fungo Rhizopus stolonifer, que pode atacar diversos hospedeiros. Este trabalho foi conduzido com o objetivo de se avaliar os efeitos do uso da taxtomina A como indutor de resistência em plantas. Como resultado preliminar a produção de taxtomina A pode ser aumentada utilizando-se como fonte de inóculo amostra de uma cultura liquida pré-estabelecida de Streptomyces scabies. Foram testados a aspersão de taxtomina A em plantas de abobrinha de moita, para se avaliar a proteção contras os vírus CMV e ZYMV. Também foi aspergida taxtomina A em bagas de uva \"Itália\" para se avaliar o desenvolvimento da podridão pós-colheita causada por R. stolonifer. As plantas de abobrinha de moita, mantidas em casa de vegetação, tratadas com taxtomina A não apresentaram sintomas de mosaico característico do CMV. O teste ELISA apresentou resultados negativos indicando a não presença de partículas virais em níveis detectáveis nas plantas. Com relação ao ZYMV, a taxtomina A não foi capaz de controlar o desenvolvimento da doença, pois apareceram sintomas em todas as plantas. A taxtomina A apresentou efeito in vitro sobre R. stolonifer retardando o crescimento do fungo principalmente nas duas maiores concentrações usadas. Porém, a aplicação de taxtomina A não foi capaz de proteger as bagas de uva contra o patógeno. Finalizando a taxtomina A foi capaz de controlar em 100% a incidência de CMV em abobrinha, porém não foi capaz de controlar o ZYMV em abobrinha e a podridão nas uvas causada por R. stolonifer. / There is an increasing demand for alternative control of diseases to reduce the use of chemical control, for example the induced resistance studies on plants with phytotoxins like thaxtomin A and other substances. The Cucumber mosaic virus (CMV) and the Zucchini yellow mosaic virus (ZYMV) cause a lot of losses on important crops and also the fungus Rhizopus stolonifer that can cause disease in different hosts. This work was carried out to evaluate the use of thaxtomin A to induce resistance. Thus, as a preliminary result, the thaxtomin A production could be increased by using as inoculum an established Streptomyces scabies liquid culture. It was tested the spraying of thaxtomin A on zucchini plants to evaluate the protection against the viruses CMV e ZYMV. On the other hand, grape berries cv \"Italia\" were also sprayed with thaxtomin A to evaluate the development of the postharvest rot caused by R. stolonifer. The zucchini plants were maintained in the greenhouse and the treatment with thaxtomin A inhibited mosaic symptom development by the CMV. The ELISA test showed negative results indicating that there was no detectable virus particles inside the plants. The thaxtomin A was not able to control the ZYMV development since all plants exhibited symptoms. The thaxtomin A showed in vitro control of R. stolonifer as it reduced the fungal growing at the two highest concentrations used. However, the thaxtomin A was not able to control the postharvest rot in grapes. Finally, the thaxtomin A was able of controlling 100% of the incidence of CMV on zucchini plants, but was not able of controlling ZYMV in zucchini and the grape berrie rot caused by R. stolonifer.