Les cytokinines sont des hormones régulant de nombreux processus physiologiques. Elles sont en particulier impliquées dans certaines interactions plantes-microorganismes pathogènes. Leur perception est assurée par les récepteurs Histidine Kinases (HK) dont la fonction est prédominante dans la transduction de signaux moléculaires et environnementaux. Ce travail a permis une caractérisation complète des cinq récepteurs aux cytokinines de type CHASE-Histidine Kinase (MdCHK) et d’un osmosenseur (MdHK1) chez le pommier, espèce d’intérêt économique soumise à l’attaque de nombreux pathogènes. La combinaison d’approches moléculaires et bio-informatiques a révélé les particularités fonctionnelles et les rôles spécifiques des MdCHK. Ces connaissances ont permis de sélectionner une souche de levures exprimant le récepteur MdCHK2 en tant que biosenseur de cytokinines. Ce nouvel outil biotechnologique optimisé pour la détection rapide de cytokinines a conduit à la mise en évidence d’une production de ces composés par Erwinia amylovora, bactérie pathogène du pommier. Le criblage de bactéries pathogènes humaines grâce au biosenseur a également révélé leur production par Staphylococcus aureus et Streptococcus agalactiae. Afin d’optimiser ce biosenseur, une approche mécanistique de MdCHK2 a été entreprise et dévoile l’importance de certains domaines dans ses spécificités de perception. / Cytokinins are hormones regulating numerous physiological processes. They are particularly involved in some plant-pathogen interactions. Their perception is ensured by Histidine Kinase (HK) receptors which play a prevailing role in transducing molecular and environmental signals. This work enabled the full characterization of the five cytokinin CHASE-Histidine Kinase receptors (MdCHK) and the osmosensor (MdHK1) in apple tree, a species of economic interest subjected to many pathogen attacks. Combining molecular and bioinformatics approaches led us to reveal the functional features and specific roles of the MdCHK. This knowledge allowed us to select a yeast expressing the MdCHK2 receptor as a cytokinin biosensor. This new biotechnological tool optimized for fast cytokinin detection led us to expose the production of such molecules in Erwinia amylovora, a pathogenic bacterium of apple. Screening human pathogenic bacteria with the biosensor also unveiled their production in Staphylococcus aureus and Streptococcus agalactiae. In order to optimize this biosensor, we initiated a mechanistic approach on MdCHK2 and showed the importance of some domains in its perception specificities.

Stellenbosch University. Faculty of AgriSciences. Dept. of Horticulture. / Thesis (PhD (Agric))--Stellenbosch University, 2006. / ENGLISH ABSTRACT: Market preferences for larger fruit have forced producers to adopt cultural practices that will ensure bigger fruit even if this comes at the expense of reduced total yields. In order to obtain acceptable fruit size there must be an adequate supply of photosynthetic carbon products especially during the cell division stage of fruit growth. Competition between fruits and between fruit and vegetative growth, as well as adverse climatic conditions, may limit the carbon supply to the fruits at this critical period and thus limit the final fruit size. Growers are showing renewed interest in the use of girdling or scoring in combination with the usual fruit thinning program to achieve growth control and increase fruit size. A new and milder chemical growth retardant, prohexadione-calcium (ProCa), is now also available for vegetative growth control of apple trees. [n this study, the effects of, and potential interaction between scoring, ProCa and fruit thinning were investigated, with respect to shoot and fruit growth, yield and photosynthetic capacity, in 'Royal Gala" 'Fuji' and 'Cripps' Pink' apple trees. ProCa decreased final extension shoot length in all three cultivars. Extension shoots were generally more sensitive than bourse shoots to scoring and ProCa. ProCa seems to be a more effective way of controlling shoot growth than scoring, with 'Fuji' and 'Cripps' Pink' being more sensitive than 'Royal Gala' to the application of ProCa. Scoring led to increased fruit growth rates during the first 40 days after full bloom (DAFB), and culminated in better fruit size at harvest. Scoring improved the total soluble solids concentration (TSS) of 'Fuji' and 'Cripps' Pink' compared to control and ProCa treatments. ProCa inhibited shoot growth effectively, but no evidence was found for improved carbon allocation to fruits and reproductive buds. Yield efficiencies of scored trees were significantly improved in all cultivars during the second season, due to better reproductive bud development after the first year of scoring. On 'Royal Gala' and 'Cripps' Pink', the efficiency of scoring to stimulate reproductive bud development on old and new spurs declined after 4 weeks after full bloom (WAFB). In 'Fuji', scoring later than I WAFB led to a decreasing positive response on old and new spurs. In contrast, reproductive bud development on long shoots increased with later scoring (6 and 8 W AFB) on all cultivars. It seems that the most beneficial time of scoring is 2-4 W AFB, as early as possible during the cell division stage offruit growth, but not before natural drop has occurred. The combination of early-season scoring and application of ProCa seems to hold potential for increasing carbon allocation to the fruit and improving fruit size and quality attributes. Scoring early in the season reduced photosynthetic capacity, and this reduction in carbon availability led to earlier cessation of shoot growth as well as shorter shoots. Later in the season, reduced fruit numbers led to a decrease in photosynthesis. The optimum temperature range for photosynthesis was found to adjust according to seasonal temperature variations. The harvest-induced reduction in sink strength changed stomatal sensitivity to higher temperature. Due to a reduced demand for carbohydrates by the plant, the maximum rate of photosynthesis (Amax) was reduced post-harvest. Following this reduction in sink strength and Am .. , stomata became more sensitive to high leaf temperatures, thus restricting water loss. Pre-harvest there was a strong demand for carbohydrates, therefore stomata were kept open at higher temperatures to ensure a high rate of C02 incorporation, but at a cost with regard to water use efficiency. The sharp increase in dark respiration (Rd) in leaves and fruit with an increase in temperature would mean that significant carbohydrate shortages could occur in trees during source limited periods early in the season, especially under warm weather conditions commonly experienced in the Western Cape region. / AFRIKAANSE OPSOMMING: Die effek van omgewings- en interne faktore op koolstofproduksie en -allokasie in appelbome (Malus domestlca Borkh.). Mark voorkeur vir groter vrugte forseer produsente om tegnieke te gebruik wat groter vrugte sal verseker selfs al beteken dit verlaagde totale opbrengste. Om aanvaarbare vruggrootte te bereik moet daar voldoende voorsiening van fotosinteties geproduseerde koolstofprodukte aan die vrug wees, veral gedurende die selverdelingstadium van vruggroei. Kompetisie tussen vrugte, en tussen vrugte en vegetatiewe groei sal die voorsiening van koolstof gedurende die kritieke stadium aan die vrug beperk en so vruggroote benadeel. Daar is dus hernude belangstelling in die gebruik van ringelering in kombinasie met die gewone vrug uitdunprogramme om groei te beheer en vruggrootte te verbeter. 'n Nuwe, sagter chemiese groeireguleerder, proheksadioon-kalsium (ProCa) is ook nou beskikbaar vir groeibeheer op appelbome. In hierdie studie is die effekte van, en moontlike interaksie tussen ringelering, ProCa en vruguitdunning ondersoek ten opsigte van loot- en vruggroei en opbrengs, sowel as die effekte op gaswisseling op 'Royal Gala', 'Fuji' en 'Cripps' Pink' appelbome. ProCa veroorsaak korter lote in aJ drie kultivars. Verlengingslote is meer sensitief as beurslote vir ringelering en ProCa. ProCa is effektiewer as ringelering om lootgroei te beheer, en 'Fuji' en 'Cripps' Pink' is sensitiewer as 'Royal Gala'. Ringelering lei tot verbeterde vruggroeitempos gedurende die eerste 40 dae na volblom en verbeterde vruggrootte by oes. Ringelering verbeter die totale oplosbare vastestotkonsentrasie (TOVS) van 'Fuji' en 'Cripps' Pink' in vergelyking met kontrole- en ProCa-behandelings, onderskeideHk. ProCa inhibeer lootgroei effektief, maar geen bewyse van verbeterde koolstomllokasie na vrugte en reproduktiewe knoppe is gevind nie. Opbrengseffektiwiteit van geringeleerde borne verhoog gedurende die tweede seisoen van ringelering weens beter reproduktiewe knopontwikkeling. Ringelering later as 4 weke na volblom (WNVB) is minder effektief om reproduktiewe knopontwikkeling op ou en nuwe spore in 'Royal Gala' en 'Cripps' Pink' te stimuleer. Die positiewe effek van ringelering op reproduktiewe knoppe op 'Fuji' verminder sodra dit later as I WNVB gedoen word. Op langlote het 'n laat ringelering (6 en 8 WNVB) die beste effek op reproduktiewe knopontwikkeling. Die mees voordelige tyd om te ringeIeer was 2-4 WNVB, so vroeg as moontlik gedurende die selverdelingstadium van vruggroei, maar nie voor natuurlike vrugval voltooi is nie. Die kombinasie van vroei! ringelering en die toediening van ProCa het potensiaal om koolstofallokasie na die vrug te verbeter, met die gepaardgaande verbetering in vruggrootte en -kwaliteit. RingeJering vroeg in die seisoen inhibeer fotosintese en hierdie verlaging in koolstof beskikbaarheid het tot gevolg dat lootgroei vroei!r gestaak word. Later in die seisoen veroorsaak verlaagde vruggetalle 'n afname in fotosintese. Die optimum temperatuur vir fotosintese verander na gelang van heersende lugtemperature. Die oes-geinduseerde verlaging in sinksterkte verander stomatale sensitiwiteit vir hoo temperature. Weens 'n verJaagde aanvraag vir koolhidrate deur die plant word fotosintese verlaag na-oes. Weens die verlaging is stomata sensitiewer vir hoer temperature en beperk dus waterverlies. V oor oes is daar 'n hoo aanvraag na koolhidrate dus word stomata oop gehou selfs by hoe temperature ten koste van watergebruiksdoeltreffendheid. Die skerp styging in donker respirasie in blare en vrugte met 'n verhoging in temperature sal beteken dat koolhidraat-tekorte kan ontstaan in bome gedurende die bron-beperkte periodes vroeg in die seisoen, veral onder warm toestande 500S wat algemeen ervaar word in die Wes Kaap.

The effects of AVG, an inhibitor of ethylene synthesis, in combination with MCP or heat treatment (HT) on quality traits of several apple cultivars after harvest (AH) or cold storage (ACS), and the involvement of ethylene in the regulation of SDH activity during the last weeks of fruit development were studied. AVG was applied to Royal Gala, Lodi, Senshu, Redchief Delicious and Red Fuji trees 4 weeks before normal harvest (H1). Control and AVG-treated (AVG) fruit were harvested at H1 and treated with MCP or air-heated. Fruit were ripened at room temperature (RT) AH or ACS. Some AVG fruit were harvested at H1 and 1 to 2 weeks after H1 (H2), or at H2 only. Ethylene production (EP), respiration rate (RR), firmness, starch index (SI), titratable acidity (TA), volatile production (VP) and AAT activity, among others, were measured AH and ACS. Peel and cortex of Gala were alcohol-fed and ester production quantified. EP and SDH presence and activity were measured at various harvest dates on control and AVG Lodi, Red Delicious and Fuji apples. AVG plus MCP was more effective in reducing HEP, RR, firmness and TA loss than either treatment alone; it did not provide further control on SI and did not repress Gala red skin color development more than AVG alone, though it consistently repressed VP. AVG plus HT was generally more effective than single treatments in reducing HEP, RR and firmness loss during storage. It was not different than the single treatments on TA, SI, and VP. The effect of AVG plus HT on fruit quality ACS was cultivar-dependent. AVG plus HT was not enough to maintain the quality of the earlyharvest cultivars, and it did not improve fruit quality of late-harvest cultivars. The effects of AVG plus MCP but not of AVG plus HT were evident at H2. Precursor availability was the major factor limiting VP, suggested by the low VP when RR was low, the increase in ester production in alcohol-fed samples, and the lack of correlation between AAT and ester production. SDH activity or expression was not affected by a reduction in ethylene production.

Memoria para optar al Título Profesional de Ingeniero Agrónomo / Se evaluó el control de Eriosoma lanigerum Hausmann infestando manzanos (Malus domestica Borkh) var. Granny Smith plantados en maceta y mantenidos en sombreadero, con aplicaciones repetidas (1 a 3) de un detergente agrícola (TS2035) en concentraciones bajas (0,5 y 1,0 % v/v), en período estival. El diseño experimental fue completamente al azar, con tres repeticiones (=1 maceta/repetición) y 8 tratamientos, incluyendo Lorsban 75 WG (estándar recomendado contra E. lanigerum; se aplicó sólo una vez) y agua pura (testigo). Se hizo una evaluación pre-, y una evaluación post-aspersiones (0,7 y 14 días post aplicación) de los individuos vivos y muertos, estimando el porcentaje de mortalidad bajo lupa estereoscópica (20x). También se evaluó el porcentaje de hojas con síntomas de fitotoxicidad. Los resultados de mortalidad se corrigieron (respecto de la inicial), se transformaron a grados Bliss y sometieron a ANDEVA factorial, seguidos del test de Tukey (p<0,05). Se encontró que la mortalidad inicial fue baja y relativamente homogénea, sin signos de fitotoxicidad, antes de las aplicaciones. Post aplicaciones, tanto el número de aplicaciones como la concentración del detergente, fueron significativos en la mortalidad final, mientras que sólo la concentración fue significativa en la ocurrencia de fitotoxicidad en el follaje, sin interacción entre los factores para ninguna de las variables medidas.

Submitted by Mendes Eduardo (dasilva@ufba.br) on 2013-08-16T20:48:15Z No. of bitstreams: 1 output.pdf: 2244230 bytes, checksum: 22d5a6ac671939479c5bc9d185de2593 (MD5) / Approved for entry into archive by Alda Lima da Silva(sivalda@ufba.br) on 2013-08-22T23:16:20Z (GMT) No. of bitstreams: 1 output.pdf: 2244230 bytes, checksum: 22d5a6ac671939479c5bc9d185de2593 (MD5) / Made available in DSpace on 2013-08-22T23:16:20Z (GMT). No. of bitstreams: 1 output.pdf: 2244230 bytes, checksum: 22d5a6ac671939479c5bc9d185de2593 (MD5) / Capes, Bagisa / Estima-se que um terço dos recursos alimentares do ser humano depende diretamente da polinização por insetos, assim o declínio de polinizadores é uma séria ameaça aos sistemas agrícolas (KEVAN, 2004). KLEIN et al (2007) analisando dados de 200 países, selecionados por uma lista da FAO com culturas que representavam 99% da produção global de alimentos, concluíram que frutas, vegetais e produção de sementes de 87 das culturas mais importantes em termos globais dependem da polinização por animais, enquanto 28 não dependem. Esses autores observaram ainda que a maioria dos cultivos poderia sofrer perdas na produção devido a limitação de polinizadores, e dentre estes 13 dependem essencialmente dos polinizadores, 30 têm alta dependência, 27 apresentam dependência moderada e 21 têm pouca dependência. A perda na produção de frutos e de sementes tem sido claramente relacionada à escassez dos serviços de polinização, principalmente em áreas de agricultura intensiva que são geralmente isoladas de áreas naturais (RICHARDS, 2001). Por isso, o desenvolvimento de programas de manejo sustentáveis dos polinizadores e de conservação dos seus habitat naturais, é de fundamental importância para garantir esses serviços (AGUILAR, 2006). Muitos produtores obtêm os serviços de polinização em seus plantios por intermédio do aluguel de colônias de Apis mellifera (KREMEN & OSFELD, 2005), pois além dessas abelhas serem facilmente transportadas entre as plantações, elas são consideradas polinizadores eficientes de diversas culturas, dentre essas a macieira (KHAN & KHAN, 2004). A macieira (Malus domestica Borkh) é uma espécie que apresenta muitos cultivares com alto grau de incompatibilidade, sendo necessário dois ou mais cultivares que permita uma polinização cruzada eficiente (SOSTER & LATORRE, 2007). O que torna os serviços de polinização ainda mais necessários para esta cultura. Portanto o declínio da diversidade e abundância de insetos polinizadores nessa cultura é um importante fator que afeta a sua produtividade (CUTHBERTSON & BROWN, 2006). A maçã pertence à família Rosaceae e subfamília Pomoideae (SOUZA & LORENZI, 2005). A origem da maçã contemporânea é incerta, contudo evidências indicam que foi originada de terras altas entre o Mar Negro, Turquia e Índia, onde se expandiu para o ocidente estabelecendo variedades que resultaram dos vários cruzamentos feitos entre diversas espécies (HOLA, 1981 apud FREITAS, 1995). Atualmente é uma das frutas que engloba maior quantidade de variedades conhecidas, em que 3 a 4 mil variedades são cultivadas em maior ou menor escala, em diferentes partes do mundo (SILVA et al, 2002). No Brasil, o cultivo da maçã é uma atividade relativamente recente, até o inicio da década de 1970, o País importava maçãs que abasteciam o mercado nacional, porém, nos últimos anos obteve um significativo crescimento com a implantação de pomares comerciais na região do sul do país (PAGANINI et al 2004), tornando-se um importante componente da renda agrícola para o País (BRAGA et al, 2001).Atualmente, a cultura da maçã é uma atividade econômica relevante, com repercussão no cenário internacional, contribuindo com cerca de 1,5% da produção mundial, ou seja, aproximadamente 887 mil toneladas (PEREZ, 2006; www.abpm.org.br). O Brasil é praticamente auto-suficiente na produção de maçã e ainda exporta cerca de 10% da produção para mercados externos exigentes, como a Europa e os Estados Unidos (PAGANINI et al 2004). Segundo BRAGA et al (2001), de acordo com informações do Ministério de Desenvolvimento, Industria e Comércio Exterior, as exportações tem se mantido crescente nos últimos anos, atingindo em 2000, 64,5 mil toneladas, com ingresso de aproximadamente 30,8 milhões de dólares no país. Em 2004 registrou-se o recorde nas exportações, na ordem de 153.043 toneladas (FIORAVANÇO, 2009) e com ingresso de 72 milhões de dólares, representando um aumento expressivo em poucos anos e podendo ser um exemplo real da possibilidade de substituição de importações e da ampliação do mercado interno e da conquista de mercado externo por produto de qualidade e competitividade (PEREZ, 2006). Na região Nordeste, a Bahia foi o primeiro a implantar a cultura da macieira (Figura 02), pela Empresa Bagisa S/A Agropecuária e Comércio no município de Ibicoara, Chapada Diamantina (Figura 03), em 2005, tendo como interesse inicial a produção no mercado regional (www.seagri.ba.gov.br). Tendo em vista a importância da polinização nas culturas agrícolas, o presente estudo se propõe a analisar a eficiência do manejo de colmeias empregadas por produtores de maçã (Malus domestica) na região da Chapada Diamantina, Bahia, mais especificamente será avaliado o efeito da distância das colmeias instaladas na área de estudo sobre a taxa de polinização e de visitação das abelhas. Pretendemos contribuir também com informações a cerca da biologia floral, dinâmica de produção de néctar, sistemas de polinização e visitantes florais dessa cultura em áreas tropicais, tendo em vista que a maioria dos estudos foi realizada em áreas temperadas e sabemos que a biologia e fenofases das variedades desta cultura podem ser influenciadas pelo clima, e ou condições físicas como umidade e temperatura. A partir dessas informações, pretendemos ainda, avaliar o déficit dos “serviços” de polinização para essa cultura na região e o papel dos recursos florais da variedade estudada na atratividade dos polinizadores. / Salvador, Bahia

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2016-04-28T14:37:35Z No. of bitstreams: 1 texto completo.pdf: 399247 bytes, checksum: 2bb5a04ca9827bc635208a00b51d1fe0 (MD5) / Made available in DSpace on 2016-04-28T14:37:35Z (GMT). No. of bitstreams: 1 texto completo.pdf: 399247 bytes, checksum: 2bb5a04ca9827bc635208a00b51d1fe0 (MD5) Previous issue date: 2014-12-16 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / O objetivo desse trabalho foi avaliar o efeito da aplicação do ácido naftaleno acético (ANA) e do inibidor de transporte de auxina ácido 2,3,5-triiodobenzóico (TIBA) na diferenciação e funcionalidade dos vasos do xilema, expressão de genes que codificam para proteínas de transporte de Ca no tonaplasto, atividade da pectinametilesterase (PME) e a ocorrência de “bitter pit” (BP), bem como sua influência sobre os atributos físico-químicos em maçãs ‘Braeburn’. Macieiras ‘Braeburn’ foram pulverizadas com duas concentrações de ANA (10 e 20 mg L-1) e TIBA (10 e 20 mg L-1) e como tratamento controle, água destilada. As aplicações foram realizadas semanalmente, durante o período de 50 a 106 dias após a plena floração (DAPF). Os frutos foram colhidos semanalmente, durante o período de 60 a 106 DAPF, e a cada 15 dias dos 107 aos 164 DAPF (até a colheita comercial), e avaliados quanto a funcionalidade do xilema e composição mineral. Os frutos colhidos aos 164 DAPF foram avaliados quanto à permeabilidade de membrana, nível de expressão de genes que codificam para bombas eletrogênicas e transportadores de Ca e atividade da PME. Seis repetições de 100 frutos foram armazenadas em atmosfera do ar a 1 °C e UR 90-95%, por dois meses, e avaliados quanto a coloração da epiderme, respiração, produção de etileno, pH, firmeza de polpa, sólidos solúveis, acidez titulável, Ca solúvel total e incidência e severidade de BP. A aplicação de ANA 20 mg L-1 proporcionou maior funcionalidade do xilema nos sistemas cortical primário e secundário das regiões proximal e distal dos frutos, quando comparado ao das plantas tratadas com TIBA 20 mg L-1. As concentrações de Ca total, aos 60, 89 e 164 DAPF, e de Ca solúvel total aos 164 DAPF, na polpa da região distal dos frutos, foram maiores no tratamento com ANA (10 e 20 mg L-1), comparadas com os demais tratamentos. Frutos tratados com ANA (10 e 20 mg L-1) tiveram redução de ~50% na atividade da PME, na polpa da região distal dos frutos, em relação ao tratamento com TIBA (10 e 20 mg L-1). Os tratamentos com TIBA (10 e 20 mg L-1) aumentaram a expressão de CAX2 e H+-PPase em comparação aos tratamentos com ANA (10 e 20 mg L-1). Após dois meses de armazenamento, os frutos tratados com ANA (10 e 20 mg L-1) apresentaram menores taxas respiratórias e de produção de etileno e exibiram menor coloração vermelha da epiderme no lado mais exposto a radiação solar na planta, em relação aos frutos tratados com TIBA. Plantas tratadas com ANA 20 mg L-1 apresentaram frutos com maiores valores de acidez titulável e sólidos solúveis do que nos demais tratamentos. Os frutos submetidos a aplicação de auxina, mantiveram níveis mais elevados de Ca solúvel total após o armazenamento, o que contribui para a manutenção da integridade da membrana plásmatica e redução na incidência e severidade de BP. De acordo com os resultados obtidos as auxinas contribuem para a redução da ocorrência de BP e para a manutenção da qualidade dos frutos em maçãs 'Braeburn'. / The aim of this study was to evaluate the effect of naphataleneacetic acid (NAA) and auxin inhibitor transport 2,3,5-triidobenzoic acid (TIBA) on xylem vessel differentiation and function , expression of genes encoding calcium (Ca) transport proteins into the vacuole, pectin methylesterase activity (PME) and bitter pit (BP) incidence, as well as, the influence on physical-chemical attributes in Braeburn apples.‘Braeburn’ trees were sprayed with two concentrations of NAA (10 and 20 mg L -1) and TIBA (10 and 20 mg L-1). The control treatment was sprayed with distilled water. The applications were carried out weekly from 50-106 days after full bloom (DAFB). The fruit were harvested weekly from 60-106 DAFB and each 15 days from 107-164 DAFB (comercial harvest) and were evaluated for xylem functionality and mineral composition. Fruits harvested at 164 DAFB were evaluated for membrane permeability, expression of genes encoding Ca pumps and carriers and PME activity. Six replications of 100 fruits were stored in a cold room at 1°C and RH 90-95% for two months. The fruit were evaluated for skin color, respiration, ethylene production, pH, firmness, total soluble solid, titratable acidity, total soluble Ca, BP incidence and severity. The use of 20 mg L-1 NAA increased the functionality of the xylem of proximal and distal regions as compared to 20 mg L -1 TIBA-treated fruit. Ca content at 60, 89 and 164 DAFB and the total soluble Ca at 164 DAFB in the distal flesh of the fruit, were higher in fruit treated with NAA (10 and 20 mg L-1) than in TIBA-treated fruits and water (control). NAA-treated fruit had 50% lower PME activity in the distal flesh of the fruits, compared to TIBA treatment (10 and 20 mg L-1). The TIBA-treated fruits (10 and 20 mg L-1) showed higher CAX2 and H+- PPase expression than the NAA treatment (10 and 20 mg L -1). After two months of storage, the NAA-treated fruit had lower respiratorion and ethylene production and showed lower red coloration of the sun exposed skin compared to TIBA-treated fruit. Plants treated with ANA 20 mg L-1 showed fruit with higher total soluble solids and titratable acidity. Fruit subjected to application of auxin maintained higher levels of soluble Ca after storage, which contributed to the main to membrane integrity, and reduced BP incidence and severity. According our results auxin reduces BP incidence and maintains 'Braeburn' fruit quality after harvest.

DIPLOMA THESIS Purification of phlorizin from Malus domestica Borkh.by solid-phase extraction and semi-preparative high-performance liquid chromatography Eva Prachařová Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany The aim of this diploma thesis was to find the best conditions for purification of a flavonoid extract from leaves of Malus domestica Borkh., and obtaining the purest fraction of phlorizin. Phlorizin may be used in the treatment of diabetes mellitus type 2 in the future, it has the ability to reduce glycemia by reducing the absorption of glucose in the small intestine and by increasing urinary glucose excretion. The first step was to find an SPE cartridge with a suitable sorbent and a suitable eluent for solid phase extraction. The DPA-6S cartridge and 100% methanol as an eluentwerefound to be the most suitable for SPE. The next step was to find the best possible conditions for semi-preparative HPLC using an ACE 5 C18 column (5 μm, C18, 150 x 10 mm i.d., 150 mm length). The mobile phase consisted of 1% (v/v) acetic acid in water (solvent A) and ethanol 100% (v/v) (solvent B), and a linear gradient elution was used (10-100% B), 0-60 min, the flow: 1mL/min. This method resulted in the 91.05% purity of phlorizin. Keywords: phlorizin, SPE,...

Philosophiae Doctor - PhD / Water availability and climate related issues are some of the greatest crop production risks to irrigated agriculture in arid regions. In South Africa, for example, the increasing frequency and severity of droughts related to climate change and the growing competition for limited water resources among different users threaten the sustainability and growth of irrigated agriculture, especially the water-intensive fruit industry. Major fruit such as apples (Malus domestica Borkh) are produced entirely under irrigation in South Africa. As a result, there has been considerable research to accurately quantify the water requirements of fruit tree orchards in order to maximize water productivity.

A rearing methodology for dogwood borer was developed, using standardized procedures at each developmental stage. These methods enabled the establishment of a laboratory colony and efficient production of synchronized cohorts of each of its lifestages throughout the year for specific experimental needs. The behavioral repertoire shown by mated female dogwood borer in an apple orchard was characterized and quantified and the diel periodicity with which those behaviors occurred was determined. Mated females were easily recognized, based on their characteristic casting flight directed toward areas below the graft union of apple trees, and were observed more frequently during the late afternoon and evening. Casting flight, probing with the ovipositor, and oviposition were the most frequent behaviors observed, but the duration of those behaviors was relatively short compared with the much lengthier periods of resting behavior that typically occurred within the canopy. Data from a previous, three-year study in two newly planted apple orchards were subjected to geostatistical analyses to examine the temporal and spatial patterns of infestation by larval dogwood borer and to gain further information about the spatial scales at which oviposition occurs. There were moderate to high degrees of aggregation of dogwood borer infestations on neighboring apple trees, with ranges of spatial dependence from 7.50–19.87 m. No directionality was observed in the spatial autocorrelation of infestation and it appears that females utilized oviposition sites equally along and across orchard rows. The aggregated nature of infestations requires that random, independent samples must be taken from a number of sample pairs at distances greater than the range of spatial dependence to ensure that sample data are not autocorrelated. Alternatively, an efficient sampling program for mapping dogwood borer infestation can be achieved by limiting sample points to distances within the range of spatial dependence. These sample points can be used in interpolating algorithms, such as kriging, to predict infestation at unsampled locations in space for use in site-specific pest management programs. The external morphology of male and female dogwood borer antennae and their sensilla were examined using light and scanning electron microscopy to characterize, measure and compare the types, number, and distribution of sensilla. Although the general shape and size of male and female antennae were similar, those from females possessed a greater number of generally smaller antennal flagellomeres. The flagellum of both male and female antennae contained seven sensillum types including auricillica, basiconica, chaetica, coeloconica, squamiformia, styloconica, and three subtypes of sensilla trichoidea. With the exception of sensilla basiconica, which were present in roughly equal numbers on male and female antennae, all other sensillum types were significantly more abundant on female antennae. The antennae of female dogwood borer appear well equipped to perceive olfactory stimuli, based on the types and number of sensilla present. Coupled gas chromatography and electroantennogram detection (GC-EAD) analyses of headspace collections from damaged and undamaged tissues from apple and dogwood trees were conducted to examine and compare the antennal responsiveness of female dogwood borer to host plant volatiles. A total of 16 and 9 compounds from apple and dogwood tissues, respectively, consistently elicited an antennal response in females. There were no differences in the response of antennae from virgin and mated females, and the amplitude of the female response to host odors was greater than that of males. Six compounds were identified from the headspace collections from apple trees, four of which (octanal, nonanal, decanal, and methyl salicylate) were identified from all apple tissues sampled. A novel compound,α-bergamotene, was identified from injured apple bark, from apple burr knots infested with dogwood borer larvae and from larval dogwood borer frass, and appears to be produced by apple trees in response to injury. Another novel compound, methyl-2,4-decadienoate, was identified from infested burr knot tissue and appears to be produced in response to an insect-plant interaction. Two compounds, hexanoic and nonanoic acid, were identified from headspace collections from dogwood trees. Numerous approaches were used to examine the behavioral response of mated female dogwood borer to host plant headspace collections and to individual compounds from those collections that elicited a strong and repeatable antennal response. Under both natural and semi-natural conditions in the field and in laboratory bioassays, neither attraction/orientation or consistent oviposition were documented and it is apparent that correlating the electrophysiological and behavioral responses of mated female dogwood borer to olfactory stimuli from their host plants will require further research on bioassay development. / Ph. D.

En raison de leur rôle central dans la physiologie et le développement des plantes, les phythormones ont depuis longtemps été considérées comme des médiateurs chimiques déterminants dans la capacité des insectes à contrôler leur environnement végétal. Les mécanismes permettant aux insectes de manipuler la balance phytohormonale permettant ainsi la régulation des apports nutritifs et la modulation des défenses végétales demeurent néanmoins pour la plupart inconnus, en particulier pour les insectes galligènes et mineurs de feuilles. L’objectif de ma thèse visait à caractériser les capacités de modulation phytohormonale par les insectes manipulateurs de plantes avec un accent particulier sur le lépidoptère mineur de feuille Phyllonorycter blancardella. Nous avons ainsi développés une caractérisation spatio-temporelle de la réponse des plantes aux attaques des larves mineuses au niveau moléculaire et biochimique. Une comparaison entre différents systèmes biologiques nous a permis d’évaluer les similitudes entre les stratégies adoptées par les les insectes galligènes et les insectes mineurs de feuilles, d’identifier l’origine possible des phytohormones impliquées dans la manipulation de la plante et le rôle des bactéries endosymbiotiques d’insectes dans ces interactions. / Because phytohormones lie at the very core of molecular mechanisms controlling the plant physiology and development, they have long been hypothesized to be involved in insect-induced plant manipulations. Insects are using phytohormones to manipulate their host plants for their own benefit, regulating nutrient provisioning and plant defenses. However, a mechanistic understanding of how phytohormones operate in plant reconfigurations by plant-manipulating insects, especially by gall-inducing and leaf-minging insects, is lacking. The objective of my Ph.D. was to provide an extensive characterization of how plant-manipulating insects modulate the plant global hormonal balance with a specific focus on the leaf-mining moth Phyllonorycter blancardella. We thus developed a time course characterization of plant transcriptomic and biochemical responses following attack by leaf-mining larvae. A comparative analysis between different biological systems allowed us to estimate similarities in strategies developed leaf-mining and gall-inducing insects, to identify the possible origin of phytohormones involved in the plant manipulation and to estimate the role of insect endosymbiotic bacteria in these interactions.