Solano-violeta (Solanum violaefolium) é uma ornamental rasteira usada pra cobrir solos de áreas sombreadas. Um vírus que induz manchas anelares nas folhas, tentativamente designado de mancha anular do S. violaefoliumm (S. violaefolium ringspot vírus - SvRSV), transmitido pelo ácaro Brevipalpus phoenicis foi encontrado nesta planta em jardins de Piracicaba, SP. Trata-se de um vírus baciliforme que se acumula no lúmen do retículo endoplasmático induzindo viroplasma citoplasmático, assemelhando-se a outros vírus do tipo citoplasmático, dos transmitidos por ácaros Brevipalpus (Acari: Tenuipalpidea). Este trabalho relata algumas de suas propriedades biológicas e a caracterização molecular parcial. SvRSV foi ser transmitido mecanicamente e pelo B. phoenicis a várias outras espécies botânicas, sempre causando lesões localizadas. Destas, Datura stramonium mostrou-se a melhor como hospedeira experimental. As propriedades físicas do SvRSV in vitro foram: temperatura de inativação - 40-45 ºC; ponto final de diluição - 10-3-10-4; longevidade in vitro- 12 dias. Posteriormente, observou-se também infestação destas plantas por B. obovatus que em ensaios preliminares transmitiu o SvRSV. Em secções ultrafinas, as partículas do SvRSV mostraram-se ligeiramente mais delgadas que as de outros vírus do tipo citoplasmático, transmitidos por Brevipalpus, e por outro lado formavam eventualmente partículas mais longas, às vezes de ca. 1 µm. Como os demais vírus, do tipo citoplasmático, transmitido por Brevipalpus, induz a formação de um viroplasma denso e vacuolado no citoplasma. Em casos favoráveis foram observadas fases do processo de morfogênese por “brotação” a partir do material do viroplasma. Dada sua labilidade não foi possível conseguir sua purificação apesar das inúmeras tentativas, usando diferentes protocolos. Logrou-se a extração de dsRNA a partir de D. stramonium e a partir dele, obter-se dois fragmentos do genoma viral, identificados como parte da proteína de movimento e da replicase, após seu sequenciamento. Foram produzidos pares de “primer” baseado nestas seqüências que amplificaram especificamente, por RT-PCR, fragmentos de DNA de tamanho esperado, a partir do RNA total extraído de lesões foliares de S. violaefolium e D. stramonium infetados. Sondas baseadas nas seqüências obtidas hibridizaram com ss- e dsRNA de lesões de D. stramonium. Ensaios preliminares de RT-PCR e hibridização não resultaram em reação com alguns outros vírus transmitidos por Brevipalpus, do tipo citoplasmático, inclusive o da leprose dos citros (CiLV-C). / Solanum violaefolium is an ornamental Solanaceae, with prostrate, trailing growth cultivated in shaded areas. Plants exhibiting necrotic ringlike spots on the leaves have been found in several gardens and parks at Piracicaba – SP. The ringspot symptoms on the leaves is caused by a vírus, named S. violaefolium ringspot virus (SvRSV), and is transmisible by mite Brevipalpus phoenicis (Acari: Tenuipalpidae). Short bacilliform particles are present within the cisternae of endoplasmic reticulum (ER) and often electron dense viroplasm is present in the cytoplasm, characterist of the cytoplasmatic type of Brevipalpus-borne virus. The present study reports some of its biological properties and partial molecular caracterization. SvRSV is easily transmitted to many plant species either by viruliferous B. phoenicis or mechanically, always causing local lesions. Datura stramonium was proved to be better as experimental host. Its physical properties in vitro were: inactivation thermal point – 40-45 ºC; final diluition point - 10-3-10-4; longevity in vitro – 12 days. Afterwards, its was observed that S. violaefolium plants were infested by B. obovatus that transmitted SvRSV in preliminary assays. Thin sections revealed that SvRSV particles are slightly thinner and sometimes appear very long. In some favorable sections intermediate steps of viral particle morphogenesis by a budding process of the dense material of the viroplasm toward the lumen of ER could be seen. Due to the fragility of the particles, several attempts to purify the virus have failed, despite many protocols tried. It was possible, however, to extract dsRNA from infected tissue of D. stramonium, and two segments of viral genome, respectively with homology to movement protein (mp) and replicase (rep) of some known viruses were obtained. Primers were designed based in these sequences, which amplified by RT-PCR, fragments of DNA of expected size from total RNA extracts from leaves lesions of infected S. violaefolium and D. stramonium. Probes based on obtained sequences hibridizated with ss- and dsRNA from lesions of S. violaefolium and D. stramonium. Preliminary assays of RT-PCR and hybridization did not result in positive reaction with other cytoplasmatic type of Brevipalpus-borne viruses, including citrus leprosis (CiLVC).

O gênero Streptomyces constitui-se em um grupo diferenciado de procariotos fitopatogênicos, por serem morfologicamente similares aos fungos e produzirem diversos importantes metabólitos secundários. A sarna comum da batata é principal doença causada pelo gênero. O principal agente causal da doença, Streptomyces scabies, produz fitotoxinas denominadas concanamicinas, porém, não foi comprovada a importância da produção das mesmas na agressividade dos isolados de S. scabies. Dessa forma, neste trabalho procurouse avaliar a reação dos cultivares mais plantados no Brasil a isolados de S. scabies exibindo diferentes níveis de produção de concanamicinas. O nível de produção de concanamicinas A e B de dez isolados de Streptomyces spp. foi avaliado por cromatografia líquida de alta pressão (HPLC) e dois isolados (produtor de alto nível da toxina; baixo produtor) foram selecionados e utilizados na avaliação da resistência de seis cultivares de batata em casa de vegetação. Vasos contendo o substrato esterilizado receberam a suspensão de esporos bacterianos junto com os tubérculos sadios no momento do plantio e após quatro meses, a severidade da doença foi avaliada. Os resultados demonstraram aumento na agressividade do isolado produtor de elevadas quantidades de concanamicinas apenas sobre os cultivares Mondial, Monalisa e Asterix. Por sua vez, os resultados da avaliação do nível de resistência ao patógeno demonstraram que os cultivares Monalisa, Ágata, Asterix e Cupido são suscetíveis e os cultivares Atlantic e Mondial mostraram-se resistentes aos dois isolados de Streptomyces spp. avaliados. Por outro lado, a fitotoxina taxtomina A é a toxina mais frequentemente produzida pelas espécies fitopatogênicas de Streptomyces spp. O estudo do mecanismo de ação desta fitotoxina poderá auxiliar no entendimento de sua contribuição no desenvolvimento da doença. A taxtomina A é uma fitotoxina não específica e causa efeitos semelhantes em plantas de diferentes famílias botânicas. Desta forma, modificações citológicas, ultraestruturais e bioquímicas induzidas pela taxtomina A foram analisadas em plântulas de sorgo. Os tecidos foram tratados com taxtomina A e avaliados após sete dias do tratamento. Observou-se desorganização em todas as células dos tecidos tratados com a taxtomina A. A membrana plasmática das células do mesofilo foliar foi retraída e destacada da parede celular em diversos pontos. Entretanto, o plasmalema não pareceu rompido. O efeito sobre a permeabilidade seletiva da membrana plasmática também foi avaliado bioquimicamente, onde foi observado aumentos na saída de eletrólitos. Os cloroplastos apresentaram deformações na superfície e perda de turgidez. Estas deformações provavelmente contribuíram para a redução do teor de clorofilas A e B. As plântulas também apresentaram elevado acúmulo de fitoalexinas do tipo deoxiantocianidinas. Desta forma, conclui-se que a taxtomina A age principalmente sobre a membrana plasmática e organelas membranosas. Em contrapartida, o acúmulo de fitoalexinas evidencia que concentrações mínimas desta toxina podem ser exploradas como eliciador da resposta de defesa em plantas. / The genus Streptomyces is a different prokaryote phytopathogen group, being morphologically similar to fungi and able to produce several important secondary metabolites. The potato common scabies is the main plant disease caused by the members of the genus. The main causal agent of the plant disease, Streptomyces scabies, is able to produce the phytoxins concanamycins. However, the importance of these toxins in the aggressiveness of S. scabies strains was not proven. This research aimed to evaluate the reaction of main potato cultivars used in Brazil to Streptomyces spp. strains, exhibiting different levels of concanamycins production. The toxin production by several strains was quantified by high performance liquid chromatography (HPLC) and two strains (higher and lower producers) were chosen and used for resistance evaluation of six potato cultivars under greenhouse. Pots containing sterile substratum received a suspension of bacterial spores and the healthy seed tubers at the same time. After four months, the level of disease was evaluated and the results showed increasing in aggressiveness for the higher concanamycins producer only in the Mondial, Monalisa and Asterix cultivars. For the two strains, the cultivars Atlantic and Mondial were resistant and the cultivars Monalisa, Cupido, Ágata and Asterix were susceptible. On the other hand, the thaxtomin A is the main phytotoxin of plant pathogenic Streptomyces spp. Therefore, the study of the thaxtomin A action mechanism is important to explain its contribution to disease development. The thaxtomin A is not a specific toxin and causes similar effect in several plants of different botanical families. Thus, the cytological, ultrastructural and biochemical modifications induced by thaxtomin A were evaluated in sorghum seedlings. Leaf tissues of seedlings were treated with high levels of thaxtomin A and evaluated 7 days after treatment. Cytological disorganization was observed in all treated cells. The plasmma membrane of the cells of immature leaves was detached from the wall in several places. However, plasmmalema did not seem degraded. The chloroplasts had showed significant deformations in the surface and volume reduction. Theses changes in chloroplasts probably contributed for the reduction of chlorophylls A and B levels. The sorghum seedlings also showed high phytoalexin accumulation (deoxiantocianydins) in response to thaxtomin A treatment. Thus, the thaxtomin A acts mainly on plasmma membrane and membranous organelles, causing plasmolises, cell disorganization, deformations in chloroplasts and chlorophyll reduction in sorghum seedlings. Finally, the accumulation of phytoalexins in response to thaxtomin A treatment points out it potential to induce resistance in plants.

Made available in DSpace on 2014-08-20T13:59:08Z (GMT). No. of bitstreams: 1 tese_cristina_huther.pdf: 966830 bytes, checksum: 5ef15a4c44099ced96bf589355808571 (MD5) Previous issue date: 2014-03-28 / The objectives of this study were: a. to compare growth and yield components of Micro-Tom tomato plants with wild plants and transformed plants for different levels of MT-sHSP23.6 expression (mitochondrial small heat shock proteins); b. to verify damages caused by UV-C application to photosynthetic activity, in the vegetative and reproductive stages, with the same genotypes previously described; c. to compare the effects of water deficit on photosynthetic activity in Micro-Tom tomato plants with the genotypes reported, in the period of floral induction. The first experiment was carried out in growth chambers, under controlled conditions. The analyses were performed at regular time intervals of 21 days. Collections started on 21st day after the transplant, a total of six collections. Leaf area, dry weight of the plants parts, and the number of green and red fruits were determined. From the results obtained we verified that the genetic transformation of this variety did not interfere significantly with these plants growth. In the second experiment that aimed to evaluate the effect of application of UV-C radiation, two trials were performed; the plants were grown in a green house, under controlled temperature and humidity. In the first trial the application of UV-C radiation was performed only once, when the plants were in vegetative stage. In the second trial the plants were exposed to two doses of UV-C radiation, with an interval of ten days, the application began when the plants were starting the reproductive stage. The application lasted two minutes with a dose of 0,5kJ.m-2. Chlorophyll index, chlorophyll a fluorescence, and gas exchanges were evaluated. At the first trial, the evaluations were performed at 16, 64, 168, and 336 hours after the application of the radiation, and in the second trial at 16, 64, and at 168 hours after first application, with the same interval after the second application of radiation. From the results we observed that the genotypes showed different behaviors in relation to the data of transient fluorescence. However, the net assimilation rate was more affected in the transformed genotypes, with greater intensity in the second trial, especially after the second application. We noticed that in both trials genetic transformation did not contributed significantly to provide better performance under these experimental conditions. In the third trial the plants were grown in a growth chamber. Fortyfour days after sowing, in floral induction period, they were submitted to water deficit for five days. Chlorophyll index, chlorophyll a fluorescence, and gas exchanges were evaluated, and at the end of the experiment we made simultaneous analysis of fluorescence decrease, modulated reflection at 820 nm, proline content in the foliar area, and dry matter of plant parts. We could verify that genotype with over-expression of MT-sHSP23.6 was more efficient than the other genotypes in photochemical reactions, allowing that water deficit data were smaller, however it was not possible to identify the relation of the over-expression of these proteins with the parameters of gas exchange. / Este trabalho teve por objetivo: a) comparar o crescimento e componentes de produção de plantas de tomateiro Micro-Tom , com plantas selvagem e plantas transformadas para diferentes níveis de expressão da MT-sHSP23.6 (mitochondrial small heat shock proteins); b) verificar as injúrias causadas pela aplicação UV-C, na atividade fotossintética, no estádio vegetativo e reprodutivo, com os mesmos genótipos descritos anteriormente; c) comparar os efeitos que o déficit hídrico apresenta sobre a atividade fotossintética em plantas de tomate Micro-Tom , no período de indução floral, nos mesmos genótipos relatados. O primeiro experimento foi conduzido em câmaras de crescimento, sob condições controladas. As análises foram realizadas com intervalos regulares de tempo de 21 dias, iniciando as coletas no 21º dia após o transplante e ao total foram realizadas seis coletas. Foram determinadas: área foliar, massa seca das partes das plantas e o número de frutos verdes e vermelhos. Através dos resultados obtidos verificou-se que a transformação genética desta variedade não interferiu de maneira significativano crescimento destas plantas. No segundo experimento, que apresentou objetivo de avaliar o efeito da aplicação da radiação UV-C, foram realizados dois ensaios, sendo queas plantas foram cultivadas em casa de vegetação, com temperatura e umidade controlada. No primeiro ensaio, a aplicação da radiação UV-C foi realizadaapenas uma só vez quando as plantas estavam no estádio vegetativo. No segundo ensaio as plantas foram submetidas a duas doses de radiação UV-C, com intervalo de 10 dias, iniciando a aplicação quando as plantas estavam em inicio de estádio reprodutivo. Aaplicação durou 2 minutos com uma dose de 0,5 kJ.m-2. Foram avaliados: índice de clorofila, fluorescência da clorofila a e trocas gasosas. No primeiro ensaio as avaliações foram realizadas após 16, 64, 168 e 336 horas após a aplicação da radiação, e no segundo as 16, 64 e 168 horas após a primeira aplicação e com o mesmo intervalo após a segunda aplicação da radiação. Por intermédio dos resultados observou-se os genótipos apresentaram comportamentos distintos em relação os dados da fluorescência transiente. Contudo a taxa assimilatória líquida foi mais afetada nos genótipos transformados e com maior intensidade no segundo ensaio, principalmente após a segunda aplicação da radiação UV-C. Dessa forma verificou-se que em ambos os ensaios, a transformação genética não contribuiu de maneira significativa para proporcionar melhor desempenho nessas condições experimentais. No terceiro experimento as plantas foram cultivadas em câmara de crescimento. Quando estas estavam com 44 dias após a semeadura, no período de indução floral, as mesmas foram submetidas ao déficit hídrico por cinco dias. Foram avaliados: índice de clorofila, fluorescência da clorofila a e trocas gasosas e ao final do experimento foram realizadas análises simultâneas da decaída da fluorescência, reflexão modulada a820nm, teor de prolina nas folhas área foliar e massa seca das partes das plantas. Pode-se verificar que o genótipo com superexpressão da MT-sHSP23.6 foi mais eficiente que os demais genótipos nas reações fotoquímicas, permitindo que os dados do déficit hídrico fossem menores, contudo não foi possível evidenciar relação da superexpressão dessa proteína com os parâmetros das trocas gasosas.

Abstract The plant can be considered a superorganism that consists of the plant per se and numerous populations of pro- and eukaryotic microorganisms. The interactions between the plant and endophytic microorganisms colonizing plant internal tissues are typically commensalistic or mutualistic. However, information on the role of endophytes in plant defense is limited because pathways are only partly known and systemic responses are typically not seen. The aim of this thesis was to study the priming capacities of endophytic Methylobacterium sp. IMBG290 on potato (Solanum tuberosum L.). Priming of plants by non-pathogenic bacteria allows the host to save energy and to reduce time needed for development of defense reaction during a pathogen attack. Priming phenomenon was demonstrated for Methylobacterium sp. IMBG290 as an activation of salicylic acid and jasmonate/ethylene-dependent defense pathways after challenge inoculation with the pathogen. Moderate activation of plant antioxidant system may also contribute to resistance induction by the strain. The viable but nonculturable state is presumably a survival strategy observed for the majority of bacterial endophytes. Pathogen attack or environmental changes can activate these quiescent forms. Thus Methylobacterium+ sp. IMBG290 became cultivable upon plant inoculation by nonpathogenic bacteria. I observed that the composition of the endophyte community changed in response to Methylobacterium sp. IMBG290 inoculation in shoot tissues and correlated with potato disease resistance and growth promotion. Therefore, the activation of endophytic bacterial populations as a putative mechanism of plant disease resistance was proposed. Endophytes have a high agricultural potential. Growth- and resistance-promoting capacities of Methylobacterium sp. IMBG290 on potato were highly variable depending on the cultivar, pathogen, inoculum density, and environmental conditions. Context-dependent efficacy requires more attention when designing complex microbial inoculants capable influencing positively plant growth, resistance, and nutritional properties. / Tiivistelmä Kasvia voidaan pitää superorganismina, joka koostuu kasvista itsestään ja lukuisista pro-ja eukaryottisista mikrobipopulaatioista. Kasvin ja sen sisäosia asuttavien endofyyttisten mikro-organismien väliset vuorovaikutukset ovat yleensä kommensalistisia tai mutualistisia. Endofyyttien rooli kasvin puolustuksessa on kuitenkin huonosti tunnettu, koska reitit tiedetään vain osittain eikä järjestelmällisiä vasteita usein havaita. Tämän väitöskirjatyön tavoitteena oli tutkia endofyyttisen Methylobacterium sp. IMBG290-kannan kykyä vahvistaa perunan (Solanum tuberosum L.) puolustusta. Tautia aiheuttamattomat bakteerit kykenevät vahvistamaan kasvien puolustusta, mikä auttaa isäntäkasvia säästämään energiaa ja nopeuttamaan puolustusreaktiota patogeenihyökkäyksen aikana. Methylobacterium sp. IMBG290-kannan vahvistuskyvyn osoitettiin perustuvan salisyylihappo- ja jasmonaatti/etyleeni-riippuvaisten puolustusreittien aktivoimiseen patogeeni-istutuksen jälkeen. Antioksidanttijärjestelmän lievä aktivoituminen voi myös vaikuttaa kannan aiheuttamaan vastustuskyvyn lisääntymiseen. Suurimmalle osalle bakteeriendofyyteistä ‘elävä mutta viljelemätön’-olotila on luultavasti selviytymisstrategia. Patogeenihyökkäys tai muutokset ympäristössä voivat aktivoida tällaiset hiljaiset olomuodot. Methylobacterium sp. IMBG290-kanta muuttui viljeltävissä olevaan muotoon kun kasviin istutettiin tautia aiheuttamaton bakteeri. Selvitin, että endofyytti-yhteisön koostumus muuttuu vasteena Methylobacterium sp. IMBG290-kannan istuttamiseen kasvin verson solukoissa, korreloiden lisääntyneen perunan vastustuskyvyn ja kasvun kanssa. Siksi endofyyttisten bakteeripopulaatioiden aktivoitumista esitettiin uutena kasvin puolustusmekanismina. Endofyyteillä on suuret mahdollisuudet maataloudessa. Methylobacterium sp. IMBG290-kannan kasvua ja vastustuskykyä lisäävät ominaisuudet perunalla vaihtelivat lajikkeen, patogeenin, lisätyn bakteeriympin ja ympäristöolosuhteiden mukaan. Suunniteltaessa monimutkaisia bakteeriymppejä kasvien kasvin, vastustuskyvyn ja ravintosisällön lisäämiseksi, täytyy tällainen tilanteesta riippuva tehokkuus ottaa enemmän huomioon.

Abstract Seed potato is the starting point in the potato (Solanum tuberosum L.) production chain. In order to secure potato production in a variety of production conditions, plant diseases must be controlled and the yield characteristics of the used cultivars ensured. In addition, production must be cost-effective. Characteristics particular to northern production conditions include long periods of daylight and a short growing season as well as a several months long seed potato storage period. The focus of the present study is on northern production conditions and methods, including haulm killing and sprout control, which are presumed to affect seed potato quality, as well as the initial stages of the seed potato production chain, i.e. micro- and minituber production, which could influence cost-effectiveness and propagation. Haulm killing is one of the methods used in seed potato production to regulate tuber size. It is often carried out on unsenesced plants. The present results, however, indicate that cultivar properties have a greater effect on the sprouting and crop yield of seed potatoes than production-phase haulm killing or temperature sum accumulation. Nevertheless, haulm killing carried out three weeks after flowering (75 DAP) accelerated emergence. When the effect of haulm killing methods on seed potatoes was compared with natural haulm senescence, haulm killing was shown to increase disease pressure. Black scurf (Rhizoctonia Solani) was present in seed tubers whose haulm had been destroyed by mechanical or mechanical-chemical haulm killing. Naturally senesced haulm had less black scurf, and crop quantity and starch content developed to a level typical of the cultivar. Storage periods lasting several months make controlling seed potato sprouting more challenging. Therefore, use of the plant hormone gibberellic acid (GA) in sprout control was investigated. GA treatments at lower concentration (100 mM) increased the number of tubers in the cultivar Fambo. Thus, the timing of haulm killing and in the case of Fambo, GA treatment influenced the characteristics of seed potatoes. Conventionally, the first tuber generation is produced using microplants to produce minitubers in greenhouses. This production method is, however, labour-intensive, and energy and investment costs are high. With the aim of increasing production efficiency in northern production conditions, the production of minitubers in the laboratory using a novel bioreactor technology and in growth rooms using the hydroponic technique was investigated. The Liquid LabTM Rocker bioreactor system was used in vitro, all the cultivars examined (Asterix, Timo, Van Gogh and Velox) produced microtubers. The quantity of tubers produced per dish varied between 30 (for the cultivar Asterix in eight weeks) and 75 (for the cultivar Velox in 11 weeks). The results showed hydroponic production of minitubers to be successful in indoor conditions: the cultivars Desiree and Van Gogh developed their first tuber three weeks faster than Asterix, and the minituber yield was 4.5 per plant for Desiree, 7.5 for Van Gogh and 4.0 for Asterix. When the results indicate that both the Liquid LabTM Rocker production method and the hydroponic production method are suitable for mass production of seed potatoes. / Tiivistelmä Siemenperuna on lähtökohta perunan (Solanum tuberosum L.) tuotantoketjussa. Jotta perunantuotanto turvataan eri tuotanto-olosuhteissa, on hallittava siemenperunan kasvitaudit, taattava sadontuotto-ominaisuudet ja tuotannon on oltava lisäksi kustannustehokasta. Pohjoisissa tuotanto-olosuhteissa erityispiirteinä ovat valoisuudeltaan pitkät päivät ja kestoltaan lyhyet kasvukaudet. Lisäksi siemenperunoiden varastointijakso kestää useita kuukausia. Tässä tutkimuksessa selvitettiin vaikuttavatko pohjoiset tuotanto-olosuhteet tai tuotannossa käytetyt varsistonhävitys tai itämisen hallinta siemenperunoiden laatuominaisuuksiin. Lisäksi selvitettiin siemenperunoiden ensimmäisen mukulasukupolven (mikro- ja minimukula) tuotantoa eri tekniikoilla. Siemenperunatuotannossa varsistonhävitystä käytetään mukulakoon säätelykeinona. Varsistonhävitys tehdään usein tuleentumattomaan kasvustoon. Saatujen tulosten perusteella lajikeominaisuudet vaikuttivat itämiseen ja sadontuotto-ominaisuuksiin enemmän kuin varsistonhävitys tai mukuloihin kerääntynyt lämpösumma. Varsistonhävitys kolme viikkoa kukinnasta (75 päivää istutuksen jälkeen) nopeutti kuitenkin siemenperunoiden taimettumista. Vertailtaessa varsistonhävitysmenetelmien vaikutusta siemenperunaan verranteena kasvuston luontainen tuleentuminen, varsistonhävitys lisäsi kasvitautipainetta. Mekaanis-kemiallisesti ja mekaanisesti varsistonhävityissä satomukuloissa tuli esiin seittirupea (Rhizoctonia solani). Luontaisesti tuleentuneen kasvuston sadoissa oli seittirupea vähemmän ja myös sadon määrä ja tärkkelyspitoisuus kehittyivät lajikkeelle luontaiselle tasolle. Useiden kuukausien varastointijakso vaikeuttaa siemenperunoiden itämisen hallintaa. Kun tutkittiin gibberelliinihapon (GA) käyttöä itämisen hallintaan, alhaisemman konsentraation (100mM) GA –käsittely lisäsi Fambo –lajikkeen mukulalukumäärää. Tulosten perusteella varsistonhävityksen ajoittamisella ja GA –käsittelyllä (Fambo –lajike) vaikutettiin siemenperunoiden ominaisuuksiin. Siemenperunan ensimmäisen mukulasukupolven eli minimukuloiden tuottaminen tapahtuu perinteisesti kasvihuonekasvatuksena mikrokasveista. Tuotantotapa on työvoima-, energia- ja invetointikustannuksia vaativaa. Tuotannon tehostamiseksi pohjoisissa tuotanto-olosuhteissa tutkittiin mikro- ja minimukuloiden tuotantoa eri teknologioilla. Mikromukuloita tuotettiin bioreaktorimenetelmällä laboratoriossa ja minimukuloita hydroponisella menetelmällä kasvatushuoneissa. Tulokset osoittavat, että kaikki tutkitut lajikkeet (Asterix, Timo, Van Gogh ja Velox) tuottivat bioreaktorissa mikromukuloita. Mikromukuloiden määrä vaihteli 30:sta (Asterix, 8 viikon kasvatus) 75:een (Velox, 11 viikon kasvatus). Myös minimukuloiden hydroponinen tuotanto sisätiloissa on mahdollista; kaikki lajikkeet muodostivat mukuloita, Desiree ja Van Gogh 3 viikkoa nopeammin kuin Asterix. Desiree tuotti minimukuloita 4.5 kpl/kasvi, Van Gogh 7.5 ja Asterix 4.0. Tulokset osoittavat, että molemmat menetelmät (bioreaktori ja hydroponinen) soveltuvat mikro- ja minimukuloiden massatuotantoon.

Five techniques were used to predict the potential biogeography of the four alien plant species, Acacia longifolia, Acacia mearnsii, Opuntia ficus-indica and Solanum sisymbrifolium. Prediction was based on five environmental factors, median annual rainfall, co-efficient of variation for rainfall, mean monthly maximum temperature for January, mean monthly minimum temperature for July and elevation. A geographical information system was used to manage the data and produce the predictive maps. The models were constructed with presence and absence data and then validated by means of an independent data set and chisquared tests. Of the five models used, three (the range, principal components analysis and discriminant function analysis) were linear while the other two (artificial neural networks and fuzzy logic) were non-linear. The two non-linear techniques were chosen as a plant's response to its environment is commonly assumed to be non-linear. However, these two techniques did not offer significant advantages over the linear methods. The principal components analysis was particularly useful in ascertaining the variables that were important in determining the distribution of each species. Artifacts on the predictive maps were also proved useful for this purpose. The techniques that produced the most statistically accurate validation results were the artificial neural networks (77% correct median prediction rate) and the discriminant function analysis (71% correct median prediction rate) while the techniques that performed the worst were the range and the fuzzy classification. The artificial neural network, discriminant function analysis and principal component analysis techniques all show great potential as predictive distribution models.

High temperature limit successful potato cultivation in the lowlands of tropical regions. One effect of high temperature may be an increase in gibberellin activity that is inhibitory to tuberization. Paclobutrazol blocks gibberellin biosynthesis and reduces its level in the plant. The effect of paclobutrazol on potato was examined under non-inductive conditions in a greenhouse and under field conditions in the hot tropical lowlands of eastern Ethiopia. Paclobutrazol was applied as a foliar spray or soil drench at rates equivalent to 0, 2, 3, and 4 kg a. i. per ha. Paclobutrazol increased chlorophyll a and b content, and photosynthetic efficiency, enhanced early tuber initiation, delayed physiological maturity, and increased tuber fresh mass, dry matter content, specific gravity and crude protein content. It reduced the number of tubers per plant and extended the tuber dormancy period. Paclobutrazol reduced shoot growth, and plant height, and increased the partitioning of assimilates to the tubers while reducing assimilate supply to the leaves, stems, roots and stolons. Stomatal conductance and the rate of transpiration were reduced. In addition, paclobutrazol treatment increased tuber N, Ca and Fe content while reducing P, K and Mg content. Growth analyses indicated that paclobutrazol decreased leaf area index, crop growth rate, and total biomass production. It increased specific leaf weight, tuber growth rate, net assimilation rate, and partitioning coefficient (harvest index). Microscopic observations showed that leaves of treated plants developed thicker epicuticular wax layers. The epidermal, palisade and spongy mesophyll cells were larger. It increased the thickness of the cortex and the size of vascular bundles and pith cells of the stem. It also increased the width of the cortex and favoured the formation of more secondary xylem vessels, resulting in thicker roots. Deposition of starch grains in the stem pith cells, and cortical cells of the stem and root, were stimulated in response to paclobutrazol treatment. In most instances the method of application did not affect the efficiency of paclobutrazol. The effect of cultivar and reproductive growth on growth, photosynthetic efficiency, water relations, dry matter production, tuber yield and quality of potato was also the subject of investigation. Non-flowering, flowering and fruiting plants of cultivars Al-624, Al-436, CIP-388453-3(A) and CIP-388453-3(B) were evaluated under field conditions of a sub-humid tropical highland of eastern Ethiopia. Cultivars exhibited differences with respect to leaf stomatal conductance, rate of transpiration, net photosynthesis, biomass production and allocation, tuber yield, tuber size distribution, specific gravity, dry matter content and nutrient composition. Fruiting plants had higher leaf stomatal conductance, and higher rates of transpiration and photosynthesis rates. The leaf area index, tuber growth rate, and partitioning coefficient (harvest index) of the fruiting plants were reduced, but crop growth rates and net assimilation rates were higher. Without affecting total dry matter production, fruit development reduced the amount partitioned to the leaves, stems, roots, and tubers. Fruit development reduced total and marketable tuber mass and tuber numbers. The effect of MCPA and paclobutrazol were studied under greenhouse and field conditions. Single foliar sprays were applied during the early and full bud development stages at rates of 0, 250, 500, and 750 g a.i. ha-1. Both MCPA and paclobutrazol greatly reduced the number of flowers and completely inhibited berry set. MCPA did not affect the number, yield, dry matter content and specific gravity of tubers. Without affecting the number of tubers, paclobutrazol increased tuber yield, dry matter content and specific gravity. / Thesis (DPhil (Horticultural Science))--University of Pretoria, 2007. / Plant Production and Soil Science / unrestricted

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-01-12T10:57:12Z No. of bitstreams: 1 texto completo.pdf: 1369974 bytes, checksum: 9fd8ea4315b0b8b8c2670673f4f5c850 (MD5) / Made available in DSpace on 2017-01-12T10:57:12Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1369974 bytes, checksum: 9fd8ea4315b0b8b8c2670673f4f5c850 (MD5) Previous issue date: 2004-07-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / As espécies Sclerolobium paniculatum Vogel var. subvelutinum Benth. e Solanum lycocarpum A. St.-Hil. são típicas do Cerrado sensu stricto que sobressaem em áreas degradadas por desmatamentos e pelo fogo, enquanto a espécie Cecropia pachystachya Trécul se destaca em Matas de Galeria perturbadas pela formação de clareiras; ambas as espécies apresentam comportamento pioneiro. Este trabalho visou verificar o pioneirismo em espécies de Cerrado sensu stricto e de Matas de Galeria do Bioma Cerrado, através da análise da germinação e do crescimento inicial dessas espécies, assim como foram avaliados os bancos de sementes dos solos de áreas do Cerrado e de Matas de Galeria, com e sem perturbações. O estudo do comportamento germinativo das sementes revelou que, uma vez aplicados os tratamentos para superação da dormência, todas as espécies apresentam elevado percentual de germinação, independentemente das condições de luz. A escarificação mecânica e o corte do tegumento aceleram a germinação de Sclerolobium paniculatum var. subvelutinum e Solanum lycocarpum, enquanto Cecropia pachystachya apresentou dormência fisiológica superada por ácido giberélico e nitrato de potássio. As três espécies apresentam comportamento similar quanto à germinação em diferentes condições de luz, uma vez superada a barreira da dormência. O desenvolvimento das variáveis alométricas e a produção de biomassa indicaram a necessidade de elevados percentuais de luz para o desenvolvimento destas, confirmando seu caráter pioneiro. O estudo do banco de sementes da Cascalheira, do Cerrado sensu stricto com e sem perturbação, das Matas do Capetinga e do Gama, com e sem perturbação, mostrou que,quanto maior o grau de perturbação, maior o estoque do banco de sementes formado por diásporos com características de gramíneas, em detrimento de espécies arbóreas. Constatou-se a existência de um gradiente no Cerrado sensu stricto no qual a relação sementes viáveis/sementes totais do banco foi: Cerrado não-perturbado > Cerrado perturbado por fogo e desmatamento > Cascalheira. Também no caso da Mata de Galeria do Capetinga houve uma nítida distinção entre área perturbada (com clereira) e não (ou menos) perturbada (interior da mata), sendo: Mata do Capetinga não perturbada > Mata do Capetinga perturbada. Contudo, na Mata de Galeria do Gama, por se tratar de uma mata mais conservada do que a do Capetinga, a relação sementes viáveis/sementes totais do banco foi semelhante para ambas as condições (borda e interior da mata), sendo possível a recuperação das áreas degradadas através do banco de sementes. / Sclerolobium paniculatum Vogel var. subvelutinum Benth. and Solanum lycocarpum A. St.-Hil. are typical species of the Cerrado sensu stricto that occur mostly in degraded lands by deforestations and fire, while Cecropia pachystachya Trécul occurs mostly in Gallery forest gaps. The objective of this work was to detect the pioneer behavior of these three species, through the analysis of germination and initial growth. Moreover, seed banks in the Cerrado sensu stricto and Gallery forest areas, with and without disturbances, were evaluated. The study of the seeds germinative behavior showed that once applied adequate treatments to overcome dormancy, all the species presented high germination levels, independent of light conditions. Mechanic scarification and seed tegument cutting accelerated germination of Sclerolobium paniculatum var. subvelutinum and Solanum lycocarpum, while physiological dormancy of Cecropia pachystachya was surpassed by giberelic acid and potassium nitrate. The three species showed a similar behavior related to germination at various light conditions, since the dormancy barrier was surpassed. The pioneering character was confirmed since alometric variables and biomass production was associated with high light percentages. The seed banks studies showed that a higher number of grass diaspora was found in the more degraded areas, replacing the forest trees species seed bank. These results suggested a gradient in relation to viable seeds/total seeds in the seed bank of the Cerrado sensu stricto: not disturbed Cerrado > Cerrado disturbed by fire and deforestation > Gravel pit. In the Capetinga gallery forest, that is a forest disturbed by fire, there was a clear distinction between a degraded area (large gap) and a less disturbed one. However, in the Gama gallery forest, that is a less disturbed forest, the relation viable seeds/total seeds was similar between disturbed and undisturbed areas (interior of the forest), allowing to state that the recovery of disturbed areas, in this forest, could occur naturally through the seed bank. / Não foi localizado o cpf do autor.

Orientador: Leandro Borges Lemos / Banca: Dirceu Maximino Fernandes / Banca: Domingos Fornasieri Filho / Banca: Paulo Cesar Tavares de Melo / Banca: Newton do Prado Granja / Resumo: A cultura da batata tem grande importância econômica para o Brasil e o Estado de São Paulo, sendo Vargem Grande do Sul e Itapetininga os principais pólos produtores paulistas. Nestes, além da cultura da batata, a do feijoeiro destaca-se por sua importância econômica e social. Geralmente os produtores utilizam o feijoeiro em sucessão à batata e objetivando obter alta produtividade de grãos, realizam a adubação de semeadura, desprezando a possível influência residual da adubação da batata. O trabalho objetivou avaliar a influência da adubação de plantio na cultura da batata, da fertilização residual da batata e de semeadura sobre o feijoeiro em sucessão. Para isso foram avaliados por meio de sua contribuição nas características agronômicas e nutricionais das plantas, das alterações nos atributos químicos do solo, bem como a combinação recomendada da adubação de plantio e de semeadura para a batata e o feijão quanto ao retorno econômico. Foram conduzidos dois experimentos, sendo o primeiro em Itapetininga (SP) num Nitossolo Vermelho Distroférrico típico, textura argilosa e o segundo em São Manuel (SP) num Latossolo Vermelho-Amarelo Distrófico típico, textura média. Em ambos foi utilizado o delineamento experimental de blocos casualizados em esquema de parcelas subdivididas, com quatro repetições. As parcelas foram constituídas das adubações de plantio da batata (0, 1.000, 2.000, 3.000 e 4.000 kg ha-1 de 8-28-16) e as subparcelas pela adubação de semeadura do feijoeiro (0 e 330 kg ha-1 de 8-28-16 em Itapetininga e 0, 150, 300 e 450 kg ha-1 de 8-28-16 em São Manuel). Foram utilizadas as cultivares de batata e de feijão Agata e Pérola, respectivamente. Para o solo de textura argilosa a produtividade total e comercial de tubérculos aumentou até a dose próxima de 3.000 kg ha-1 de 8-28-16. A produtividade...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Potato crop has great economical importance for Brazil and São Paulo State. In São Paulo State, Vargem Grande do Sul and Itapetininga are the main producing areas, where beans crop also plays a major economical and social role. Usually, growers plant beans crop after potato crop, aiming high grain yield. They make sowing fertilization and ignore the possible residual influence of potato fertilization. Therefore, the purpose of this research was to evaluate the planting fertilization influence on the potato crop, the residual potato fertilization influence on beans crop in sequence to potato crop, through its contribution in the plant nutrition and agronomic characteristics, and changes in electrical conductivity and chemical soil attributes. This study also tries to find the best economic combination of sowing and planting fertilization recommended for potato and beans crop. Two experiments were carried out, the first one being in Itapetininga (SP) in a typical "Nitossolo Vermelho Distroférrico", clay texture, and the second one in São Manuel (SP) in a typical "Latossolo Vermelho-Amarelo Distrófico", sandy loam texture. The experimental design was in randomized blocks with split plot analyses, with four replicates. The plot treatments consisted of potato fertilization (0, 1,000, 2,000, 3,000 and 4,000 kg ha-1 of 8-28-16) and the split plot of beans sowing fertilization (0 and 330 kg ha-1 of 8-28-16 in Itapetininga, and 0, 150, 300 and 450 kg ha-1 of 8-28-16 in São Manuel). Agata and Pérola varieties for potato and beans, respectively, were used in the experiments. For the clay texture the total and commercial tuber yields increased until near 3,000 kg ha-1 of 8-28-16. No grain yield response to potato fertilization residual was observed. The beans crop fertilization reduced the grain yield. The use of 8-28-16 fertilization increased the P, K and Ca and CE contents in the soil...(Complete abstract click electronic access below) / Doutor

The subject of my thesis was to genetically modify a potato for increased resistance against its pathogens and pests. In developing a resistant plant, it is quite common to use the same type of molecules that plants use themselves in their defense reactions. In this work I used the gene SPI-2 originating from a honeycomb moth (Galleria mellonella). The protein SPI-2 is a member of serine protease inhibitors. Since the previous attempts of the team to detect the protein in transformed plants haven't been successful, the basic form of the gene was modified by adding a Kozak sequence near the start codon, which should have increased the translation initiation and hence increase the level of the protein. Two constructs were prepared for the transformation: SPI-2-T a SPI-2-Y. They differ by one amino acid, which slightly changes their inhibitory activity. First, the construct SPI-2-T was used for a transient transformation of tobacco Nicotiana benthamiana by agroinfiltration of its leaves. Then both constructs were used for a stable transformation of Solanum tuberosum cv. Desireé. The detection of the protein has not been successful, although the inserted gene was transcribed and his sequence was verified by sequencing. It is therefore most likely that the protein has a low stability in the cytoplasm....