Made available in DSpace on 2014-06-11T19:28:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-04-07Bitstream added on 2014-06-13T20:18:28Z : No. of bitstreams: 1 000754808.pdf: 1123345 bytes, checksum: 945c6c9f352406de0091406c4f189c94 (MD5) / A mandioquinha-salsa (Arracacia xanthorrhiza Bancroft) é uma hortaliça originária da região andina, Venezuela, Colômbia, Equador, Peru e Bolívia. A família das apiáceas compreende também outras hortaliças importantes, como cenoura, salsa, coentro, aipo, dentre outras. A mandioquinha-salsa é propagada de forma vegetativa através dos propágulos, e isso pode favorecer a ocorrência de um número considerável de patógenos que causam degenerescência, principalmente os vírus. Está comprovado que alguns vírus são fator limitante em culturas de propagação vegetativa. Portanto, o presente trabalho teve como objetivos avaliar por meio de métodos sorológico e molecular 241 amostras oriundas de diferentes localidades, a fim de fazer um estudo de ocorrência e predominância das espécies virais e verificar a variabilidade biológica dos isolados encontrados por meio de transmissão por extratos vegetais. A detecção foi realizada através de Enzyme linked immunossorbent assay (ELISA) utilizando antissoro comercial anti-poty para o gênero Potyvirus. Para a análise da variabilidade, foram desenhados 2 oligonucleotídeos para amplificar a região codificadora da proteína capsidial para o gênero Cheravirus, e para a detecção de potyvirus e nepovirus, oligonucleotídeos foram obtidos da literatura. No estudo foi possível detectar a espécie Arracacha motlle virus do gênero Potyvirus, coletadas nos estados de Goiás, Minas Gerais, Brasília e Paraná. Os demais vírus testados não foram identificados em nenhuma das amostras analisadas. Além desses testes de identificação, ainda foi realizada microscopia eletrônica de transmissão em seis amostras, das quais duas foi possível a identificação de potyvirus. O teste de gama de hospedeiras foi realizado e mostrou que o AMoV tem gama de hospedeiras bastante restrita, sendo possível infectar somente três de nove espécies testadas... / Arracacha (Arracacia xanthorrhiza Bancroft) is a vegetable crop of the Andean region, formed by Venezuela, Colombia, Ecuador, Peru and Bolivia. The Apiacea family also have other important vegetable crops such as carrots, parsley, cilantro, celery, among others. Arracacha is propagated vegetatively and this can favor the occurrence of a considerable number of pathogens that cause degeneration, especially viruses. Some viruses are a limiting factor in vegetatively propagated crops. Therefore, the present study aimed to evaluate by serological and molecular methods 241 samples from different localities in order to verify the occurrence and predominance of the viruses species and verify the biological variability by sap inoculation. The detection was performed using enzyme-linked immunossorbent assay (ELISA) technique, using commercial antiserum against-potyvirus. For variability analysis, primers were designed to amplify the coding region of the coat protein for the 4 genus Cheravirus, and oligonucleotides were obtained from literature for the detection of the genus potyvirus and nepovirus, . It was possible to detect Arracacha mottle virus, from genus Potyvirus, in the states of Goiás, Minas Gerais, Brasilia and Paraná. The others viruses tested were not identified in the collected samples. Six samples were analysed by electron microscopy, and the potyvirus were identified only in two samples. The host range was restricted, infecting only three species of nine tested. The nucleotide identity ranged from 96% to 99%, between collected samples and sequence from GenBank (acess DQ925486), showing low genetic variability among them

Made available in DSpace on 2015-12-10T14:24:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-08-07. Added 1 bitstream(s) on 2015-12-10T14:30:35Z : No. of bitstreams: 1 000851956.pdf: 1446006 bytes, checksum: d0fd0cae1f3cd364c6ad457c27e8870b (MD5) / A cultura do pimentão (Capsicum annuum L.) tem elevada importância econômica e social para o Equador. Vários fatores têm contribuído para a baixa produtividade dessa cultura no país e dentre eles destacam-se as doenças de diversas etiologias. Poucas informações sobre a ocorrência de doenças nessa cultura estão disponíveis nas para as condições equatorianas. Neste trabalho foi realizado levantamento da ocorrência da doença pata seca e de viroses na cultura do pimentão, em plantios comerciais, durante anos de 2013 e 2014; a identificação dos agente(s) causal(ais) da pata seca; avaliações do eficácia da pulverização da aplicação de produtos fitossanitários (sulfato de cobre + fosetil alumínio; iprodione + clorotalonil; Trichoderma harzianum + T. koingii) em condições de campo, no controle da doença nos híbridos Nathalie e Quetzal e a sensibilidade in vitro do(s) patógeno(s) dessa doença a fungicidas (tebuconazole, benomil, clorotalonil, fosetil alumínio, iprodione, metalaxil + propanocarb e tebuconazole). A pata seca foi constatada em um 79,2% das propriedades, com incidência entre 5% a 53,6%, em quanto as viroses, foram detectada em 62,5% das propriedades, com incidência entre 9,6% a 61,2%. Os fungos S. clerotium rolfsii e Fusarium spp. foram isolados com maior freqüência das plantas com sintomas de pata seca. Entretanto, S.rolfsii foi mais virulento, sendo considerado o principal agente causal da doença pata seca. Não foi observada diferença dos produtos fitossanitários, aplicados no colo das plantas, na incidência de pata seca, nos dois ensaios realizados em campo. A aplicação de iprodione + clorotalonil propiciou uma maior quantidade de frutos comerciais nos dois ensaios e uma maior produção em um dos ensaios. Os Isolados de S. rolfsii apresentaram maior sensibilidade aos fungicidas tebuconazole, benomil e iprodione, ... / Sweet pepper (Capsicum annuum L.) has high economic and social importance for Ecuador. Several factors have contributed to the low productivity of this crop in the country and among them there are the diseases of different etiologies. Little information on the occurrence of diseases in this culture is available to the Ecuadorian conditions. This study was carried out survey of the occurrence of the disease pata seca and viruses in sweet pepper in commercial plantations, during the years 2013 and 2014; the identification of the agent (s) causal (es) of pata seca; evaluations of the effectiveness of spray application of pesticides (copper sulfate + fosetyl aluminum, iprodione + chlorothalonil; Trichoderma harzianum + T. koingii) in field conditions in controlling the disease in hybrid Nathalie and Quetzal and in vitro sensitivity ( s) pathogen (s) of the disease to fungicides (tebuconazole, benomyl, chlorothalonil, fosetyl aluminum, iprodione, metalaxyl + propanocarb and tebuconazole). The pata seca was found in a 79.2% of the properties, with an incidence of 5% to 53.6%, as in the viruses were detected in 62.5% of the properties, with an incidence of 9.6% to 61.2%. The fungus Sclerotium rolfsii and Fusarium spp. were isolated more frequently plants with symptoms of pata seca. However, S. rolfsii was more virulent, being considered the main causative agent of the disease pata seca. There was no difference of pesticides applied in the base of the plants, the incidence of pata seca in the two trial carried out in the field. The application of iprodione + chlorothalonil provided a larger amount of fruits in the two ...

Made available in DSpace on 2014-06-11T19:35:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-11-18Bitstream added on 2014-06-13T19:24:10Z : No. of bitstreams: 1 000754099.pdf: 1729116 bytes, checksum: 2b7d02470d50c8efac136fc81e002d7e (MD5) / Espécies de vírus dos gêneros Potyvirus, Carlavirus e Allexivirus são comumente encontradas na cultura do alho (Allium sativum L.) e em decorrência da sua propagação vegetativa, há o acúmulo de vírus de um ciclo para outro, formando um complexo entre espécies dos gêneros citados. Uma das principais causas da redução da produtividade ocorre devido à infecção por vírus, e a termoterapia associada à cultura de tecido pode ser um método eficiente para limpeza clonal e obtenção de alho semente livre de vírus. O estudo da degenerescência do alho livre de vírus é, portanto, importante para se conhecer o número de ciclos em que o alho semente, inicialmente sadio, pode ser multiplicado no campo, sem que ocorra a redução da produtividade. Neste estudo foi verificado que após três anos a produtividade foi 13,75% superior ao alho 100% infectado por vírus. Os bulbilhos aéreos também podem ser utilizados para a multiplicação de sementes e tem como vantagem o baixo custo quando comparado à utilização de bulbos provenientes da cultura de tecido. Entretanto, essa técnica só se torna viável com a utilização de matrizes isentas de vírus, pois a transmissão de vírus para os bulbilhos aéreos, provenientes de matrizes infectadas, podem chegar a 83,33% no caso dos potyvirus, 20% para carlavirus e 70% para allexivirus, segundo dados obtidos neste trabalho. Realizar uma diagnose precisa dos vírus que infectam o alho torna-se essencial para que não ocorram falhas durante a indexação. Pôde-se observar que para a detecção dos potyvirus, utilizando o teste de ELISA indireto, deve-se utilizar folhas novas, entre 21 e 63 dias após o plantio, pois a concentração viral nesse período é a mais elevada. Diferentes técnicas como o DIBA colorimétrico, DIBA quimioluminescente, ELISA e PCR em tempo real foram avaliadas para detecção do LYSV, aos 21 dias após a inoculação... / Virus species of genera Potyvirus, Carlavirus and Allexivirus are commonly infecting garlic plants (Allium sativum L.). Due to the vegetative propagation, the accumulation of viruses might occur from one cycle to another. The infection of viruses might induce yield losses, and the technique of thermotherapy associated with meristem tip culture is an efficient method to obtain virus-free seeds. The study of degeneration of seeds free of virus is important to know the number of cycles in which garlic may be multiplied in the field without reduction of productivity. In this study it was observed that after three years, the productivity increased 13,75% compared to 100% infected seeds. Currently, aerial bulbils may be used for multiplication of seeds with low cost, compared to meristem tip culture. However, this is a viable technique only if the matrix is free of viruses, because the transmission to aerial bulbils, from infected plants can reach 83.33 % for potyviruses, 20% for carlavirus and 65% for allexivirus, data obtained in this work. An accurate diagnosis in viruses that infect garlic is important to avoid mistakes during indexing material. It was 4 observed that the best time to detect potyvirus, through ELISA test is between 21 and 63 days after planting on young leaves, as the virus concentration is higher in this period. Whem using different techniques, such as the colorimetric DIBA, chemiluminescent DIBA, ELISA, and real-time PCR, it was possible to perform the detection of LYSV 21 days after inoculation. ELISA test detected LYSV only at 21 days after inoculation, whereas using real time PCR, in 5 days after inoculation it was possible to detect the virus due to its high sensitivity. The other tests also detected the virus 21 days after infection

Orientador: Marcos Antonio Machado / Resumo: A morte súbita dos citros (MSC) causou a morte ou erradicação de aproximadamente quatro milhões de plantas de laranja doce na principal região citrícola do Brasil. Embora sua etiologia ainda não esteja completamente resolvida, seus sintomas e distribuição (especial e temporal) indicam uma provável doença viral. Trabalhos anteriores associaram a MSC ao vírus da tristeza dos citros (CTV) e ao CSDaV (Citrus sudden death-associated virus), no entanto, os resultados obtidos destes trabalhos não são conclusivos. Afim de estudar as populações de vírus presentes em plantas de citros afetadas pela MSC, este trabalho realizou uma análise comparativa, através do sequenciamento de alta performance do transcriptoma e dos pequenos RNAs de plantas sintomáticas e assintomáticas para MSC. Os dados revelaram uma infecção viral mista, incluindo o CTV como vírus mais predominante, seguido do CSDaV, um pararetrovírus endógeno de citros (CitPRV) e dois possíveis novos vírus, putativamente denominados de Citrus jingmen-like virus (CJLV) e Citrus virga-like virus (CVLV). As análises de correlação com a MSC indicaram uma provável associação de plantas sintomáticas com o CitPRV, enquanto que os dois novos vírus mostraram estar mais associados com plantas assintomáticas. A associação mais evidente foi observada entre plantas sintomáticas e um genótipo específico de CSDaV, o que nos conduziu a um estudo mais específico de variabilidade genética de 31 isolados de CSDaV, obtidos de plantas MSC-sintomática... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Please read the abstract in the section 00front of this document / Dissertation (Magister Istitutiones Agrariae)--University of Pretoria, 2006. / Plant Production and Soil Science / unrestricted

Bibliography: pages 175-184. / The occurrence of maize dwarf mosaic virus (MDMV) in field grown maize was investigated. For this purpose, maize showing mosiac symptoms was collected from different maize growing areas in South Africa by Prof. M.B. von Wechmar. These samples from Transvaal, Orange Free State and Natal were then investigated for the presence of MDMV and possible strains of this virus. Three virus isolates were purified and partially characterised. These isolates were serologically compared together with a fourth isolate SCMV 4975, obtained from the U.S., to establish strain relationships.

Two isolates of peanut stripe virus (PStV), stripe and blotch, were compared ultrastructurally in peanut (Arachis hypogaea L. 'Florigiant') at several stages of leaf expansion. Ultrathin sections of young leaves infected with either isolate of PStV revealed pinwheel inclusions attached to the cell wall near plasmodesmata. The cytoplasm of infected cells were highly vesiculated. Virus particles amassed in crystalline arrays were observed in blotch infected cells. Virus particles were observed along the arms of pinwheel inclusions. Scroll inclusions appeared in PStV infected cells at a later stage of leaf expansion. In more mature leaves, pinwheel and scroll inclusions occurred in the cytoplasm in association with mitochondria. Virus particles were observed free in the cytoplasm as well as concentrated in linear arrays along the inner surface of the tonoplast. Membrane and organelle degradation was evident in cells infected with either isolate of the virus. Numerous cytoplasmic inclusions and virus particles were observed in cells from light green areas of the leaf. Cells from dark green areas did not contain cytoplasmic inclusions and contained few if any virus particles. Particle measurements show stripe and blotch isolates to have a mean length of 753 nm and 747 nm for leaf dip preparations and 746 nm and 745 nm for partially purified preparations, respectively. Both isolates had a modal length of 750 nm, regardless of the extraction procedure. The relative virus titer of each isolate was determined in peanut leaves at five stages of leaf expansion and in dark green and light green areas of infected leaves. Virus titer increased significantly from the closed to the fully expanded stage, at which time the virus titer peaked and then decreased slightly. Virus titer was consistently higher in leaves infected with the blotch isolate at all expansion stages. Virus titer was also higher in cells from light green areas of the leaf than from dark green areas of the leaf, regardless of isolate. / M.S.

No description available.

Responses from pedigree selection for resistance to gibberella ear rot were assessed in four maize (Zea mays L.) populations, two selected after inoculation of Fusarium graminearum (Schwabe) macroconidia into the silk channel and two selected after inoculation into developing kernels. Responses were significant in both populations selected for silk resistance and in one of the populations selected for kernel resistance. Selection was more effective in later generations and genetic gains were associated with among-family selection but not with within-family selection. Results obtained here indicate that responses to selection could be more efficiently obtained by applying high selection intensities in advanced generations, by managing earlier generations as bulks and by reducing the number of plants per family. In another experiment, a wide sample of Argentine maize germplasm was evaluated for silk and kernel resistance to gibberella ear rot and to fusarium ear rot (caused by F. verticillioides (Saccardo) Nirenberg [=F. moniliforme (Sheldon)]. Several entries exhibited disease resistance in comparison with local check hybrids, particularly for fusarium ear rot, the most prevalent ear rot in Argentina. Results obtained in this study suggested the presence of general mechanisms controlling silk and kernel resistance to both diseases. In a supplementary study, viral diseases were surveyed in maize fields from the provinces of Ontario and Quebec in 1999 and 2000. Barley yellow dwarf was found in 1999. Sugarcane mosaic, maize dwarf mosaic and wheat streak mosaic were found in 2000. These diseases were not important for grain-maize planted in May, the most prevalent kind of maize crop in these provinces. Some of these diseases, such as sugarcane maize mosaic and maize dwarf mosaic were found important only in maize fields planted during or after the month of June, and this is of commercial relevance only for sweet corn.

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: The aim of this study was to combine two common diagnostic tools: serological kits and genetic fingerprinting to identify cherry leaf-roll nepovirus (CLRV), and to establish a marker system to characterize walnut germplasm. The detection of plant viruses is difficult. Restrictions are imposed for quarantine purposes on the importation of plant material from foreign countries. Modern techniques such as a PCR based screening method for CLRV are required to ensure material do not harbour viruses. A primer pair was designed to amplify a 430 bp non-coding homologous region. For the choice of primers, consensus sequences were considered and areas where the sequence data shared 98.5% homology, were chosen. The sensitivity of this detection method was 100-fold higher when compared to the ELISA. The PCR fragment was verified by nucleotide sequencing. AFLP technology was used to identify polymorphic fragments for 6 walnut cultivars and a rootstock, and SCARs were developed from AFLP specific bands. The AFLP technique distinguished all the walnut cultivars and the rootstock. However, conversion of AFLP fragments to SCAR markers for the development of a simple robust technique for cultivar discrimination, was not successful. Using 27 AFLP primer combinations, polymorphic fragments as high as 47.8% were scored. The reason for the lack of efficient conversion was as the result of the AFLP technique. The SCAR primers were generated from sequences internal to the AFLP primers but the specificity of the markers was in the AFLP primers not the internal sequence. In this study using AFLP, walnut cultivars were found to be closely related. The AFLP primer pairs used, provided polymorphic fragments. From these fragments, 7 SCAR markers were developed. It was expected that these SCARs derived from the AFLP markers would detect slight differences between cultivars. The Paradox SCAR marker was the only one that could divide the cultivars into two groups. When Chandler SCAR products were digested with the restriction enzyme Rsal, the same banding pattern as that of Paradox SCAR products was observed. / AFRIKAANSE OPSOMMING: Die doel van hierdie studie was om twee algemene opsporingstegnieke te kombineer: serologiese toetsstelle en genetiese vingerafdrukke om cherry leaf-roll nepovirus (CLRV) te eien en om In merkersisteem te ontwikkel wat okkerneut kiemplasma kan karakteriseer. Die opsporing van plant virusse is baie moeilik. As gevolg van kwarantyn vereistes, word daar beperkinge geplaas word op die invoer van plant materiaal vanuit die buiteland. Moderne tegnieke soos hierdie een wat op PKR berus, word benodig om te verseker dat CLRV nie in plantmateriaal teenwoordig is nie. In Stel inleiers is ontwerp wat In 430 bp nie-koderende homoloë area amplifiseer. Hiervoor is konsensus volgordes bestudeer en slegs die volgordes wat 98,5% homologie getoon het, is gekies. In vergelyking met ELISA was die sensitiwiteit van hierdie deteksie metode 100 maal beter. DNA volgordebepaling is op die resulterende fragment gedoen om die PKR produk te verifieer. AFLP tegnologie is gebruik om polimorfiese fraqmente vir 6 okkerneut kultivars en 'n onderstok te identifiseer en SCARs is uit hierdie fragmente ontwikkel. Die AFLP tegniek kon tussen al die okkerneut kultivars en die onderstok onderskei. Die omskakeling van die AFLP fragmente in SCAR merkers om sodoende In eenvoudige kragtige tegniek vir kultivar onderskeiding te ontwikkel, was egter nie suksesvol nie. Met die gebruik van 27 AFLP inleier kombinasies, kon polimorfiese fragmente van so hoog as 47.8% verkry word. Die rede hoekom omskakeling onsuksesvol was lê by die aard van die AFLP tegniek. Die SCAR inleiers is ontwikkel uit volyordes intern tot die AFLP inleiers, maar die spesifisiteit van die merkers het juis in die AFLP inleiers gelê en nie in die interne volgordes nie. In hierdie studie, met die gebruik van AFLP, is gevind dat okkerneut kultivars baie naby verwant is. Die AFLP inleierstelle wat gebruik is, het polimorfiese fragmente gelewer. Uit hierdie fragmente is 7 SCAR merkers ontwikkel. Daar is verwag dat die SCARs wat uit die AFLP merkers ontwikkel is, klein verskille tussen kultivars sou opspoor. Dit was egter net die Paradox SCAR merker wat die kultivars in twee groepe kon verdeel. Restriksie ensiem vertering met Rsalop die Chandler SCAR produkte het dieselfde bandpatrone as die van die Paradox SCAR produkte gelewer.