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1

Cole, Anthony Blaine Thomas. "Investigations into the hypersensitive response of Nicotiana species to virus infections /." free to MU campus, to others for purchase, 2001. http://wwwlib.umi.com/cr/mo/fullcit?p3012960.

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2

Chen, Pengyin. "Genetics of reactions to soybean mosaic virus in soybean." Diss., Virginia Polytechnic Institute and State University, 1989. http://hdl.handle.net/10919/54781.

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The genetic interactions among 9 soybean [Glycine max (L.) Merr.] cultivars and 6 strains of soybean mosaic virus (SMV) were investigated. The objectives were to identify genes and/or alleles conditioning resistant and necrotic reactions to SMV and to determine the genetic relationships among resistance genes from cultivars exhibiting differential responses to the SMV strains. Seven SMV-resistant (R) cultivars (‘PI 486355’, ‘Suweon 97’, ‘PI 96983’, ‘Ogden’, ‘York’, ‘Marshall’, and ‘Kwanggyo’) were crossed in all combinations among each other and with susceptible (S) cultivars ‘Essex’ and ‘Lee 68’. F₂ populations and F₂-derived F₃ lines were inoculated in field with the SMV type strain Gl and in the greenhouse with the virulent strains G4, G5, G6, G7, and G7A. All F₂ populations from R x S and necrotic (N) x S crosses having PI 96983, Ogden, York, Marshall, and Kwanggyo as either resistant or necrotic parents segregated 3R:1S and 3N:1S, respectively. F₂-derived F₃ progenies from R x S crosses exhibited an F₂ genotypic ratio of 1 homogeneous R : 2 segregating (3R:1S) : l homogeneous S. The results indicate that each of these five resistant parents has a single, dominant or partially dominant gene conditioning the resistant and necrotic reactions to SMV. No segregation for SMV reaction was evident in F₂ and F₃ generations from R x R, N x N, and S x S crosses among the five differential cultivars, indicating that the resistance genes in the five cultivars are alleles at a common locus. The alleles in PI 96983 and Ogden were previously labeled Rsy and rsyt, respectively. Gene symbols, Rsyy, Rsym, and Rsyk are proposed for the resistance genes in York, Marshall, and Kwanggyo, respectively. It is also proposed that the gene symbol rsyt be changed to Rsyt to more accurately reflect its genetic relationship to the susceptible allele. The R x S crosses with PI 486355 and Suweon 97 as resistant parents segregated 15R:1S in the F₂ and 7 (all R) : 4 (3R:1S) : 4 (15R:1S) : 1 (all S) in the F₃, indicating that each has two independent genes for resistance to SMV. The F₂ plants of PI 486355 x Suweon 97 showed no segregation for SMV reaction, suggesting that they have at least one gene in common. The crosses among all 7 resistant parents produced no susceptible segregates when inoculated with strain G1. It is concluded that the 7 resistant cultivars each have a gene or allele at the Rsy locus. Data from the experiments furnished conclusive evidence that the necrotic reaction in segregating populations is highly associated with plants that are heterozygous for the resistance gene.
Ph. D.
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3

Martin, Pierre. "Genetic studies on resistance to alfalfa mosaic virus (AMV) and tolerance to white clover mosaic virus (WCMV) in red clover (Trifolium pratense L.)." Thesis, McGill University, 1989. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=61820.

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4

Al-Kaff, Nadia Saleh Ahmed. "Biological and molecular diversity of cauliflower mosaic virus." Thesis, University of East Anglia, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.240834.

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5

Burger, Johan Theodorus. "The characterisation of Ornithogalum mosaic virus." Doctoral thesis, University of Cape Town, 1991. http://hdl.handle.net/11427/21824.

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Bibliography: pages 155-179.
Ornithogalum mosaic virus (OMV) is the most serious pathogen of commercially grown Ornithogalum and Lachenalia species in South Africa. Although omithogalum mosaic disease was first reported as early as 1940, attempts to purify or characterise the virus(es) were not successful. The extremely mucilaginous nature of omithogalum and lachenalia plant extracts severely hampered virus purification from these hosts. No alternative propagation host for OMV is known: a virus purification protocol for systemically infected ornithogalum and lachenalia was therefore developed. This method eliminated the mucilage in leaf extracts by hemicellulase digestion. Physicochemical characterisation of purified particles suggested that a single virus was present: it had elongated, filamentous particles with a modal length in the range 720- 760 nm; a single major coat protein of Mᵣ30 000, and a single genomic ssRNA of Mᵣ2.90 x 10⁶ daltons. Oligo(dT)-cellulose chromatography confirmed that the genomic RNA was polyadenylated.
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6

Alabi, Olufemi Joseph. "Studies on epidemiology, molecular detection and genetic diversity of selected viruses infecting cassava and wine grapes." Pullman, Wash. : Washington State University, 2009. http://www.dissertations.wsu.edu/Dissertations/Fall2009/o_alabi_110409.pdf.

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7

Gomez, Luengo Rodolfo Gustavo. "Proteins and serological relationships of maize mosaic virus isolates and replication of the virus in Maize (Zea Mays L.) protoplasts /." The Ohio State University, 1987. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487327695621001.

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8

Király, Lóránt. "Interactions between cauliflower mosaic virus isolates and nicotiana species that determine systemic necrosis /." free to MU campus, to others for purchase, 1997. http://wwwlib.umi.com/cr/mo/fullcit?p9841160.

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9

Burbano, Villavicencio Roberto Carlos. "Identificação de genótipos de Saccharum spp. resistentes ao amarelinho (Sugarcane yellow leaf virus) e ao mosaico (Sugarcane mosaic virus) e associação a marcadores moleculares /." Jaboticabal, 2019. http://hdl.handle.net/11449/183546.

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Orientador: Luciana Rossini Pinto
Coorientador: Marcos Cesar Gonçalves
Banca: Dilermando Perecin
Banca: Paula Macedo Nobile
Banca: Antonio de Góes
Banca: Ivan Antônio dos Anjos
Resumo: O vírus do amarelinho (Sugarcane yellow leaf virus, SCYLV) e o vírus do mosaico (Sugarcane mosaic virus, SCMV) são duas importantes viroses que afetam os canaviais dos países produtores de cana-de-açúcar no mundo. As principais características da resistência a essas viroses, as metodologias de avaliação em campo, quantificação viral e as fontes de resistência foram estudadas neste trabalho. Para atingir esse objetivo foi estabelecido um painel com 98 genótipos do gênero Saccharum spp. provenientes do banco ativo de germoplasma do Centro de Cana - IAC (Instituto Agronômico de Campinas). A resposta dos genótipos ao SCYLV e SCMV foi avaliada em campo utilizando uma escala diagramática de notas de sintomas e a concentração viral do SCYLV foi determinada mediante DAS-ELISA e RT-qPCR. Os genótipos do painel susceptíveis ao SCMV foram amostrados e uma análise de sequenciamento da sequência parcial do gene que codifica a capa proteica foi feita para determinar a estirpe predominante no ensaio. Adicionalmente, e com o intuito de identificar marcadores moleculares associados com resistência ao SCYLV e SCMV, foi realizado um estudo de análise de associação entre marcas moleculares e notas de severidade de sintomas. O painel foi genotipado com 955 marcas polimórficas usando AFLP e SSR e submetido a análise de regressão linear simples. Um total de 29 genótipos foram categorizados como resistentes para o SCYLV e 72 para SCMV, considerando que a estirpe predominante causadora dos s... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Sugarcane yellow leaf virus (SCYLV) and Sugarcane mosaic virus (SCMV) are two important viruses affecting the sugarcane producing countries worldwide. The main resistance characteristics of these viruses, symptoms expression phenotyping, virus titer and sources of resistance were studied in this research. To achieve this goal, a panel with 98 genotypes of Saccharum spp. genus was established from the active germplasm bank of the IAC Sugarcane Research Centre (Instituto Agronômico de Campinas). Genotypes responses to SCYLV and SCMV was evaluated in the field using a diagrammatic scale of symptoms and SCYLV virus titer was measured by DAS-ELISA and RT-qPCR. Genotypes with SCMV symptoms were sampled and the partial sequence of the coat protein gene analyzed by sequencing and restriction fragment polymorphism to determine the predominant strain in the plot. In order to identify molecular markers associated to SCYLV and SCMV resistance, an association study between molecular markers and symptoms severity was performed. The panel was genotyped with 955 polymorphic markers using AFLP and SSR and subjected to simple regression analysis. A total of 29 and 72 genotypes were categorized as SCYLV and SCMV resistant, respectively. Our study suggests that the predominant strain causing mosaic symptoms was SCMV-RIB1. The main source of resistance to these viruses probably comes from Saccharum spontaneum accessions and, in smaller proportion, from Saccharum robustum. To SCYLV, the... (Complete abstract click electronic access below)
Doutor
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10

Choi, Chang Won. "Soybean mosaic virus-soybean interactions : molecular, biochemical, physiological, and immunological analysis of resistance responses of soybean to soybean mosaic virus /." Diss., This resource online, 1991. http://scholar.lib.vt.edu/theses/available/etd-07282008-134858/.

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11

Salgueiro, Sancha P. "Molecular studies on pea enation mosaic virus." Thesis, University of East Anglia, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.317586.

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12

Rabelo, Luiz Cláudio. "Seleção de estirpe fraca do Zucchini Yellow Mosaic Vírus (ZYMV) e controle dos mosaicos comum (Papaya ringspot vírus) e amarelo (ZYMV) por dupla premunização em abobrinha-de-moita." Universidade de São Paulo, 2002. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-18102002-170255/.

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No presente trabalho são apresentados os resultados da seleção de estirpes fracas do vírus do mosaico amarelo da abobrinha (Zucchini yellow mosaic virus – ZYMV) e a sua eficiência para o controle do mosaico amarelo em abobrinha-de-moita (Cucurbita pepo cv. Caserta), através da premunização. Mostram também os resultados da eficiência da dupla premunização, com a estirpe fraca do ZYMV e uma estirpe fraca do Papaya ringspot virus – type Watermelon, denominada PRSV-W-1, no controle dos mosaico amarelo e comum e na produção das plantas de abobrinha-de-moita, em condições de campo. A procura de estirpes fracas do ZYMV foi realizada através das seguintes abordagens: a) a partir de lesões locais em plantas indicadoras mantidas em casa-de-vegetação; b) a partir de lesões locais em plantas indicadoras mantidas a 15 °C e c) a partir de lesões locais em plantas indicadoras inoculadas com suspensão parcialmente purificada do vírus, exposta à luz ultravioleta. Apenas uma estirpe fraca (“Mild”) do vírus, denominada ZYMV-M, foi obtida em abobrinha-de-moita diretamente inoculada com suspensão parcialmente purificada do vírus, exposta à luz ultravioleta, durante 30 minutos. Essa estirpe fraca mostrou-se altamente estável, com base na sintomatologia, após 12 transferências sucessivas em plantas de abobrinha-de-moita, durante 15 meses. Plantas de abobrinha-de-moita premunizadas com a estirpe ZYMV-M e superinoculadas (desafiadas) com estirpes severas originárias de Atibaia, SP (ZYMV-AT), de Iacri, SP (ZYMV-IA) e de Vargem Paulista, SP (ZYMV-VP), não exibiram sintomas severos da doença em testes em casa-de-vegetação. A mesma proteção foi observada em plantas duplamente premunizadas com as estirpes ZYMV-M e PRSV-W-1 e desafiadas com uma mistura de estirpes severas desses dois vírus, em condições de casa-de-vegetação. Teste de proteção em campo, com plantas de abobrinha-de-moita premunizadas apenas com a estirpe ZYMV-M, ou duplamente premunizadas com as estirpes ZYMV-M e PRSV-W-1, também mostraram a eficiência dessa tecnologia para o controle dos mosaicos amarelo e comum, com ganhos significativos na produção de frutos comerciais. As plantas premunizadas apenas com a estirpe ZYMV-M, e duplamente premunizadas, tiveram médias de produções de semelhante, com média de 1,85 kg e 1,70 kg de frutos comercias/planta, respectivamente. Os ganhos de produções dessas plantas, em relação às plantas inoculadas com as estirpes severas desses vírus foram de 101 % e 85 %, respectivamente. Diante desses resultados, tornam-se necessários estudos para avaliar a eficiência da dupla premunização para o controle dos mosaicos amarelo e comum, em outras espécies de cucurbitáceas suscetíveis ao ZYMV e PRSV-W.
Due to the present high incidence of Zucchini yellow mosaic virus (ZYMV) and the damage it causes to cucurbit crops, studies were carried out to select mild strains of the virus and evaluate their efficiency for the control of the disease by cross protection in zucchini squash (Cucurbita pepo L. cv. Caserta). Studies were also done to evaluate the efficiency of double cross protection for the control of ZYMV and Papaya ringspot virus – type W (PRSV-W) in zucchini squash under greenhouse and field conditions. Searching for mild strains was carried out as follows: a) from local lesions caused by ZYMV on indicator hosts maintained under green house conditions; b) from local lesions caused on indicator hosts maintained at 15 °C; and c) from local lesions caused on indicator hosts inoculated with suspension of purified ZYMV, exposed to ultra-violet light. Only one very mild strain of the virus, named ZYMV-M, was obtained in zucchini squash plant directly inoculated with suspension of purified ZYMV exposed to ultra-violet light for 30 minutes. This mild strain remained stable for a period of 15 months, after 12 successive transfers in zucchini squash plants. Experiments carried out under greenhouse showed that zucchini squash plants protected with ZYMV-M, at the cotyledonal stage, did not show severe symptoms of the disease after double challenge inoculation with severe strains of the virus, obtained from three regions of the State of São Paulo. Plants inoculated with the mild strain ZYMV-M and a mild strain of PRSV-W, named PRSV-W-1, were also protected against superinfection with severe strains of both viruses. Field test carried out with zucchini squash protected with ZYMV-M and doubly protected with ZYMV-M and PRSV-W-1, showed that this technology was effective for the control of the mosaic diseases caused by severe strains of these viruses. The yield of marketable fruits from plants protected with ZYMV-M, or doubly protected, were 1.85 kg and 1.70 kg of fruits/plant, respectively. These yields were, respectively, 101 % and 85 % higher than the yield of marketable fruits from plants inoculated with a mixture of severe strains of both viruses. Studies are necessary to evaluate the efficiency of double cross protection for the control of ZYMV and PRSV-W in other cucurbit species susceptible to these viruses.
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13

Donahue, Patrick J. "Inheritance of reactions to gray leaf spot and maize dwarf mosaic virus in maize and their associations with physiological traits." Diss., Virginia Polytechnic Institute and State University, 1989. http://hdl.handle.net/10919/54518.

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Gray leaf spot, caused by Cercospora zeae-maydis, can be a yield-limiting factor in maize where continuous minimum tillage practices are followed. Commercial corn hybrids were evaluated for response to gray leaf spot for seven years at two Virginia locations (Shenandoah and Wythe Counties) and one year at a third location in Virginia (Montgomery County). Yield losses, when comparing resistant to susceptible classes, were approximately 2,000 kg ha⁻¹ at Wythe County in 1982, 750 kg ha⁻¹ at Shenandoah County in 1984, and 2,150 kg ha⁻¹ at Montgomery County in 1988. The inheritance of reaction to gray leaf spot was studied using a 14 inbred diallel in Montgomery and Wythe Counties, Virginia in 1987 and 1988 planted in randomized complete block designs. Resistance was found to be highly heritable and controlled by additive gene action. Inbreds producing high yielding, resistant, and agronomically superior hybrids were identified (B68, NC250, Pa875, Va14, Va17, and Va85); and several hybrids between these lines had high levels of resistance, high yield, and good general agronomic characters (B68 x KB1250, KB1250 x Pa875, and NC250 x Pa875). Currently available inbreds could be used to produce hybrids with higher levels of resistance than hybrids currently available to growers, and these could serve as a basis for gray leaf spot breeding programs. Lesion size measurements were not correlated with disease scores. Late-season photosynthesis rates were associated positively with resistance. The hybrids of some inbreds were found to produce high levels of pigment (believed to be anthocyanins) around the gray leaf spot lesions. These did not limit the size of the individual lesion later in the season. Some pigment(s)-producing genotypes were found to be resistant when the pigment character was expressed. This type of resistance must prevent or inhibit infection of the leaf but not later colonization, once established. Maize dwarf mosaic virus (MDMV) also limits maize production in some areas where johnsongrass (Sorghum halepense L.) is a problem. Resistance to MDMV was found to be mainly additive and highly heritable. However, a strong specific combining ability component was found, indicating that the background of the material receiving resistance genes may have a strong effect on the expression of resistance. Inbreds capable of producing high-yielding, resistant, and agronomically acceptable hybrids are available (B68, NC250, A632, Pa875, Va17, and Va85); and several hybrids between these lines have high levels of resistance, high yield, and good general agronomic characters (B68 x KB1250, KB1250 x Pa875, and NC250 x Pa875).
Ph. D.
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14

Ubalijoro, Eliane. "Characterization of resistance to lettuce mosaic virus in Lactuca sativa." Thesis, McGill University, 1994. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=22821.

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Lettuce mosaic virus (LMV) is an economically important pathogen with worldwide distribution. LMV infection in L. sativa can cause significant yield losses. Resistance to LMV in L. sativa is conferred by the recessive gene mo. We attempted to position the mo gene on the L. sativa map. The ultimate goal is a better understanding of plant-virus interactions. To do so, Random Amplified Polymorphic DNA (RAPD) markers were screened in the near isogenic lines (NILs) Vanguard and Vanguard 75. These NILs differ in the presence of the mo gene in Vanguard 75. Polymorphic markers were screened for linkage to mo in two F$ sb2$ populations segregating for resistance to LMV. The F$ sb2$ populations used were derived from 2 crosses, the first one between the L. sativa cultivars Dwarf 2 (resistant to LMV via the presence of mo) and Saffier and the second one between two breeding lines 87-25M-1 (momo) and 87-1090M-1 (MoMo). In order to develop a highly stringent antibody detection system to phenotype plants infected with LMV, a plasmid construct was developed which overproduces LMV coat protein. This construct will be used in the future to produce enough recombinant LMV coat protein for antibody production. To further characterize mo, a selection of cultivars resistant and susceptible to LMV according to the literature were subjected to various temperature changes to determine the environmental influences on virus movement.
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15

Mukoko, Olivia Zvinofa. "Breeding beans (Phaseolus vulgaris L.) for resistance to bean common mosaic virus in Zimbabwe." Thesis, University of Cambridge, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.240145.

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16

Ma, Guorong. "Genetic analysis of soybean reactions to soybean mosaic virus." Diss., Virginia Tech, 1995. http://hdl.handle.net/10919/40253.

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The soybean [Glycine max (L.) Merr.] mosaic disease, caused by soybean mosaic virus (SMV), is one of the most important soybean diseases in many areas of the world. This research, conducted in four separate studies, was designed to identify and characterize new sources of genes for resistance to SMV and to investigate the interaction of soybean resistance genes and SMV strains.
Ph. D.
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17

Otim-Nape, George William. "Epidemiology of the African cassava mosaic geminivirus disease (ACMD) in Uganda." Thesis, University of Reading, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.357982.

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18

Maruthi, M. N. "Bemisia tabaci and geminivirus variability in relation to cassava mosaic disease." Thesis, University of Greenwich, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.369144.

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19

Ali, Akhtar. "Pathology and molecular comparison of a range of pea seed-borne mosaic virus isolates." Title page, contents and summary only, 1999. http://web4.library.adelaide.edu.au/theses/09ACP/09acpa398.pdf.

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Copies of author's previously published articles inserted. Bibliography: leaves 128-143. This thesis describes the development of serological and nucleic acid based diagnostic methods for pea-seed borne mosaic virus (PSbMV), the isolation of specific effects on infected pea plants, the collection and biological comparison of new PSbMV isolates from Pakistan, the cloning and sequencing of specific parts of the genome of selected isolates, nucleotide and amino acid sequence comparisons between selected isolates, and the development of a ribonuclease protection assay (RPA) for identifying genomic differences among the PSbMV isolates. It is the first comparison of a range of geographically different isolates of PSbMV on the basis of both biological and molecular properties.
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20

Yu, Weichang. "CAMV gene VI protein : a virulence factor and the host responses in Arabidopsis /." free to MU campus, to others for purchase, 2002. http://wwwlib.umi.com/cr/mo/fullcit?p3075411.

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21

Balcı, Evrim Doğanlar Sami. "Genetic characterization of cucumber mosaic virus(CMV)resistance in tomato and pepper." [s.l.]: [s.n.], 2005. http://library.iyte.edu.tr/tezler/master/biyoloji/T000388.pdf.

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22

Gunduz, Irfan. "Genetic Analysis of Soybean Mosaic Virus Resistance in Soybean." Diss., Virginia Tech, 2000. http://hdl.handle.net/10919/26439.

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This research was conducted to analyze the genetics of soybean mosaic virus (SMV) resistance in soybean [Glycine max (L.) Merr.] and to determine allelic relationships of SMV resistance genes and their interactions with SMV strain groups. In the first part of this study, the inheritance of SMV resistance in OX670 and 'Harosoy' was studied to determine the source and identity of the resistance gene/genes in OX670. Other researchers reported that OX670 possesses a single gene at a locus independent of Rsv1 and assigned the gene symbol Rsv2. Rsv2 was presumably derived from the cultivar 'Raiden'. However, later work showed that Raiden contains a single resistance gene at the Rsv1 locus, raising the possibility that the resistance gene in OX670 was not from Raiden. Harosoy and its derivatives make up much of the remaining pedigree of OX670. Results from crosses of OX670 with susceptible cultivars indicate that it contains two independent genes for SMV resistance. One is allelic to the Rsv1 locus, expresses resistance to SMV-G1 and G7 and is derived from Raiden. The other is allelic to the Rsv3 locus, expresses resistance to SMV-G7 but susceptibility to SMV-G1 and is derived from Harosoy. Therefore the Rsv2 locus does not appear to exist in OX670 or its ancestors. The presence of Rsv1 and Rsv3 makes OX670 resistant to all SMV strains from G1 through G7. The second study was conducted to investigate the inheritance and allelomorphic relationships of resistance gene(s) in 'Tousan 140' and 'Hourei', which were reported to carry single independent resistance genes when inoculated with the Japanese SMV strain C. Both of these lines exhibit resistance to strains SMV-G1 through G7. This inheritance study shows that Tousan 140 and Hourei each possess two resistance genes. One of the genes in Hourei confers resistance to SMV-G1 and G7 strains; the other gene confers susceptibility to SMV-G1 but resistance to SMV-G7. Allelism tests indicate that one of the genes in both Hourei and Tousan 140 is allelic to Rsv1, and the other is allelic to Rsv3. The two genes in Tousan 140 were separated into individual lines, R1 and R2. R1, most probably containing Rsv1, exhibited resistance to SMV-G1 through G3 but was susceptible to SMV-G5 through G7. Line R2, most likely possesses Rsv3 gene, was susceptible to SMV-G1 through G3 but resistant to SMV-G5 through G7. Therefore, presence of these two genes makes Tousan 140 resistant to SMV-G1 through G7. The objective of the third study was to investigate inheritance and allelomorphic relationships of SMV resistance in PI88788. PI88788 exhibits resistance to SMV-G1 through G7. Genetic analysis of our data reveals that SMV resistance in PI88788 is conferred by a single gene at a locus tentatively labeled 'Rsv4'. Expression of this gene in the homozygous state decreased accumulation rate and prevented vascular movement of SMV. In the heterozygous state vascular movement of the SMV was delayed but not prevented.
Ph. D.
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23

Yu, Yong Gang. "Molecular genetic analysis of host resistance to soybean mosaic virus." Diss., Virginia Tech, 1994. http://hdl.handle.net/10919/37253.

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24

Parsons, Stephen H. "Comparing orchid transformation using agrobacterium tumefaciens and particle bombardment." Virtual Press, 1995. http://liblink.bsu.edu/uhtbin/catkey/941350.

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The Wheeler Orchid Collection is home to some of the most endangered species of orchids in the world. This fantastic reservoir of endangered species has been enhanced and broadened by its function as a plant rescue station for the U.S. customs service. Unfortunately, this responsibility increases the risk of bringing orchids, which harbor contageous diseases, into the greenhouse where sap transmitted diseases such as the Tobacco Mosaic Virus (TMV), can run rampant. Although manipulation of orchid characteristics is typically done by classical plant breeding techniques, genetic engineering is emerging as a useful technique for the introduction of desirable traits into the orchid genome. Through the use of genetic engineering techniques it may be possible to mitigate the symptoms associated with this destructive virus. Virus resistance may be achieved through the expression of either the sense or antisense viral coat protein gene in orchid tissues if an efficient means of orchid transformation is developed. In this research two transformation protocols were examined for their ability to efficiently transform orchid tissue. The first transformation protocol explored utilized the native ability of Aq bacterium tumefaciens to incorporate DNA into host plants to achieve transformation. The second mechanism explored was particle bombardment transformation.Many strains of A. tumefaciens were employed using direct exposure of Cattleya_ orchid protocorm and callus tissue. Particle bombardment using DNA coated 0.5 um diameter tungsten particles and high pressure helium tank acceleration was employed. The particle bombardment procedure employed the pG35barB plasmid which confers herbicide resistance to the herbicide basta when integrated and expressed in plant tissues.GUS fluorescence assays and PCR analysis indicate that T-DNA is present in orchid tissues, while Southern blot analysis was unable to display that integration had occurred. Particle bombardment yielded herbicide resistant orchid tissues which have yet to be analyzed by Southern blot analysis to confirm integration due to limited tissue quantities.
Department of Biology
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25

Sheih, Tianna. "Development of a Dengue Fever Vaccine from Recombinant DENV2 Protein and Tobacco Mosaic Virus." Scholarship @ Claremont, 2016. http://scholarship.claremont.edu/scripps_theses/810.

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Dengue fever is a rapidly growing concern to human health and is currently the most prevalent mosquito-borne viral disease worldwide. Although there are several vaccine candidates being tested in clinical trials, there are no vaccines publicly available to prevent this disease. Plant-based vaccines are rapidly becoming viable alternatives to traditional animal-based vaccines because they are safe, easy to manufacture, and more cost-efficient. The purpose of this project is to develop a vaccine against the dengue virus by producing a recombinant DENV2 protein, engineered by Dr. David Lo and his lab at University of California Riverside, in Nicotiana benthamiana plants through Tobacco Mosaic Virus (TMV) infection. Initial attempts to ligate the complete DENV2 epitope, a combination of hybrid flagellin sequences and the envelope protein from dengue viral serotype 2, into the pJL TRBO vector were incompatible with established protocols. However, a proof of concept test that replaced the DENV2 envelope protein with Green Fluorescent Protein (GFP) successfully inserted the new sequence into pJL TRBO. In the future, the DENV2 envelope protein sequence will be re-inserted into the construct and updated protocols will be repeated for DENV2 protein expression. The recombinant DENV2 proteins will be extracted from the plants after signs of infection become apparent and tested for their ability to induce an immunogenic response that produces pathogen-specific antibodies.
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26

Masiri, Jongkit Murphy John F. "The nature of cucumber mosaic virus-induced symptoms in bell pepper (Capsicum annuum L.)." Auburn, Ala., 2009. http://hdl.handle.net/10415/1977.

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27

Syme, Jennifer. "Characterization of Arabidopsis thaliana (Columbia) infected with turnip mosaic virus (TuMV)." Thesis, McGill University, 1996. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=24043.

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The response of Arabidopsis thaliana (Columbia) to infection with turnip mosaic virus (TuMV) was characterized at the level of: disease symptom expression, cell content and protein composition. Visual symptoms observed were chlorotic and mottled leaf colouring, severely stunted growth, distortion of leaf blades and delayed bolting. All plants died before seed cases dehisced. Electron microscopy revealed three types of cylindrical inclusion bodies: pinwheels, scrolls and laminated aggregates, in the cytoplasm of infected plants similar to those observed in other plants infected with TuMV. Inoculation of Arabidopsis with TuMV resulted in quantitative changes in several proteins in both soluble and membrane proteins, as revealed by electrophoresis on 12% polyacrylamide gels. Antibodies were made to both infected membrane and soluble proteins. Western blots of infected and uninfected, soluble and membrane proteins probed with antibodies revealed quantitative changes in the same proteins identified by polyacrylamide gels. A CNBr 4B activated sepharose column was used to make infection-specific antibodies to infected soluble proteins. No infection-specific host proteins were detected in Arabidopsis infected with TuMV.
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28

Tah, Tapashree Schoelz James E. "Chloroplast GFP expression in tobacco plants agroinfiltrated with tobacco mosaic virus based vectors." Diss., Columbia, Mo. : University of Missouri--Columbia, 2009. http://hdl.handle.net/10355/6604.

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Title from PDF of title page (University of Missouri--Columbia, viewed on Feb 19, 2010). The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. Thesis advisor: Dr. James E. Schoelz. Includes bibliographical references.
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29

Trevisan, Flavio. "Transformação genética de maracujazeiro (Passiflora edulis f. flavicarpa) para resistência ao vírus do endurecimento dos frutos." Universidade de São Paulo, 2005. http://www.teses.usp.br/teses/disponiveis/11/11144/tde-29092005-134710/.

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O objetivo do trabalho foi estudar uma forma alternativa para o controle do endurecimento dos frutos do maracujazeiro, pela produção de plantas transgênicas contendo o gene da proteína capsidial do Passionfruit woodness virus - PWV. O vetor de expressão foi construído utilizando-se os plasmídeos pCambia 2300 e pCambia 2301, que contêm o gene de seleção nptII, para resistência ao antibiótico canamicina. O plasmídeo pCambia 2301 contém também o gene repórter uidA (GUS). Os plasmídeos foram introduzidos em Agrobacterium tumefaciens, estirpes EHA 105 e LBA 4404, pelo método do choque térmico. Os explantes para transformação genética constituíram-se de discos de folhas jovens (6 mm de diâmetro), das variedades IAC 275 e IAC 277, coletados de plantas mantidas em sob fotoperíodo de 16 h luz, a 27 °C. Os explantes foram inoculados com suspensão bacteriana (5x108 UFC/mL) por 20 min e transferidos para placa de Petri contendo o meio de cultura MS + thidiazuron (TDZ - 0,25 mg/L) + nitrato de prata (AgNO3 - 4 mg/L) + acetoseringona (1 µM/L). O co-cultivo foi realizado à temperatura de 24 °C, em ausência de luz, por um período de 3 dias. Para seleção e regeneração de plantas os explantes foram transferidos para meio de cultura de seleção MS + TDZ (0,25 mg/L) + AgNO3 (4 mg/L) + canamicina (100 mg/L) + cefotaxime (500 mg/L). A incubação foi realizada a 27 °C, em ausência de luz, por um período de 4 - 6 semanas. As gemas adventícias desenvolvidas foram transferidas para o meio de cultura MSM + 10% de água de coco e incubadas sob fotoperíodo de 16 h de luz. A transformação genética foi identificada pelo teste histoquímico GUS e por PCR. Obteve-se um total de 22 plantas PCR positivas. Destas, 8 foram analisadas por Southern blot para confirmação da integração do transgene. A transcrição e expressão do transgene foram analisadas por Northern e Western blot, respectivamente. As plantas transgênicas avaliadas foram multiplicadas e inoculadas com 3 diferentes estirpes do PWV. A linhagem T2 apresentou resistência a infecção dos três isolados utilizados.
The main purpose of this work was to study an alternative way to control the Passionfruit woodiness virus - PWV through the production of transgenic plants which contained the Passionfruit woodness virus coat protein gene. The binary vector was built by using pCambia 2300 and pCambia 2301 plasmids, which contain the selection gene nptII. The pCambia 2301 plasmid also contains the reporter gene uidA (GUS). The plasmids were introduced into Agrobacterium tumefaciens, EHA 105 and LBA 4404 strains, via thermal shock method. The explants for the genetic transformation were young leaf disks (6 mm of diameter) of IAC 275 and IAC 277 varietys, extracted from plants kept under 16 h photoperiod, at 27 °C. The explants were inoculated with a bacterial suspension (5x108 UFC/mL) for 20 min and then transferred to Petri dishes containing cocolture medium MS + thidiazuron (TDZ - 0,25 mg/L) + silver nitrate (AgNO3 - 4 mg/L) + acetosyringone (1 µM/L). The co-culture was performed at 24 °C t, in the dark, for a three-day period. For the selection and regeneration of plants, the explants were transferred to the selection culture medium MS + TDZ (0,25 mg/L) + AgNO3 (4 mg/L) + kanamycin (100 mg/L) + cefotaxime (500 mg/L). The incubation was performed at 27 °C, in dark, for 4 - 6 weeks. The adventitious buds developed were then transferred to the culture medium MSM + 10% coconut water and kept incubated under 16 h photoperiod. The genetic transformation was identified through GUS and PCR tests. There were 22 PCR positive plants. Out of those, 8 were Southern blot analyzed for the confirmation of transgenc integration. The transgene transcription and expression were determined by Northern and Western blot respectively. The transgenic plants were then multiplied and inoculated with 3 different strains of PWV, and the line 2 showed resistance to the three strains used.
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30

Reyes, Castro Guillermo. "Studies on cocoyam (Xanthosoma spp.) in Nicaragua, with emphasis on Dasheen mosaic virus /." Uppsala : Dept. of Plant Biology and Forest Genetics, Swedish University of Agricultural Sciences, 2006. http://epsilon.slu.se/200607.pdf.

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31

Giampan, José Segundo. "Infectividade e proteção de três estirpes fracas do Papaya ringspot virus em plantas de melancia." Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-25022003-134805/.

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Este trabalho teve como objetivo avaliar a infectividade de estirpes fracas do Papaya ringspot virus - type W (PRSV-W) em plantas de melancia (Citrullus lanatus), em função da origem da estirpe fraca, da concentração e da espécie doadora do inóculo e da idade da planta-teste de melancia, inoculada mecanicamente. Também foi avaliado o efeito protetor dessas estirpes em plantas de melancia em casa de vegetação e em campo. A seleção de estirpes fracas do PRSV-W foi feita a partir de bolhas de folhas de melancia infectadas naturalmente em campo. A infectividade da estirpe fraca selecionada foi comparada com a das estirpes fracas PRSV-W-1 e PRSV-W-2, previamente selecionadas de bolhas de folhas de abobrinha de moita (Cucurbita pepo) 'Caserta' com mosaico. Como controle foi utilizada uma estirpe severa do vírus obtida de abobrinha de moita (PRSV-W-C). A avaliação do efeito da concentração e da espécie da planta fonte do inóculo na infectividade de plantas de melancia foi feita com extratos de 4, 8, 12 e 16 discos de folhas de abobrinha de moita e de melancia, infectadas separadamente com as estirpes fracas e severa, e diluídos em 2 mL de tampão fosfato. O efeito da idade da planta-teste de melancia na infectividade das estirpes fracas foi estudado comparando-se plantas inoculadas em quatro estádios de desenvolvimento, a partir do estádio cotiledonar, com inóculos das diferentes estirpes do PRSV-W extraídos de 12 discos foliares/2 mL de tampão. O efeito protetor da estirpe fraca obtida de bolhas de folhas de melancia com mosaico foi avaliado em plantas premunizadas e desafiadas com a estirpe PRSV-W-C, em casa de vegetação e em campo. Plantas de melancia premunizadas com as estirpes fracas PRSV-W-1 e PRSV-W-2 e plantas não protegidas também foram avaliadas no teste em campo. Foram avaliadas a proteção, com base nos sintomas, a produção e o conteúdo de açúcares (grau brix) dos frutos colhidos das plantas premunizadas e não premunizadas. Uma estirpe fraca do vírus, denominada PRSV-W-3, foi selecionada de bolhas de folhas de melancia com mosaico. Em todos os testes de infectividade em plantas de melancia, independente da concentração e da planta fonte do inóculo e do estádio de desenvolvimento da planta-teste inoculada, a estirpe fraca PRSV-W-3 apresentou taxas de infectividade semelhantes as das estirpes PRSV-W-1 e PRSV-W-2, chegando a 100% em alguns casos. A infectividade da estirpe severa PRSV-W-C foi de 100% em todos os testes. Aparentemente, a infectividade das três estirpes fracas foi mais diretamente afetada pela intensidade de fricção das folhas no momento da inoculação mecânica do que pelas variáveis estudadas. A estirpe fraca PRSV-W-3 protegeu as plantas de melancia contra a infecção e/ou manifestação da estirpe PRSV-W-C em casa de vegetação. Em campo, todas as plantas de melancia premunizadas com as três estirpes fracas também ficaram protegidas contra a estirpe severa utilizada no desafio. A produção das plantas premunizadas não diferiu estatisticamente entre si, nem mesmo daquelas inicialmente sadias infectadas em campo. O conteúdo de açúcares e a aparência da polpa dos frutos também foram semelhantes em todos os tratamentos.
The purpose of this work was to evaluate the infectivity of three mild strains of Papaya ringspot virus - type W (PRSV-W) on watermelon (Citrullus lanatus). The effect of the origin of the mild strain, the concentration of the inoculum, the species of the source of the inoculum and the age of the test-plant on the infectivity of mechanically inoculated watermelon were also evaluated. The protective effect of these mild strains on preimmunized watermelon plants was evaluated under greenhouse and field conditions. Mild strains were selected from blisters formed on mosaic leaves of naturally infected watermelon plants. The infectivity of the selected mild strain was compared with that of mild strains PRSV-W-1 and PRSV-W-2, which were previously obtained from blisters formed on mosaic leaves of zucchini squash (Cucurbita pepo cv. Caserta). A severe strain isolated from zucchini squash (PRSV-W-C) was used as control. The effect of the concentration and the species source of inoculum of the mild strains on the infectivity of watermelon plants was studied with inoculum extracted from 4, 8, 12 and 16 leaf discs of zucchini squash and watermelon plants, separately infected with the mild and severe strains, diluted in 2 mL of phosphate buffer. Four stages of development of watermelon plants, starting at the cotyledonal stage, were tested for the infectivity with the mild strains. Inocula were prepared with extracts of 12 leaf discs diluted in 2 mL of phosphate buffer. The protective effect of the mild strain selected from blisters on mosaic leaves of watermelon plants was evaluated on preimmunized plants challenge inoculated with severe strain PRSV-W-C, under greenhouse and field conditions. Watermelon plants preimmunized with mild strains PRSV-W-1 and PRSV-W-2 and unprotected plants were also included in the field trial. Protection was evaluated based on plant simptons, yield and sugar content in the fruits. One mild strain, named PRSV-W-3, was obtained from blisters on mosaic leaves of watermelon plants. The rate of infection of watermelon plants with mild strain PRSV-W-3 was similar to that with mild strains PRSV-W-1 and PRSV-W-2 in all infectivity tests, independently of the concentration of the inoculum, species source of the inoculum and stage of development of the inoculated test-plant, reaching 100% in some cases. Rate of infectivity with severe strain PRSV-W-C was always 100%. Apparently, the infectivity of the mild strains on watermelon was more directly related with the intensity of the abrasion produced by mechanical inoculation than with the above studied variables. The selected mild strain PRSV-W-3 protected watermelon plants against superinfection with the severe strain in the greenhouse tests. Protection was also effective under field conditions. Yield of plants preimmunized with all three mild strains and unprotected plants were statistically similar. The sugar content and the quality of the pulp of the fruits were similar for all treatments.
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32

Nogueira, Diêgo Rodrigues Soares. "Produção e avaliação de anti-soro policlonal visando a detecção do Pepper yellow mosaic virus." Universidade Federal de Viçosa, 2014. http://locus.ufv.br/handle/123456789/4436.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Pepper yellow mosaic virus (PepYMV) naturally infects sweet pepper and tomato plants, leading to severe losses since its first report in Brazil (Brasília, DF, 2002). Molecular and serological methods can be used to detect the pathogen. Serological methods for viral detection require the use of a high quality antiserum, offering good sensitivity and specificity. Traditionally, purified viral particles are used as immunogens. However, the purification process is very laborious and the final preparation may have unsatisfactory purity and/or concentration. Thus, the aim of this work was to produce a polyclonal antiserum against the recombinant capsid protein (CP) of PepYMV to allow its use both for diagnosis and for studies of the interaction of the PepYMV CP with other viral proteins and host factors. The coding sequence of the capsid protein gene of PepYMV was cloned into an expression vector (pRSET-A) and transformed into Escherichia coli strain BL21::DE3 for in vitro expression. The recombinant protein, fused to a histidine tag, was purified under denaturing conditions by affinity chromatography using a Ni-NTA column. The purified recombinant protein was dialyzed under renaturing conditions. Its integrity and identity were confirmed by polyacrylamide gel electrophoresis and mass spectrometer analyses. New Zealand rabbits were immunized with increasing amounts of the recombinant protein. The sensitivity and specificity of the antisera were analyzed by Western blot and indirect ELISA assays. The antisera raised against the recombinant CP showed good specificity and sensibility, proving to be a reliable tool for the detection of PepYMV.
O Pepper yellow mosaic virus (PepYMV), agente causal do mosaico amarelo do pimentão e do tomateiro, desde seu primeiro relato no Brasil no ano de 2002 em Brasília, DF, vem se disseminando em regiões produtoras de pimentão e tomate causando perdas substanciais ao produtores dessas culturas. Para identificação dessa enfermidade algumas ferramentas são utilizadas, dentre elas, podemos destacar os métodos moleculares e sorológicos, sendo estes últimos mais utilizados por apresentarem alta especificidade e sensibilidade, além de possuir um custo relativamente baixo. Para a produção de anti-soro, tradicionalmente, utilizam-se partículas virais concentradas como imunógenos. No entanto, o processo de purificação é muito trabalhoso e pode apresentar pureza e concentrações insatisfatórias. Desta forma, o objetivo deste trabalho foi produzir um anti-soro policlonal, a partir da proteína capsidial recombinante do PepYMV, que permita sua utilização tanto para diagnose quanto para estudos de interação da proteína capsidial do PepYMV com outras proteínas virais e fatores do hospedeiro. A sequência do gene da proteína capsidial do PepYMV foi clonada em vetor de expressão (pRSET-A) e transformada em Escherichia coli, linhagem BL21::DE3, para expressão in vitro. A proteína expressa fusionada a uma cauda de histidina foi purificada sob condições desnaturantes por cromatografia de afinidade em coluna de resina Ni-NTA. Em seguida a proteína purificada foi dialisada sob condições renaturantes e sua integridade e identidade foram confirmadas por gel de poliacrilamida a 12% e análise de espectrometria de massa. Dois coelhos da raça Nova Zelândia foram imunizados com quantidades crescentes da proteína recombinante dialisada adicionados do adjuvante de Freud incompleto na proporção 1:1. A sensibilidade e a especificidade do anti-soro foram testados por Western blot e ELISA indireto. O anti-soro produzido apresentou boa especificidade e sensibilidade, provando ser uma ferramenta confiável para a diagnose do PepYMV.
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33

Hajimorad, Mohammad Reza. "Variation in alfalfa mosaic virus with special reference to its immunochemical properties." Title page, contents and summary only, 1990. http://web4.library.adelaide.edu.au/theses/09PH/09phh154.pdf.

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Includes Appendix listing other publications by the author. Includes bibliographical references (leaves 134-181). Alfalfa mosaic virus was isolated from lucerne (Medicago sativa) plants with a variety of disease symptoms. Experiments showed that each isolate was biologically distinct and that the host range and symptomatology of each isolate was affected by the environmental condition.
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34

Ligat, Julio S. "Pathology and distribution in the host of pea seed-borne mosaic virus." Title page, contents and summary only, 1993. http://web4.library.adelaide.edu.au/theses/09PH/09phl723.pdf.

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Includes bibliographical references (leaves 82-92). Five isolates of pea seed-borne mosaic virus were compared by host range and symptomatology on 16 pisum sativum cultivars lines, 21 lines of Lathyrus and Lens spp. and several indicator species
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35

Torok, Valeria Anna. "Biological and molecular variation among isolates of pea seed borne mosaic virus." Title page, contents and abstract only, 2001. http://web4.library.adelaide.edu.au/theses/09PH/09pht686.pdf.

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Corrigendum inserted at the back. Includes bibliographical references (leaves 133-158). Ch. 1. General introduction -- ch. 2. General materials and methods -- ch. 3. Biological characterisation of Australian PSbMV isolates -- ch. 4. Developing nucleic acid based diagnostics for PSbMV -- ch. 5. Detection of PSbMV isolates by RT-PCR and RFLP analysis -- ch. 6. Developing an internal control for PSbMV RT-PCR -- ch. 7. Molecular analysis of the PSbMV VPG -- ch. 8. PSbMV sequence and phylogenetic analysis -- ch. 9. General discussion Sixteen pea seed borne mosaic virus (PSbMV) isolates were collected between 1995 and 1998. These isolates were biologically distinct yet serologically indistinguishable. The conclusion is that PSbMV is widespread and occurs at a low incidence in Australia. Reports sequence information on new isolates of PSbMV which has allowed genomic regions to be identified which distinguish PSbMV pathotypes and isolates; and, to the development of PSbMV nucleic acid hybridisation and RT-PCR assays.
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36

Jimenez, Garcia Emilio. "ETIOLOGY, PATHOLOGY AND CHARACTERIZATION OF VIRUSES FROM BEANS GROWING IN THE SONORA DESERT OF MEXICO (COWPEA, CHLOROTIC MOTTLE)." Diss., The University of Arizona, 1985. http://hdl.handle.net/10150/187907.

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Survey of crops of common bean (Phaseolus vulgaris L.) in Sonora, Mexico revealed the presence of two isometric viruses and one flexuous rod virus on the basis of host reaction, particle morphology, serology and physico-chemical properties. The isometric viruses were identified as Bean Southern Mosaic Virus (BSMV) and Cowpea Chlorotic Mottle Virus (CCMV); the flexuous rod virus was identified as Bean Common Mosaic Virus (BCMV). Using bean cultivar differentials, two strains of the potyvirus BCMV were identified, NY-15 and a previously undescribed strain designated YV-1. Host range, serological tests, and RNA electrophoresis indicated that the Sonoran BSMV cultures are similar to BSMV-strain A. Serology and RNA-electrophoresis indicated that the Sonoran CCMV isolates are identical to CCMV-strain A. BSMV and CCMV were always isolated as a mixture from seed lots and from field collected bean tissue. BCMV occurred alone or in mixed infections with BSMV and CCMV. BCMV was seed transmitted with an average efficiency of 58 percent. The BSMV-CCMV mixture was transmitted with an efficiency of 6 percent. BSMV and CCMV were seed transmitted together, but separate transmission of BSMV or CCMV was not detected. Commercial seed lots from two major bean growing regions of Sonora (Hermosillo Coast, Sonora River) were contaminated with the BSMV-CCMV mixture but not with BCMV. The average contamination level was 13 percent. Two common weeds present in Sonoran agricultural areas were found to be potential alternate hosts of CCMV. Both Sisymbrium irio L. and Melilotus indica L. were infected systemically, although the infection in M. indica was latent. Potential losses due to Sonoran bean viruses were measured in greenhouse experiments with the cultivar Pinto 111. BCMV strains caused a 29.4 to 60.1% reduction, whereas BSMV-CCMV mixtures induced a 22.5 to 74.6% yield reduction. A synergism occurred between the BSMV-CCMV mixture and BCMV resulting in more severe symptoms and a yield reduction of 92.7%. Synergistic effects were also observed between BSMV and CCMV. Actual yield reduction resulted from impaired flower production and, consequently, reduced pod production. Significant effects on plant tissue production, flower fertilization and seed quality were not observed. Cowpea chlorotic mottle virus infected mung bean (Vigna radiata (L.) Wilczek) a previously unreported host. Infection of mung bean by BSMV was only possible when CCMV was present in the inoculum. Both BSMV and CCMV could be isolated from symptomatic plants infected with the BSMV-CCMV mixture, however, symptoms on mung bean were unchanged from infection by CCMV alone.
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37

Srivatsavai, Venkata Suresh Kumar Huettel Robin Norton. "Identification, distribution and vector biology of brome mosaic virus of wheat in Alabama." Auburn, Ala., 2005. http://hdl.handle.net/10415/1266.

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38

Ndunguru, Joseph. "Molecular characterization of cassava mosaic geminiviruses in Tanzania." Thesis, University of Pretoria, 2005. http://hdl.handle.net/2263/30648.

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Cassava (Manihot esculenta Crantz) is a basic staple food crop in Tanzania. Cassava mosaic disease (CMD) caused by cassava mosaic geminiviruses (CMGs) constitutes a major limiting factor to cassava production in the country. This study was undertaken to characterize the CMGs occurring in Tanzania using molecular techniques and to map their geographical distribution to generate information on which the formulation of control measures can be based. Using Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) for analysis of CMGs DNA-A genomes, different CMGs were found to be associated with CMD. Higher molecular diversity was observed among East African cassava mosaic viruses (EACMVs) than African cassava mosaic viruses (ACMVs), which was confirmed later by complete nucleotide sequence analysis. In addition to EACMV and ACMV isolates, two isolates of EACMV Cameroon virus (EACMCV) were found in Tanzania. These were confirmed to be strains of EACMCV Cameroon, originally described in Cameroon, West Africa and here named EACMCV- [TZ1] and EACMCV-[TZ7]. They had high (92%) overall DNA-A nucleotide sequence identity and EACMCV-[TZ1] was widespread in the southern part of the country. A subgenomic DNA form of CMG that appeared to be truncated was identified in a CMD-infected cassava plant. It was confirmed upon sequence analysis to be a defect of EACMV DNA-A and had a capacity of attenuating symptoms when coinoculated with wild-type EACMV. In addition, this study revealed for the first time the presence of two novel non-geminivirus single-stranded DNA (ssDNA) sub-genomic molecules associated with CMG infection. They were shown to be dependent on CMG for replication and movement within the plants, confirming their status as satellite molecules named here as satDNA-II and satDNA-III. When present in coinfection with CMGs, they enhance symptoms and can break high levels of resistance in a cassava landrace. Finally a simple, inexpensive technique is described of archiving, transporting and recovering plant DNA for downstream geminivirus characterisation.
Thesis (PhD)--University of Pretoria, 2007.
Microbiology and Plant Pathology
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39

Sassi, Giovanna. "Relative quantification of host gene expression and protein accumulation upon turnip mosaic potyvirus infection in tobacco." Thesis, McGill University, 2004. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=81433.

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Turnip mosaic virus (TuMV) infects a variety of crops, worldwide, including the economically relevant Brassicacea family. It was previously demonstrated that TuMV infection in tobacco protoplasts leads to an overall decrease of host protein. However, it remains unclear whether this phenomenon is due to the repression of plant gene transcription during the infection period or due to viral inhibition of host translation. In this study, quantification of various transcripts and protein products from infected tobacco was performed via real-time RT-PCR and ELISA. In comparison to the gamma-tubulin endogenous control, gene expression for the tobacco H3, HSP70 and granule-bound starch synthase was affected by TuMV infection with time.
Tobacco protein accumulation in whole leaf tissues was also significantly affected by increase of virus particles.
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40

Plante, Daniel 1970. "Interaction of the turnip mosaic potyvirus VPg with the plant translation apparatus." Thesis, McGill University, 2000. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=37812.

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An interaction was recently detected between the potyviral protein, genome-linked (VPg) and the Arabidopsis thaliana translation initiation factor eIF(iso)4E (Wittmann et al., 1997).
Here, experiments were undertaken to address biological aspects of the VPg-eIF4E interaction. First, coimmunoprecipitation experiments performed with purified recombinant proteins have shown that VPg not only associates with eIF4E, as was previously published, but also with the larger eIF4F complex, of which eIF4E is a subunit. These results were confirmed by ELISA-type binding assays. It was also shown that there is no direct interaction between VPg and the other subunit of eIF4F, namely eIF4G. Finally, with the same experimental system, it was shown that the presence of eIF4G does not influence the binding affinity of VPg and eIF4E.
The interaction of VPg with the plant translation apparatus suggests that potyviral infection may alter the host protein expression profile. This hypothesis was investigated with the use of a protoplast system. We have shown that the global rates of protein synthesis in protoplasts transfected with an infectious TuMV cDNA clone dropped shortly after transfection, by as much as an estimated 70%. Recovery to normal levels occurred within 48 hours.
Evidence was obtained that the interaction between VPg and eIF4E is instrumental in this transient down-regulation of protein expression: protoplasts transfected with a mutant TuMV cDNA clone, the VPg of which has no affinity for eIF4E, failed to exhibit the drop in protein synthesis observed with the wild-type clone.
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41

Montesclaros, Luz B. "Mapping of molecular markers surrounding the Tu gene conferring resistance to turnip mosaic virus in Lactuca sativa L." Thesis, McGill University, 1996. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=23923.

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In lettuce (Lactuca sativa), the dominant gene Tu confers resistance to turnip mosaic virus (TuMV) infection. In order to eventually clone and characterize the Tu gene using a map-based cloning strategy, the chromosome region in which Tu is located needs to be saturated with molecular markers. Random polymorphic DNA (RAPD) markers were screened using bulked segregant analysis. Nine new RAPD markers, UBC431$ rm sb{420}, UBC431 sb{940}, UBC434 sb{360}, UBC434 sb{1000}, UBC439 sb{520}, UBC448 sb{685:750}, UBC135 sb{240}, OP108 sb{410} and OP108 sb{1305},$ were identified as linked to Tu. Each marker was mapped relative to Tu using F$ sb2$ individuals previously known to be recombinant in the area surrounding the Tu locus. Three new markers, UBC431$ rm sb{420}, UBC439 sb{520} and UBC135 sb{240}$ are within a 5 cM area of Tu. As the number of DNA markers on the map increased map expansion and difficulties in determining a unique order were encountered. To increase the confidence in the estimate of genetic distances, a population of 500 F$ sb2$ plants was screened in order to identify more recombinant individuals around the Tu locus. The population was screened using markers UBC431$ sb{420}$ and UBC135$ sb{240}.$ Thirty-three recombinants were identified in an interval of 6.6 cM. Two markers, UBC346$ sb{1067}$ and OP108$ sb{634},$ tightly flanking Tu were converted to sequence characterized amplified regions (SCAR 346 and SCAR L08). No polymorphism was detected among the SCARs generated. The area surrounding Tu now includes 24 RAPD markers in an interval of 44 cM.
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42

Robbins, Marjorie. "The location of Tu on the genetic map of Lactuca sativa and the identification of random amplified polymorphic DNA markers flanking and tightly linked to Tu /." Thesis, McGill University, 1993. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=69684.

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In Lactuca sativa, the dominant gene Tu confers resistance to infection by turnip mosaic virus (TuMV). Tu and Dm5/8, a gene for resistance to Bremia lactucae, are linked in L. sativa. The area surrounding Dm5/8 on the genetic map of L. sativa contains restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers. The orientation of Tu relative to Dm5/8 was not known. Locating Tu would indicate which markers are on the map of lettuce close to Tu. To locate Tu on the L. sativa genetic map, F$ sb3$ families from recombinant F$ sb2$ in the Dm5/8 area of a cross between TuMV-resistant (Cobbham Green) and susceptible (Calmar) cultivars were inoculated with TuMV and phenotyped for Tu by indirect enzyme-linked immunosorbent assay. Polyclonal antibodies for immunodetection were produced using turnip mosaic virus coat protein expressed in E. coli. Phenotypic ratios within F$ sb3$ families were used to determine individual F$ sb2$ genotypes for Tu. With these genotypes, Tu was located on the genetic map of L. sativa relative to data present for Dm5/8 and surrounding markers, between OPM18 and OPY13. Using bulked segregant analysis, bulks created for the Dm5/8 locus were screened for genetic polymorphisms by the RAPD technique. Five new RAPD markers, UBC346, UBC517, UBC563, UBC599, and UBC675 were found linked to Tu after mapping relative to F$ sb2$ genotypes for Tu and other RAPD markers. The resulting three-point mapping information indicates that Tu is flanked by two markers, OPM18/OPL08 and UBC346, at respective genetic distances of 0.4 and 0.7 cM.
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43

Moreira, Alécio Souza. "Epidemiologia comparativa de três viroses em abobrinha de moita (Cucurbita pepo L.)." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-08022011-092407/.

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A abobrinha de moita (Cucurbita pepo L.), espécie pertencente à família Cucurbitaceae, apresenta significativa participação na produção mundial e brasileira desta família de plantas. Porém, como em toda cultura de importância econômica, as cucurbitáceas apresentam problemas fitossanitários causados por diferentes agentes etiológicos. Na produção brasileira de abóboras, já foi constatada a presença de 8 vírus, dentre eles os potyvirus PRSV-W (Papaya ringspot virus-type W) e ZYMV (Zucchini yellow mosaic virus) e o tospovirus ZLCV (Zucchini lethal chlorosis virus) têm sido considerados os mais importantes pela predominância em diversas regiões produtoras de cucurbitáceas no país e pelos consideráveis prejuízos que geralmente causam à produção. Considerando que a maioria dos estudos epidemiológicos existentes sobre estas três viroses são poucos e fragmentados, percebe-se que não há estudos que abordam em conjunto todos os parâmetros epidemiológicos de tais vírus. Assim, os objetivos deste trabalho foram estudar o progresso temporal e espacial destas três viroses e verificar a relação entre a epidemiologia das mesmas em campo de abobrinha de moita, além de melhor entender o patossistema em que a clorose letal (causada pelo ZLCV) está inserida Para isso, foram conduzidos experimentos com abobrinha de moita Caserta nos campos experimentais dos Departamentos de Fitopatologia e Nematologia (DFN) e Departamento de Genética (DGN) da Esalq/USP. Primeiramente foram conduzidos em 2009 dois experimentos simultâneos nas áreas do DFN e DGN para estudar a epidemiologia isolada da clorose letal acompanhada da dinâmica populacional do tripes Frankliniella zucchini, vetor deste vírus. Em seguida, em épocas diferentes 3 experimentos foram conduzidos em 2010 com o intuito de comparar as epidemiologias da clorose letal e dos mosaicos comum e amarelo, estes últimos causados por PRSV-W e ZYMV, respectivamente. Nos experimentos apenas com a clorose letal no ano de 2009, o modelo monomolecular foi o que melhor se ajustou aos dados de incidência e através das análises espaciais detectou-se agregação da doença ao final dos dois experimentos. Nos 3 experimentos realizados em 2010, variações na incidência, no ajuste do modelo e no comportamento espacial de cada virose foram freqüentes. Para a clorose letal, o modelo monomolecular ofereceu melhor ajuste apenas na 3ª época de plantio. Nas duas primeiras o modelo gompertz apresentou maior coeficiente de determinação. No comportamento espacial, novamente agregação da doença foi detectada ao final do ciclo da cultura. Para o mosaico amarelo, os modelos que melhores se ajustaram na 1ª, 2ª e 3ª épocas de plantio, foram o logístico e o monomolecular (este nas duas últimas), respectivamente. O padrão espacial desta doença foi ao acaso quando da baixa incidência da doença e agregado nos casos em que elevada incidência ocorreu. Já o mosaico comum apresentou a menor incidência nas 3 épocas. O modelo logístico foi o que melhor se ajustou em todas as épocas e a doença apresentou comportamento espacial ao acaso nos três experimentos. Nos levantamentos do tripes vetor do ZLCV, verificou-se preferência do mesmo por plantas sintomáticas e consideráveis correlações do número de insetos coletados com a incidência da clorose letal.
The zucchini squash (Cucurbita pepo L.) belonging to the Cucurbitaceae family, has a good share of world and brazilian output. However, as in every plants of economically important, cucurbits have problems caused by different etiological agents. In the Brazilian production of zucchini squash, already confirmed the presence of 8 viruses, including the potyviruses PRSV-W (Papaya ringspot virus-type W) and ZYMV (Zucchini yellow mosaic virus) and the tospovirus ZLCV (Zucchini lethal chlorosis virus) have been considered the most important viruses by the predominance in several cucurbits producing regions in the Brazil and the considerable damage on production. Whereas the most of the existing epidemiological studies about these three viruses are few and fragmented, it is clear that there are no studies that deal together all the epidemiological parameters of such viruses. The objectives of this work were to study the temporal and spatial progress of these three viruses and the relation between the epidemiology of these viruses in the same field of zucchini squash in addition to better understand the lethal chlorosis pathosystem (caused by ZLCV). Trials were carried out with zucchini squash \'Caserta in the experimental fields of the Departments of Plant Pathology and Nematology (DPP) and Department of Genetics (DGN) at Esalq/USP. The firsts were conducted in 2009 simultaneously in DPP and DGN to study the epidemiology of lethal chlorosis only and to study the population dynamic of thrips Frankliniella zucchini, the vector of this virus. In 2010 three experiments were carried out in different growing seasons in order to compare the epidemiology of the lethal chlorosis, yellow mosaic and common mosaic caused by ZLCV, PRSV-W and ZYMV, respectively. In the experiments with the lethal chlorosis in 2009, the monomolecular model was the best fit to the incidence data and spatial analysis indicated aggregation of the disease at the end of both experiments. In three experiments carried out in 2010, variations in incidence, in the fit of the model and in the spatial distribution of each virus were frequents. For lethal chlorosis, the monomolecular model provided a better fit only in the 3rd growing season. In the first and second growing seasons Gompertz model had the best coefficient of determination. In the spatial distribution, aggregation of disease was detected at the end of the crop cycle again. For yellow mosaic, the models that best fit in the 1st, 2nd and 3rd planting dates were the logistic and monomolecular (this in the last two) respectively. The spatial pattern of this disease were randomly when the disease incidence was low and aggregated when the disease incidence was high. The common mosaic had the lowest incidence in all three seasons. The logistic model was the best fit in all growing seasons and the disease showed a spatial random distribuctions in all experiments. The thrips vector of ZLCV prefer symptomatic plants and good correlations between the number of insects collected with the incidence of lethal chlorosis was found.
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44

Teixeira, Ana Paula Matoso. "Identificação de marcadores moleculares ligados a gene de resistência ao vírus do mosaico (PRSV-W) em melão (Cucumis melo L.)." Universidade de São Paulo, 2004. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-10112004-153914/.

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A importância da cultura do meloeiro é crescente no Brasil, sobretudo na região Nordeste, tanto pelo volume comercializado como por ser estabelecida geralmente em pequenas propriedades. Diversas enfermidades acometem esta cultura, destacando-se as viroses. Dentre estas, o mosaico, causado pelo Papaya ringspot virus - estirpe melancia (PRSV-W) é das mais importantes. Dentre as estratégias de controle desta doença, o emprego de cultivares resistentes apresenta-se como um método prático e eficiente. O objetivo deste trabalho foi identificar marcadores moleculares do tipo AFLP ligados ao gene Prv1 de resistência a PRSV-W em melão que futuramente pudessem ser utilizados em seleção assistida por marcadores. Para isto, foram analisadas duas linhagens quase-isogênicas (LQI-R e LQI-S) do tipo Amarelo CAC contrastantes para resistência ao vírus e uma linhagem do tipo Charentais doadora do gene de resistência. A LQI resistente foi obtida através de cruzamento entre a linhagem doadora do gene (LRD) e a linhagem recorrente (LQI-S), seguido de cinco retrocruzamentos de plantas resistentes com a linhagem recorrente. A porcentagem do genoma do parental recorrente recuperado na LQI-R foi de aproximadamente 98,44%. Polimorfismos entre as linhagens resistentes e a suscetível foram considerados marcadores candidatos ligados ao gene de reistência Prv1. Para análise de co-segregação entre gene e marcadores candidatos, foi utilizada uma população RC1F1, fenotipada para resistência a PRSV-W, obtida a partir do cruzamento entre as LQIs. Para cálculo da distância entre o gene e os marcadores foi utilizada fórmula de Kosambi para porcentagens de indivíduos recombinantes maiores que 1% e para porcentagens menores admitiu-se ser a distância em centiMorgans equivalente a esta porcentagem. A técnica AFLP em conjunto com a utilização de linhagens quase-isogênicas mostrou-se eficiente na detecção de marcadores moleculares em melão. Para digestão do DNA, foram utilizadas três combinações diferentes de enzimas de restrição (EcoRI/MseI, HindIII/MseI e PstI/MseI), sendo avaliados perfis eletroforéticos gerados a partir da amplificação com 474 combinações diferentes de iniciadores. Aproximadamente 28.700 fragmentos foram analisados, sendo verificada diversidade genética de 8,6% (2462 fragmentos polimórficos) entre as linhagens quase-isogênicas e a linhagem doadora Charentais. Apenas três fragmentos mostraram-se polimórficos na análise da população RC1F1, estando ligados ao gene de resistência a PRSV-W, de acordo com o teste de co-segregação. Os fragmentos EA270 e HF155 mostramse ligados entre si e estão localizados a uma distância aproximada de 40,9 cM do gene Prv1, enquanto o fragmento EK190 mostrou-se ligado ao gene a uma distância de 0,526 cM. Pela sua proximidade ao gene Prv1, o marcador EK190 pode ser utilizado em programas de seleção assistida por marcadores moleculares visando o melhoramento de linhagens com resistência ao vírus PRSV-W.
The growing importance of melon in Brazil is due to the increased production, especially in the Northern region, where crops are established in small properties. Several diseases affect melons. Among the viruses, the mosaic, caused by Papaya ringspot virus – type watermelon (PRSV-W) is the most important. The use of resistant cultivars is a practical and effective method of disease control. The objective of this work was to identify AFLP markers linked to the Prv1 gene that confers resistance to PRSV-W, that in the future could be used in marker assisted selection. Two near isogenic lines (LQI-R and LQI-S) of the Amarelo CAC type that differ with respect to the presence of Prv1 and one Charentais type line donor of the resistance gene were analyzed. The resistant LQI was obtained through the crossing between the donor line (LRD) and the recurrent line (LQI-S), followed by five backcrosses between resistant plants and the recurrent line. The percentage of recurrent parental genome recovered in the LQI-R was approximately 98.44%. Polymorphisms between resistant and susceptible lines were considered as candidate markers linked to the Prv1 resistance gene. An RC1F1 population obtained from a cross between the LQIs lines and screened for resistance to PRSV-W was used in co-segregation analyses. The distance between markers and resistance gene was calculated using the Kosambi equation for recombination fractions higher than 1%. For lower values, the percentage of recombinants was considered equal to the distance in centiMorgans. The AFLP technique combined with the use of nearisogenic lines seemed to be efficient in detecting molecular markers in melon. DNA digestion was performed with three combinations of different enzymes (EcoRI/MseI, HindIII/MseI and PstI/MseI), and electrophoretic profiles of fragments obtained from 474 combinations of different primers were evaluated. Approximately 28,700 fragments were analyzed. Genetic diversity was estimated as 8.6% (2,462 polymorphic fragments) between near-isogenic lines and the donor Charentais line. Only three fragments were found to be polymorphic and linked to the resistance gene. The markers EA270 and HF155 are linked to each other and located 40.9 cM of the Prv1 gene. The fragment EK190 is linked to the same gene with a distance of 0.526 cM. Because EK190 fragment is very close to the resistance gene, it is a suitable marker to be used in marker-assisted selection aiming to develop melon cultivars resistant to PRSV-W.
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45

Hutchinson, Chad M. "Agrobacterium tumefaciens mediated transformation of orchid tissue with the sense and antisense coat protein genes from the odontoglossum ringspot virus." Virtual Press, 1992. http://liblink.bsu.edu/uhtbin/catkey/834608.

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This research was an attempt to use a dicot transformation vector to transform a monocot. The initial purpose of this thesis was to transform orchids with the sense and antisense coat protein genes from the Odontoglossum ringspot virus (ORSV) in an effort to mitigate viral symptoms in transgenic plants using the transformation vector, Agrobacterium tumefaciens. However, it soon became apparent that much time would be needed to develop a transformation protocol. The transformation vectors used included the Agrobacterium tumefaciens disarmed strain LBA4404 with the binary plasmid pB1121, the disarmed strain At699 with the binary plasmid pCNL65, and the wild-type strain Chry5. The marker gene on the binary plasmids of both disarmed strains was p-glucuronidase (GUS).Several transformation protocols were used in an effort to determine if this transformation system would work on orchids. Transformation was not achieved even though a number of experimental conditions were varied. These included using two different types of orchid tissue, callus and protocorms; using two different species of orchids, Cattleya Chocolate Drop x Cattleytonia Kieth Roth and Cymbidium maudidum; varying the time the plant tissue was exposed to the bacteria from 1 hour to 96 hours; performing experiments with and without the wound signal molecule acetosyringone; and exposing the tissue to the virulent strains of A. tumefaciens mentioned previously.This research also developed GUS assay conditions necessary to decrease the number of false positives due to bacterial contamination. These conditions included chloramphenicol in the GUS assay buffer.
Department of Biology
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46

Narita, Nobuyoshi 1961. "Epidemiologia do "Cowpea aphid borne mosaic virus" (CABMV) em maracujazeiros na região produtora da Alta Paulista, SP /." Botucatu : [s.n.], 2007. http://hdl.handle.net/11449/103210.

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Resumo: Dos vírus que infectam o maracujazeiro no Brasil, atualmente o Cowpea aphid borne mosaic virus (CABMV), é considerado fator limitante à cultura. Dependendo da velocidade de disseminação e idade com que as plantas são infectadas no campo, a cultura torna-se comercialmente improdutiva. O presente estudo teve como objetivo, avaliar a diversidade e a dinâmica populacional dos afídeos na região da Alta Paulista, SP e a possibilidade de transmissão do vírus pela semente. Assim, quatro locais (Leste e Oeste da cidade de Marília e Municípios de Ocauçú e Guaimbê) foram monitorados durante 24 meses com armadilhas amarelas de água do tipo Moericke. Constatou-se nas quatro regiões a predominância do gênero Aphis. Outras espécies coletadas foram Myzus persicae, Geopenphigus flocculosus, Brevicoryne brassicae, Rhopalosiphum spp, Dysaphis spp e Lipaphis erysimi. A flutuação populacional de formas aladas do gênero Aphis, caracterizou-se por apresentar maiores revoadas em maio, junho, agosto e setembro. As espécies de Aphis (A. fabae, A. gossypii, A. spiraecola) devem ser os principais vetores do CABMV na região. Plantios novos, ao lado de plantações infectadas, tornam-se infectadas em três meses. Nos testes de transmissão através de sementes, do total de 13056 semeadas oriundas de plantas doentes, germinaram 10592, e em avaliações visuais dois meses após a germinação, não foram observadas plantas sintomáticas, indicando a não transmissão pela semente.
Abstract: From the viruses were described infecting passionfruit plants in Brasil, and the Cowpea aphid borne mosaic virus (CABMV), is considered the most hazardous. Depending on the spread velocity of aphids and the age that the plants are infected, the crops doesn’t produce commercial fruits. The present study was designed to evaluate the diversity and dynamic population of aphids in the Alta Paulista, SP region and aspects of seed transmission. For this, four regions (East and West of Marília city, Guaimbê and Ocauçú) were monitored for 24 months using yellow water Moerick trap. The predominance of the genus Aphis was observed in the four evaluated areas. Other species founded in the area were: Myzus persicae, Geopenphigus flocculosus, Brevicoryne brassicae, Rhopalosiphum spp, Dysaphis spp and Lipaphis erysimi. The population curve of alate Aphis spp showed the highest frequency of flights during May, June, August and September. The Aphis spp (A. fabae, A. gossypii, A. spiraecola) probably is the most important vector of the CABMV in the region. New crops near old infected plants, were infected in three months. To evaluate properties of seed transmission, from 13056 collected from infected plants, 10592 were germinated and evaluated during two months for the presence of visual symptoms. No plants with simptoms were observed indicating no seed transmission.
Orientador: Marcelo Agenor Pavan
Coorientador: Valdir Atsushi Yuki
Banca: Renate Krause Sakate
Banca: Aloisio Costa Sampaio
Banca: Alexandre Levi R. Chaves
Banca: Hugo Kuniyuki
Doutor
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47

Schuck, Heather A. "Differentially expressed genes of Sophrolaeliacattleya Ginny Champion "Riverbend" in response to the odontoglossum ringspot virus." Virtual Press, 2000. http://liblink.bsu.edu/uhtbin/catkey/1164841.

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Due to the rapid destruction of native orchid habitats it has become necessary to house many endangered orchid species in greenhouse environments where enhanced spread of viral disease occurs due to the close contact between plants. This research was concerned with the construction of a library of genes whose expression is induced in response to viral challenge. In uncovering the genes that are activated during plant-pathogen interactions, it may be possible to manipulate these pathways to develop virus resistant orchids. Furthermore, this research will contribute additional information for the existing framework of plant-pathogen interactions of all plant species.In order to construct a library of genes expressed in response to viral infection, suppression subtractive hybridization was performed using the PCR-Select cDNA Subtraction Kit (CLONTECH, Palo Alto, CA) on Sophrolaeliacattleya Ginny Champion 'Riverbend' clones. RNA was isolated from plants that had been inoculated with the Odontoglossum ringspot virus (ORSV) and from control plants that had not been inoculated with ORSV. Following reverse transcription-PCR (RT-PCR) to obtain cDNA, cDNAs of the tester population (those cDNAs containing differentially expressed messages in response to ORSV) and the driver population (reference cDNAs from uninfected plants) were obtained. The two different cDNA populations are mixed together and hybridized. The sequences common to both populations were subtracted, leaving only the differentially expressed sequences available for PCR amplification.A library containing these genes was constructed, and one clone, chosen at random, was sequenced. Based on homology comparisons to known genes, we have cloned a gene that may contain a nucleotide binding site similar to that of the tobacco N gene, important for plant resistance to pathogens. In the near future, this clone will be used to construct probes for use in northern analysis to determine the timing and localization of the products of this gene. This information will aid in characterizing the function of the orchid N-gene and identifying other members of this signal cascade. In addition, many other clones await sequencing and similar characterization.
Department of Biology
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48

Masli, Aryananda. "Search for restriction fragment length polymorphism of Phaseolus vulgaris in relation to the immune gene to bean common mosaic virus." Thesis, University of North Texas, 1991. https://digital.library.unt.edu/ark:/67531/metadc798405/.

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A technique involving Restriction Fragment Length Polymorphism (RFLP) was used to observe the DNA fragment polymorphism between a bean cultivar with I/I genotype and a bean cultivar with i/i genotype. The I gene encodes immunity to bean common mosaic virus (BCMV).
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49

Ozumit, Alen. "Interaction between turnip mosaic potyvirus (TuMV) cylindrical inclusion protein and Arabidopsis thaliana histone H3 protein." Thesis, McGill University, 2003. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=79060.

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Turnip mosaic potyvirus (TuMV) is a single-stranded RNA plant virus. One of its proteins, the cylindrical inclusion (CI) protein, was hypothesized to interfere with host transcription via interaction with histone H3 protein. Interaction between CI and histone H3 was previously observed in Dr. Fortin's laboratory. Based on previous studies that demonstrated the importance of the H3 tail domain in gene regulation and chromosome arrangement, it was hypothesized that CI would interact with the tail rather than the globular domain. The objective of this project was to identify which histone H3 domains CI protein interacts with. The full-length, globular, and tail domains of histone H3 DNA were expressed in E. coli and purified. Based on in vitro interaction experiments, the CI protein was observed to interact with the globular domain of histone H3.
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50

Freitas, Debora Maria Sansini. "Novas observações sobre a proteção com estirpes fracas do Papaya ringspot virus - type W e do Zucchini yellow mosaic virus em plantas de abobrinha-de-moita." Universidade de São Paulo, 2007. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-17082007-095734/.

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O Papaya ringspot virus - type W (PRSV-W) e o Zucchini yellow mosaic virus (ZYMV) são as duas espécies de potyvírus que mais causam danos em cucurbitáceas no estado de São Paulo e no Brasil. Uma boa alternativa de controle para as doenças causadas por esses vírus é o emprego da premunização. O principal objetivo desse trabalho foi estudar a competição por sítios de replicação como possível mecanismo de proteção entre a estirpe fraca PRSV-W-1 e a estirpe severa PRSV-W-C em plantas de abobrinha-de-moita. Além disso, procurou-se também conhecer o período mínimo necessário para a proteção de plantas de abobrinha-de-moita premunizadas com a estirpe fraca ZYMV-M, só e em dupla premunização com a estirpe fraca PRSV-W-1, para fornecer subsídios para o uso em condições de campo. O estudo da competição por sítios de replicação como mecanismo de proteção foi feito inoculando-se plantas de abobrinha-de-moita ‘Caserta‘ com a estirpe fraca PRSV-W-1 e desafiando-as com a estipe severa PRSV-W-C. As inoculações de proteção foram feitas nas folhas cotiledonares e as de desafio na folha nova verdadeira, e vice-versa, aos 3, 6 e 9 dias após a primeira inoculação. Plantas infectadas com a estirpe fraca e não desafiadas e plantas infectadas com a estirpe severa foram usadas como controles. As avaliações foram feitas com base na manifestação dos sintomas 30 dias após o desafio. Também foi feito teste de recuperação da estirpe desafiante e detecção desta por RT-PCR, com primers específicos, aos 8 dias após o desafio. Os resultados sugerem que, independente do local onde foi realizada a inoculação de proteção (folha cotiledonar ou folha nova expandida), de uma maneira geral parece haver alguns sítios livres para a superinfecção com a estirpe severa. Quando esta foi inoculada aos três dias após a proteção, ela se estabeleceu em algumas plantas, moveu-se sistemicamente e sobrepôs a estirpe fraca, uma vez que as plantas exibiram sintomas severos. Com o passar do tempo (seis e nove dias após a proteção), todas as plantas ficaram protegidas contra a expressão dos sintomas severos da estirpe desafiante, porém esta foi capaz de se estabelecer nas folhas inoculadas e até mesmo mover sistemicamente em algumas plantas. No caso da proteção com a estirpe fraca ZYMV-M, só ou em mistura com a estirpe fraca PRSV-W-1, contatou-se que plantas de abobrinha-de-moita premunizadas e desafiadas sete dias depois ficaram protegidas contra a infecção e/ou manifestação dos sintomas das respectivas estirpes severas usadas no desafio.
Papaya ringspot virus - type W (PRSV-W) and Zucchini yellow mosaic virus (ZYMV) are two potyviruses associated with severe yield losses on cucurbit crops in the State of São Paulo and other parts of the Brazilian territory. Preimmunization with mild strains has proved to be a good alternative for the control of both viruses in susceptible cultivars. The main purpose of this work was to evaluate the competition for infectable sites as a possible mechanism of cross protection between the mild strain PRSV-W-1 and the severe strain PRSV-W-C in zucchini squash (Cucurbita pepo cv. Caserta). Protective inoculation with the mild strain was done at the cotyledon and the challenge inoculation with the severe strain was applied on the first true expanded leaf, and viceversa. Different plants were challenged at three, six and nine days, respectively. No challenge protected plants and healthy plants infected with the severe strain were used as controls. Evaluations were based on the expression of the symptoms at 30 days after challenge inoculation. Attempts to recover the challenge strain from challenge inoculated and new developed leaves were also done at eight days after challenge inoculation. RT-PCR with specific pairs of primers was also used to detect both stains in some of these samples. Regardless the leaf on which the protective strain was applied (cotyledon or first true expanded leaf) there appear to be some infectable sites available for superinfection with the severe strain. When the challenge inoculation was done at three days after preimmunization, the severe strain was able to superinfect some plants, move systemically and overcome the mild strain, since these plants expressed severe symptoms. All plants become protected against the expression of the symptoms induced by the severe strain when the challenge inoculation was done at six and nine days after preimmunization. However, the severe strain was still detected in the inoculated and upper leaves of few test-plants, eight days after challenge inoculation. In addition to this, it was also determined the period of time necessary to protect zucchini squash plants with a mild strain of ZYMV, named ZYMV-M, alone and in mixture with mild strain PRSV-W-1. The results indicated that single and double preimmunized plants were protected against infection and/or expression of the homologous severe strain seven days after protective inoculation.
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