No description available.

No description available.

Cassava mosaic disease (CMD) is caused by whitefly-transmitted geminiviruses and is a major constraint to cassava production in Africa. Field surveys were conducted in three (Bushbuckridge, Mariti and Tonga) cassava growing areas of Limpopo and Mpumalanga provinces in South Africa during two seasons (2004/2005 and 2005/2006). Results showed that a higher percentage (27.1%) of CMD infection was due to the use of infected planting materials compared to whitefly borne-infections (10.4%). Disease symptoms were generally mild. There was no change in disease incidence over the survey period. Molecular characterization of cassava mosaic geminiviruses (CMG’s), using differential primer PCR, restriction fragment length polymorphisms (RFLP’s), phylogenetic and recombination analysis and screening for satellite DNA’s. Differential primer PCR and RFLP’s showed that African cassava mosaic virus (ACMV) was the most prevalent virus in South Africa and that mixed infections were a common occurrence. Phylogenetic analysis and RFLP’s showed the presence of a ‘new’ strain of ACMV in South Africa. EACMV isolates from this study showed more frequent recombination compared to ACMV isolates. None of the samples tested positive for satellite DNA’s. Phylogenetic analysis of Bemisia tabaci using the mitochondrial cytochrome oxidase gene sequences revealed a ‘new’ sister clade of B. tabaci that is closely related to the previously identified southern African clade and the presence of the Q biotype that groups with Q biotypes of North African/Mediterranean origin. Good cultural practices, introduction of resistant cultivars and continuous monitoring are required to reduce the impact of CMD in South Africa.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-05-02T12:52:56Z No. of bitstreams: 1 texto completo.pdf: 718578 bytes, checksum: eabef42b1aa937ca451902a3e9e0adbf (MD5) / Made available in DSpace on 2017-05-02T12:52:56Z (GMT). No. of bitstreams: 1 texto completo.pdf: 718578 bytes, checksum: eabef42b1aa937ca451902a3e9e0adbf (MD5) Previous issue date: 2004-05-12 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A família Geminiviridae, caracterizada pela morfologia de partículas icosaédricas geminadas e genoma composto por DNA de fita simples circular, é dividida em quatro gêneros, de acordo com o tipo de inseto vetor, gama de hospedeiros, organização do genoma e relacionamento filogenético. Os begomovírus possuem dois componentes genômicos, são transmitidos por mosca-branca e infectam dicotiledôneas. A partir de 1994 houve relatos sucessivos de begomovírus em tomateiro em diversos estados brasileiros. O seqüenciamento parcial do genoma de alguns destes vírus revelou que há uma grande diversidade de espécies, provavelmente como resultado da disseminação de vírus nativos pelo biótipo B da mosca-branca Bemisia tabaci. O objetivo do presente trabalho foi a caracterização biológica e molecular de um isolado de begomovírus detectado em tomateiros do município de São Joaquim de Bicas, MG, denominado MG-Bi2. A caracterização biológica consistiu no teste de gama de hospedeiros, realizado por meio de inoculação via extrato foliar tamponado e biobalística. Os resultados indicaram que o isolado infecta plantas das famílias Solanaceae e Amaranthaceae e que a transmissão é altamente dependente do método de inoculação. Para a caracterização molecular, procedeu-se à amplificação do genoma viral (DNA-A e -B) utilizando a DNA polimerase do bacteriófago Φ29, seguida de clonagem e determinação de suas seqüências parciais de nucleotídeos. A comparação de seqüências e análise filogenética demonstraram relacionamento próximo com o genoma de Sida micrantha mosaic virus (SimMV), indicando que o isolado MG-Bi2 pertence a esta espécie, e reforçando a hipótese de que plantas daninhas e silvestres servem como reservatório natural para os geminivírus que infectam plantas cultivadas como o tomateiro. / The Geminiviridae family, characterized by a particle morphology of twinned incomplete icosahedra and a single-stranded, circular DNA genome, is divided into four genera according to the type of insect vector, host range, genomic organization and phylogeny. Begomoviruses have two genomic components and are transmitted by whiteflies to dicotyledoneous plants. Since 1994 there have been successive reports of begomovirus infection in tomatoes in several states of Brazil. Partial genome sequencing of several isolates revealed a high degree of genetic diversity, probably due to the dissemination of indigenous viruses by the B biotype of the whitefly Bemisia tabaci. The objective of the present work was the biological and molecular characterization of a begomovirus isolate detected in tomatoes in the city of São Joaquim de Bicas, MG, named MG-Bi2. Biological characterization consisted on a host range study using sap inoculation and particle bombardment. Results indicated that the isolate infects species of Solanaceae and Amaranthaceae, and that transmition is highly dependent on the inoculation method. For the molecular characterization, the full-length genome (DNA-A and -B) was amplified using the DNA polimerase of bacteriophage Φ29, cloned and partially sequenced. Sequence comparisons and phylogenetic analyses demonstrated a close relationship with Sida micrantha mosaic virus (SimMV), indicating that the isolate MG-Bi2 is a member of this species, and reinforcing the hypothesis that weeds and wild plants could act as a natural reservoir of indigenous begomoviruses which can infect crop plants such as the tomato. / Dissertação importada do Alexandria

The whitefly Bemisia tabaci (Gennadius) (Hemiptera) complex is the sole arthropod vector of the genus, Begomovirus (family, Geminiviridae), which causes debilitating diseases of plants, worldwide. Virus-vector specificity is conferred through co-evolved, whitefly vector-viral capsid protein-protein interactions. Membrane-bound receptors are thought to facilitate virion passage across the gut-hemolymph and hemolymph-salivary gland interfaces, and virion circulation is expected to elicit innate defense and stress-related proteins. Our goal was to select and validate genes involved in whitefly-mediated transmission. Whitefly expressed sequence tags (ESTs) from a previous study were re-annotated, taking advantage of newly available insect EST, UniGene, and Protein sequences. Six whitefly genes and transcripts, actin, cyclophilin, GBLP, GAPDH 3, knottin, and whitefly endosymbiont HSP60, representing three gene ontology (GO) categories, were analyzed using PCR or RT- PCR, respectively, followed by cloning and DNA sequencing. Analysis confirmed the presence of all six whitefly genes and five transcripts, with the knottin transcript being undetectable.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2015-11-09T13:25:54Z No. of bitstreams: 1 texto completo.pdf: 5304872 bytes, checksum: c7e0a87885d9d7ca4c2ef6d55611b704 (MD5) / Made available in DSpace on 2015-11-09T13:25:54Z (GMT). No. of bitstreams: 1 texto completo.pdf: 5304872 bytes, checksum: c7e0a87885d9d7ca4c2ef6d55611b704 (MD5) Previous issue date: 2014-02-26 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Vírus pertencentes à família Geminiviridae possuem genoma circular de fita simples e infectam uma ampla gama de hospedeiros causando perdas significativas, principalmente em países tropicais e subtropicais. A família é dividida em sete gêneros (Begomovirus, Becurtovirus, Curtovirus, Eragrovirus, Mastrevirus, Topocuvirus e Turncurtovirus) de acordo com o tipo de inseto vetor, gama de hospedeiros, organização genômica e filogenia. Begomovírus são transmitidos por mosca-branca e causam epidemias em culturas economicamente importantes em todo o mundo. Plantas silvestres/não-cultivadas possuem um papel epidemiológico importante, agindo como reservatórios de begomovírus e como hospedeiros promíscuos onde recombinação pode ocorrer. Além disso, resultados anteriores sugerem que populações de begomovírus presentes em plantas não-cultivadas possuem maior variabilidade genética do que aquelas presentes em plantas cultivadas. A maioria dos begomovírus do Novo Mundo (NM) possuem dois componentes genômicos denominados DNA-A e DNA-B, e seu genoma tem a capacidade de evoluir rapidamente via mutação, pseudo-recombinação e recombinação. Neste contexto, este trabalho objetivou: (i) avaliar o efeito do hospedeiro na variabilidade genética de populações de begomovírus; e (ii) estudar o efeito de recombinação na evolução de begomovírus no Novo Mundo. Para o primeiro objetivo, duas plantas leguminosas cultivadas (feijão-comum, Phaseolus vulgaris, e fava, P. lunatus) e uma não-cultivada (Macroptilium lathyroides) foram intensamente amostradas em duas regiões no Brasil entre 2005 e 2012. Um total de 212 genomas (DNA-A) foram clonados e sequenciados, e populações dos begomovírus Bean golden mosaic virus (BGMV) e Macroptilium yellow spot virus (MaYSV) foram obtidas a partir dos três hospedeiros. Os resultados indicam que a variabilidade genética presumida dos hospedeiros não afetou a variabilidade viral. O MaYSV (N = 99) apresentou maior variabilidade genética que o BGMV (N = 147), com a população de BGMV (porém não de MaYSV) estruturada por hospedeiro e região geográfica. Para o segundo objetivo, conjuntos de dados incluindo todas as sequências- referência de DNA-A e DNA-B de begomovírus do NM foram obtidas do GenBank. As análises indicaram que begomovírus sul-americanos não exibem monofilia geográfica, com evidência de pelo menos dois eventos independentes de introdução nesta região. Recombinação foi detectada como um importante mecanismo evolutivo agindo sobre a evolução do DNA-A. Encontrou-se evidência de elevada variabilidade genética em begomovírus presentes na América Central e Caribe, e em menor grau em begomovírus na América do Sul (AS). Novos eventos de introdução podem acelerar a evolução desses patógenos na AS, favorecendo a recombinação e aumentando a adaptabilidade e/ou a virulência dos begomovírus nativos. / Viruses belonging to the family Geminiviridae have circular ssDNA genomes and infect a broad range of plant species causing devastating diseases, mainly in subtropical and tropical countries. The family is divided into seven genera (Begomovirus, Becurtovirus, Curtovirus, Eragrovirus, Mastrevirus, Topocuvirus and Turncurtovirus) according to the type of insect vector, host range, genome organization and phylogeny. Begomoviruses are whitefly- transmitted and are among the most damaging pathogens causing epidemics in economically important crops worldwide. Wild/non-cultivated plants play a crucial epidemiological role, acting as begomovirus reservoirs and as "mixing vessels" where recombination can occur. Furthermore, previous results suggest that begomovirus populations in non-cultivated hosts are more genetically variable then those infecting cultivated hosts. Most New World (NW) begomoviruses have two genomic components designated as DNA-A and DNA-B, and their genomes have the capacity to evolve quickly via mutation, reassortment and recombination. In this context, this work aimed: (i) to assess the effect of the host on the standing genetic variability of begomovirus populations; and (ii) to study the effect of recombination on the evolution of New World begomoviruses. For the first objective, cultivated (common bean, Phaseolus vulgaris, and lima bean, P. lunatus) and non-cultivated (Macroptilium lathyroides) legume hosts were intensively sampled from two regions across Brazil between 2005 and 2012. A total of 212 full-length DNA-A components were cloned and sequenced, and populations of the begomoviruses Bean golden mosaic virus (BGMV) and Macroptilium yellow spot virus (MaYSV) were obtained from the three hosts. Our results indicate that the presumed genetic variability of the host did not affect viral variability. MaYSV (N = 99) showed higher genetic variability than BGMV (N = 147), with the BGMV (but not the MaYSV) population being structured based on both host and geography. For the second objective, datasets including all DNA-A and DNA-B reference sequences of NW begomoviruses were obtained from GenBank. Our analyses indicate that South American begomoviruses do not exhibit geographic monophyly, with evidence of at least two independent introduction events in this region. Recombination was detected as a very important evolutionary mechanism acting on DNA-A evolution. We found evidence of greater genetic variability in begomoviruses from Central America and the Caribbean compared to South America (SA). Additional introduction events may impact the evolution of begomoviruses in SA by favoring recombination and introducing new virulence features to indigenous South American begomoviruses.

No description available.

Doenças causadas por virus do gênero Begomovirus (família Geminiviridae) são incomuns em espécies de passifloras. Nos últimos dez anos, entretanto, foram encontrados no Brasil begomovirus infectando passifloras em pomares nos municipios de São Fidelis (RJ), Paragominas (PA), Patos de Minas (MG) e Araguari (MG). Estes isolados foram transmitidos mecanicamente para plantas de Nicotiana benthamiana, que apresentaram sintomas de mosaico e deformação foliar. Também foi possível a transmissão para plantas de Sida rhombifolia através de inoculação por biobalística com o DNA amplificado do isolado de São Fidelis. Estas plantas demonstraram sintomas de mosaico amarelo e deformação foliar. O DNA total extraído de plantas infectadas foi amplificado por RCA, sendo que o componente A (DNA-A) dos isolados de Paragominas e Patos de Minas foram sequenciados diretamente por \"primer walking\". O DNA-A dos isolados de São Fidelis e Araguari foram clonados e sequenciados. As sequências de nucleotideos dos isolados de Paragominas e de São Fidelis apresentaram 90% de similaridade ao Sida mottle virus (SiMoV), enquanto a sequência de nucleotídeos do isolado de Araguari apresentou 96% de similaridade ao Sida micrantha mosaic virus (SimMV). Assim esses isolados encontrados em maracujazeiro podem ser considerados estirpes do SiMoV e SimMV, respectivamente. Não foi possivel a transmissão desses isolados através de Bemisia tabaci biótipo B, no entanto, os insetos foram capazes de adquirir o vírus. O isolado de São Fidelis foi detectado separadamente na região onde se encontra a glandula salivar (cabeça e protórax) e na região posterior do inseto, indicando que o vírus transpos a barreira do mesenteron e circulou pela hemolinfa do inseto. Alves (2008) obteve uma forma atenuada do isolado de São Fidelis através de inoculações mecanicas em plantas de N. benthamiana. O DNA-A desta forma atenuada foi sequênciado e apresentou 90% de identidade ao isolado do qual se originou. A forma atenuada do begomovirus foi capaz de proteger plantas de maracujazeiro contra a estirpe severa do vírus. / Diseases caused by begomoviruses (family Geminiviridae) are hardly found in Passiflora species. In the last years, however, begomovirus infected passionflowers were found in orchards in the counties of São Fidelis (state of Rio de Janeiro), Paragominas (Pará), Araguari and Patos de Minas (Minas Gerais). These isolates were mechanically transmitted to Nicotiana benthamiana plants, which showed variable symptoms of mosaic and leaf distortion. Another susceptible host is Sida rhombifolia, which was biolistic inoculated with amplified DNA of São Fidelis isolate, and showed symptoms of yellow mosaic and leaf distortion. Total DNA extracted from field infected passiflora was amplified by RCA, and the DNA-A of Paragominas and Patos de Minas isolates were directly sequenced by primer walking. The A component of São Fidelis and Araguari isolates were cloned and also completely sequenced. The complete nucleotide sequence of DNA-A of Araguari isolate shared 96% identity with that of Sida micrantha mosaic virus (SimMV), whereas the DNA-A of Paragominas and São Fidelis isolates shared 90% identity with that of Sida mottle virus (SiMoV). These viruses may be consider as strains of SiMoV and SimMV, respectively. It was not possible to transmit these isolates by Bemisia tabaci biótipo B, although the insects were able to acquire the virus. São Fidelis isolate could be detected separately at salivary gland region and posterior region of the insect, indicating that the virus could cross the digestive tract and circulate in the hemolymph. Alves (2008) obtained a mild strain of São Fidelis isolate by mechanical inoculation in N. benthamiana plants. The mild isolate was able to protect passionflower against the severe isoalte of this begomovirus.

Les épidémies virales constituent une menace majeure pour les cultures dans de nombreuses régions tropicales et subtropicales du monde. En Afrique de l'Ouest et Centrale, malgré la caractérisation d'une multitude de nouvelles espèces de géminivirus, les connaissances sur l'épidémiologie des maladies virales émergentes restent incomplètes, notamment concernant la diversité et l'identification précise des espèces virales incriminées. Paradoxalement, si l'on considère que la plupart des maladies émergentes causées par les géminivirus sont liées à des virus infectant naturellement les plantes sauvages, et qui se seraient adaptés aux plantes cultivées introduites, la diversité des populations virales dans leurs écosystèmes naturels reste très largement méconnue. De plus, le caractère non pérenne de la majorité des plantes maraîchères suggère l'existence de plantes hôtes alternatives ou réservoirs, qui permettent aux populations virales de se maintenir lors des périodes d'intercultures. Par conséquent, pour lutter contre ces nouvelles maladies virales dévastatrices, il est primordial d'acquérir de meilleures connaissances de la diversité des populations virales des plantes cultivées et sauvages, de leurs répartitions géographiques, de leurs liens phylogénétiques, de leurs pathogénicités et des principaux facteurs épidémiologiques à ''échelle des agro-écosystèmes. Ainsi, les travaux entrepris dans cette thèse ont porté sur l'analyse d’échantillons de plantes maraichères et non cultivées provenant de 48 localités réparties sur le territoire du Burkina Faso et de Côte d'Ivoire. Nos principaux résultats confirment à la fois la non-exhaustivité de notre connaissance de la diversité des géminivirus présent sur les cultures maraîchères, avec l'existence d’au moins cinq espèces de géminivirus impliqués dans la maladie du ToLCD-TYLCD, d'un gradient Nord-Sud de complexité des espèces de géminivirus au Burkina Faso, la prédominance du PepYVMLV sur les solanées cultivées et la présence de nombreuses plantes alternatives ou réservoirs. Au regard de la découverte d'un nouveau composant d'ADN-B en association avec le PepYVMLV, les principaux paramètres du pouvoir pathogène du PepYVMLV en infection simple ou mixte avec le composant d'ADN-B ont été évalués en conditions contrôlées. Nos résultats démontrent que même si le PepYVMLV n'est pas un bégomovirus bipartite strict, l'ADN-B représente à la fois un activateur fort de la virulence, de l'accumulation du virus dans la plante et de la transmission par son insecte vecteur Bemisia tabaci, qui semblent concourir à sa prédominance sur le terrain. Une diffusion plus large de ce composant d'ADN-B représenterait une menace majeure pour la culture de la tomate au Burkina Faso, en Afrique et plus largement dans le monde. L'élargissement des analyses par l'utilisation d'une approche métagénomique à non seulement permis de confirmer la présence d'une majorité des virus décrits préalablement avec les approches classiques, mais également la caractérisation de nouvelles espèces virales. L'analyse des réseaux d'associations virus-plantes hôtes et virus-virus ont montré des liens forts entre les populations virales associés à des groupes de genres de plantes cultivées et non cultivées soulignant la nécessité de considérer les agro-écosystèmes dans leur totalité pour lutter contre les maladies à géminivirus. L'ensemble de nos résultats soulignent l'importance des travaux d'épidémiosurveillance et d'inventaire des populations virales sur les plantes cultivées et sauvages afin de comprendre le fonctionnement des communautés virales à l'échelle des agroécosystèmes et l'impact des perturbations anthropiques sur les processus d’émergence de nouvelles maladies. / Geminiviruses have emerged to become one of the largest and most economically important groups of plant-infecting viruses, and geminivirus-induced diseases are a major threat to worldwide vegetable production, particularly in tropical and subtropical regions of the world. Importantly, the accumulated body of work on some of the most important geminiviral associated diseases clearly demonstrate the role of geminiviruses associated with wild plants on the emergence of disease on imported crops. Moreover, recent metagenomic data suggested that the vast majority of viruses characterized from crops represent only a small fraction of the phytoviruses in general. It is therefore of prime interest to obtain a better knowledge of viral diversity infecting crops and wild plants, the main epidemiological parameters involved in their emergence and their dynamic at the scale of agro-ecological systems. In this work, a survey of solanaceous crop fields and their surrounding uncultivated plants from 48 localities in Burkina Faso and Ivory Coast was performed. The sample analysis using classical molecular biology tools both confirm the incompleteness of our knowledge of the geminivial diversity and the existence of numerous alternative wild host plants. At least five species of begomovirus and mastrevirus were found in association with the ToLCD-TYLCD. A North to South increasing gradient of complexity of viruses populations was uncover with PepYVMLV being the most prevalent on cultivated solanaceous plants. The discovery of the association of a newly described DNA-B component with the PepYVMLV also lead to the study of the epidemiological parameters of this co-infection. Despite this association being relaxed, it was demonstrated that the virulence of the disease, the viral accumulation and the transmission by Bemisia tabaci were increased with the presence of the co-infection with the DNA-B component. All these factors are probably associated with the success of this association on the field. Because of the extreme severity of the resulting disease, the diffusion of this new DNA-B component at a larger scale would represent a major threat to tomato culture in Burkina Faso, Africa and the world in general. The use of a metagenomic approach, allow the generalization of our findings to full agro-ecological settings. Besides confirming previous species discovery, species yet undescribed in Burkina Faso along with completely new begomovirus species were described. The inspection of the virus-plant and virus-virus associations networks allow to uncover strong links existing between the viral corteges associated to groups of cultivated and uncultivated plants. These findings emphasized the necessity to consider full agro-ecological settings plant diversity rather than only crops in order to understand and prevent geminiviruses associated diseases. Globally, our results highlight the necessity to carry on the ongoing plant disease monitoring work and the inventory of viral populations associated with cultivated and uncultivated plants in order to understand the functioning of natural geminiviral community and the impact of human practices on the emergence of viral disease.

Made available in DSpace on 2015-03-26T13:36:35Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2085482 bytes, checksum: 22303ee6597c676fbc153930781c403e (MD5) Previous issue date: 2007-09-28 / Geminivirus are plant virus packed as geminate semi- icosahedral particles and single-stranded circular DNA genomes. They can be either mono or bipartite and in the bipartite geminivirus, the genomic components are designated DNA-A and DNA-B. DNA-A encodes the proteins needed for replication, transactivation and encapsidation of the viral genome. DNA-B encodes two movement proteins, Movement Protein, MP, and Nuclear Shuttle Protein, NSP, both required for the establishment of a systemic infection. NSP transports the viral DNA between the nucleus and cytoplasm and acts cooperatively with MP to move the viral DNA cell-to-cell across the wall. The geminivirus are highly dependent on the host factors for proliferation and spread. NSP interacts with a plasma membrane receptor kinase, designated NIK (NSP- interacting kinase), from tomato, soybean and Arabidopsis. It has been demonstrated previously that inactivation of NIK gene enhances the susceptibility of the mutant to geminivirus infection. To elucidate the possible role of NIK in the protection mechanism to geminivirus infection, we investigated here the effects of NIK overexpression in the tomato response to geminivirus infection. The NIK gene from Arabidopsis thaliana was used to transform tomato. The primary transformants were confirmed by PCR, selected by RT-PCR according to the transgene expression, in vitro replicated and used for developmental analysis in vitro and in green house. NIK overexpression affected the overall developmental performance of transgenic lines which display delayed and stunted growth, with lower stem, less developed root and shoot systems and a leaf curly phenotype. These transgenic line phenotypes overexpression NIK were similar to that displayed by inhibition of Brassinosteroid synthesis or signaling. Infectivity assays in tomato with ToYSV-[MG-Bi2] revealed that overexpression of NIK caused attenuation of symptom, as the transgenic lines developed less severe symptoms in comparison with the untransformed lines. These results are consistent with a protective role of NIK against viral infection and suggest that the NIK-mediated signaling communicates to some level with the brassinosteroid signaling. / Geminivírus são vírus de plantas com partículas semi- icosaédricas geminadas e genoma de DNA circular de fita simples, podendo ser mono ou bissegmentados, sendo, no último caso, os dois componentes genômicos, denominados DNA-A e DNA-B. No DNA-A, encontram-se os genes que codificam as proteínas necessárias para a replicação viral e encapsidação. O DNA-B codifica duas proteínas de movimento, Movement Protein, MP e Nuclear Shuttle Protein, NSP, ambas requeridas para o estabelecimento de uma infecção sistêmica. NSP transporta o DNA viral entre o núcleo e o citoplasma e atua cooperativamente com MP para transportar o DNA viral de célula-a-célula através da parede celular. Os geminivírus são altamente dependentes de fatores do hospedeiro para sua proliferação. A proteína NSP interage com uma quinase receptora da membrana plasmática, designada NIK (NSP-Interacting Kinase) de tomate, soja e Arabidopsis. Foi demonstrado, em estudos anteriores, que a inativação do gene NIK aumenta a suscetibilidade dos mutantes à infecção viral. Visando elucidar o possível papel de NIK no mecanismo de proteção à infecção por geminivírus, este estudo propôs uma avaliação dos efeitos da superexpressão de NIK na resposta de tomate à infecção por geminivírus. Assim sendo, o gene NIK de Arabidopsis thaliana foi utilizado para transformar tomateiro. Os transformantes primários foram confirmados por PCR, selecionados por RT- PCR, de acordo com a expressão do transgene, repicados in vitro e utilizados em experimentos de análise de desenvolvimento in vitro e em casa-de-vegetação. Análises fenotípicas demonstraram que a superexpressão da proteína promoveu retardo de crescimento dos transformantes, os quais apresentaram baixa estatura, menor comprimento caulinar, menor desenvolvimento de sistema radicular e aéreo, folhas enrugadas/encarquilhadas. O fenótipo observado nas linhagens transgênicas superexpressando NIK é muito similar àquele resultante da inibição da via de biossíntese ou sinalização de brassinosteróides. Os ensaios de infectividade em tomateiro com o vírus ToYSV-[MG-Bi2] revelaram que a superexpresão de NIK causou atenuação dos sintomas uma vez que os transformantes apresentaram menor severidade de sintomas em relação ás linhagens não transformadas. Estes resultados são consistentes com o papel protetor de NIK contra a infecção viral e sugere que a sinalização mediada por NIK comunica-se, em algum nível, com a via de sinalização de brassinosteróides.