Identificação de genes candidatos para resistência ao mosaico da cana-de-açúcar a partir da técnica de cDNA-AFLP
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Data
2013-12-10
Autores
Medeiros, Cibele Nataliane Facioli [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A doença do mosaico da cana-de-açúcar, causada pelo Sugarcane mosaic virus (SCMV), é uma das principais doenças incidentes na cultura. Para compreender a base molecular dessa interação planta-patógeno o presente trabalho objetivou identificar potenciais genes candidatos à resistência ao mosaico através da técnica de cDNA-AFLP utilizando duas variedades contrastantes quanto a resistência ao SCMV (estirpe Rib-1). Plantas das variedades IACSP95-5000 (resistente) e IAC91-1099 (suscetível) obtidas por micromeristemas foram inoculadas com a estirpe Rib-1 em condições controladas de casa de vegetação para a identificação de fragmentos diferencialmente expressos (FDE) nos tempos de 24, 48 e 72 horas após a inoculação (hpi). No total, foram obtidos 772 FDEs, dos quais 392 observados na variedade resistente e 380 na suscetível. As variedades apresentaram comportamento diferencial quanto ao número de FDEs induzidos e reprimidos ao longo dos tempos de amostragens (24, 48 e 72 hpi). A variedade resistente apresentou um número maior de FDEs, correspondentes a prováveis genes induzidos no tempo de 24 hpi em relação a suscetível, diminuindo com o passar do tempo de inoculação, enquanto o número de FDEs correspondentes a prováveis genes induzidos aumentou com o tempo de inoculação na variedade suscetível. Sessenta e seis FDEs foram identificados exclusivamente na variedade resistente, sendo que 9 foram reprimidos e 57 induzidos após a inoculação com o SCMV. Destes 57, 23 foram sequenciados e destes, 10 FDEs mostraram identidade com sequências conhecidas de plantas, sendo a maioria relacionada com mecanismos de defesa da planta contra o patógeno e 3 FDEs não apresentam identidades com sequências descritas e podem exercer papel importante no mecanismo de resistência. A técnica de cDNA-AFLP foi eficiente em revelar alterações do perfil de transcrição dentro...
The sugarcane mosaic disease caused by the Sugarcane Mosaic Virus (SCMV) is a major disease incident in this crop. To understand the molecular basis of the plant pathogen interaction, the present study aimed to identify potential candidate genes for mosaic resistance by cDNA-AFLP technique using two contrasting varieties for SCMV (Rib-1 strain) resistance. Plants of the varieties IACSP95-5000 (resistant) and IAC91-1099 (susceptible) obtained by micro-meristem were inoculated with the Rib-1 strain under greenhouse controlled conditions to the identification of differentially expressed fragments (DEF) at the sampling time points of 24, 48 and 72 hours after inoculation (hpi). In total, 772 DEFs were obtained, of which 392 were observed in the resistant variety and 380 in the susceptible. The varieties showed differential behavior in the number of induced and repressed DEFs during the sampling time points (24, 48 and 72 hpi). The resistant variety produced a higher number of DEFs at 24 hpi (probably corresponding to induced genes) decreasing over the time of inoculation, when compared to the susceptible, in which the number of DEFs increased over the hpi. Sixty-six DEFs were identified exclusively in the resistant variety, of which 9 were repressed and 57 induced after inoculation with SCMV. Of these 57, 23 were sequenced and 10 of them showed identity with known plant sequences, mostly related to mechanisms of plant defense against pathogen, while 3 DFEs have no identities with sequences described in the bank and probably may play an important role in the resistance mechanism. The cDNA-AFLP technique was effective in revealing changes in the transcription profile within and between contrasting varieties when challenged by the mosaic virus. The total set of DFEs can be an important tool in studies to understand the resistance mechanism, as well as for the development of molecular markers to be ...
The sugarcane mosaic disease caused by the Sugarcane Mosaic Virus (SCMV) is a major disease incident in this crop. To understand the molecular basis of the plant pathogen interaction, the present study aimed to identify potential candidate genes for mosaic resistance by cDNA-AFLP technique using two contrasting varieties for SCMV (Rib-1 strain) resistance. Plants of the varieties IACSP95-5000 (resistant) and IAC91-1099 (susceptible) obtained by micro-meristem were inoculated with the Rib-1 strain under greenhouse controlled conditions to the identification of differentially expressed fragments (DEF) at the sampling time points of 24, 48 and 72 hours after inoculation (hpi). In total, 772 DEFs were obtained, of which 392 were observed in the resistant variety and 380 in the susceptible. The varieties showed differential behavior in the number of induced and repressed DEFs during the sampling time points (24, 48 and 72 hpi). The resistant variety produced a higher number of DEFs at 24 hpi (probably corresponding to induced genes) decreasing over the time of inoculation, when compared to the susceptible, in which the number of DEFs increased over the hpi. Sixty-six DEFs were identified exclusively in the resistant variety, of which 9 were repressed and 57 induced after inoculation with SCMV. Of these 57, 23 were sequenced and 10 of them showed identity with known plant sequences, mostly related to mechanisms of plant defense against pathogen, while 3 DFEs have no identities with sequences described in the bank and probably may play an important role in the resistance mechanism. The cDNA-AFLP technique was effective in revealing changes in the transcription profile within and between contrasting varieties when challenged by the mosaic virus. The total set of DFEs can be an important tool in studies to understand the resistance mechanism, as well as for the development of molecular markers to be ...
Descrição
Palavras-chave
Cana-de-açúcar - Doenças e pragas, Cana-de-açúcar - Resistência a doenças e pragas, Virus de plantas, Genes, Expressão gênica, Sugarcane
Como citar
MEDEIROS, Cibele Nataliane Facioli. Identificação de genes candidatos para resistência ao mosaico da cana-de-açúcar a partir da técnica de cDNA-AFLP. 2013. iv, 46 p. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal, 2013.