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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2022.tde-13092022-113950
Document
Author
Full name
Nicole Orsi
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2022
Supervisor
Committee
Vieira, Maria Lucia Carneiro (President)
Camargo, Luis Eduardo Aranha
Pinheiro, Daniel Guariz
Title in Portuguese
Perfil transcricional de genótipos de feijoeiro-comum contrastantes para a resposta à infecção por Meloidogyne incognita
Keywords in Portuguese
Meloidogyne incognita
Phaseolus vulgaris
Genes diferencialmente expressos
Resistência moderada
RNA-Seq
Transcriptoma
Abstract in Portuguese
O feijoeiro-comum (Phaseolus vulgaris L., Fabaceae) é uma cultura anual herbácea, de importância econômica e social, cujos grãos proveem uma das principais fontes de proteína de origem vegetal. Problemas fitossanitários causam severos impactos sobre o cultivo de feijoeiro. Dentre os patógenos mais frequentes e danosos à cultura do feijoeiro estão os do gênero Meloidogyne, chamados 'nematoides das galhas' (RKN), com destaque para M. incognita, amplamente disseminado nas regiões produtoras brasileiras. É consenso que a base genética e molecular da resistência/suscetibilidade de plantas aos RKNs é complexa e o seu entendimento demanda o emprego de variadas abordagens. Por exemplo, a análise do transcritoma pode ser uma excelente estratégia para a identificação de genes envolvidos na resposta do feijoeiro a este patógeno. O objetivo do presente trabalho foi avaliar o perfil transcricional de genótipos de feijoeiro contrastantes para a resposta à M. incognita (IAC-Tybatã, resistente vs. Branquinho, suscetível) e, assim, identificar os genes diferencialmente expressos em dois tempos durante a interação (4 e 10 dias após a inoculação, DAI). No total, foram 8 tratamentos, com 3 repetições, gerando 24 bibliotecas de RNA que foram construídas e sequenciadas. Foi obtido um total de 587 milhões de reads. Em média, considerando ambos os genótipos, o alinhamento das sequências ao genoma de referência produziu 537 milhões de reads (~92%). Foram encontrados 4.862 e 4.835 genes diferencialmente expressos em IAC-Tybatã (R) e Branquinho (S), respectivamente. Foi realizada a anotação funcional de 6.938 genes (89%) pela plataforma Blast2GO, com o maior número de genes atribuídos às categorias funcionais "atividade catalítica e ligação", "processos metabólicos e celulares" e "membrana". Usando o banco de dados KEGG, as categorias funcionais "biossíntese de metabólitos secundários", "processos celulares" e "sinalização" apresentaram-se mais enriquecidas em IAC-Tybatã (R) comparativamente a Branquinho (S), onde a categoria "metabolismo de carboidratos" se mostrou mais enriquecida. Mudanças nos níveis de expressão gênica em resposta à infecção por M. incognita foram identificadas em ambos os genótipos. Por exemplo, genes que codificam proteínas putativas de resistência a doenças com domínios LRR, proteínas quinases, citocromos, fatores de transcrição myb e WRKY e receptores de membrana mostraram-se superexpressos no genótipo resistente. Por outro lado, genes envolvidos nas vias hormonais, com destaque para a via da auxina, mostraram-se superexpressos no genótipo suscetível, e genes que codificam fatores de transcrição (WRKY, myb e MYC2), proteínas com domínios ANK, proteínas com repetições tetratricopeptídicas e beta glucosidases mostraram-se reprimidos. Um modelo foi proposto na tentativa de elucidar as respostas do feijoeiro-comum à infecção por M. incognita.
Title in English
Transcriptional profiling of common bean contrasting genotypes in response to Meloidogyne incognita infection
Keywords in English
Meloidogyne incognita
Phaseolus vulgaris
Differentially expressed genes
Moderate resistance
RNA-Seq
Transcriptome
Abstract in English
The common bean (Phaseolus vulgaris L., Fabaceae) is an annual herbaceous crop of economic and social importance whose grains provide one of the main sources of plant proteins. Phytosanitary problems may cause severe impacts on common bean cultivation. Of the most common and harmful pathogens to the common bean crop are those of Meloidogyne genus, known as 'root-knot nematodes' (RKN), predominantly M. incognita, which is widespread in Brazilian agricultural areas. It is a consensus that the genetic and molecular basis of plant resistance/susceptibility to RKNs is complex and its understanding requires the use of diverse approaches. For instance, the transcriptome analysis may be an excellent strategy for the identification of genes involved in the common bean response to this pathogen. The objective of the present study was to evaluate the transcriptional profile of common bean genotypes with contrasting responses against M. incognita (IAC-Tybatã, resistant vs. Branquinho, susceptible) in order to identify differentially expressed genes at two different time points during this interaction (4 and 10 days after the inoculation, DAI). In total, there were 8 treatments, with 3 replicates, generating 24 RNA libraries that were constructed and sequenced. We have obtained 587 million reads. On average, taking into account both genotypes, the sequence alignment to the reference genome produced 537 million reads (~92%). A total of 4,862 and 4,835 differentially expressed genes were found in IAC-Tybatã and Branquinho, respectively. The functional annotation of 6,938 genes (89%) was performed using the Blast2GO platform, with the largest number of genes assigned to the functional categories "catalytic activity and binding", "metabolic and cellular processes" and "membrane". Using KEGG database resource, the functional categories "biosynthesis of secondary metabolites" and "cellular processes" and "signaling" were the most enriched in IAC-Tybatã (R) in comparison to Branquinho, in which "carbohydrate metabolism" was the most enriched category. Genes showing changes in expression levels under M. incognita infection were identified in both genotypes. For instance, genes coding for disease resistance proteins with LRR domains, protein kinases, cytochromes, myb and WRKY transcription factors and receptors-like kinases were overexpressed in the resistant genotype. On the other hand, genes involved in hormonal pathways, namely auxin pathway, were overexpressed in the susceptible genotype, and genes coding for transcription factors (WRKY, myb and MYC2), proteins with ANK domains, proteins with tetratricopeptide repeats and beta glucosidases were repressed. A tentative model was proposed to explain the responses of common bean to M. incognita infection.
 
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Release Date
2024-07-05
Publishing Date
2022-09-14
 
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