Navegando por Autor "Alves, Murilo S."
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Item Complete inventory of soybean NAC transcription factors: Sequence conservation and expression analysis uncover their distinct roles in stress response(Gene, 2009-09-01) Pinheiro, Guilherme L.; Marques, Carolina S.; Costa, Maximiller D.B.L.; Reis, Pedro A. B.; Alves, Murilo S.; Carvalho, Claudine M.; Fietto, Luciano G.; Fontes, Elizabeth P. B.We performed an inventory of soybean NAC transcription factors, in which 101 NAC domain-containing proteins were annotated into 15 different subgroups, showing a clear relationship between structure and function. The six previously described GmNAC proteins (GmNAC1 to GmNAC6) were located in the nucleus and a transactivation assay in yeast confirmed that GmNAC2, GmNAC3, GmNAC4 and GmNAC5 function as transactivators. We also analyzed the expression of the six NAC genes in response to a variety of stress conditions. GmNAC2, GmNAC3 and GmNAC4 were strongly induced by osmotic stress. GmNAC3 and GmNAC4 were also induced by ABA, JA and salinity but differed in their response to cold. Consistent with an involvement in cell death programs, the transient expression of GmNAC1, GmNAC5 and GmNAC6 in tobacco leaves resulted in cell death and enhanced expression of senescence markers. Our results indicate that the described soybean NACs are functionally non-redundant transcription factors involved in response to abiotic stresses and in cell death events in soybean.Item Differential expression of four soybean bZIP genes during Phakopsora pachyrhizi infection(Functional & Integrative Genomics, 2015-05-27) Alves, Murilo S.; Soares, Zamira G.; Vidigal, Pedro M. P.; Barros, Everaldo G.; Poddanosqui, Adriana M. P.; Aoyagi, Luciano N.; Abdelnoor, Ricardo V.; Marcelino-Guimarães, Francismar C.; Fietto, Luciano G.Asian soybean rust (ASR), caused by the obligate biotrophic fungus Phakopsora pachyrhizi, is one of most important diseases in the soybean (Glycine max (L.) Merr.) agribusiness. The identification and characterization of genes related to plant defense responses to fungal infection are essential to develop ASR-resistant plants. In this work, we describe four soybean genes, GmbZIP62, GmbZIP105, GmbZIPE1, and GmbZIPE2, which encode transcription factors containing a basic leucine zipper (bZIP) domain from two divergent classes, and that are responsive to P. pachyrhizi infection. Molecular phylogenetic analyses demonstrated that these genes encode proteins similar to bZIP factors responsive to pathogens. Yeast transactivation assays showed that only GmbZIP62 has strong transactivation activity in yeast. In addition, three of the bZIP transcription factors analyzed were also differentially expressed by plant defense hormones, and all were differentially expressed by fungal attack, indicating that these proteins might participate in response to ASR infection. The results suggested that these bZIP proteins are part of the plant defense response to P. pachyrhizi infection, by regulating the gene expression related to ASR infection responses. These bZIP genes are potential targets to obtain new soybean genotypes resistant to ASR.Item Early responsive to DEHYDRATION 15, a new transcription factor that integrates stress signaling pathways(Plant Signaling & Behavior, 2011-12-01) Alves, Murilo S.; Fontes, Elizabeth P.B.; Fietto, Luciano G.The Early Responsive to Dehydration (ERD) genes are defined as those genes that are rapidly activated during drought stress. The encoded proteins show a great structural and functional diversity, with a particular class of proteins acting as connectors of stress response pathways. Recent studies have shown that ERD15 proteins from different species of plants operate in cross-talk among different response pathways. In this mini-review, we show the recent progress on the functional role of this diverse family of proteins and demonstrate that a soybean ERD15 homolog can act as a connector in stress response pathways that trigger a programmed cell death signal.Item Expressão gênica do colágeno em ferida cutânea de equinos tratada com plasma rico em plaquetas(Pesquisa Veterinária Brasileira, 2014-03-25) Souza, Maria V. de; Pinto, José de O.; Costa, Marcela B.M. da; Alves, Murilo S.; Silva, Micheline O. da; Martinho, Karina O.; Fietto, Luciano G.O plasma rico em plaquetas (PRP) é um produto derivado da centrifugação do sangue total, cuja utilização concentra-se em melhorar a reparação de diferentes tecidos, tendo em vista os fatores de crescimento nele contido. Entretanto, os benefícios da terapia no contexto clínico ainda não estão totalmente esclarecidos. Objetivou-se avaliar a expressão dos genes dos colágenos tipos I e III durante diferentes fases do processo de cicatrização da pele tratada com PRP. Foram utilizados oito equinos machos castrados, mestiços, hígidos, com idade entre 16 e 17 (16,37±0,52) anos. Três feridas em formato quadrangular (6,25cm2) foram confeccionadas nas regiões glúteas direita e esquerda de todos os animais. Doze horas após indução das lesões, 0,5mL do PRP foi administrado em cada uma das quatro extremidades das feridas (T=grupo tratado), de uma das regiões glúteas, escolhida aleatoriamente. A região contralateral foi utilizada como controle (NT=grupo não tratado). As feridas foram submetidas à limpeza diária com água Milli Q, e amostras foram obtidas com biópsias utilizando-se Punch de 6mm de diâmetro. Seis biópsias de pele foram obtidas a primeira no dia de indução das lesões (T0), e as demais com 1 (T1) 2 (T2) 7 (T3) e 14 (T4) dias após a realização das feridas. A sexta biópsia (T5) foi realizada após o completo fechamento da pele. A avaliação da expressão dos genes dos colágenos tipos I e III foi realizada pela técnica qRT-PCR e os dados analisados pelo teste de Bonferroni, t de Student, t pareado e análise de regressão (p<0,05). Diferenças (p<0,05), entre grupos, foram observadas para a expressão de ambos os colágenos nos T1 a T4, sendo maior nos animais do grupo T. O pico de expressão dos colágenos tipos I e III ocorreu no T5 para ambos os grupos, mas a maior expressão foi diferente (p<0,05) do tempo zero a partir do T3. Nos animais do grupo tratado a expressão dos colágenos começou a estabilizar no T5, enquanto que nos equinos do NT os valores permaneceram elevados. A administração local de uma única dose do PRP em ferida cutânea na região glútea de equinos, resulta em maior expressão gênica local dos colágenos tipos I e III. Entretanto, essa expressão não altera o tempo máximo de fechamento macroscópico da ferida.Item A novel transcription factor, ERD15 (Early Responsive to Dehydration 15), connects Endoplasmic Reticulum stress with an osmotic stress-induced cell death signal(The Journal of Biological Chemistry, 2011-04-11) Alves, Murilo S.; Reis, Pedro A. B.; Dadalto, Silvana P.; Faria, Jerusa A. Q. A.; Fontes, Elizabeth P. B.; Fietto, Luciano G.As in all other eukaryotic organisms, endoplasmic reticulum (ER) stress triggers the evolutionarily conserved unfolded protein response in soybean, but it also communicates with other adaptive signaling responses, such as osmotic stress-induced and ER stress-induced programmed cell death. These two signaling pathways converge at the level of gene transcription to activate an integrated cascade that is mediated by N-rich proteins (NRPs). Here, we describe a novel transcription factor, GmERD15 (Glycine max Early Responsive to Dehydration 15), which is induced by ER stress and osmotic stress to activate the expression of NRP genes. GmERD15 was isolated because of its capacity to stably associate with the NRP-B promoter in yeast. It specifically binds to a 187-bp fragment of the NRP-B promoter in vitro and activates the transcription of a reporter gene in yeast. Furthermore, GmERD15 was found in both the cytoplasm and the nucleus, and a ChIP assay revealed that it binds to the NRP-B promoter in vivo. Expression of GmERD15 in soybean protoplasts activated the NRP-B promoter and induced expression of the NRP-B gene. Collectively, these results support the interpretation that GmERD15 functions as an upstream component of stress-induced NRP-B-mediated signaling to connect stress in the ER to an osmotic stress-induced cell death signal.