Transcription factors expressed in soybean roots under drought stress.

To gain insight into stress-responsive gene regulation in soybean plants, we identified consensus sequences that could categorize the transcription factors MYBJ7, BZIP50, C2H2, and NAC2 as members of the gene families myb, bzip, c2h2, and nac, respectively. We also investigated the evolutionary relationship of these transcription factors and analyzed their expression levels under drought stress. The NCBI software was used to find the predicted amino acid sequences of the transcription factors, and the Clustal X software was used to align soybean and other plant species sequences. Phylogenetic trees were built using the Mega 4.1 software by neighbor joining and the degree of confidence test by Bootstrap. Expression level studies were carried out using hydroponic culture; the experiments were designed in completely randomized blocks with three repetitions. The blocks consisted of two genotypes, MG/BR46 Conquista (drought-tolerant) and BR16 (drought-sensitive) and the treatments consisted of increasingly long dehydration periods (0, 25, 50, 75, and 100 min). The transcription factors presented domains and/or conserved regions that characterized them as belonging to the bzip, c2h2, myb, and nac families. Based on the phylogenetic trees, it was found that the myb, bzip and nac genes are closely related to myb78, bzip48 and nac2 of soybean and that c2h2 is closely related to c2h2 of Brassica napus. Expression of all genes was in general increased under drought stress in both genotypes. Major differences between genotypes were due to the lowering of the expression of the mybj7 and c2h2 genes in the drought-tolerant variety at some times. Over-expression or silencing of some of these genes has the potential to increase stress tolerance

Molecular, anatomical and physiological properties of a genetically modified soybean line transformed with rd29A:AtDREB1A for the improvement of drought tolerance.

We evaluated the molecular, anatomical and physiological properties of a soybean line transformed to improve drought tolerance with an rd29A:AtDREB1A construct. This construct expressed dehydration- responsive element binding protein DREB1A from the stress-inducible rd29A promoter. The greenhouse growth test included four randomized blocks of soybean plants, with each treatment performed in triplicate. Seeds from the non-transformed soybean cultivar BR16 and from the genetically modified soybean P58 line (T2 generation) were grown at 15% gravimetric humidity for 31 days. To induce water deficit, the humidity was reduced to 5% gravimetric humidity (moderate stress) for 29 days and then to 2.5% gravimetric humidity (severe stress). AtDREB1A gene expression was higher in the genetically modified P58 plants during water deficit, demonstrating transgene stability in T2 generations and induction of the rd29A promoter. Drought-response genes, including GmPI-PLC, GmSTP, GmGRP, and GmLEA14, were highly expressed in plants submitted to severe stress. Genetically modified plants had higher stomatal conductance and consequently higher photosynthetic and transpiration rates. In addition, they had more chlorophyll. Overexpression of AtDREB1A may contribute to a decrease in leaf thickness; however, a thicker abaxial epidermis was observed. Overexpression of AtDREB1A in soybean appears to enhance drought tolerance

Soja resistente à ferrugem: participação de compostos voláteis nas respostas de defesa?

A ferrugem asiática da soja (FAS), causada por Phakopsora pachyrhizi, é uma grande ameaça à sojicultura brasileira. A avaliação das atividades de enzimas marcadoras de indução de resistência pode auxiliar na elucidação de mecanismos envolvidos em respostas de defesa das plantas. Proteína total e atividades de enzimas marcadoras da indução de resistência foram determinadas em extratos de folhas de plantas dos genótipos PI561356 (resistente, PI) e Embrapa 48 (susceptível, E48), inoculadas ou não com P. pachyrhizi. Respostas muito discrepantes entre os dois genótipos e entre os tempos de coleta (12, 72 e 168 h após inoculação) foram observadas, em especial para lipoxigenases e B-1, 3-glucanases. Diferentemente de E48, plantas de PI responderam tardiamente à inoculação pelo fungo com a redução da concentração da proteína total e de atividades enzimáticas. Os dados indicam que as vias que envolvem ação dessas duas enzimas devem participar ativamente da defesa das plantas de soja de E48, mas não de PI. Uma hipótese para explicar a resistência de PI é que voláteis produzidos por plantas inoculadas atuam como moduladores da resposta de defesa

Enzimas marcadoras de indução de resistência diferencialmente reguladas em soja resistente e suscetível à ferrugem-asiática-da-soja.

O objetivo deste trabalho foi avaliar, por meio de enzimas marcadoras, a indução de resistência à ferrugem-asiática-da-soja em genótipos de soja contrastantes quanto à suscetibilidade a Phakopsora pachyrhizi. Aproteína total e as atividades de cinco enzimas marcadoras da indução de resistência (lipoxigenases, peroxidases, fenilalanina amônia-liase, quitinases e ?-1, 3-glucanases) foram avaliadas em extratos de folhas de plantas de soja dos genótipos Embrapa 48 (suscetível) e PI 561356 (resistente), submetidas à inoculação ou não com o patógeno. Foram observadas respostas de defesa discrepantes entre os dois genótipos e entre os tempos de coleta (12, 72 e 168 horas após inoculação). A resposta de indução dessas enzimas assemelha-se à defesa bifásica, para Embrapa 48, e é consistente com o observado para outros patossistemas. No entanto, o genótipo PI 561356 respondeu com diminuição da concentração de proteína total e das atividades enzimáticas, o que indica redução do metabolismo geral das plantas infectadas. Há um importante mecanismo de resistência do genótipo PI 561356, ainda não relatado, embasado em vias que envolvem essas enzimas marcadoras e em mecanismos que utilizam menor concentração de proteínas, como os de via metabólica de resposta em cascata. Differentially regulated induced resistance marker enzymes in soybean genotypes resistant and susceptible to Asian soybean rust. The objective of this work was to evaluate induced resistance to Asian soybean rust by means of enzyme activities in soybean genotypes contrasting as to their susceptibility to Phakopsora pachyrhizi. Total protein and the activities of five induced resistance marker enzymes (lipoxygenases, peroxidases, phenylalanine ammonia-lyase, chitinases and ?-1, 3-glucanases) were evaluated in leaf extracts of soybean plants of the genotypes Embrapa 48 (susceptible) and PI 561356 (resistant), inoculated or not with the pathogen. Discrepant defense responses were obtained between the two genotypes and among the leaf harvest times (12, 72, and 168 hours after inoculation). The induction response of these enzymes resembles the biphasic defense in Embrapa 48 and is consistent with that observed in other pathological systems. However, the genotype PI 561356 responded with a decrease in total protein concentration and in enzymatic activities, indicating a general reduction in the metabolism of the infected plants. There is an important mechanism of resistance for the genotype PI 561356, not yet reported, which is grounded on the metabolic ways involving these induced resistance marker enzymes and on the mechanisms that use lower concentrations of total protein, such as the ones with metabolic pathways in response cascade

Enhanced Graph Rewriting Systems for Complex Software Domain

International audienceMethodologies for correct by construction reconfigurations can efficiently solve consistency issues in dynamic software architecture. Graph-based models are appropriate for designing such architectures and methods. At the same time, they may be unfit to characterize a system from a non functional perspective. This stems from efficiency and applicability limitations in handling time-varying characteristics and their related dependencies. In order to lift these restrictions, an extension to graph rewriting systems is proposed herein. The suitability of this approach, as well as the restraints of currently available ones, are illustrated, analysed and experimentally evaluated with reference to a concrete example. This investigation demonstrates that the conceived solution can: (i) express any kind of algebraic dependencies between evolving requirements and properties; (ii) significantly ameliorate the efficiency and scalability of system modifications with respect to classic methodologies; (iii) provide an efficient access to attribute values; (iv) be fruitfully exploited in software management systems; (v) guarantee theoretical properties of a grammar, like its termination