A novel expression cassette for the efficient visual selection of transformed tissues in florists' chrysanthemum (Chrysanthemum morifolium Ramat.).

Constructs carrying visual reporter genes coupled with efficient promoters could facilitate the process of identification and selection of stable transformants in recalcitrant crops. Here, a novel construct utilizing a ribulose-1,5-bisphosphate carboxylase (RbcS) promoter combined with the green fluorescent protein (GFP) reporter gene to initiate very high expression of GFP in florist's chrysanthemum (Chrysanthemum morifolium Ramat.) was described. Based on this expression cassette, a new regeneration protocol using leaf discs as explants was developed for the Agrobacterium-mediated transformation of Chrysanthemum genotype ‘1581’, and a transformation efficiency of 7% was obtained. The expression of two different GFP constructs targeted to either cytosol or plastids was compared in transgenic lines. Both GFP constructs were expressed at such a high level that the green fluorescence dominated red fluorescence in the leaf tissues, allowing easy observation and microdissection of transformed tissues even without a GFP filter. Under normal light, plants with GFP targeted to plastids had a light green phenotype deriving from the high GFP expression. Quantitative reverse transcriptional PCR analysis showed that the plastid targeted construct with intron had significantly higher steady state transcript levels of GFP mRNA. This novel expression cassette may allow direct visual selection of transformed tissues independent of antibiotic selection in a wide range of plant specie

Roland Barthes: la conquista del discurso amoroso

El texto póstumo de Roland Barthes proporciona claves para la mejor interpretación de su libro Los fragmentos de un discurso amoroso, escrito en 1977. Se trata de entender la constelación del amor, la muerte, la madre, la escritura y el lenguaje, constelación que está montada sobre el discurso amoroso de Los fragmentos..

Nieves Rodríguez Valle, Los refranes en el “Quijote”: poética cervantina. El Colegio de México, México, 2014; 426 pp. (Serie Estudios de Lingüística y Literatura, 62).

Se reseña el libro:Los refranes en el “Quijote”: poética cervantina

A voz do narrador, o olhar dos habitantes da Corte e o corpo de Precisoa

El narrador caracteriza a Preciosa llena de virtudes y dotada de una misteriosa condición excepcional. Los pobladores de la Corte contribuyen con su mirada y sus comentarios a la composición del personaje. Aunque Preciosa se somete al poder de esta sociedad y a la de los gitanos, sus actos y su discurso establecerán un contrapunto dialógico con la voz del narrador y la apropiación de los dos grupos.O narrador caracteriza Preciosa como cheia de virtudes e dotada de uma misteriosa condição excepcional. Os habitantes da Corte contribuem com seu olhar e seus comentários para a composição do personagem. Ainda que Preciosa se submeta ao poder desta sociedade e dos ciganos, seus atos e seu discurso estabelecerão um contraponto dialógico com a voz do narrador e a apropriação dos dois grupos

A novel expression cassette for the efficient visual selection of transformed tissues in florist's chrysanthemum (Chrysanthemum morifolium Ramat.)

Constructs carrying visual reporter genes coupled with efficient promoters could facilitate the process of identification and selection of stable transformants in recalcitrant crops. Here, a novel construct utilizing a ribulose-1,5-bisphosphate carboxylase (RbcS) promoter combined with the green fluorescent protein (GFP) reporter gene to initiate very high expression of GFP in florist's chrysanthemum (Chrysanthemum morifolium Ramat.) was described. Based on this expression cassette, a new regeneration protocol using leaf discs as explants was developed for the Agrobacterium-mediated transformation of Chrysanthemum genotype ‘1581’, and a transformation efficiency of 7% was obtained. The expression of two different GFP constructs targeted to either cytosol or plastids was compared in transgenic lines. Both GFP constructs were expressed at such a high level that the green fluorescence dominated red fluorescence in the leaf tissues, allowing easy observation and microdissection of transformed tissues even without a GFP filter. Under normal light, plants with GFP targeted to plastids had a light green phenotype deriving from the high GFP expression. Quantitative reverse transcriptional PCR analysis showed that the plastid targeted construct with intron had significantly higher steady state transcript levels of GFP mRNA. This novel expression cassette may allow direct visual selection of transformed tissues independent of antibiotic selection in a wide range of plant species.Key words: Florist's chrysanthemum (Chrysanthemum morifolium), genetic transformation, GFP gene, RbcS promoter, RbcS targeting signal

Binational collaboration in recovery of endangered species: the Mexican wolf as a case study

The goal of this inductive study was to identify factors that facilitate and inhibit binational collaboration in the recovery of endangered species in the northern Mexico borderlands, focusing on the Mexican wolf (Canis lupus baileyi). A conceptual model was developed using qualitative techniques, providing the basis for design of a mail survey. The target population included participants with experience in recovery efforts for over a dozen species at risk in the region. Long interviews were recorded with 44 participants from Mexico and the United States. Thematic hierarchical analysis was used to develop a conceptual model of how interviewees talked about factors influencing binational collaboration. Issues were classified in five thematic clusters: project, organization, people, resources, culture/history. The survey was used to conduct a needs assessment, measuring respondents' attitudes about the relative priority of issues identified in the conceptual model. High priority needs were identified from each thematic cluster: (a) equitable participation in project design and implementation, (b) continuity of personnel, (c) coordination of federal, state and local efforts, (d) increased funding, managed with accountability, and (e) exchange visits to facilitate understanding of diverse perspectives. Responses to almost half the survey items indicated accord among the sample of respondents, providing a basis for shared common ground. The nature of discord was within the range of "manageable", with no clear polarization of attitudes measured. This exploratory data analysis suggested that the structure of the conceptual model developed from the Mexican wolf case study was generally a valid basis for future deductive analysis and reflection by practitioners. For 82% of 22 statements of need, priorities of participants in the Mexican wolf recovery efforts did not differ significantly from other respondents. Nationality (of respondents) significantly affected priority rankings for only 18% of the need statements. Significant effects of five demographic variables indicated that interactive effects should be examined in future multivariate analyses to determine how respondents' attitudes on issues related to priority rankings. Recommendations were provided for a more efficient and effective approach to collaborative problem-solving, engaging reflective practitioners from the private and public sectors in principled negotiation processes to better understand diverse perspectives