A chromosomal insertion toolbox for promoter probing, mutant complementation and pathogenicity studies in Ralstonia solanacearum

We describe here the construction of a delivery system for stable and directed insertion of gene constructs in a permissive chromosomal site of the bacterial wilt pathogen Ralstonia solanacearum. The system consists of a collection of suicide vectors the Ralstonia chromosome (pRC) series that carry an integration element flanked by transcription terminators and two sequences of homology to the chromosome of strain GMI1000, where the integration element is inserted through a double recombination event. Unique restriction enzyme sites and a GATEWAY cassette enable cloning of any promoter::gene combination in the integration element. Variants endowed with different selectable antibiotic resistance genes and promoter::gene combinations are described. We show that the system can be readily used in GMI1000 and adapted to other R. solanacearum strains using an accessory plasmid. We prove that the pRC system can be employed to complement a deletion mutation with a single copy of the native gene, and to measure transcription of selected promoters in monocopy both in vitro and in planta. Finally, the system has been used to purify and study secretion type III effectors. These novel genetic tools will be particularly useful for the construction of recombinant bacteria that maintain inserted genes or reporter fusions in competitive situations (i.e., during plant infection).

Aprendizaje léxico y desarrollo de la gramática: vocabulario verbal, aceleración morfológica y complejidad sintáctica

This paper studies the initial development of certain language components. More precisely, we analyse the relation between three aspects that are closely involved in the grammar of the verb: morphological productivity, syntactic complexity, and verb vocabulary learning. The study is based on data about the relationship between lexical development and grammatical development, and also on proposals that a critical mass of vocabulary is needed in order to develop a grammatical component. The sample comprised six subjects who are monolingual or bilingual in Catalan andlor Spanish. Results show a morphological spurt some time afer the learning of a certain quantity of verbs. Moreover, syntactic complexity is only evident some months after this morphological spurt

La productividad de la morfología verbal a los años dos años de edad

Este trabajo se centra en estudiar la adquisición de la morfología verbal en una lengua con un paradigma verbal morfológicamente rico. Se predice que los niños de dos años no mostrarán productividad morfológica ante verbos desconocidos. La muestra se compone de 12 sujetos de edades comprendidas entre los 22 y los 31 meses. El procedimiento, de tipo experimental, se caracteriza por entrenar a los niños con verbos inventados. Los resultados muestran que los sujetos solamente utilizan los nuevos verbos con la forma morfológica con la que se han presentado. También se observa la omisión de determinados constituyentes de la oración. Estos resultados se discuten en relación con la asunción de discontinuidad de las competencias lingüísticas infantiles