RESUMEN
The isolation of cultivable E. cecorum from the environment of poultry houses remains a challenge. Environmental samples (dust wipes, equipment swabs, pooled feces) and samples from culled bird vertebras were collected from an infected broiler flock on d 37 posthatching. To isolate the bacterium from the cultivable microbiota, suspensions from the environmental samples were streaked onto a blood agar base medium supplemented with 5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid cyclohexylammonium salt (X-Gluc), colistin sulfate, and nalidixin. The chromogenic reaction facilitated the isolation of E. cecorum from contaminated surfaces and pooled feces. Isolates from both the environment and vertebras were confirmed using MALDI-TOF and PCR analysis. Colony appearance and antimicrobial susceptibility tests revealed no phenotypic differences among the isolates. It remained unclear whether the isolates originated from the same clone. However, the principle of isolating the pathogen by streaking on a chromogenic agar may motivate researchers to investigate the transmission routes of infectious isolates, potentially leading to the optimization of biosecurity measures.
Asunto(s)
Pollos , Aves de Corral , Animales , Pollos/microbiología , Agar , EnterococcusRESUMEN
The ß-glucuronidase (GUS) reporter gene system is an important technique with versatile uses in the study of flower development in a broad range of species. Transcriptional and translational GUS fusions are used to characterize gene and protein expression patterns, respectively, during reproductive development. Additionally, GUS reporters can be used to map cis-regulatory elements within promoter sequences and to investigate whether genes are regulated post-transcriptionally. Gene trap/enhancer trap GUS constructs can be used to identify novel genes involved in flower development and marker lines useful in mutant characterization. Flower development studies primarily have used the histochemical assay in which inflorescence tissue from transgenic plants containing GUS reporter genes are stained for GUS activity and examined as whole-mounts or subsequently embedded into wax and examined as tissue sections. In addition, quantitative GUS activity assays can be performed on either floral extracts or intact flowers using a fluorogenic GUS substrate. Another use of GUS reporters is as a screenable marker for plant transformation. A simplified histochemical GUS assay can be used to quickly identify transgenic tissues.
Asunto(s)
Flores , Glucuronidasa , Glucuronidasa/genética , Glucuronidasa/metabolismo , Regiones Promotoras Genéticas , Genes Reporteros , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Flores/genética , Flores/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
Carotenoids comprise one of the major groups of pigments in flowers. Because carotenoids are physiologically indispensable pigments for all photosynthetic plants, their catabolism must be discretely regulated in photosynthetic organs and non-photosynthetic organs such as petals or fruits. In the chrysanthemum, carotenoid cleavage dioxygenase 4a (CmCCD4a), which is dominantly expressed in petals, cleaves carotenoid, leading to a white flower. CmCCD4a-5 was recently identified as a new member of the CmCCD4a family, but its detailed expression profile in plant tissues has not yet been established. In this study, we sequenced a 1094-bp region upstream of CmCCD4a-5 and assessed its petal-specific promoter activity. To evaluate the activity of this gene, we constructed two types of transgenic Arabidopsis thaliana that possessed, respectively, a fusion gene of a 1090-bp or 505-bp segment of the upstream region plus the ß-d-glucuronidase (GUS) gene (1090bUR::GUS and 505bUR::GUS). GUS activity in the 505bUR::GUS strain was observed mainly in the anthers/pollen in flower buds, whereas GUS activity of the 1090bUR::GUS strain was observed in immature petals of the flower buds. Among the cis-acting elements located between positions -505 and -1090, no elements that have previously been reported to enhance the expression in petals or to suppress it in anthers/pollen were detected by PLACE analysis, indicating the existence of unknown cis-element(s). A semiquantitative reverse transcription-polymerase chain reaction analysis revealed that CmCCD4a-5 transcription was prominent in petals but was undetectable in roots, stems and leaves.