RESUMO
Barley and Cereal yellow dwarf viruses (B/CYDVs), Wheat spindle streak mosaic (WSSMV), Soil-borne wheat mosaic virus (SBWMV) and Wheat streak mosaic virus (WSMV) constitute the most economically important group of wheat viruses. In this paper, a multiplex reverse transcription polymerase chain reaction (M-RT-PCR) method was developed for the simultaneous detection and discrimination of eight viruses: five strains of B/CYDVs, WSSMV, SBWMV and WSMV. The protocol uses specific primer sets for each virus producing five distinct fragments 295, 175, 400, 237, and 365 bp, indicating the presence of two strains of BYDVs, -PAV, -MAV, CYDV-RPV and two unassigned Luteoviridae BYDV-SGV and -RMV, respectively. This system also readily detected WSSMV, SBWMV and WSMV specific amplicons at 154, 219 and 193 bp, respectively. The amplification specificity of these primers was tested against a range of field samples from different parts of United States. The protocol also utilizes fluorescently tagged primers that can streamline the detection of each virus through capillary electrophoresis. This study fulfills the need for a rapid and specific wheat virus diagnostic tool that also has the potential for investigating the epidemiology of these viral diseases.
Assuntos
Doenças das Plantas/virologia , Vírus de Plantas/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Primers do DNA/genética , Eletroforese Capilar , Sensibilidade e Especificidade , Triticum/virologia , Estados UnidosRESUMO
[reaction: see text] L-alpha-(1-Cyclobutenyl)glycine (1-Cbg) was targeted as a potentially translatable analogue of isoleucine and valine and as a useful building block for peptides. An enantioselective synthesis was executed in which the key step was diastereoselective addition of 1-cyclobutenylmagnesium bromide to the sulfinimine 2b derived from (S)-t-butanesulfinimide and tert-butyl glyoxylate. 1-Cbg was found to substitute efficiently for isoleucine and valine, but not leucine, in the translation of green fluorescent protein in vitro.