ABSTRACT
The development of a multi-method for the detection of seven allergens based on liquid chromatography and triple-quadrupole tandem mass spectrometry in multiple reaction mode is described. It is based on extraction of the allergenic proteins from a food matrix, followed by enzymatic digestion with trypsin. The chosen marker peptides were implemented into one method that is capable of the simultaneous detection of milk, egg, soy, hazelnut, peanut, walnut and almond. This method has been used to detect all seven allergenic commodities from incurred reference bread material, which was baked according to a standard recipe from the baking industry. Detected concentrations ranged from 10 to 1000 µg/g, demonstrating that the mass spectrometric based method is a useful tool for allergen screening.
Subject(s)
Allergens/analysis , Chromatography, Liquid/methods , Food Analysis/methods , Tandem Mass Spectrometry/methods , Allergens/chemistry , Amino Acid Sequence , Animals , Bread/analysis , Eggs/analysis , Milk/chemistry , Molecular Sequence Data , Nuts/chemistry , Peptides/analysis , Peptides/chemistry , Soy Foods/analysisABSTRACT
Transgenic tobacco plants that express a gene encoding a defective mutant of the tobacco mosaic virus (TMV) movement protein which are known to be resistant to several tobamoviruses were inoculated with viruses from different taxonomic groups to determine the breadth of resistance. There were significant delays in the time of appearance of disease symptoms and/or there was reduced systemic accumulation of virus in upper leaves of plants inoculated with tobacco rattle tobravirus, tobacco ringspot nepovirus, alfalfa mosaic alfamovirus, peanut chlorotic streak caulimovirus, and cucumber mosaic cucumovirus. Conversely, tobacco plants that express a gene encoding the functional tobacco mosaic virus wild-type movement protein accelerated symptom development, enhanced the severity of symptom formation, and/or increased the accumulation of these viruses and, additionally, TMV. Our results indicate that there are similar functions among the movement proteins of a number of plant viruses despite the apparent lack of sequence similarity between them.
Subject(s)
Mosaic Viruses/physiology , Nicotiana/virology , Plants, Toxic , Viral Proteins/physiology , Mosaic Viruses/genetics , Mosaic Viruses/pathogenicity , Nepovirus/pathogenicity , Plant Diseases , Plant Viral Movement Proteins , Plants, Genetically Modified , Nicotiana/immunologyABSTRACT
Fifteen field isolates of satellite tobacco mosaic virus (STMV) were collected from wild Nicotiana glauca plants within a 20-km2 area in southern California. Analyses of the entire length of the satellite genomic RNA of each isolate by RNase protection assays showed that there were 10 different genotypes within the 15 isolates, including some with regions of extreme divergence from the STMV type strain. This indicates a very high level of genetic diversity between natural STMV populations. Isolates from adjacent plants (within 3 m) were not always identical, but were more similar than those from more distant locations. Mapping of the genomic sites of heterogeneity revealed that in all cases the 3' half of the genome, which comprises the untranslated region, was more conserved than were the 5' coding regions.