The cuticle, covering the surface of all primary plant organs, plays important roles in plant development and protection against the biotic and abiotic environment. In contrast to vegetative organs, very little molecular information has been obtained regarding the surfaces of reproductive organs such as fleshy fruit. To broaden our knowledge related to fruit surface, comparative transcriptome and metabolome analyses were carried out on peel and flesh tissues during tomato (Solanum lycopersicum) fruit development. Out of 574 peel-associated transcripts, 17% were classified as putatively belonging to metabolic pathways generating cuticular components, such as wax, cutin, and phenylpropanoids. Orthologs of the Arabidopsis (Arabidopsis thaliana) SHINE2 and MIXTA-LIKE regulatory factors, activating cutin and wax biosynthesis and fruit epidermal cell differentiation, respectively, were also predominantly expressed in the peel. Ultra-performance liquid chromatography coupled to a quadrupole time-of-flight mass spectrometer and gas chromatography-mass spectrometry using a flame ionization detector identified 100 metabolites that are enriched in the peel tissue during development. These included flavonoids, glycoalkaloids, and amyrin-type pentacyclic triterpenoids as well as polar metabolites associated with cuticle and cell wall metabolism and protection against photooxidative stress. Combined results at both transcript and metabolite levels revealed that the formation of cuticular lipids precedes phenylpropanoid and flavonoid biosynthesis. Expression patterns of reporter genes driven by the upstream region of the wax-associated SlCER6 gene indicated progressive activity of this wax biosynthetic gene in both fruit exocarp and endocarp. Peel-associated genes identified in our study, together with comparative analysis of genes enriched in surface tissues of various other plant species, establish a springboard for future investigations of plant surface biology.
The degradation of the warfare agent sulfur mustard (HD) adsorbed onto KF/Al(2)O(3) sorbents is described. These processes were explored by MAS NMR, using (13)C-labeled sulfur mustard (HD*) and LC-MS techniques. Our study on the detoxification of this blister agent showed the formation of nontoxic substitution and less-toxic elimination products (t(1/2) = 3.5-355 h). Interestingly, the reaction rates were found to be affected by MAS conditions, i.e., by a centrifugation effect. The products and the mechanisms of these processes are discussed.
SARS-CoV-2, the etiologic
agent of the COVID-19 pandemic, emerged
as the cause of a global crisis. Rapid and reliable clinical diagnosis
is essential for effectively controlling transmission. The gold standard
assay for SARS-CoV-2 identification is the highly sensitive real-time
quantitative polymerase chain reaction (RT-qPCR); however, this assay
depends on specialized reagents and may suffer from false results.
Thus, additional assays based on different approaches could be beneficial.
Here, we present a novel method for SARS-CoV-2 identification based
on mass spectrometry. The approach we implemented combines a multistep
procedure for the rational down-selection of a set of reliable markers
out of all optional
in silico
derived tryptic peptides
in viral proteins, followed by monitoring of peptides derived from
tryptic digests of purified proteins, cell-cultured SARS-CoV-2, and
nasopharyngeal (NP) swab matrix spiked with the virus. The marker
selection was based on specificity to SARS-CoV-2 and on analytical
parameters including sensitivity, linearity, and reproducibility.
The final assay is based on six unique and specific peptide markers
for SARS-CoV-2 identification. The simple and rapid (2.5 h) protocol
we developed consists of virus heat inactivation and denaturation,
tryptic digestion, and identification of the selected markers by liquid
chromatography coupled to high-resolution mass spectrometry (LC-MS/MS).
The developed assay enabled the identification of 10
4
PFU/mL
SARS-CoV-2 spiked into buffer. Finally, the assay was successfully
applied to 16 clinical samples diagnosed by RT-qPCR, achieving 94%
concordance with the current gold standard assay. To conclude, the
novel MS-based assay described here is specific, rapid, simple, and
is believed to provide a complementary assay to the RT-qPCR method.
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