In vivo synthesis and purification of radioactivep-hydroxybenzylglucosinolate inSinapis alba L.

Abstract

Glucosinolates are amino acid‐derived, natural plant products found in the order Capparales. Radiolabelled glucosinolates for use in biological studies concerning metabolism and transport are not commercially available. We have developed a method for the synthesis and isolation of radiolabelled p‐hydroxybenzylglucosinolate (p‐OHBG) produced in vivo by Sinapis alba, which has the tyrosine‐derived p‐OHBG as its major glucosinolate. The method is based on administration of radiolabelled tyrosine to jasmonate‐treated primary leaves, which have high de novo biosynthesis of p‐OHBG. After incubation, the plant material was extracted with methanol and the glucosinolates purified by a procedure including two ion exchange chromatographic steps. The method provides an attractive alternative to chemical synthesis of radiolabelled glucosinolates and is applicable to other plants producing a single or one major glucosinolate. Copyright © 2000 John Wiley & Sons, Ltd.

Artificial Fluorescent Glucosinolates (F-GSLs) Are Transported by the Glucosinolate Transporters GTR1/2/3

“…LCMS has also been used to measure GTR-mediated glucosinolate import in other heterologous hosts, including yeast [ 25 ], cotton cells [ 26 ], and insect cells (unpublished data) [ 27 ]. In one example, glucosinolate import was detected by monitoring the influx of radiolabeled glucosinolates that were generated by feeding radiolabeled tyrosine to CYP79A1 overexpressing Arabidopsis plants [ 7 , 28 ].…”

Glycine Conjugates in a Lepidopteran Insect Herbivore—The Metabolism of Benzylglucosinolate in the Cabbage White Butterfly, Pieris rapae

“…When most of the liquid had been taken up by the leaves, MS medium (50 μL) was added to each well. After 12 h incubation, 2 was isolated38 and converted into its protonated form 39. In the purified preparation obtained after feeding of [U‐ 14 C]phenylalanine, 14 C‐labelled 2 accounted for 97 % of the radioactivity present, as shown by HPLC analysis (system 1, detectors 1 and 2).…”

Desulfation Followed by Sulfation: Metabolism of Benzylglucosinolate in Athalia rosae (Hymenoptera: Tenthredinidae)

“…Metabolism of [ 14 C]‐labelled benzylglucosinolate : [1,1′,2′,3′,4′,5′,6′‐ 14 C 7 ]Benzylglucosinolate was prepared by feeding L ‐[U‐ 14 C]phenylalanine to transgenic A. thaliana 35S:CYP79A235 as described in ref. 36 and isolated as its potassium salt37 ( 14 C‐labelled 1 , Scheme , 3.5 m M , 13.96 Bq nmol −1 ). Last‐instar larvae ( n =4) were each fed one leaf of A. thaliana that had been treated with [ 14 C]‐labelled benzylglucosinolate (6 μL).…”