Wasternack, C.; Hause, B. Jasmonsäure – ein universelles Pflanzenhormon: Blütenduft, Abwehr, Entwicklung Biologie in unserer Zeit 44, 164 - 171, (2014) DOI: 10.1002/biuz.201410535
Jasmonsäure (JA) und ihre Metaboliten kommen in allen niederen und höheren Pflanzen vor. Sie sind universell wirksame, aus Lipiden gebildete Signalstoffe bei der Abwehr von biotischem und abiotischem Stress sowie in der pflanzlichen Entwicklung. Rezeptor und Komponenten von JA–Signalketten wurden identifiziert. In der Entwicklung von Blüten, Früchten, Samen, Trichomen oder in der Abwehr von Insekten und Pathogenen treten ähnliche JA-vermittelte Signalproteine auf, die eine Feinregulation der Prozesse erlauben und eine Verbindung (cross-talk) zu anderenPflanzenhormonen aufweisen.
Ziegler, J.; Abel S. Analysis of amino acids by HPLC/electrospray negative ion tandem mass spectrometry using 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) derivatization Amino Acids 46, 2799-2808, (2014) DOI: 10.1007/s00726-014-1837-5
A new method for the determination of amino acids is presented. It combines established methods for the derivatization of primary and secondary amino groups with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) with the subsequent amino acid specific detection of the derivatives by LC–ESI–MS/MS using multiple reaction monitoring (MRM). The derivatization proceeds within 5 min, and the resulting amino acid derivatives can be rapidly purified from matrix by solid-phase extraction (SPE) on HR-X resin and separated by reversed-phase HPLC. The Fmoc derivatives yield several amino acid specific fragment ions which opened the possibility to select amino acid specific MRM transitions. The method was applied to all 20 proteinogenic amino acids, and the quantification was performedusing l-norvaline as standard. A limit of detection as low as 1 fmol/μl with a linear range of up to 125 pmol/μl could be obtained. Intraday and interday precisions were lower than10 % relative standard deviations for most of the amino acids. Quantification usingl-norvaline as internal standard gave very similar results compared to the quantificationusing deuterated amino acid as internal standards. Using this protocol, it was possible to record the amino acid profiles of only a single root from Arabidopsis thaliana seedlings and to compare it with the amino acid profiles of 20 dissected root meristems (200 μm).
Grubb, C. D.; Zipp, B. J.; Kopycki, J.; Schubert, M.; Quint, M.; Lim, E.-K.; Bowles, D. J.; Pedras, M. S. C.; Abel, S. Comparative analysis of Arabidopsis UGT74 glucosyltransferases reveals a special role of UGT74C1 in glucosinolate biosynthesis Plant J. 79, 92–105, (2014) DOI: 10.1111/tpj.12541
The study of glucosinolates and their regulation has provided a powerful framework for the exploration of fundamental questions about the function, evolution, and ecological significance of plant natural products, but uncertainties about their metabolism remain. Previous work has identified one thiohydroximate S-glucosyltransferase, UGT74B1, with an important role in the core pathway, but also made clear that this enzyme functions redundantly and cannot be the sole UDP-glucose dependent glucosyltransferase (UGT) in glucosinolate synthesis. Here, we present the results of a nearly comprehensive in vitro activity screen of recombinant Arabidopsis Family 1 UGTs, which implicate other members of the UGT74 clade as candidate glucosinolate biosynthetic enzymes. Systematic genetic analysis of this clade indicates that UGT74C1 plays a special role in the synthesis of aliphatic glucosinolates, a conclusion strongly supported by phylogenetic and gene expression analyses. Finally, the ability of UGT74C1 to complement phenotypes and chemotypes of the ugt74b1-2 knockout mutant and to express thiohydroximate UGT activity in planta provides conclusive evidence for UGT74C1 being an accessory enzyme in glucosinolate biosynthesis with a potential function during plant adaptation to environmental challenge.