Overexpression of an isoprenyl diphosphate synthase in spruce leads to unexpected terpene diversion products that function in plant defense

Spruce (Picea spp.) and other conifers employ terpenoid-based oleoresin as part of their defense against herbivores and pathogens. The short-chain isoprenyl diphosphate synthases (IDS) are situated at critical branch points in terpene biosynthesis, producing the precursors of the different terpenoid classes. To determine the role of IDS and to create altered terpene phenotypes for assessing the defensive role of terpenoids, we overexpressed a bifunctional spruce IDS, a geranyl diphosphate and geranylgeranyl diphosphate synthase in white spruce (Picea glauca) saplings. While transcript level (350-fold), enzyme activity level (7-fold), and in planta geranyl diphosphate and geranylgeranyl diphosphate levels (4- to 8-fold) were significantly increased in the needles of transgenic plants, there was no increase in the major monoterpenes and diterpene acids of the resin and no change in primary isoprenoids, such as sterols, chlorophylls, and carotenoids. Instead, large amounts of geranylgeranyl fatty acid esters, known from various gymnosperm and angiosperm plant species, accumulated in needles and were shown to act defensively in reducing the performance of larvae of the nun moth (Lymantria monacha), a conifer pest in Eurasia. These results show the impact of overexpression of an IDS and the defensive role of an unexpected accumulation product of terpenoid biosynthesis with the potential for a broader function in plant protection

Hydrogen Spectroscopy with a Lamb-shift Polarimeter - An Alternative Approach Towards Anti-Hydrogen Spectroscopy Experiments

A Lamb-shift polarimeter, which has been built for a fast determination of the polarization of protons and deuterons of an atomic-beam source and which is frequently used in the ANKE experiment at COSY-J\"ulich, is shown to be an excellent device for atomic-spectroscopy measurements of metastable hydrogen isotopes. It is demonstrated that magnetic and electric dipole transitions in hydrogen can be measured as a function of the external magnetic field, giving access to the full Breit-Rabi diagram for the $2^2S_{1/2}$ and the $2^2P_{1/2}$ states. This will allow the study of hyperfine structure, $g$ factors and the classical Lamb shift. Although the data are not yet competitive with state-of-the-art measurements, the potential of the method is enormous, including a possible application to anti-hydrogen spectroscopy.Comment: 6 pages, 7 figures, accepted by European Physical Journal

The reaction 2H(p,pp)n in three kinematical configurations at E_p = 16 MeV

We measured the cross sections of the $^2$H(p,pp)n breakup reaction at E$_p$=16 MeV in three kinematical configurations: the np final-state interaction (FSI), the co-planar star (CST), and an intermediate-star (IST) geometry. The cross sections are compared with theoretical predictions based on the CD Bonn potential alone and combined with the updated 2$\pi$-exchange Tucson-Melbourne three-nucleon force (TM99'), calculated without inclusion of the Coulomb interaction. The resulting excellent agreement between data and pure CD Bonn predictions in the FSI testifies to the smallness of three-nucleon force (3NF) effects as well as the insignificance of the Coulomb force for this particular configuration and energy. The CST also agrees well whereas the IST results show small deviations between measurements and theory seen before in the pd breakup space-star geometries which point to possible Coulomb effects. An additional comparison with EFT predictions (without 3NF) up to order N$^3$LO shows excellent agreement in the FSI case and a rather similar agreement as for CD Bonn in the CST and IST situations.Comment: 20 pages, 11 figure

The Polarized H and D Atomic Beam Source for ANKE at COSY-J\"ulich

A polarized atomic beam source was developed for the polarized internal storage-cell gas target at the magnet spectrometer ANKE of COSY-J\"ulich. The intensities of the beams injected into the storage cell, measured with a compression tube, are $7.5\cdot 10^{16}$ hydrogen atoms/s (two hyperfine states) and $3.9\cdot 10^{16}$ deuterium atoms/s (three hyperfine states). For the hydrogen beam the achieved vector polarizations are $p_{\rm z}\approx\pm0.92$. For the deuterium beam, the obtained combinations of vector and tensor ($p_{\rm zz}$) polarizations are $p_{\rm z}\approx\pm 0.90$ (with a constant $p_{\rm zz}\approx +0.86$), and $p_{\rm zz}=+0.90$ or $p_{\rm zz}=-1.71$ (both with vanishing $p_{\rm z}$). The paper includes a detailed technical description of the apparatus and of the investigations performed during the development.Comment: 18 pages, 26 figures, 4 table

11-keto-alpha-boswellic acid, a novel triterpenoid from Boswellia spp. with chemotaxonomic potential and antitumor activity against triple-negative breast cancer cells

Boswellic acids, and particularly 11-keto-boswellic acids, triterpenoids derived from the genus Boswellia (Burseraceae), are known for their anti-inflammatory and potential antitumor efficacy. Although boswellic acids generally occur as α-isomers (oleanane type) and β-isomers (ursane type), 11-keto-boswellic acid (KBA) was found only as the β-isomer, β-KBA. Here, the existence and natural occurrence of the respective α-isomer, 11-keto-α-boswellic acid (α-KBA), is demonstrated for the first time. Initially, α-KBA was synthesized and characterized by high-resolution mass spectrometry (HR-MS) and nuclear magnetic resonance (NMR) spectroscopy, and a highly selective, sensitive, and accurate high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) method was developed by Design of Experiments (DoE) using a pentafluorophenyl stationary phase. This method allowed the selective quantification of individual 11-keto-boswellic acids and provided evidence for α-KBA in Boswellia spp. oleogum resins. The contents of α-KBA as well as further boswellic acids and the composition of essential oils were used to chemotaxonomically classify 41 Boswellia oleogum resins from 9 different species. Moreover, α-KBA exhibited cytotoxicity against three treatment-resistant triple-negative breast cancer (TNBC) cell lines in vitro and also induced apoptosis in MDA-MB-231 xenografts in vivo. The respective β-isomer and the acetylated form demonstrate higher cytotoxic efficacies against TNBC cells. This provides further insights into the structure-activity relationship of boswellic acids and could support future developments of potential anti-inflammatory and antitumor drugs