Fungal elicitors induce a transient release of active oxygen species from cultured spruce cells that is dependent on Ca2+ and protein-kinase activity

Cell-wall components from the ectomycorrhizal fungi Amanita muscaria and Hebeloma crustuliniforme and from the spruce pathogen Heterobasidion annosum elicited a transient release of active oxygen species from cultured spruce cells (Picea abies (L.) Karst.). Since the detection of active oxygen was suppressed by catalase, H₂O₂ was assumed to be the prevailing O₂ species. On the other hand, superoxide dismutase enhanced the concentration of detectable H₂O₂ indicating that the superoxide anion was formed before dismutating to H₂O₂. The elicitors induced the formation of active oxygen in a dose-dependent manner. Interestingly, elicitors from mycorrhizal fungi had a lower H₂O₂-inducing activity than equal amounts of cell-wall preparations from the pathogen H. annosum. In Ca²⁺-depleted medium the production of active oxygen by elicitor-treated spruce cells was suppressed. Additionally, the ionophore A 23187 induced active oxygen formation in a medium with Ca²⁺ but not in a Ca²⁺-depleted medium. Furthermore, the protein-kinase inhibitor staurosporine inhibited the oxidative burst. At a concentration of 34 nM the effect was diminished to 50%. From these results it is suggested that the release of active oxygen species from cultured spruce cells triggered by cell-wall-derived fungal elicitors depends on external Ca²⁺ and a protein-kinase activity. In these respects the effect shows similarities with the well-studied respiratory burst of mammalian neutrophils.Abbreviations EGTA ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid - KP_i potassium phosphate This work was supported by grants from Deutsche Forschungsgemeinschaft and Fonds der Chemischen Industrie.     

Cytoplasmic acidification as an early phosphorylation-dependent response of tobacco cells to elicitors

Elicitor-induced cytoplasmic pH changes of tobacco (Nicotiana tabacum L. cv. Xanthi) cells grown in suspension cultures were explored under a variety of conditions by using a flexible technique based on the distribution of [¹⁴C] benzoic acid between the intracellular and extracellular compartments. Comparison of data obtained by this technique and by ³¹P-nuclear magnetic resonance spectrometry qualifies the benzoic acid distribution method as a convenient and reliable way to probe cytoplasmic pH variations. Various elicitors shown to induce several defense-related responses in tobacco cells, namely oligogalacturonides of degree of polymerization 7–20, pectolyase from Aspergillus japonicus, Phytophthora megasperma crude elicitors and purified cryptogein, triggered cytoplasmic acidifications differing in intensity and kinetics according to the signal molecule. In contrast, no changes in cytoplasmic protons and external pH were observed in cells treated with short galacturonide oligomers, or with soybean-specific hepta -glucoside from P. megasperma, which are devoid of elicitor activity in tobacco cells. The oligogalacturonide-induced cytoplasmic acidification was inhibited by two structurally unrelated protein kinase inhibitors, staurosporine and 6-dimethylaminopurine, which both reduced the external alkalinization response to the elicitor. The protein phosphatase inhibitor calyculin A alone behaved as an elicitormimicking molecule in triggering cytoplasmic acidification, again associated with extracellular alkalinization. These results indicate that the increase in the cytoplasmic concentration of protons may be considered as a common early intracellular response of tobacco cells to elicitors, associated with the extracellular alkalinization response and controlled by protein phosphorylation.Abbreviations BA(H) benzoic acid (protonated form) - 6-DMAP 6-dimethylaminopurine - DP degree of polymerization - Mes 2-(N-morpholino)ethanesulfonic acid - OG oligogalacturonide - pHc cytoplasmic pH - ³¹P-NMR nuclear magnetic resonance spectroscopy of ³¹P atoms The authors thank P. Albersheim (CCRC, Athens, Georgia, USA) for providing the purified oligogalacturonides and the hepta -glucoside and P. Ricci (INRA, Antibes, France) for providing the purified cryptogein.     

Initial signalling processes induced by elicitors of ectomycorrhiza-forming fungi in spruce cells can also be triggered by G-protein-activating mastoparan and protein phosphatase-inhibiting cantharidin

The first responses in spruce [Picea abies (L.) Karst.] cells induced by elicitors (N-acetylglucosamine oligomers) from ectomycorrhizal fungi have been described as follows: efflux of Cl⁻ and K⁺, influx of Ca²⁺, extracellular alkalinization, phosphorylation of a 63-kDa protein (pp63), dephosphorylation of a 65-kDa protein (pp65) and synthesis of H₂O₂ (Salzer et al. 1996, Planta 198: 118–126). In order to obtain new insights into the triggering mechanism and the sequence of these rapid responses we used compounds which are known to activate or block specific steps within an elicitor-induced signal transduction cascade in plant cells. Comparable to elicitors the two protein phosphatase inhibitors, cantharidin and calyculin A, as well as mastoparan, an activator of trimeric G-proteins, were able to induce the release of Cl⁻ and K⁺ from spruce cells and the alkalinization of the medium. Half-maximal activation of the alkalinization occurred at 133 nM calyculin A, 2.3 μM cantharidin and 1.6 μ mastoparan. The structural analogue of mastoparan, Mas 17, which has no G-protein-stimulating properties, was unable to trigger the above-mentioned reactions. In addition, cantharidin and calyculin A induced an increased synthesis of H₂O₂ in spruce cells which was prolonged in comparison to the elicitor-induced transient formation of H₂O₂. Also, the cantharidin-induced release of K⁺ was more pronounced and longer lasting than that caused by elicitors from the ectomycorrhizal fungus Hebeloma crustuliniforme (Bull. ex Fries.) and N-acetylglucosamine oligomers. Furthermore, cantharidin, calyculin A and mastoparan induced the phosphorylation of pp63. Remarkably, the protein kinase inhibitor, staurosporine, inhibited all the rapid responses described above, no matter whether they were triggered by fungal elicitors or by the protein phosphatase inhibitors. These results indicate that in the initial signalling events in spruce cells, essential protein phosphorylations occur either as an (auto) phosphorylation of a membrane-bound receptor kinase prior to the activation of a G-protein or (and) immediately downstream of the activated G-protein in a phosphorylation cascade and are the basic requirements for the ion fluxes following downstream. Received: 24 April 1998 / Accepted: 23 July 1998     

Drone bees fixate the queen with the dorsal frontal part of their compound eyes

Summary Free flying drone bees (Apis mellifera carnia () were filmed from the side while chasing the honey bee queen or landing on a hive.Drones use the upper frontal part of the field of view of their eyes while chasing the queen. They fixate with the lower frontal part of the field of view of their eyes during landing. Drones keep a particular distance from the queen while chasing her.The diameter of ommatidial lenses in drones shows a maximum in the region where the queen is fixated.Niedersächsisches Landesinstitut für Bienenforschung und Bienenwirtschaftliche Betriebslehre, Wehrstrasse 4, D-3100 Celle, Federal Republic of GermanyLehrstuhl Entwicklungsphysiologie, Fakultät für Biologie, Universität Tübingen, Auf der Morgenstelle, D-7400 Tübingen, Federal Republic of GermanyWe thank Walter F. Gelinski for kind cooperation and Professor W. Engels for support.     

Studien zum L-Cyclus der Bakterien

Ohne ZusammenfassungAuszug aus dem ersten Teil einer Dissertation Studien zum Gestaltwandel der Bakterien. (Nach Untersuchungen an Proteus vulgaris und einem gram-negativen unbeweglichen Stäbchen) der Mathematisch-Naturwissenschaftlichen Fakultät der Universität Tübingen 1956.     

Temperatursensitive Mutanten des Tabakmosaikvirus

Ohne ZusammenfassungVerwendete Abkürzungen TMV Tabakmosaikvirus - ts temperatursensitiv - tr temperaturresistent - def defekt - HP Hüllprotein - RNS Ribonukleinsäure - RNase Ribonuklease - EDTA Äthylendiamintetraessigsäure Dissertation der Math.-Naturw. Fakultät der Universität Tübingen.     

Die Beziehungen zwischen Elektrorezeptoren,Elektrischem Organ,Seitenlinienorganen und Nervensystem bei den Mormyridae (Teleostei,Pisces)

Ohne Zusammenfassung

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The interrelations between electroreceptors, electric organ, lateral-line organ and nervous system in Mormyridae (Teleostei, Pisces)

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.

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Der Mathematisch-Naturwissenschaftlichen Fakultät der Universität Tübingen im Jahre 1968 als Habilitationsarbeit vorgelegt.     

A rapid induction by elicitors of the mRNA encoding CCD-1, a 14 kDa Ca2+-binding protein in wheat cultured cells

Intracellular Ca²⁺ has been implicated in the signal transduction processes during the development of the plant defense system against fungal pathogens. From wheat cultured cells that had been treated with the elicitor derived from Typhula ishikariensis, the ccd-1 gene encoding a 14 kDa Ca²⁺-binding protein with an acidic amphiphilic feature was isolated. The ccd-1-encoded protein (CCD-1) shares homology to the C-terminal half domain of centrin, a Ca²⁺-binding protein conserved in eukaryotes. Unlike typical eukaryotic centrins, CCD-1 contains only one Ca²⁺-binding loop, which corresponds to the one in the fourth EF-hand from the N-terminus of centrin. The recombinant CCD protein expressed in Escherichia coli bound to a phenyl-Sepharose column in the presence of Ca²⁺ and was eluted out by EGTA. It also showed a Ca²⁺-dependent electrophoretic mobility shift on the non-denaturing polyacrylamide gel. The ccd-1 mRNA expression was rapidly induced by treatment with fungal and chitosan oligosaccharide elicitors, implying that it might have a role in transducing Ca²⁺ signals provoked by the elicitors. The expression of the ccd-1 mRNA was induced by treatment with A23187, and the induction was suppressed by La³⁺ or 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid (BAPTA). This study suggests the involvement of intracellular Ca²⁺ in the elicitor-induced mRNA expression of a novel class of Ca²⁺-binding proteins conserved in higher plants.     

Equilibria in structured populations

The existence of a stable positive equilibrium state for the density of a population which is internally structured by means of a single scalar such as age, size, etc. is studied as a bifurcation problem. Using an inherent birth modulus n as a bifurcation parameter it is shown for very general nonlinear model equations, in which vital birth and growth processes depend on population density, that a global unbounded continuum of nontrivial equilibrium pairs (n, ) bifurcates from the unique (normalized) critical point (1, 0). The pairs are locally positive and conditions are given under which the continuum is globally positive. Local stability is shown to depend on the direction of bifurcation. For the important case when density dependence is a nonlinear expression involving a linear functional of density (such as total population size) it is shown how a detailed global bifurcation diagram is easily constructed in applications from the graph of a certain real valued function obtained from an invariant on the continuum. Uniqueness and nonuniqueness of positive equilibrium states are studied. The results are illustrated by several applications to models appearing in the literature.This research was done while the author was on leave at the Lehrstuhl für Biomathematik, Universität Tübingen, Auf der Morgenstelle 10, 7400 Tübingen 1, Federal Republic of Germany     

The inhibitory effect of Ca2+ on the flavonoid production of Tetrastigma hemsleyanum suspension cells induced by metal elicitors

Tetrastigma hemsleyanum suspension cells were treated with four metal salts to screen suitable elicitors for the promotion of plant cell biomass and flavonoid production. The effects of calcium ions (Ca²⁺) on induction were also studied. It was found that the most effective elicitors were 50 μM of the heavy metal ion copper (Cu²⁺) and 100 μM of the rare earth element cerium (Ce³⁺). The maximal biomass levels under respective treatments over a 16-d culture period increased by 1.3- and 1.6-fold, and the total flavonoid content was 1.8- and 1.6-fold greater than the control, respectively. Reducing the exogenous Ca²⁺ concentration or adding Ca²⁺ antagonists (1 mM ethylene glycol-bis(2-aminoethylether)-N,N,N′,N-tetraacetc acid (EGTA) or 1 mM verapamil) strengthened inductive effects of metal elicitors and enhanced flavonoid production. However, 0.5 μM of the calcium ionophore A23187 showed contrary results. The increase in exogenous Ca²⁺ concentration in the presence of A23187 suppressed H₂O₂ bursts and peroxidase activity caused by metal elicitors. The results suggest that Ca²⁺ plays an inhibitory role in the plant cell response to metal elicitors. This suppression could have been caused by Ca²⁺ preventing the cells from absorbing metal ions and then easing the induction, or because the decrease of Ca²⁺ concentration worked as an induction signal. Therefore, reducing the Ca²⁺ concentration in culture medium, or adding Ca²⁺ antagonists could be used to improve flavonoid production and cell growth in combination with induction by metal elicitors during in vitro culture of T. hemsleyanum suspension cells.     

Untersuchungen über Beziehungen zwischen Ascorbinsäure und Photosynthese

Zusammenfassung An der einzelligen GrünalgeChlorella pyrenoidosa und an Systemen mit isolierten Spinat-Chloroplasten wurde das Problem einer möglichen Beziehung zwischen Ascorbinsäure und Photosynthese (unter Ausschluß der Photophosphorylierung) bearbeitet.Eine Erhöhung des intracellulären Ascorbinsäure-Spiegels derChlorella pyrenoidosa konnte bei Dauerlichtkulturen erst bei einer Ascorbinsäure-Konzentration des Außenmediums von5·10^–2 Mol/l nachgewiesen werden. An Synchronkulturen ergab sich eine Phase im Entwicklungscyclus der Algen (18. Std), in der bereits bei einer Ascorbinsäure-Konzentration des Außenmediums von 1·10^–3 Mol/l eine signifikante Aufnahme erzielt werden konnte.In Versuchen anChorella-Zellen mit künstlich erhöhtem intracellulären Ascorbinsäure-Spiegel wurde weder bei Gaswechsel-Messungen noch bei der¹⁴CO₂-Fixierung eine Wirkung der Ascorbinsäure auf die Photosynthese festgestellt. Untersuchungen an Synchronkulturen ergaben keine Korrelation zwischen Ascorbinsäure-Gehalt und Photosynthese-Leistung im Verlauf der Zellentwicklung.Der Ascorbinsäure-Gehalt von Spinat-Chloroplasten nach Isolierung in nicht-wäßrigem Medium wurde bestimmt.Versuche an Systemen mit isolierten Chloroplasten ergaben keine Abhängigkeit der Hill-Reaktion mit NADP⁺ als Elektronenacceptor von der Ascorbinsäure. Ein positiver Effekt der Ascorbinsäure zeigte sich nur bei gealterten Chloroplasten-Bruchstücken sowie bei deren Gewinnung bei p_H 8,0. Er wurde im Sinne einer Stabilisator Wirkung der Ascorbinsäure gedeutet.Die Untersuchungen dieser Arbeit und die anderer Autoren zu einer möglichen Beziehung zwischen Ascorbinsäure und Photosynthese wurden hinsichtlich der Versuchsproblematik und der Ergebnisse diskutiert.Aus den Versuchsergebnissen an Synchronkulturen vonChlorella pyrenoidosa wurde auf eine mögliche Funktion der Ascorbinsäure für die Vorbereitung oder Auslösung der Teilung hingewiesen.Mit 9 TextabbildungenDissertation der Mathematisch-Naturwissenschaftlichen Fakultät der Universität Tübingen 1963.     

Terminal versus segmental development in the Drosophila embryo: the role of the homeotic gene fork head

Summary Mutations of the homeotic gene fork head (fkh) of Drosophila transform the non-segmented terminal regions of the embryonic ectoderm into segmental derivatives: Pre-oral head structures and the foregut are replaced by post-oral head structures which are occasionally associated with thoracic structures. Posterior tail structures including the hindgut and the Malpighian tubules are replaced by post-oral head structures associated with anterior tail structures. The fkh gene shows no maternal effect and is required only during embryogenesis. The phenotypes of double mutants indicate that fkh acts independently of other homeotic genes (ANT-C, BX-C, spalt) and caudal. In addition, the fkh domains are not expanded in Polycomb (Pc) group mutant embryos. Ectopic expression of the homeotic selector genes of the ANT-C and BX-C in Pc group mutant embryos causes segmental transformations in terminal regions of the embryo only in the absence of fkh gene activity. Thus, fkh is a region-specific homeotic rather than a selector gene, which promotes terminal as opposed to segmental development. Offprint requests to: Institut für Biologie II (Genetik), Universität Tübingen, Auf der Morgenstelle 28, D-7400 Tübingen, Federal Republic of Germany     

Calcium-regulated appressorium formation of the entomopathogenic fungusZoophthora radicans

Summary The fungusZoophthora radicans (Zygomycetes: Entomophthorales) requires external Ca²⁺ for appressorium formation but not for conidial germination. The number of appressoria formed depends on the Ca²⁺ concentration of the medium. At low [Ca²⁺] (100 pM) nuclear division and germ tube growth are significantly reduced compared to higher Ca²⁺ concentrations (10 and 1,000 M). By contrast, neither external K⁺ nor external Cl^– is needed for germination or appressorium formation. Treatment of conidia with a Ca²⁺-antagonist, Nd³⁺, and a Ca²⁺-channel blocker, nifedipine, inhibits appressorium formation, showing that a Ca²⁺ influx is required for appressorium formation. Furthermore, the partial yet saturating inhibition by nifedipine and complete inhibition by Nd³⁺ indicates that at least two kinds of Ca²⁺ channels are involved in appressorium formation. A contribution of intracellular Ca²⁺ to the signal transduction chain for the formation of appressoria is demonstrated by the inhibitory effect of the intracellular Ca²⁺ antagonist TMB-8. The calmodulin antagonists R24571, TFP, W-7, and W-5 inhibit appressorium formation at concentrations which have no effect on germination. The data presented in this paper are consistent with the hypothesis that a Ca²⁺/calmodulin system is involved in regulating appressorium formation. However, since the direct effects of the drugs were not specifically tested on their proposed binding sites, we leave room for alternative hypotheses that have yet to be formulated.Abbreviations A-9-C 9-anthracenecarboxylic acid - DAPI 4,6 diamino-2-phenylindole - EGTA ethylene glycol bis(-aminoethylether)-N,N-tetraacetic acid - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - H-7 N-(2-methylamino)ethyl-5-isoquino-linesulphonamide dihydrochloride - IC₅₀ concentration of inhibitor that causes 50% inhibiton - R24571 (calmidazolium) 1-[bis-(4-chlorophenyl)methyl]-3-[2,4-dichloro--(2,4-dichlorobenzyloxy)phenethyl]-imidazolium chloride - TEA tetraethylammonium - TFP (trifluoperazine) 10-[3-(4-methylpiperazine-1-yl)-propyl]-2-trifluomethylphenothiazine - TMB-8 8-(diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride - W-5 N-(6-aminohexyl)-1-naphthalene-sulfonamide - W-7 N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide     

On the determination of relative fitness from frequencies of genotypes in subsequent generations

Summary In part I of this paper (Theoret. Appl. Genetics 40, p. 11–17) an algebraic treatment of reproduction and selection processes in populations of diploids was given. Here the statistical properties of a special fitness estimator, which is due to Hayman, are considered. Confidence intervals and tests of significance concerning hypotheses about fitnesses are established and checked by computer simulation studies in part III.

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Zusammenfassung Im ersten Teil dieser Arbeit^* wurde eine alge-Darstellung von Reproduktions- und Selektionsvorgängen in Populationen diploider Organismen gegeben. Im zweiten Teil werden die statistischen Eigenschaften einer speziellen, von Hayman stammenden Schätzfunktion für die Fitness betrachtet. Es werden verschiedene Möglichkeiten zur Be-rechnung von Konfidenzintervallen und zur Durch-führung von Signifikanztesten angegeben und mit Hilfe von Simulationsstudien geprüft.

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Gekürzte Fassung einer der Mathem.-naturwiss. Fakultät der Universität Tübingen eingereichten Dissertationsschrift (Teil II und III).     

Requirements for different Ca2+ pools in the activation of rabbit platelets: I. Release reaction and protein phosphorylation

We examined the role of Ca²⁺, both extracellular and intracellular in origin, in the release reaction and protein phosphorylation in rabbit platelets stimulated with platelet activating factor (acetylglyceryl ether phosphorylcholine), thrombin, or ionophore A23187. In the presence of extracellular Ca²⁺, 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8), a putative antagonist of intracellular Ca²⁺ transport, blocked platelet activating factor-initiated serotonin release at a half-maximal inhibitor concentration of 40 μM, compared to 350 μM for thrombin-induced release and greater than 500 μM, for A23187-induced release. Platelet activating factor-induced phosphorylation of two platelet proteins of M_r=41 000 (P7P) and 20 000 (P9P) was inhibited by TMB-8, an effect which was additive to that caused by removing extracellular Ca²⁺. TMB-8 demonstrated only minor to non-existant inhibitory effects on phosphorylation in thrombin- or A23187-stimulated platelets. In contrast to P9P phosphorylation, phosphorylation of P7P caused by platelet activating factor was more dependent on a TMB-8 sensitive step than on the availability of extracellular Ca²⁺. Experiments with buffers containing fixed concentrations of free Ca²⁺ revealed that both processes (release and phosphorylation), when stimulated by platelet activating factor and thrombin, had the same threshold requirement (1–3 μM) for extracellular free Ca²⁺. These studies provide evidence that stimulation of rabbit platelets by platelet activating factor is more dependent on a TMB-8-sensitive intracellular Ca²⁺ source than is stimulation caused by thrombin. Furthermore, our data indicate that activation of different intracellular processes involved in platelet secretion (such as P7P and P9P phosphorylation) may require Ca²⁺ from different pools.     

Requirement for different Ca2+ pools in the activation of rabbit platelets: II. Phospholipase activity

The effects of extracellular Ca²⁺ concentration and the putative antagonist of intracellular Ca²⁺ movement, 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) on platelet phospholipase activity and thromboxane B₂ synthesis were examined in rabbit platelets stimulated by platelet activating factor, thrombin and ionophore A23187. TMB-8 markedly inhibited the platelet activating factor-induced decrease in [¹⁴C]arachidonate content in platelet phsophatidylacholine and phosphatidylinositol, while showing minimal effects on thrombin-induced phospholipase activation. A23187 stimulation of these processes was inhibited to an intermediated degree by TMB-8. In contrast, extracellular Ca²⁺ removal inhibited phospholipase activity to a similar degree with all three stimuli. Moreover, the threshold concentration of extracelullar Ca²⁺ for phospholiphase activation, as measured by thromboxane B₂ synthesis, was similar for platelet activating factor- and thrombin-stimulated platelets. The data provide evidence that, while platelet activating factor and thrombin may, to some extent, have similar requirements for extracellular Ca²⁺, they utilize a TMB-8 sensitive step to different degrees during activation of platelet phospholipase.     

The role of Ca2+ in elicitation of phytoalexin synthesis in cell culture of onion (Allium cepa L.)

Summary Treatment of Allium cepa L. cellsuspension cultures with a biotic elicitor derived from the fungus Botrytis cinerea, resulted in phytoalexin synthesis. Two phytoalexins, 5-octylcyclopenta-1,3-dione and 5-hexyl-cyclopenta-1,3-dione, were accumulated in cultured onion cells. Removal of extracellular Ca²⁺ by the calcium chelator ethylene glycol bis(b-aminoethyl ether) N,N-tetraacetic acid abolished the elicitor-mediated phytoalexin synthesis. The calcium channel blockers, verapamil and 8-N,N-(dimethylamino)octyl-3,4,5-trimethoxybenzoate caused similar effects, whereas the addition of the Ca²⁺ ionophore A23187 enhanced the accumulation of phytoalexins in the absence of the elicitor. Increase in the cytoplasmic Ca²⁺ concentration in elicitor-treated onion cells was observed as monitored by the fluorescent calcium indicator indo-1. These observations suggest that Ca²⁺ acts as a second messenger in the regulation of phytoalexin synthesis in cultured onion cells.Abbreviations A23187 4-bromo-calcium ionophore - cAMP adenosine 3,5-cyclic monophosphate - [Ca²⁺]_cyt cytoplasmic Ca²⁺ concentration - EGTA ethylene glycol bis(b-aminoethyl ether) N,N-tetraacetic acid - EtOH ethanol - Et₂O diethyl ether - fr.wt fresh weight - HR hypersensitive response - PIPES piperazine N,N-bis-(2-ethanesulfonic acid) - TMB-8 [8-N,N-(dimethylamino)] octyl-3,4,5-trimethoxy-benzoate - Tsl tsibulin