Vorkommen Und Verbreitung Von Pyrrolizidinalkaloiden in Pflanzen

KIEL Biochemical Ecology and Molecular Evolution Pyrrolizidinalkaloiden in in PflanzenPyrrolizidinalkaloiden Vorkommen und Verbreitung von von Verbreitung und Vorkommen Botanischesund Institut GartenBotanischer Universität Kiel UniversitätKiel DietrichOber  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution The The Model System:Pyrrolizidine (PAs)Alkaloids u motivation: Our H • 3 C senecionine O nesadn h ucinadeouinof evolution and function the understanding • • fseii dpain oPsb insects PAsby to adaptations specific of PA biosynthesis in plants in PAbiosynthesis O H O CH N H O + 3 CH O N 3 -oxide O necine necine base necic acid ato fascinating of part • nms casesmost in • • se alkaloidsester lnsadseilzdinsects specialized and plants toxic osiuieyproducedconstitutively , interactions t etbae n insects) and vertebrates (to between  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution yi jacobaeaeTyria tteKiel fjord at the   Dietrich Dietrich Ober Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution aveo eti efbettles leaf certain of larvae Lepidoptera ete (Coleoptera) Beetles rshpes(Orthoptera)Grasshoppers Aphids e.g. West-african e.g. eti speciescertain aiu oh n butterflies and moths various net ihseii dpain oP-otiigplants PA-containing to adaptations specific with Insects H 3 C O O H O CH N H O + 3 CH O Zonocerus 3 O laodacmlto nteinsect in the Alkaloidaccumulation ann ooaino h insect the of coloration Warning laodbsdpheromonesAlkaloid-based A dpnetmorphologyPA-dependent PA-dep.behaviour  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution Toxic principle of pyrrolizidine alkaloids pyrrolizidine of principle Toxic The The twofacespyrrolizidineof alkaloids H 3 C O toxicafter bioactivation - lipophilic- etayP PA PA tertiary O PA tert. H CH N H O 3 CH O 3 O H 3 C O nontoxic- hydrophilic- O N N PA -ox -oxide H O CH N H O + 3 CH O 3 O  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution Toxic principle of pyrrolizidine alkaloids pyrrolizidine of principle Toxic caterpillar hemolymph gut ina nonadapted nonadapted herbivor  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution Toxic principle of pyrrolizidine alkaloids pyrrolizidine of principle Toxic N passiveresorption - partialexcretion - unspecificreduction - PA -ox caterpillar hemolymph bioactivationbycyt.P450sinhemolymph Bioactivation gut TOXIC N tert. PA tert. PA PA -ox ina nonadapted herbivor  Dietrich Dietrich Ober tert. PA KIEL Biochemical Ecology and Molecular Evolution Sequestration of toxic pyrrolizidine pyrrolizidine alkaloids toxic of Sequestration (order of grasshoppers, locusts, and crickets) Orthoptera genus: genus: Zonocerus e. g. e. Zonocerus variegatus Zonocerus  izuWangLinzhu (order of butterflies and moths) Lepidoptera family: Nymphalidae (butterflies) Nymphalidae(butterflies) family: family: moths) (certain Arctiidae e. g. g. e. Tyriajacobaeae  Dietrich Dietrich Ober  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution Sequestration of toxic pyrrolizidine pyrrolizidine alkaloids toxic of Sequestration N PA -ox caterpillar hemolymph gut N tert. N PA PA tert. PA -ox PA -ox inan adapted herbivor  Dietrich Dietrich Ober tert. PA KIEL Biochemical Ecology and Molecular Evolution Sequestration of toxic pyrrolizidine pyrrolizidine alkaloids toxic of Sequestration The The twofacespyrrolizidineof alkaloids H 3 C O toxicafter bioactivation - lipophilic- etayP PA PA tertiary O PA tert. H CH N H O 3 CH O 3 O passivereduction N O H NADPH+ senecionine + 2 -oxygenase inthegut H NADP 2 O H + 3 C inan O nontoxic- hydrophilic- adapted O N N PA -ox -oxide H O CH N H O + 3 herbivor CH O 3 O  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution Biosynthesis arginine biosynthesis primarymetabolism amino acid amino biosynthesis protein spermidine putrescine polyamine isoleucine pool HSS pyrrolizidine alkaloid H akoestructurebackbone (senecionine (senecionine 3 C homospermidine O ytei fa of synthesis O N H NH H secondarymetabolism O CH N H O 2 + N 3 N -oxide) CH H O 2 3 O H Ober & Hartmann Ober (2000) & Hartmann PlantBiolMol 3 3 3 C CH CH diversification of the backbone structure O O O O O O seneciphylline H H O CH CH N O CH N H O O N CH H OH O + + 3 3 3 senkirkine 2 retrorsine retrorsine CH O CH O O H O 3 3  O O O Dietrich Dietrich Ober N N -oxide -oxide KIEL Biochemical Ecology and Molecular Evolution vlto fhmsemdn ytaeb eeduplicatio gene by homospermidine synthase Evolutionof euaoyproteinregulatory deoxyhypusine) ciaino a of activation frainof (formation DHS dhs gene biosynthesis protein homospermidine omto of formation eeduplicationgene activation NO DHS HSS dhs dhs hss hss gene gene n biosynthesis protein homospermidine pyrrolizidine omto of formation alkaloids Ober Ober et al. JBC(2003)  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution

Occurrence of pyrrolizidine alkaloids pyrrolizidine of Occurrence

modified from PhylogenyAngiosperm modified from Group III (2009)

Occurrence of pyrrolizidine alkaloids pyrrolizidine of Occurrence

modified from PhylogenyAngiosperm modified from Group III (2009)

eudicots oeeudicots core basal basal dicots Commelinales Poales Dioscoreales/Liliales/Pandanales Zingiberales Asparagales Arecales Alismatales Acorales Saxifragales Santalales Ranunculales/Proteales Caryophyllales Lamiales Boraginales Sapindales Fagales Cucurbitales Rosales Dipsacales Ericales Brassicales Fabales Solanales Gentianales Garryales Cornales Malvales Myrtales Oxalidales Malpighiales Geraniales Apiales Asterales Aquifoliales rosids monocots asterids eea ie independently times several („ ovretevolutionconvergent oyhltcoriginpolyphyletic Santalaceae Poaceae Apocynaceae Heliotropiaceae / Boraginaceae Orchidaceae Sapotaceae Asteraceae Fabaceae Convolvulaceae  Dietrich Dietrich Ober “) KIEL Biochemical Ecology and Molecular Evolution hlgntcte fDS n S-oigcN sequences cDNA HSS-coding and DHS- of tree Phylogenetic H oigcDNAcoding DHS S oigcDNA HSS coding Bootstrap Bootstrap proportions: 1000 replicates Tree Likelihood Maximum Kaltenegger et (2013) al. PlantCell  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution iseseii xrsino HSS of expression Tissue-specific HSS highly variable between different plant plant lineages different between variable HSShighly specific highly is HSSexpression Heliotropium Phalaenopsis Senecio Eupatorium Cynoglossum Symphytum efepidermisleaf tipsroot budsflower ru fclsi h root in the cells of group root in the cells cortex pericycle and root: endodermis endodermis root: only (Asteraceae) (Asteraceae) (Boraginaceae, Boraginales) (Heliotropiaceae, Boraginales) (orchid) (Boraginaceae, Boraginales)  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution iseseii xrsino HSS of expression Tissue-specific efcosscinof section cross leaf imle et al. Plant(2012) Physiol. Niemüller eitoimindicumHeliotropium   Dietrich Dietrich Ober Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution nlsso S-vlto ihnteConvolvulaceae the HSS-Evolutionwithin Analysisof MorningGloryFamily prxmtl 1600-1700 species approximately • A aebe detected been PAs have • eog oteodrSolanales order the to belongs • fqiedvreceia structures diverse chemical quite of nyi niiulurltdspecies in individual unrelated only  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution hlgntcte fDS n S-oigcN sequences cDNA HSS-coding and DHS- of tree Phylogenetic H oigcDNAcoding DHS S oigcDNA HSS coding Bootstrap Bootstrap proportions: 1000 replicates Tree Likelihood Maximum Kaltenegger et (2013) al. PlantCell  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution nlsso S-vlto ihnteConvolvulaceae the HSS-Evolutionwithin Analysisof H oigcDNAcoding DHS S oigcDNA HSS coding pseudogenization only PAs n eeduplication gene one and eelossgene vn ihnteCnovlca lineage Convolvulaceae the within event fteHSrltdgn oyi eea Afe species PA-free in several copy gene HSS-related the of Kaltenegger et (2013) al. PlantCell  Dietrich Dietrich Ober PAs KIEL Biochemical Ecology and Molecular Evolution hlgn fteBoraginales the of Phylogeny Boraginales Boraginales 0.03 II I Hoplestigmataceae Namaceae Coldeniaceae Codonaceae Hydrophyllaceae Heliotropiaceae Cordiaceae Ehretiaceae hlgn ae nfu hools sequences chloroplast four on based Phylogeny Solanales Solanales (outgroup) Lennoaceae Wellstediaceae P A ? P A ? PA ! oiidfo egn et al. Cladistics(2013) Weigend modified from Boraginaceae ( no e.g. PAs described s. s. str. in in Phacelia  Dietrich Dietrich Ober PA ! ) KIEL Biochemical Ecology and Molecular Evolution inr&Miwl (1984-2000)Eisner & Meinwald • Fabaceae within PAbiosynthesis • erhfraP-pcfcHSSwas not successfull a PA-specific for search PAoccurrence PAs eeaofgenera ttes ornatrixUtetheisa r sdby used are mating high A r transferredPAsare PA ♀ ♂ Crotalaria sdsrbdfrtetocoeyrelated closely two the for described is pcaie insectsspecialized and Lotononis aigsuccessmating low ♂ PA NO like otenx generation next the to female the to male the from PA drvdpheromones-derived ins datg by advantage fitness ln-eie toxinsplant-derived with rtlraretusaCrotalaria ttes ornatrixUtetheisa   Dietrich Dietrich Ober Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution oprsno ouae n non-nodulated and nodulated of Comparison Fabaceae within PAbiosynthesis PAcontent aasgettendlst etest fPAbiosynthesis of site the be to nodules the suggest data C. spectabilis C. 40 days after infection after days 40 spectabilisC. Irmer et (2015) Irmer al. PNAS  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution oprsno ouae n non-nodulated and nodulated of Comparison Fabaceae within PAbiosynthesis HSS HSS expression S sepesdecuieyi h nodulesthe in exclusively expressed HSS is

infection relative transcript level of HSS / UBQ

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root 40 days after infection after days 40 spectabilisC. Irmer et (2015) Irmer al. PNAS  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution in Fabaceae within PAbiosynthesis Crotalaria PAs are a link betweenlink a PAsare pcaie insectspecialized • plant • bacteriasymbiotic • PAs  Dietrich Dietrich Ober KIEL Biochemical Ecology and Molecular Evolution Molecular Evolution of Evolutionof Molecular ChemicalDiversity hn o ! you Thank Evolutionary Biology IMPRS  Dietrich Dietrich Ober