TY - GEN A1 - Jantzen, Friederike A1 - Wozniak, Natalia Joanna A1 - Kappel, Christian A1 - Sicard, Adrien A1 - Lenhard, Michael T1 - A high‑throughput amplicon‑based method for estimating outcrossing rates T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - Background: The outcrossing rate is a key determinant of the population-genetic structure of species and their long-term evolutionary trajectories. However, determining the outcrossing rate using current methods based on PCRgenotyping individual offspring of focal plants for multiple polymorphic markers is laborious and time-consuming. Results: We have developed an amplicon-based, high-throughput enabled method for estimating the outcrossing rate and have applied this to an example of scented versus non-scented Capsella (Shepherd’s Purse) genotypes. Our results show that the method is able to robustly capture differences in outcrossing rates. They also highlight potential biases in the estimates resulting from differential haplotype sharing of the focal plants with the pollen-donor population at individual amplicons. Conclusions: This novel method for estimating outcrossing rates will allow determining this key population-genetic parameter with high-throughput across many genotypes in a population, enabling studies into the genetic determinants of successful pollinator attraction and outcrossing. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 745 KW - Outcrossing KW - Mixed mating KW - Outcrossing rate KW - Capsella KW - Amplicon sequencing Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-435657 SN - 1866-8372 IS - 745 ER - TY - JOUR A1 - Jantzen, Friederike A1 - Wozniak, Natalia Joanna A1 - Kappel, Christian A1 - Sicard, Adrien A1 - Lenhard, Michael T1 - A high‑throughput amplicon‑based method for estimating outcrossing rates JF - Plant Methods N2 - Background: The outcrossing rate is a key determinant of the population-genetic structure of species and their long-term evolutionary trajectories. However, determining the outcrossing rate using current methods based on PCRgenotyping individual offspring of focal plants for multiple polymorphic markers is laborious and time-consuming. Results: We have developed an amplicon-based, high-throughput enabled method for estimating the outcrossing rate and have applied this to an example of scented versus non-scented Capsella (Shepherd’s Purse) genotypes. Our results show that the method is able to robustly capture differences in outcrossing rates. They also highlight potential biases in the estimates resulting from differential haplotype sharing of the focal plants with the pollen-donor population at individual amplicons. Conclusions: This novel method for estimating outcrossing rates will allow determining this key population-genetic parameter with high-throughput across many genotypes in a population, enabling studies into the genetic determinants of successful pollinator attraction and outcrossing. KW - Outcrossing KW - Mixed mating KW - Outcrossing rate KW - Capsella KW - Amplicon sequencing Y1 - 2019 U6 - https://doi.org/10.1186/s13007-019-0433-9 SN - 1746-4811 VL - 15 IS - 47 PB - BioMed Central CY - London ER - TY - GEN A1 - Eldridge, Tilly A1 - Łangowski, Łukasz A1 - Stacey, Nicola A1 - Jantzen, Friederike A1 - Moubayidin, Laila A1 - Sicard, Adrien A1 - Southam, Paul A1 - Kennaway, Richard A1 - Lenhard, Michael A1 - Coen, Enrico S. A1 - Østergaard, Lars T1 - Fruit shape diversity in the Brassicaceae is generated by varying patterns of anisotropy T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Fruits exhibit a vast array of different 3D shapes, from simple spheres and cylinders to more complex curved forms; however, the mechanism by which growth is oriented and coordinated to generate this diversity of forms is unclear. Here, we compare the growth patterns and orientations for two very different fruit shapes in the Brassicaceae: the heart-shaped Capsella rubella silicle and the near-cylindrical Arabidopsis thaliana silique. We show, through a combination of clonal and morphological analyses, that the different shapes involve different patterns of anisotropic growth during three phases. These experimental data can be accounted for by a tissue level model in which specified growth rates vary in space and time and are oriented by a proximodistal polarity field. The resulting tissue conflicts lead to deformation of the tissue as it grows. The model allows us to identify tissue-specific and temporally specific activities required to obtain the individual shapes. One such activity may be provided by the valve-identity gene FRUITFULL, which we show through comparative mutant analysis to modulate fruit shape during post-fertilisation growth of both species. Simple modulations of the model presented here can also broadly account for the variety of shapes in other Brassicaceae species, thus providing a simplified framework for fruit development and shape diversity. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 986 KW - Brassicaceae KW - Capsella KW - arabidopsis KW - fruit shape KW - modelling KW - anisotropic growth Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-438041 SN - 1866-8372 IS - 986 SP - 3394 EP - 3406 ER - TY - JOUR A1 - Eldridge, Tilly A1 - Langowski, Lukasz A1 - Stacey, Nicola A1 - Jantzen, Friederike A1 - Moubayidin, Laila A1 - Sicard, Adrien A1 - Southam, Paul A1 - Kennaway, Richard A1 - Lenhard, Michael A1 - Coen, Enrico S. A1 - Ostergaard, Lars T1 - Fruit shape diversity in the Brassicaceae is generated by varying patterns of anisotropy JF - Development : Company of Biologists N2 - Fruits exhibit a vast array of different 3D shapes, from simple spheres and cylinders to more complex curved forms; however, the mechanism by which growth is oriented and coordinated to generate this diversity of forms is unclear. Here, we compare the growth patterns and orientations for two very different fruit shapes in the Brassicaceae: the heart-shaped Capsella rubella silicle and the near-cylindrical Arabidopsis thaliana silique. We show, through a combination of clonal and morphological analyses, that the different shapes involve different patterns of anisotropic growth during three phases. These experimental data can be accounted for by a tissue level model in which specified growth rates vary in space and time and are oriented by a proximodistal polarity field. The resulting tissue conflicts lead to deformation of the tissue as it grows. The model allows us to identify tissue-specific and temporally specific activities required to obtain the individual shapes. One such activity may be provided by the valve-identity gene FRUITFULL, which we show through comparative mutant analysis to modulate fruit shape during post-fertilisation growth of both species. Simple modulations of the model presented here can also broadly account for the variety of shapes in other Brassicaceae species, thus providing a simplified framework for fruit development and shape diversity. KW - Brassicaceae KW - Capsella KW - Arabidopsis KW - Fruit shape KW - Modelling KW - Anisotropic growth Y1 - 2016 U6 - https://doi.org/10.1242/dev.135327 SN - 0950-1991 SN - 1477-9129 VL - 143 SP - 3394 EP - 3406 PB - Company of Biologists Limited CY - Cambridge ER - TY - THES A1 - Jantzen, Friederike T1 - Genetic basis and adaptive significance of repeated scent loss in selfing Capsella species T1 - Genetische Grundlage und Adaptive Signifikanz von wiederholtem Duftverlust bei selbstenden Capsella Arten N2 - Floral scent is an important way for plants to communicate with insects, but scent emission has been lost or strongly reduced during the transition from pollinator-mediated outbreeding to selfing. The shift from outcrossing to selfing is not only accompanied by scent loss, but also by a reduction in other pollinator-attracting traits like petal size and can be observed multiple times among angiosperms. These changes are summarized by the term selfing syndrome and represent one of the most prominent examples of convergent evolution within the plant kingdom. In this work the genus Capsella was used as a model to study convergent evolution in two closely related selfers with separate transitions to self-fertilization. Compared to their outbreeding ancestor C. grandiflora, the emission of benzaldehyde as main compound of floral scent is lacking or strongly reduced in the selfing species C. rubella and C. orientalis. In C. rubella the loss of benzaldehyde was caused by mutations to cinnamate:CoA ligase CNL1, but the biochemical basis and evolutionary history of this loss remained unknown, together with the genetic basis of scent loss in C. orientalis. Here, a combination of plant transformations, in vitro enzyme assays, population genetics and quantitative genetics has been used to address these questions. The results indicate that CNL1 has been inactivated twice independently by point mutations in C. rubella, leading to a loss of benzaldehyde emission. Both inactivated haplotypes can be found around the Mediterranean Sea, indicating that they arose before the species´ geographical spread. This study confirmed CNL1 as a hotspot for mutations to eliminate benzaldehyde emission, as it has been suggested by previous studies. In contrast to these findings, CNL1 in C. orientalis remains active. To test whether similar mechanisms underlie the convergent evolution of scent loss in C. orientalis a QTL mapping approach was used and the results suggest that this closely related species followed a different evolutionary route to reduce floral scent, possibly reflecting that the convergent evolution of floral scent is driven by ecological rather than genetic factors. In parallel with studying the genetic basis of repeated scent loss a method for testing the adaptive value of individual selfing syndrome traits was established. The established method allows estimating outcrossing rates with a high throughput of samples and detects successfully insect-mediated outcrossing events, providing major advantages regarding time and effort compared to other approaches. It can be applied to correlate outcrossing rates with differences in individual traits by using quasi-isogenic lines as demonstrated here or with environmental or morphological parameters. Convergent evolution can not only be observed for scent loss in Capsella but also for the morphological evolution of petal size. Previous studies detected several QTLs underlying the petal size reduction in C. orientalis and C. rubella, some of them shared among both species. One shared QTL is PAQTL1 which might map to NUBBIN, a growth factor. To better understand the morphological evolution and genetic basis of petal size reduction, this QTL was studied. Mapping this QTL to a gene might identify another example for a hotspot gene, in this case for the convergent evolution of petal size. N2 - Blütenduft stellt einen wichtigen Kommunikationsweg für Pflanzen mit ihrer Außenwelt dar, jedoch ging die Emission von Duftstoffen während des Übergangs von Bestäuber-vermittelter Auskreuzung zur Selbstung verloren oder wurde stark reduziert. Dieser Wandel des Fortpflanzungssystems wurde nicht nur vom Duftverlust begleitet, sondern auch von einer Reduktion anderer Merkmale, die Bestäuber anlocken, so zum Beispiel auch einer verringerten Blütenblattgröße. Diese veränderten Merkmale werden unter dem Begriff Selbstungssyndrom zusammengefasst und können mehrfach in der Evolution von Angiospermen beobachtet werden. Der Übergang von Auskreuzung zu Selbstung und die damit einhergehende Veränderung von Blüten stellt eines der bekanntesten Beispiele für konvergente Evolution im Pflanzenreich dar. In dieser Arbeit wurde die Gattung Capsella als Modell für die Untersuchung konvergenter Evolution genutzt, da sie zwei eng verwandte selbstbestäubende Arten enthält, die unabhängig voneinander den Übergang zur Selbstbefruchtung vollzogen. Verglichen mit dem auskreuzendem Vorfahr C. grandiflora, ist die Emission von Benzaldehyd als Hauptbestandteil des Blütenduftes in den selbstbestäubenden Arten C. rubella und C. orientalis verloren gegangen oder stark reduziert. Der Verlust von Benzaldehyd wurde in C. rubella durch Mutationen in der Cinnamat:CoA Ligase CNL1 verursacht, die biochemische Grundlage und evolutionäre Geschichte dieses Verlustes waren jedoch unbekannt, ebenso wie die genetische Grundlage für den Duftverlust von C. orientalis. In der vorliegenden Arbeit wurde eine Kombination aus transformierten Pflanzen, in vitro Enzymassays, Populationsgenetik und quantitativer Genetik genutzt, um diese Fragen zu beantworten. Die Ergebnisse sprechen dafür, dass in C. rubella CNL1 zweimal unabhängig voneinander durch Punktmutationen inaktiviert wurde und diese zu einem Verlust der Benzaldehydemission führten. Beide inaktive Haplotypen können in Populationen rund um das Mittelmeer detektiert werden, was dafür spricht, dass die Mutationen auftraten, bevor sich die Art geografisch ausbreiten konnte. Diese Arbeit bestätigt, dass CNL1 ein sogenannter Hotspot für die Eliminierung der Benzaldehydemission ist, wie es schon von vorhergehenden Arbeiten angedeutet wurde. Im Gegensatz dazu verbleibt CNL1 in C. orientalis funktionsfähig. Um herauszufinden, ob bei dieser Art ähnliche Mechanismen der konvergenten Evolution zum Duftverlust führten, wurde ein QTL-Kartierungsansatz gewählt. Die Ergebnisse sprechen dafür, dass hier die Duftreduktion auf einem evolutionär anderen Weg erreicht wurde als in C. rubella, eventuell spiegelt dies wider, dass die Evolution von Blütenduft eher durch ökologische als genetische Faktoren angetrieben wird. Parallel zur Aufklärung der genetischen Basis von wiederholtem Duftverlust wurde eine Methode zum Testen des adaptiven Wertes einzelner Merkmale des Selbstungssyndromes etabliert. Diese Methode erlaubt es, Auskreuzungsraten mit einem hohen Probendurchsatz abzuschätzen, und es konnte gezeigt werden, dass sie erfolgreich Auskreuzungsereignisse detektiert, die von Insekten vermittelt wurden. Diese neue Methode bringt einige Vorteile gegenüber bisher angewandten Protokollen mit sich; sie ist wesentlich schneller und eine Vielzahl an Proben lässt sich mit deutlich geringerem Aufwand analysieren. Dies ermöglicht eine Korrelation der Auskreuzungsrate mit unterschiedlichen Merkmalen, indem, wie hier demonstriert, quasi-isogene Linien verwendet werden. Denkbar wäre auch eine Kombination mit anderen morphologischen oder ökologischen Parametern, um deren Einfluss auf die Auskreuzungsrate zu untersuchen. Konvergente Evolution kann nicht nur beim Duftverlust in der Gattung Capsella beobachtet werden, sondern auch bei der morphologischen Evolution der Petalengröße. Vorhergehende Studien fanden mehrere QTL, die der Reduktion der Petalengröße in C. rubella und C. orientalis unterliegen, manche von ihnen sind sogar in beiden Spezies zu finden. Einer der gemeinsamen QTL ist PAQTL1, der vielleicht auf NUBBIN zurückgeführt werden kann, einem Wachstumsfaktor. Um die morphologische Evolution und die genetische Grundlage der Reduktion der Petalengröße besser zu verstehen, soll dieser QTL weiter charakterisiert werden, da ihm eventuell ein Hotspot-Gen für die konvergente Evolution der Petalengröße unterliegen könnte. KW - Capsella KW - selfing syndrome KW - floral scent KW - outcrossing rate KW - benzaldehyde KW - CNL1 KW - Capsella KW - Selbstungssyndrom KW - Blütenduft KW - Auskreuzungsrate KW - Benzaldehyd KW - CNL1 Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-435253 ER - TY - GEN A1 - Jantzen, Friederike A1 - Lynch, Joseph H. A1 - Kappel, Christian A1 - Höfflin, Jona A1 - Skaliter, Oded A1 - Wozniak, Natalia Joanna A1 - Sicard, Adrien A1 - Sas, Claudia A1 - Adebesin, Funmilayo A1 - Ravid, Jasmin A1 - Vainstein, Alexander A1 - Hilker, Monika A1 - Dudareva, Natalia A1 - Lenhard, Michael T1 - Retracing the molecular basis and evolutionary history of the loss of benzaldehyde emission in the genus Capsella T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - The transition from pollinator-mediated outbreeding to selfing has occurred many times in angiosperms. This is generally accompanied by a reduction in traits attracting pollinators, including reduced emission of floral scent. In Capsella, emission of benzaldehyde as a main component of floral scent has been lost in selfing C. rubella by mutation of cinnamate-CoA ligase CNL1. However, the biochemical basis and evolutionary history of this loss remain unknown, as does the reason for the absence of benzaldehyde emission in the independently derived selfer Capsella orientalis. We used plant transformation, in vitro enzyme assays, population genetics and quantitative genetics to address these questions. CNL1 has been inactivated twice independently by point mutations in C. rubella, causing a loss of enzymatic activity. Both inactive haplotypes are found within and outside of Greece, the centre of origin of C. rubella, indicating that they arose before its geographical spread. By contrast, the loss of benzaldehyde emission in C. orientalis is not due to an inactivating mutation in CNL1. CNL1 represents a hotspot for mutations that eliminate benzaldehyde emission, potentially reflecting the limited pleiotropy and large effect of its inactivation. Nevertheless, even closely related species have followed different evolutionary routes in reducing floral scent. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 775 KW - benzaldehyde KW - Capsella KW - cinnamate-CoA ligase KW - evolution KW - floral scent KW - selfing syndrome KW - shepherd’s purse Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-437542 SN - 1866-8372 IS - 775 SP - 1349 EP - 1360 ER - TY - JOUR A1 - Streubel, Susanna A1 - Fritz, Michael Andre A1 - Teltow, Melanie A1 - Kappel, Christian A1 - Sicard, Adrien T1 - Successive duplication-divergence mechanisms at the RCO locus contributed to leaf shape diversity in the Brassicaceae JF - Development : Company of Biologists N2 - Gene duplication is a major driver for the increase of biological complexity. The divergence of newly duplicated paralogs may allow novel functions to evolve, while maintaining the ancestral one. Alternatively, partitioning the ancestral function among paralogs may allow parts of that role to follow independent evolutionary trajectories. We studied the REDUCED COMPLEXITY (RCO) locus, which contains three paralogs that have evolved through two independent events of gene duplication, and which underlies repeated events of leaf shape evolution within the Brassicaceae. In particular, we took advantage of the presence of three potentially functional paralogs in Capsella to investigate the extent of functional divergence among them. We demonstrate that the RCO copies control growth in different areas of the leaf. Consequently, the copies that are retained active in the different Brassicaceae lineages contribute to define the leaf dissection pattern. Our results further illustrate how successive gene duplication events and subsequent functional divergence can increase trait evolvability by providing independent evolutionary trajectories to specialized functions that have an additive effect on a given trait. KW - Plant development KW - Gene duplication KW - Leaf shape KW - Morphological evolution KW - Capsella KW - Arabidopsis Y1 - 2018 U6 - https://doi.org/10.1242/dev.164301 SN - 0950-1991 SN - 1477-9129 VL - 145 IS - 8 PB - Company of Biologists CY - Cambridge ER -