Evolutionary genetics of carpel development using California poppy (Eschscholzia californica) as a new model species
Final Report Abstract
The project on the evolution of carpel development using the basal eudicot model species Eschscholzia californica (California poppy) firstly installed California poppy as model plant for evo-devo studies by showing that analysis of candidate's gene functions by Virus-induced Gene Silencing is highly efficient and reliable. Consequently, the method was used to unravel the function of several genes required for California poppy carpel development, such as the poppy's CRABS CLAW and two AGAMOUS orthologs. Other Arabidopsis thaliana carpel development orthologs are still being analyzed functionally using VIGS, as plant transformation of California poppy still is problematic. While we can regenerate plants that have been cocultured with transgenic Agrobacterium tumefaciens strains and that are able to regenerate on selective media, we are still lacking final proof by Southern Blot hybridization that the transgenes have integrated into the poppy's genome. Thus, VIGS and analysis of mutagenized plants are currently our methods of choice to identify gene functions and regulatory networks governing poppy development. A BAC library was constructed to allow mutant identification and efficient promoter isolation and this BAC library will most likely also be utilized for future poppy genome sequencing. Several mutants could be identified already, due to the lack of hands only few of them could be analyzed further, such as the winzig mutant or seirene. Due to technical advances in the field we have now a high quality transcriptome of premeiotic gynoecia of poppy produced by Laser Capture Microdissection followed by RNAseq. This will yield an even larger number of putative carpel development genes in an unbiased approach that can be analyzed by VIGS to better understand how carpel development is controlled in basal eudicot species. We have been able to address, with one exception, all aims of the project. Meanwhile, in my lab the sequences are used produced by genome sequencing of phylogenetically informative species to unravel the phylogenetic history of genes required for carpel development in A. thaliana. It is shown that the regulatory network orchestrating carpel development in A. thaliana was not recruited from a pre-existing network but has been sequentially assembled over time. Further, we are working on the interaction network and non-cell autonomous action of CRC. A PhD student is currently carrying out VIGS with a number of putative poppy carpel genes to elucidate their function. My lab will pursue many interesting lines of research that are derived from the funded project.
Publications
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(2006) Conservation and divergence in the AGAMOUS Subfamily of MADS-Box Genes: Evidence of Independent Sub- and Neofunctionalization Events. Evol Dev 8: 30-45
Zahn LM, Leebens-Mack J, Arrington JM, Hu Y, Landherr L, dePamphilis C, Becker A, Theissen G, Ma H
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(2006) EST database for early flower development in California poppy (Eschscholzia californica Cham., Papaveraceae) tags over 6000 genes from a basal eudicot. Plant Mol Biol 62: 351- 369
Carlson J, Leebens-Mack JH, Wall PK, Zahn LM, Mueller LM, Landherr LL, Hu Y, Nut DC, Arrington JM, Choirean S, Becker A, Field D, Tanksley SD, Ma H, dePamphilis CW
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(2006) Petaloidy and petal identity MADS-box genes in the balsaminoid genera Impatiens and Marcgravia. Plant J 47: 501-518
Geuten K, Becker A, Kaufmann K, Caris P, Janssens S, Viaene T, Theissen G, Smets E
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(2007) Highly efficient virus-induced gene silencing (VIGS) in California poppy {Eschscholzia californica Cham.): An evaluation of VIGS as a strategy to obtain functional data from non-model plants. Annals Bot 100: 641-649
Wege S, Scholz A, Gleissberg S, Becker A
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(2009) The CRABS CMIA^ortholog from the basal eudicot California poppy, {Eschschoizia californica), EcCRC, is involved in floral meristem termination and gynoecium development. Plant Journal 58: 682-693
Orashakova S, Lange M, Lange S, Wege S, Becker A
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(2010) Diverse functions ofthe AGL6 MADS domain transcription factor lineage afler a core eudicot duplication suggests functional diversification. BMC Plant Biol 10:148
Viane T, Vekemans D, Becker A, Melzer S, Geuten
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(2010) Floral homeotic C function genes repress specific B function genes in the carpel whorl of the basal eudicot California poppy {Eschscholzia califomica). BMC Evodevo 1:13
Yellina A, Orashakova S, Lange S, Erdmann E, Leebens-Mack J, Becker A
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(2010) VIGS - genomics goes functional. Trends in Plant Sciences 15(1): 1-4
Lange M, Becker A
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(2013) Duplicated STM-like KNOX l genes act in floral meristem activity in Eschscholzia californica (Papaveraceae). Development Genes and Evolution, Volume 223, Issue 5, pp 289-301
Stammler A, Meyer SS, Plant AR, Townsley BT, Becker A, Gleissberg S
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(2013) The seirena B class floral homeotic mutant of California Poppy (Eschscholzia californica) reveals a function of the enigmatic PI motif in the fonmation of specific multimeric MADS domain protein complexes. Plant Cell 25(2):438-53
Lange M, Orashakova S, Lange S, Melzer R, Theißen G, Smyth DR, Becker A
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(2013) Virus-Induced Gene Silencing (VIGS) in Plants: An Overview of Target Species and the Virus-Derived Vector Systems. Methods in molecular biology (Clifton, NJ) 01/2013; 975:1-14
Lange M, Yellina AL, Orashakova S, Becker A
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(2013) Virus-Induced Gene Silencing of the Alkaloid-Producing Basal Eudicot Model Plant Eschscholzia californica (California Poppy). Methods in molecular biology (Clifton, N.J.) 01/2013; 975:83-98
Tekleyohans DG, Lange S, Becker A
