Germline transformation of Shepherd's purse (Capsella bursa-pastoris) by the ‘floral dip’ method as a tool for evolutionary and developmental biology

Bartholmes, Conny; Nutt, Pia; Theißen, Günter

doi:10.1016/j.gene.2007.10.033

Cited by 22 publications

(14 citation statements)

References 29 publications

(36 reference statements)

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“…Further, as transformation technology improves for many species, including more and more cases of drip/dip transformation (e.g. Bartholmes et al. , 2008), it may become possible to use recipient species other than A. thaliana .…”

Section: Discussionmentioning

confidence: 99%

An assessment of transgenomics as a tool for identifying genes involved in the evolutionary differentiation of closely related plant species

et al. 2011

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Summary Transgenomics is the process of introducing genomic clones from a donor species into a recipient species and then screening the resultant transgenic lines for phenotypes of interest. This method might allow us to find genes involved in the evolution of phenotypic differences between species as well as genes that have the potential to contribute to reproductive isolation: potential speciation genes.More than 1,100 ~20 Kbp genomic clones from Leavenworthia alabamica were moved into Arabidopsis thaliana by transformation. After screening a single primary transformant for each line, clones associated with mutant phenotypes were tested for repeatability and cosegregation.We found 84 clones with possible phenotypic effects of which eight were repeatedly associated with the same phenotype. One clone, 11_11B, cosegregated with a short fruit phenotype. Further study showed that 11_11B affects seed development, with as much as one-third of the seeds aborted in some fruit.Transgenomics is a viable strategy for discovering genes of evolutionary interest. We identify methods to reduce false positives and false negatives in the future. 11_11B can be viewed as a potential speciation gene, illustrating the value of transgenomics for studying the molecular basis of reproductive isolation.

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Section: Discussionmentioning

confidence: 99%

An assessment of transgenomics as a tool for identifying genes involved in the evolutionary differentiation of closely related plant species

et al. 2011

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“…Plant material, experimental design and analysis of floral visitation All wild-type (1947-wt) and floral homeotic (1947-Spe) plants used in this study trace back to one wild-type and one Spe plant, respectively; these were originally isolated from natural populations of C. bursa-pastoris growing in and near vineyards close to Gau-Odernheim, 30 km southwest of the city of Mainz (Rheinhessen, Germany) Nutt et al 2006;Bartholmes et al 2008). In this study, the term 'floral phenotype' is defined either as 'wild-type' if second whorl floral organs are petals or as 'Spe' or 'mutant' if these organs are stamens.…”

Section: Methodsmentioning

confidence: 99%

Floral visitation and reproductive traits of Stamenoid petals, a naturally occurring floral homeotic variant of Capsella bursa-pastoris (Brassicaceae)

Ziermann

Ritz

Hameister

et al. 2009

Planta

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Homeotic changes played a considerable role during the evolution of flowers, but how floral homeotic mutants initially survive in nature has remained enigmatic. To better understand the evolutionary potential of floral homeotic mutants, we established as a model system Stamenoid petals (Spe), a natural variant of Capsella bursa-pastoris (Brassicaceae). In the flowers of Spe plants, petals are transformed into stamens, whereas all other floral organs are unaffected. In contrast with most other homeotic mutants, the Spe variant occurs in relatively stable populations in the wild. In order to determine how the profound change in floral architecture influences plant performance in the wild, we performed common garden experiments running over 3 years. Here, we show that Spe and wild-type plants attract the same assemblage of floral visitors: mainly hoverflies, wild bees and thrips. However, floral visitation is about twice as frequent in wild-type plants as in Spe plants. Nevertheless, the numbers of seeds per fruit were about the same in both variants. Wild-type plants produced more flowers, fruits and seeds per plant than Spe plants, whereas the germination capacity of Spe seeds was higher than that of the wild-type. Determination of volatile composition revealed monoterpenes and 3,4-dimethylbenzaldehyde, which were detected only in wild-type flowers, presumably because they are produced only by petals. Our data indicate that the similar fitness of Spe and wild-type C. bursa-pastoris in the field results from complex compensation between plant architecture and germination capacity. In contrast, flower structure and floral visitation are only of minor importance, possibly because C. bursa-pastoris is mainly self-pollinating.

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“…Several protocols for floral-dip transformation of various Brassicaceae species (Clough and Bent, 1998;Curtis and Nam, 2001;Tague, 2001;Martinez-Trujillo et al, 2004;Bartholmes et al, 2008) were used as a guideline to optimize this technique for L. campestre. The Agrobacterium tumefaciens strains GV3101/pMP90 (Van Larebeke et al, 1974;Koncz and Schell, 1986), LBA4404 (Hoekema et al, 1983) and AGL1 (Lazo et al, 1991) containing binary plasmids pGPTV-Bar (http://biotech.unl.edu/pgptv-bar) or pFGC5941 (http://www.chromdb.org/rnai/pFGC5941.html) were grown at 28°C on YEB medium (5 g/l of each, beef extract, peptone, and sucrose, 1 g/l yeast extract, 0.5 g/l MgSO 4 ) complemented with 50 lg ml À1 kanamycin and 50 lg ml À1 rifampicin (all strains) and additionally with 25 lg ml À1 gentamycin (GV3101).…”

Section: Floral-dip Transformationmentioning

confidence: 99%

“…The most widely used technique to transform A. thaliana is the floral-dip method (Clough and Bent, 1998;Bechtold et al, 2000;Desfeux et al, 2000). Success with this protocol (with slight variations) has also been demonstrated for several other Brassicaceae species (Qing et al, 2000;Curtis and Nam, 2001;Tague, 2001;Wang et al, 2003;Bartholmes et al, 2008;Lu and Kang, 2008). Nevertheless, a previous attempt to transform L. campestre via floral dip proved unsuccessful (Eriksson, 2009).…”

Section: Introductionmentioning

confidence: 99%

Conservation of fruit dehiscence pathways between Lepidium campestre and Arabidopsis thaliana sheds light on the regulation of INDEHISCENT

Lenser

Theißen

2013

The Plant Journal

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SUMMARYThe mode of fruit opening is an important agronomic and evolutionary trait that has been studied intensively in the major plant model system Arabidopsis thaliana. Because fruit morphology is highly variable between species, and is also often the target of artificial selection during breeding, it is interesting to investigate whether a change in fruit morphology may alter the developmental pathway leading to fruit opening. Here we have studied fruit development in Lepidium campestre, a Brassicaceae species that forms silicles instead of siliques. Transgenic L. campestre plants with altered expression levels of orthologs of A. thaliana fruit developmental genes (ALCATRAZ, FRUITFULL, INDEHISCENT and SHATTERPROOF1,2) were found to be defective in fruit dehiscence, and anatomical sections revealed similar changes in tissue patterning as found in respective A. thaliana mutants. Gene expression analyses demonstrated a high degree of conservation in gene regulatory circuits, indicating that, despite great differences in fruit morphology, the process of fruit opening remains basically unchanged between species. Interestingly, our data identify ALCATRAZ as a negative regulator of INDEHISCENT in L. campestre. By mutant analysis, we found the same regulatory relationship in A. thaliana also, thereby shedding new light on how ALCATRAZ drives separation layer formation.

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Germline transformation of Shepherd's purse (Capsella bursa-pastoris) by the ‘floral dip’ method as a tool for evolutionary and developmental biology

Cited by 22 publications

References 29 publications

An assessment of transgenomics as a tool for identifying genes involved in the evolutionary differentiation of closely related plant species

An assessment of transgenomics as a tool for identifying genes involved in the evolutionary differentiation of closely related plant species

Floral visitation and reproductive traits of Stamenoid petals, a naturally occurring floral homeotic variant of Capsella bursa-pastoris (Brassicaceae)

Conservation of fruit dehiscence pathways between Lepidium campestre and Arabidopsis thaliana sheds light on the regulation of INDEHISCENT

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