5 results on '"Aimee Walmsley"'
Search Results
2. The Soil Microbiome Reduces Striga Infection of Sorghum by Modulation of Host-Derived Signaling Molecules and Root Development
- Author
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Dorota Kawa, Benjamin Thiombiano, Mahdere Shimels, Tamera Taylor, Aimee Walmsley, Hannah E Vahldick, Marcio FA Leite, Zayan Musa, Alexander Bucksch, Francisco Dini-Andreote, Alexander J Chen, Jiregna Daksa, Desalegn Etalo, Taye Tessema, Eiko E Kuramae, Jos M Raaijmakers, Harro Bouwmeester, and Siobhan M Brady
- Abstract
Sorghum bicoloris one of the most important cereals in the world and a staple crop for smallholder famers in sub-Saharan Africa. However approximately 20% of sorghum yield is annually lost on the African continent due to infestation with the root parasitic weedStriga hermonthica.Existing Striga management strategies often show an inconsistent to low efficacy. Hence, novel and integrated approaches are needed as an alternative strategy. Here, we demonstrate that the soil microbiome suppresses Striga infection in sorghum. We associate this suppression with microbiome-mediated induction of root endodermal suberization and aerenchyma formation, and depletion of haustorium inducing factors (HIFs), root exudate compounds that are critical for the initial stages of Striga infection. We further identify microbial taxa associated with reduced Striga infection with concomitant changes in root cellular anatomy and differentiation as well as HIF degradation. Our study describes novel microbiome-mediated mechanisms of Striga suppression, encompassing repression of haustorium formation and induction of physical barriers in the host root tissue. These findings open new avenues to broaden the effectiveness of Striga management practices.
- Published
- 2023
3. Probing strigolactone perception mechanisms with rationally designed small-molecule agonists stimulating germination of root parasitic weeds
- Author
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Dawei Wang, Zhili Pang, Haiyang Yu, Benjamin Thiombiano, Aimee Walmsley, Shuyi Yu, Yingying Zhang, Tao Wei, Lu Liang, Jing Wang, Xin Wen, Harro J. Bouwmeester, Ruifeng Yao, and Zhen Xi
- Subjects
Lactones ,Multidisciplinary ,Seeds ,General Physics and Astronomy ,Humans ,Plant Weeds ,Germination ,Perception ,General Chemistry ,Striga ,Heterocyclic Compounds, 3-Ring ,General Biochemistry, Genetics and Molecular Biology - Abstract
The development of potent strigolactone (SL) agonists as suicidal germination inducers could be a useful strategy for controlling root parasitic weeds, but uncertainty about the SL perception mechanism impedes real progress. Here we describe small-molecule agonists that efficiently stimulate Phelipanchce aegyptiaca, and Striga hermonthica, germination in concentrations as low as 10−8 to 10−17 M. We show that full efficiency of synthetic SL agonists in triggering signaling through the Striga SL receptor, ShHTL7, depends on the receptor-catalyzed hydrolytic reaction of the agonists. Additionally, we reveal that the stereochemistry of synthetic SL analogs affects the hydrolytic ability of ShHTL7 by influencing the probability of the privileged conformations of ShHTL7. Importantly, an alternative ShHTL7-mediated hydrolysis mechanism, proceeding via nucleophilic attack of the NE2 atom of H246 to the 2′C of the D-ring, is reported. Together, our findings provide insight into SL hydrolysis and structure-perception mechanisms, and potent suicide germination stimulants, which would contribute to the elimination of the noxious parasitic weeds.
- Published
- 2021
4. BIBAC-GW-based vectors for generating reporter lines for site-specific genome editing in planta
- Author
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Rurika Oka, Damar Tri Anggoro, Mariliis Tark-Dame, Aimee Walmsley, Mara de Sain, Maike Stam, Plant Development & (Epi)Genetics (SILS, FNWI), Systems Biology, Intelligent Sensory Information Systems (IVI, FNWI), and Molecular Plant Pathology (SILS, FNWI)
- Subjects
DNA, Bacterial ,Genetic Markers ,0301 basic medicine ,Transgene ,Genetic Vectors ,Arabidopsis ,Gene Expression ,Mutagenesis (molecular biology technique) ,Computational biology ,Biology ,Genome ,03 medical and health sciences ,Transformation, Genetic ,Genome editing ,Genes, Reporter ,Gene Order ,Transgenes ,Vector (molecular biology) ,Molecular Biology ,Selectable marker ,Gene Editing ,Cloning ,Genetics ,Transcription activator-like effector nuclease ,Base Sequence ,Plants, Genetically Modified ,030104 developmental biology ,Genome, Plant - Abstract
When generating transgenic plants, one of the objectives is to achieve stable expression of the transgene. Transgene silencing can be avoided by single copy integration of the transgene. Binary systems that predominantly result in single copy integrations, such as BIBAC vectors, are also single-copy in E. coli, the organism in which the T-DNA to be delivered to the plant is assembled. Although a low-copy number is important for stable maintenance of large DNA fragments in E. coli, it hampers cloning into the vector due to a low DNA yield. Here we describe BIBAC vectors to which Gateway site-specific recombination sites are added. These sites provide a fast and easy introduction of sequences of interest into any vector. Our Gateway-compatible BIBAC vectors are available with two selectable markers for plants - resistance to Basta (BIBAC-BAR-GW) and DsRed fluorescence in the seed coat (BIBAC-RFP-GW).Using the BIBAC-BAR-GW vector we have generated different fluorescence-based reporter constructs that, when delivered to plant cells, can be used to study and optimize precise, template-dependent site-specific genome editing by CRISPR-Cas9, TALENs or ZFP-nuclease complexes, and oligonucleotide-directed mutagenesis. We have generated 59 reporter lines in A. thaliana with our reporter constructs, and for the lines carrying single T-DNA integrations (32 out of 59) we have determined the integrity of the integrations, their genomic locations and the expression level of the reporters. Similarly to its original counterpart, BIBAC-BAR-GW generates single T-DNA integrations in Arabidopsis with 50% efficiency, and 90% of those are intact. The reporter constructs in the independent transgenic lines exhibit only an up to 3-fold difference in expression level. These features combined with an easy manipulation of the vector due to the added Gateway sites make the BIBAC-GW vectors an attractive tool for generating transgenic plants.
- Published
- 2017
5. Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
- Author
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Mariliis Tark-Dame, Rechien Bader, Damar Tri Anggoro, Aimee Walmsley, Rurika Oka, Mara de Sain, Maike Stam, Blaise Weber, Plant Development & (Epi)Genetics (SILS, FNWI), Molecular Plant Pathology (SILS, FNWI), and Plant Hormone Biology (SILS, FNWI)
- Subjects
0106 biological sciences ,0301 basic medicine ,Transfer DNA ,Bacterial artificial chromosome ,General Immunology and Microbiology ,biology ,Agrobacterium ,General Chemical Engineering ,General Neuroscience ,Transgene ,Genetic Vectors ,Computational biology ,Genetically modified crops ,Plants, Genetically Modified ,Gateway cassette ,biology.organism_classification ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Transformation (genetics) ,Transformation, Genetic ,030104 developmental biology ,Genetics ,010606 plant biology & botany ,Southern blot - Abstract
When generating transgenic plants, generally the objective is to have stable expression of a transgene. This requires a single, intact integration of the transgene, as multi-copy integrations are often subjected to gene silencing. The Gateway-compatible binary vector based on bacterial artificial chromosomes (pBIBAC-GW), like other pBIBAC derivatives, allows the insertion of single-copy transgenes with high efficiency. As an improvement to the original pBIBAC, a Gateway cassette has been cloned into pBIBAC-GW, so that the sequences of interest can now be easily incorporated into the vector transfer DNA (T-DNA) by Gateway cloning. Commonly, the transformation with pBIBAC-GW results in an efficiency of 0.2–0.5%, whereby half of the transgenics carry an intact single-copy integration of the T-DNA. The pBIBAC-GW vectors are available with resistance to Glufosinate-ammonium or DsRed fluorescence in seed coats for selection in plants, and with resistance to kanamycin as a selection in bacteria. Here, a series of protocols is presented that guide the reader through the process of generating transgenic plants using pBIBAC-GW: starting from recombining the sequences of interest into the pBIBAC-GW vector of choice, to plant transformation with Agrobacterium, selection of the transgenics, and testing the plants for intactness and copy number of the inserts using DNA blotting. Attention is given to designing a DNA blotting strategy to recognize single- and multi-copy integrations at single and multiple loci.
- Published
- 2018
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