Progress 10/01/18 to 10/30/18
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Dr. Michum has left the University of Missouri and work on her project here as ended. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Dr. Michum has left the University of Missouri and work on her project here as ended.
Publications
|
Progress 02/21/14 to 10/30/18
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Dr. Michum has left the University of Missouri and work on her project here as ended.
Publications
|
Progress 10/01/17 to 09/30/18
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Dr. Michum has left the University of Missouri and work on her project here as ended. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Dr. Michum has left the University of Missouri and work on her project here as ended.
Publications
|
Progress 10/01/16 to 09/30/17
Outputs
Target Audience:Scientists studying plant developmental biology, peptide signaling, and plant-nematode interactions. Additional target audiences include commodity groups, growers, and industry. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided training to one post-doctoral research associate. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?We will perform genomic complementation assays using newly identified candidate genes. We also expect that genes involved in nematode CLE effector trafficking and/or signaling may exhibit altered expression at nematode feeding sites. The expression patterns of identified genes will be studied by qPCR and/or promoter-GUS experiments. Nematode CLEs play important roles in feeding site initiation and maintenance. Previously, we demonstrated that knockingout nematode CLE receptors in Arabidopsis results in the development of fewer females and smaller syncytia. Thus, we expect that mutations in plant genes affecting CLE protein trafficking and/or signaling will compromise feeding site formation. On the other hand, overexpression of the candidate genes may promote infection. Available mutant lines of the identified genes will be requested from the Arabidopsis seed stock center or other research labs. Overexpression lines will be generated. Cyst nematode infection assays will be performed using established methodology.
Impacts
What was accomplished under these goals?
We 1) examined the expression profile of this gene at nematode feeding sites, 2) overexpressed the wild-type and mutated version in plants, 3) conducted nematode infection assays on mutant plants, 4) carried out a complementation experiment with a genomic clone carrying this gene, and 5) knocked out the gene using CRISPR/Cas. Unfortunately, all of these experiments suggested that the mapped causal gene was not the gene underlying the observed phenotype. For this reason, we have reanalyzed our mapping by sequencing data. Two additional candidate genes have been identified and further testing is underway.?
Publications
|
Progress 10/01/15 to 09/30/16
Outputs
Target Audience:Scientists studying plant developmental biology, peptide signaling, and plant-nematode interactions. Additional target audiences include commodity groups, growers, and industry. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided training to one post-doctoral research associate. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?We will 1) examine the expression profile of this gene at nematode feeding sites, 2) overexpress the wild-type and mutated version in plants, and 3) conduct nematode infection assays on mutant plants, 4) carry out a complementation experiment with a genomic clone carrying this gene, and 5) knock out the gene using CRISPR/Cas.
Impacts
What was accomplished under these goals?
Several mutant lines have been identified in our preliminary screening. One mutant harbors a mutation in a gene encoding a plant transcription factor and represents a potentially novel component of plant and nematode CLE signaling. We generated the transgenic plants and other resources to further characterize the potential role of this gene in the plant-nematode interaction.
Publications
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Mitchum, MG. Sainsbury Lab Symposium Induced Plant Development, University of Cambridge, April 4-6, 2016, Modulation of CLE Signaling Systems for Nematode Parasitism
|
Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Scientists studying plant developmental biology, peptide signaling, and plant-nematode interactions. Additional target audiences include commodity groups, growers, and industry. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided training to one post-doctoral research associate and one undergraduate student in Plant Sciences. How have the results been disseminated to communities of interest?Results were presented at professional scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?We expect that genes involved in nematode CLE effector trafficking and/or signaling may exhibit altered expression at nematode feeding sites. The expression patterns of identified genes will be studied by qPCR and/or promoter-GUS experiments. Promoter sequences will be amplified by PCR and cloned into pBI101 to generate promoter-GUS fusion constructs. Arabidopsis plants will be transformed by floral dipping. Transgenic Arabidopsis plants will be infected with BCN and GUS expression will be monitored at different time points following nematode infection. Nematode CLEs play important roles in feeding site initiation and maintenance. Previously, we demonstrated that knocking out nematode CLE receptors in Arabidopsis results in the development of fewer females and smaller syncytia. Thus, we expect that mutations in plant genes affecting CLE protein trafficking and/or signaling will compromise feeding site formation. On the other hand, overexpression of the candidate genes may promote infection. Available mutant lines of the identified genes will be requested from the Arabidopsis seed stock center or other research labs. Overexpression lines will be generated. Cyst nematode infection assays will be performed using established methodology.
Impacts
What was accomplished under these goals?
Several mutant lines have been identified in our preliminary screening and more mutant lines are expected during the proposed funding period. Using a mapping by sequencing approach, we have identified the causal genes in three of the mutant lines to demonstrate proof-of-concept in 1) our ability to map the causal genes using this approach, and 2) in the approach itself, both by rediscovering plant/nematode CLE receptors and identifying novel signaling components. Two of the mutants harbor mutations in the previously identified nematode CLE receptors, CLV1 and CRN. However, a third mutant harbors a mutation in a gene encoding a plant transcription factor and represents a potentially novel component of plant and nematode CLE signaling.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Mitchum, MG. American Society of Plant Biology Meeting, Minneapolis, MN, July, 2015, Signaling in Plant-Nematode Interactions Plenary Speaker
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Wang J, Baum TJ, Davis EL, Wang X, Mitchum MG. A translocation signal for delivery of nematode CLE effectors to the extracellular space of host plant cells. Plant Biology 2015, July 25-30, 2015, Minneapolis, MN.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Wang J, Baum TJ, Davis EL, Wang X, Mitchum MG. A translocation signal for delivery of nematode CLE effectors to the extracellular space of host plant cells. Interdisciplinary Plant Group Symposium, Plants Between a Rock and a Hard Place: The Interface Between Abiotic and Biotic Stress Responses, May 27-29, 2015, Columbia, MO.
|
Progress 02/21/14 to 09/30/14
Outputs
Target Audience:Scientists studying plant developmental biology, peptide signaling, and plant-nematode interactions. Additional target audiences include commodity groups, growers, and industry. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided training to one post-doctoral research associate and one undergraduate student in Plant Sciences. How have the results been disseminated to communities of interest?Results were presented at professional scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?Next steps include mapping the mutations and characterizing the genes underlying these mutations for a role in plant-nematode interactions.
Impacts
What was accomplished under these goals?
We recently demonstrated that the nematode CLE VD fused to the AtCLV3 CLE domain, under the control of the native CLV3 promoter, fully complements the clv3-2 mutant phenotype. A fully complemented line was selected for mutagenesis. After a pilot mutagenesis experiment with different concentrations of EMS, 15mM was chosen to achieve an embryo lethality rate of 50%. EMS mutagenesis was conducted on more than 10,000 M0 seeds. Mutagenized seeds were planted in soil at a density of 20-30 seeds/pot. Seeds of M1 plants from each pot were pooled to produce 359 M2 pools. Approximately 200 M2 seedlings from each pool have been screened for full or partial restoration of the clv3-2 mutant phenotype based on the silique phenotype (>2 carpels). We expect the causal mutations in these lines to be potentially related to nematode CLE peptide trafficking or signaling.
Publications
|
|