An assessment of heavy ion irradiation mutagenesis for reverse genetics in wheat (Triticum aestivum L.)

Fitzgerald, Timothy L., Powell, Jonathan J., Stiller, Jiri, Weese, Terri L., Abe, Tomoko, Zhao, Guangyao, Jia, Jizeng, McIntyre, C. Lynne, Li, Zhongyi, Manners, John M. and Kazan, Kemal (2015) An assessment of heavy ion irradiation mutagenesis for reverse genetics in wheat (Triticum aestivum L.). PLoS One, 10 2: . doi:10.1371/journal.pone.0117369


Author Fitzgerald, Timothy L.
Powell, Jonathan J.
Stiller, Jiri
Weese, Terri L.
Abe, Tomoko
Zhao, Guangyao
Jia, Jizeng
McIntyre, C. Lynne
Li, Zhongyi
Manners, John M.
Kazan, Kemal
Title An assessment of heavy ion irradiation mutagenesis for reverse genetics in wheat (Triticum aestivum L.)
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2015-02-26
Year available 2015
Sub-type Article (original research)
DOI 10.1371/journal.pone.0117369
Open Access Status DOI
Volume 10
Issue 2
Total pages 23
Place of publication San Francisco, CA United States
Publisher Public Library of Science
Language eng
Subject 1300 Biochemistry, Genetics and Molecular Biology
1100 Agricultural and Biological Sciences
Abstract Reverse genetic techniques harnessing mutational approaches are powerful tools that can provide substantial insight into gene function in plants. However, as compared to diploid species, reverse genetic analyses in polyploid plants such as bread wheat can present substantial challenges associated with high levels of sequence and functional similarity amongst homoeologous loci. We previously developed a high-throughput method to identify deletions of genes within a physically mutagenized wheat population. Here we describe our efforts to combine multiple homoeologous deletions of three candidate disease susceptibility genes ( TaWRKY11, TaPFT1 and TaPLDβ1 ). We were able to produce lines featuring homozygous deletions at two of the three homoeoloci for all genes, but this was dependent on the individual mutants used in crossing. Intriguingly, despite extensive efforts, viable lines possessing homozygous deletions at all three homoeoloci could not be produced for any of the candidate genes. To investigate deletion size as a possible reason for this phenomenon, we developed an amplicon sequencing approach based on synteny to Brachypodium distachyon to assess the size of the deletions removing one candidate gene (TaPFT1) in our mutants. These analyses revealed that genomic deletions removing the locus are relatively large, resulting in the loss of multiple additional genes. The implications of this work for the use of heavy ion mutagenesis for reverse genetic analyses in wheat are discussed.
Formatted abstract
Reverse genetic techniques harnessing mutational approaches are powerful tools that can provide substantial insight into gene function in plants. However, as compared to diploid species, reverse genetic analyses in polyploid plants such as bread wheat can present substantial challenges associated with high levels of sequence and functional similarity amongst homoeologous loci. We previously developed a high-throughput method to identify deletions of genes within a physically mutagenized wheat population. Here we describe our efforts to combine multiple homoeologous deletions of three candidate disease susceptibility genes (TaWRKY11, TaPFT1 and TaPLDß1). We were able to produce lines featuring homozygous deletions at two of the three homoeoloci for all genes, but this was dependent on the individual mutants used in crossing. Intriguingly, despite extensive efforts, viable lines possessing homozygous deletions at all three homoeoloci could not be produced for any of the candidate genes. To investigate deletion size as a possible reason for this phenomenon, we developed an amplicon sequencing approach based on synteny to Brachypodium distachyon to assess the size of the deletions removing one candidate gene (TaPFT1) in our mutants. These analyses revealed that genomic deletions removing the locus are relatively large, resulting in the loss of multiple additional genes. The implications of this work for the use of heavy ion mutagenesis for reverse genetic analyses in wheat are discussed.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Alliance for Agriculture and Food Innovation
Official 2016 Collection
 
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