ArticlePlant physiology2009
Deletion-based reverse genetics in Medicago truncatula.
Article in Plant physiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- Genomics of predictive radiation mutagenesis in oilseed rape: modifying seed oil composition.Plant biotechnology journal · 2024Article
- Review
- Critical role for uricase and xanthine dehydrogenase in soybean nitrogen fixation and nodule development.The plant genome · 2023Article
- Review
- Biological and genomic analysis of a symbiotic nitrogen fixation defective mutant inFrontiers in plant science · 2023Article
- Rapid identification of mutations caused by fast neutron bombardment in Medicago truncatula.Plant methods · 2021Article
- Screening populations for copy number variation using genotyping-by-sequencing: a proof of concept using soybean fast neutron mutants.BMC genomics · 2019Article
- Expanding Avenue of Fast Neutron Mediated Mutagenesis for Crop Improvement.Plants (Basel, Switzerland) · 2019Review
- The MtDMI2-MtPUB2 Negative Feedback Loop Plays a Role in Nodulation Homeostasis.Plant physiology · 2018Article
- FNBtools: A Software to Identify Homozygous Lesions in Deletion Mutant Populations.Frontiers in plant science · 2018Article
- An Array-based Comparative Genomic Hybridization Platform for Efficient Detection of Copy Number Variations in Fast Neutron-induced Medicago truncatula Mutants.Journal of visualized experiments : JoVE · 2017Article
- Identification of Homogentisate Dioxygenase as a Target for Vitamin E Biofortification in Oilseeds.Plant physiology · 2016Article
- Analysis of Large Seeds from Three Different Medicago truncatula Ecotypes Reveals a Potential Role of Hormonal Balance in Final Size Determination of Legume Grains.International journal of molecular sciences · 2016Article
- A Snapshot of Functional Genetic Studies in Medicago truncatula.Frontiers in plant science · 2016Review
- Gene Overexpression Resources in Cereals for Functional Genomics and Discovery of Useful Genes.Frontiers in plant science · 2016Review
- A draft genome of field pennycress (Thlaspi arvense) provides tools for the domestication of a new winter biofuel crop.DNA research : an international journal for rapid publication of reports on genes and genomes · 2015Article
- Genome resilience and prevalence of segmental duplications following fast neutron irradiation of soybean.Genetics · 2014Article
- Article
- Article
- pENCODE: a plant encyclopedia of DNA elements.Annual review of genetics · 2014Review
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Authors and funding
4 authors.
Funding
Abstract
The primary goal of reverse genetics, the identification of null mutations in targeted genes, is achieved through screening large populations of randomly mutagenized plants. T-DNA and transposon-based mutagenesis has been widely employed but is limited to species in which transformation and tissue culture are efficient. In other species, TILLING (for Targeting Induced Local Lesions IN Genomes), based on chemical mutagenesis, has provided an efficient method for the identification of single base pair mutations, only 5% of which will be null mutations. Furthermore, the efficiency of inducing point mutations, like insertion-based mutations, is dependent on target size. Here, we describe an alternative reverse genetic strategy based on physically induced genomic deletions that, independent of target size, exclusively recovers knockout mutants. Deletion TILLING (De-TILLING) employs fast neutron mutagenesis and a sensitive polymerase chain reaction-based detection. A population of 156,000 Medicago truncatula plants has been structured as 13 towers each representing 12,000 M2 plants. The De-TILLING strategy allows a single tower to be screened using just four polymerase chain reaction reactions. Dual screening and three-dimensional pooling allows efficient location of mutants from within the towers. With this method, we have demonstrated the detection of mutants from this population at a rate of 29% using five targets per gene. This De-TILLING reverse genetic strategy is independent of tissue culture and efficient plant transformation and therefore applicable to any plant species. De-TILLING mutants offer advantages for crop improvement as they possess relatively few background mutations and no exogenous DNA.
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