TILLING and Point Mutation Detection

Maize reverse genetics is extremely powerful because of the wealth of active transposable elements still residing in the maize genome. In addition to serving as molecular tags for mutated genes, these elements tend to knock out genes into which they insert. Once an initial, reference allele has been identified, it facilitates obtaining an allelic series of mutations within a gene that have a range of phenotypes. Such an allelic series can provide great insight into the function of the gene and its cognate protein. In addition to spontaneous mutations, this allelic series is typically the result of chemical mutagenesis to create point mutations at locations throughout the gene. Forward, “targeted” mutagenesis has long been the way to do this. Mutagenizing with ethyl methane sulfonate (EMS) and then crossing onto a line homozygous for a reference allele, any mutant progeny that arise in the F1 represent new mutations in the gene of interest. While effective, a new mutagenesis is required for each gene of interest. The capacity to screen any mutagenized population using reverse genetic techniques for any gene of interest is therefore valuable as well.


Maize Genome Ethyl Methane Sulfonate Reference Allele Allelic Series Single Base Change 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. McCallum CM, Comai L, Greene EA, Henikoff S: Targeting induced local lesions IN genomes (TILLING) for plant functional genomics. Plant Physiol 2000, 123:439–442.PubMedCrossRefGoogle Scholar
  2. Till BJ, Colbert T, Tompa R, Enns LC, Codomo CA, Johnson JE, Reynolds SH, Henikoff JG,Greene EA, Steine MN, et al.: High-throughput TILLING for functional genomics. Methods Mol Biol 2003, 236:205–220.PubMedGoogle Scholar
  3. Henikoff JG, Pietrokovski S, McCallum CM, Henikoff S: Blocks-based methods for detecting protein homology. Electrophoresis 2000, 21:1700–1706.PubMedCrossRefGoogle Scholar
  4. Rozen S, Skaletsky H: Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 2000, 132:365–386.PubMedGoogle Scholar
  5. Till BJ, Burtner C, Comai L, Henikoff S: Mismatch cleavage by single-strand specific nucleases. Nucleic Acids Res 2004, 32:2632–2641.PubMedCrossRefGoogle Scholar
  6. Perry JA, Wang TL, Welham TJ, Gardner S, Pike JM, Yoshida S, Parniske M: A TILLING reverse genetics tool and a web-accessible collection of mutants of the legume Lotus japonicus. Plant Physiol 2003, 131:866–871.PubMedCrossRefGoogle Scholar
  7. Ng PC, Henikoff S: Predicting deleterious amino acid substitutions. Genome Res 2001, 11:863–874.PubMedCrossRefGoogle Scholar
  8. Ng PC, Henikoff S: SIFT: Predicting amino acid changes that affect protein function. Nucleic Acids Res 2003, 31:3812–3814.PubMedCrossRefGoogle Scholar
  9. Liu K, Goodman M, Muse S, Smith JS, Buckler E, Doebley J: Genetic structure and diversity amongmaize inbred lines as inferred from DNA microsatellite Genetics, 2003, 165:2117–2128.PubMedGoogle Scholar
  10. Comai L, Young K, Till BJ, Reynolds SH, Greene EA, Codomo CA, Enns LC, Johnson JE, Burtner C, Odden AR, et al.: Efficient discovery of DNA polymorphisms in natural populations by Ecotilling. Plant J 2004, 37:778–786.PubMedCrossRefGoogle Scholar
  11. Margulies M, Egholm M, Altman WE, Attiya S, Bader JS, Bemben LA, Berka J, Braverman MS, Chen YJ, Chen Z, et al.: Genome sequencing in microfabricated high-density picolitre reactors. Nature 2005, 437:376–380.PubMedGoogle Scholar
  12. Seo TS, Bai X, Kim DH, Meng Q, Shi S, Ruparel H, Li Z, Turro NJ, Ju J: Four-color DNA sequencing by synthesis on a chip using photocleavable fluorescent nucleotides. Proc Natl Acad Sci U S A 2005, 102:5926–5931.PubMedCrossRefGoogle Scholar
  13. Shendure J, Porreca GJ, Reppas NB, Lin X, McCutcheon JP, Rosenbaum AM, Wang MD, Zhang K, Mitra RD, Church GM: Accurate multiplex polony sequencing of an evolved bacterialgenome. Science2005, 309:1728–1732.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science + Business Media, LLC 2009

Authors and Affiliations

  1. 1.Agronomy Dept. and Whistler Center for Carbohydrate ResearchPurdue UniversityWest LafayetteUSA

Personalised recommendations