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Diagnostics in Plant Breeding pp 145–165Cite as

TILLING and EcoTILLING

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Abstract

TILLING (Targeting Induced Local Lesions IN Genomes) combines the formation of a population with high density point-mutations provided by chemical mutagenesis with rapid mutational screening in pools of DNA. It is mainly applied as a reverse-genetic method to reveal the function of genes with known DNA sequence, but also as a non-GMO knock-out tool for molecular breeding. The steps involved are mutagenesis, the development of the TILLING population including DNA isolation and pooling, mutation detection and identification of the respective phenotype. In all steps, numerous methodological choices and modification can be applied that will influence the outcome of the results. EcoTILLING is a variation of TILLING that is used to study the allelic variation of natural populations instead of mutation populations. It has many possible applications both in scientific research to study the genetic diversity at certain genes as well as in molecular breeding to search for new allelic variation and genetic markers.

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References

  • Al-Qurainy F, Khan S (2009) Mutagenic effects of sodium azid and its application in crop improvement. World Appl Sci J 6:1589–1601

    CAS  Google Scholar 

  • Barkley NA, Wang ML (2008) Application of TILLING and EcoTILLING as reverse genetic approaches to elucidate the function of genes in plants and animals. Curr Genomics 9:212–226. doi:10.2174/138920208784533656

    Article  PubMed  CAS  Google Scholar 

  • Beetham PR, Kipp PB, Sawycky XL, Arntzen CJ, May GD (1999) A tool for functional plant genomics: chimeric RNA/DNA oligonucleotides cause in vivo gene-specific mutations. Proc Natl Acad Sci USA 96:8774–8778. doi:10.1073/pnas.96.15.8774

    Article  PubMed  CAS  Google Scholar 

  • Brady SM, Provart NJ (2007) Extreme breeding: leveraging genomics for crop improvement. J Sci Food Agric 87:925–929. doi:10.1002/jsfa.2763

    Article  CAS  Google Scholar 

  • Bui CT, Lambrinakos A, Babon JJ, Cotton RG (2003) Chemical cleavage reactions of DNA on solid support: application in mutation detection. BMC Chem Biol 3:1. doi:10.1186/1472-6769-3-1

    Article  PubMed  Google Scholar 

  • Caldwell DG, McCallum N, Shaw P, Muehlbauer GJ, Marshall DF, Waugh R (2004) A structured mutant population for forward and reverse genetics in Barley (Hordeum vulgare L.). Plant J 40:143–150. doi:10.1111/j.1365-313X.2004.02190.x

    Article  PubMed  CAS  Google Scholar 

  • Chao SM, Somers D (2012) Wheat and barley DNA extraction in 96-well plates. In: MAS wheat. http://maswheat.ucdavis.edu/protocols/general_protocols/DNA_extraction_003.htm. Accessed 3 Jan 2012

  • Chuang CF, Meyerowitz EM (2000) Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci USA 97:4985–4990. doi:10.1073/pnas.060034297

    Article  PubMed  CAS  Google Scholar 

  • Colbert T, Till BJ, Tompa R, Reynolds S, Steine MN, Yeung AT, McCallum CM, Comai L, Henikoff S (2001) High-throughput screening for induced point mutations. Plant Physiol 126:480–484. doi:10.1104/pp. 126.2.480

    Article  PubMed  CAS  Google Scholar 

  • Comai L, Henikoff S (2006) TILLING: practical single-nucleotide mutation discovery. Plant J 45:684–694. doi:10.1111/j.1365-313X.2006.02670.x

    Article  PubMed  CAS  Google Scholar 

  • Comai L, Young KJ, Codomo CA, Enns LC, Johnson JE, Burtner C, Odden AR, Henikoff S, Reynolds SH, Greene EA (2004) Efficient discovery of DNA polymorphisms in natural populations by Ecotilling. Plant J 37:778–786. doi:10.1111/j.1365-313X.2003.01999.x

    Article  PubMed  CAS  Google Scholar 

  • Cooper J, Till B, Laport R, Darlow M, Kleffner J, Jamai A, El-Mellouki T, Liu S, Ritchie R, Nielsen N, Bilyeu K, Meksem K, Comai L, Henikoff S (2008) TILLING to detect induced mutations in soybean. BMC Plant Biol 8:9. doi:10.1186/1471-2229-8-9

    Article  PubMed  Google Scholar 

  • Cotton RG, Rodrigues NR, Campbell RD (1988) Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations. Proc Natl Acad Sci USA 85:4397–4401. doi:10.1073/pnas.85.12.4397

    Article  PubMed  CAS  Google Scholar 

  • Cross MJ, Waters DLE, Lee LS, Henry RJ (2008) Endonucleolytic mutation analysis by internal labeling (EMAIL). Electrophoresis 29:1291–1301. doi:10.1002/elps.200700452

    Article  PubMed  CAS  Google Scholar 

  • Desai NA, Shankar V (2003) Single-strand-specific nucleases. FEMS Microbiol Rev 26:457–491. doi:10.1111/j.1574-6976.2003.tb00626.x

    Article  PubMed  CAS  Google Scholar 

  • Dibya D (2010) High-throughput mutation screening on a TILLING platform using the AdvanCE™ FS96 system. Advanced Analytical, Ames

    Google Scholar 

  • Gady AL, Hermans FW, Van de Wal MH, van Loo EN, Visser RG, Bachem CW (2009) Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations. Plant Methods 5:13. doi:10.1186/1746-4811-5-13

    Article  PubMed  Google Scholar 

  • Galeano CH, Gomez M, Rodriguez LM, Blair MW (2009) CEL I nuclease digestion for SNP discovery and marker development in common bean (Phaseolus vulgaris L.). Crop Sci 49:381–394. doi:10.2135/cropsci2008.07.0413

    Article  CAS  Google Scholar 

  • Garvin MR, Gharett AJ (2007) DEco-TILLING: an inexpensive method for single nucleotide polymorphism discovery that reduces ascertainment bias. Mol Ecol Notes 7:735–746. doi:10.1111/j.1471-8286.2007.01767.x

    Article  CAS  Google Scholar 

  • Gelvin SB (2003) Agrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool. Microbiol Mol Biol Rev 67:16–37. doi:10.1128/MMBR.67.1.16-37.2003

    Article  PubMed  CAS  Google Scholar 

  • Gilchrist EJ, Haughn GW, Ying CC, Otto SP, Zhuang J, Cheung D, Hamberger B, Aboutorabi F, Kalynyak T, Johnson L, Bohlmann J, Ellis BE, Douglas CJ, Cronk QCB (2006a) Use of Ecotilling as an efficient SNP discovery tool to survey genetic variation in wild populations of Populus trichocarpa. Mol Ecol 15:1367–1378. doi:10.1111/j.1365-294X.2006.02885.x

    Article  PubMed  CAS  Google Scholar 

  • Gilchrist EJ, O’Neil NJ, Rose AM, Zetka MC, Haughn GW (2006b) TILLING is an effective reverse genetics technique for Caenorhabditis elegans. BMC Genomics 7:262. doi:10.1186/1471-2164-7-262

    Article  PubMed  Google Scholar 

  • Gottwald S, Bauer P, Komatsuda T, Lundqvist U, Stein N (2009) TILLING in the two-rowed barley cultivar “Barke” reveals preferred sites of functional diversity in the gene HvHox1. BMC Res Notes 2:258. doi:10.1186/1756-0500-2-258

    Article  PubMed  Google Scholar 

  • Greene EA, Codomo CA, Taylor NE, Henikoff JG, Till BJ, Reynolds SH, Enns LC, Burtner C, Johnson JE, Odden AR, Comai L, Henikoff S (2003) Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis. Theor Appl Genet 164:731–740

    CAS  Google Scholar 

  • Hecker KH, Taylor PD, Gjerde DT (1999) Mutation detection by denaturing DNA chromatography using fluorescently labeled polymerase chain reaction products. Anal Biochem 272:156–164. doi:10.1006/abio.1999.4171

    Article  PubMed  CAS  Google Scholar 

  • Heckmann AB, Lombardo F, Miwa H, Perry JA, Bunnewell S, Parniske M, Wang TL, Downie JA (2006) Lotus japonicus nodulation requires two GRAS domain regulators, one of which is functionally conserved in a non-legume. Plant Physiol 142:1739–1750. doi:10.1104/pp. 106.089508

    Article  PubMed  CAS  Google Scholar 

  • Henikoff S, Comai L (2003) Single-nucleotide mutations for plant functional genomics. Annu Rev Plant Biol 54:375–401. doi:10.1143/annurev.arplant.54.031902.135009

    Article  PubMed  CAS  Google Scholar 

  • Hermann S, Brumbley S, McIntyre CL (2006) Analysing diversity in sugarcane resistance gene analogues. Australas Plant Pathol 35:631. doi:10.1071/AP06066

    Article  CAS  Google Scholar 

  • Himelblau E, Gilchrist EJ, Buono K, Bizzell C, Mentzer L, Vogelzang R, Osborn T, Amasino RM, Parkin IAP, Haughn GW (2009) Forward and reverse genetics of rapid-cycling Brassica oleracea. Theor Appl Genet 118:953–961. doi:10.1007/s00122-008-0952-7

    Article  PubMed  Google Scholar 

  • Hirochika H (1997) Retrotransposons of rice: their regulation and use for genome analysis. Plant Mol Biol 35:231–240. doi:10.1023/A:1005774705893

    Article  PubMed  CAS  Google Scholar 

  • Hohmann U, Jacobs G, Jung C (2005) An EMS mutagenesis protocol for sugar beet and isolation of non-bolting mutants. Plant Breed 124:317–321. doi:10.1111/j.1439-0523.2005.01126.x

    Article  Google Scholar 

  • Horst I, Welham T, Kelly S, Kaneko T, Sato S, Tabata S, Parniske M, Wang TL (2007) TILLING mutants of Lotus japonicus reveal that nitrogen assimilation and fixation can occur in the absence of nodule-enhanced sucrose synthase. Plant Physiol 144:806–820. doi:10.1104/pp. 107.097063

    Article  PubMed  CAS  Google Scholar 

  • Howard JT, Ward J, Watson JN, Roux KH (1999) Heteroduplex cleavage analysis using S1 nuclease. Biotechniques 27:18–19

    PubMed  CAS  Google Scholar 

  • Huang J, Kirk B, Favis R, Soussi T, Paty P, Cao W, Barany F (2002) An endonuclease/ligase based mutation scanning method especially suited for analysis of neoplastic tissue. Oncogene 21:1909–1921. doi:10.1038/sj.onc.1205109

    Article  PubMed  CAS  Google Scholar 

  • Igarashi H, Nagura K, Sugimura H, Kulinski J, Besack D, Oleykowski CA, Godwin AK, Yeung AT (2000) CEL I enzymatic mutation detection. Biotechniques 29:44–48

    Google Scholar 

  • Iida S, Terada R (2004) A tale of two integrations, transgene and T-DNA: gene targeting by homologous recombination in rice. Curr Opin Biotechnol 15:132–138. doi:10.1016/j.copbio.2004.02.005

    Article  PubMed  CAS  Google Scholar 

  • Kadaru SB, Yadav AS, Fjellstrom RG, Oard JH (2006) Alternative ecotilling protocol for rapid, cost-effective single-nucleotide polymorphism discovery and genotyping in rice (Oryza sativa L.). Plant Mol Biol Rep 24:3–22. doi:10.1007/BF02914042

    Article  CAS  Google Scholar 

  • Kahn SD (2011) On the future of genomic data. Science 331:728–729. doi:10.1126/science.1197891

    Article  PubMed  CAS  Google Scholar 

  • Klug A (2010) The discovery of Zinc figures and their applications in gene regulation and genome manipulation. Annu Rev Biochem 79:213–231. doi: 10.1146/annurev-biochem-010909-095056

    Article  PubMed  CAS  Google Scholar 

  • Krysan PJ, Young JC, Jester PJ, Monson S, Copenhaver G, Preuss D, Sussman MR (2002) Characterization of T-DNA insertion sites in Arabidopsis thaliana and the implications for saturation mutagenesis. OMICS 6:163–174. doi:10.1089/153623102760092760

    Article  PubMed  CAS  Google Scholar 

  • Kuhn DN, Borrone J, Meerow AW, Motamayor JC, Brown JS, Schnell RJ (2005) Single-strand conformation polymorphism analysis of candidate genes for reliable identification of alleles by capillary array electrophoresis. Electrophoresis 26:112–125. doi:10.1002/elps.200406106

    Article  PubMed  CAS  Google Scholar 

  • Kutyavin IV (2000) 3′-Minor groove binder-DNA probes increase sequence specificity at PCR extension temperatures. Nucleic Acids Res 28:655–661. doi:10.1093/nar/28.2.655

    Article  PubMed  CAS  Google Scholar 

  • Lababidi S, Mejlhede N, Rasmussen SK, Backes G, Al-Said W, Baum M, Jahoor A (2009) Identification of barley mutants in the cultivar “Lux” at the Dhn loci through TILLING. Plant Breed 128:332–336. doi:10.1111/j.1439-0523.2009.01640.x

    Article  CAS  Google Scholar 

  • Le Signor C, Savois V, Aubert G, Verdier J, Nicolas M, Pagny G, Moussy F, Sanchez M, Baker D, Clarke J, Thompson R (2009) Optimizing TILLING populations for reverse genetics in Medicago truncatula. Plant Biotechnol J 7:430–441. doi:10.1111/j.1467-7652.2009.00410.x

    Article  PubMed  Google Scholar 

  • Lefebvre V, Goffinet B, Chauvet JC, Caromel B, Signoret P, Brand R, Palloix A (2001) Evaluation of genetic distances between pepper inbred lines for cultivar protection purposes: comparison of AFLP, RAPD and phenotypic data. Theor Appl Genet 102:741–750. doi:10.1007/s001220051705

    Article  CAS  Google Scholar 

  • Li Q, Liu Z, Monroe H, Culiat CT (2002a) Integrated platform for detection of DNA sequence variants using capillary array electrophoresis. Electrophoresis 23:1499–1511. doi:10.1002/1522-2683(200205)23:10<1499::AID-ELPS1499>3.0.CO;2-X

    Article  PubMed  CAS  Google Scholar 

  • Li X, Lassner M, Zhang Y (2002b) Deleteagene: a fast neutron deletion mutagenesis-based gene knockout system for plants. Comp Funct Genomics 3:158–160. doi:10.1002/cfg.148

    Article  PubMed  CAS  Google Scholar 

  • Lu A-L, Hsu I-C (1992) Detection of single DNA base mutations with mismatch repair enzymes. Genomics 14:249–255. doi:10.1016/S0888-7543(05)80213-7

    Article  PubMed  CAS  Google Scholar 

  • Mashal RD, Koontz J, Sklar J (1995) Detection of mutations by cleavage of DNA heteroduplexes with bacteriophage resolvases. Nat Genet 9:177–183. doi:10.1038/ng0295-177

    Article  PubMed  CAS  Google Scholar 

  • May BP, Liu H, Vollbrecht E, Senior L, Rabinowicz PD, Roh D, Pan X, Stein L, Freeling M, Alexander D, Martienssen R (2003) Maize-targeted mutagenesis: a knockout resource for maize. Proc Natl Acad Sci USA 100:11541–11546. doi:10.1073/pnas.1831119100

    Article  PubMed  CAS  Google Scholar 

  • McCallum CM, Comai L, Greene EA, Henikoff S (2000) Targeted screening for induced mutations. Nat Biotechnol 18:455–457

    Article  PubMed  CAS  Google Scholar 

  • Mejlhede N, Kyjovska Z, Backes G, Burhenne K, Rasmussen SK, Jahoor A (2006) EcoTILLING for the identification of allelic variation in the powdery mildew resistance genes mlo and Mla of barley. Plant Breed 125:461–467. doi:10.1111/j.1439-0523.2006.01226.x

    Article  CAS  Google Scholar 

  • Nataraj AJ, Olivos-Glander I, Kusukawa N, Highsmith WE (1999) Single-strand conformation polymorphism and heteroduplex analysis for gel-based mutation detection. Electrophoresis 20:1177–1185. doi:10.1002/(SICI)1522-2683(19990101)20:6<1177::AID-ELPS1177>3.0.CO;2-2

    Article  PubMed  CAS  Google Scholar 

  • Nieto C, Piron F, Dalmais M, Marco CF, Moriones E, Gómez-Guillamon ML, Truniger V, Gómez P, Garcia-Mas J, Aranda MA, Bendahmane A (2007) EcoTILLING for the identification of allelic variants of melon eIF4E, a factor that controls virus susceptibility. BMC Plant Biol 7:34. doi:10.1186/1471-2229-7-34

    Article  PubMed  Google Scholar 

  • Nolan PM, Hugill A, Cox RD (2002) ENU mutagenesis in the mouse: application to human genetic disease. Brief Funct Genomic Proteomic 1:278–289. doi:10.1093/bfgp/1.3.278

    Article  PubMed  CAS  Google Scholar 

  • Okagaki RJ, Neuffer MG, Wessler SR (1991) A deletion common to two independently derived waxy mutations of maize. Genetics 128:425–431

    PubMed  CAS  Google Scholar 

  • Oleykowski CA, Mullins CRB, Godwin AK, Yeung AT (1998) Mutation detection using a novel plant endonuclease. Nucleic Acids Res 26:4597–4602. doi:10.1093/nar/26.20.4597

    Article  PubMed  CAS  Google Scholar 

  • Owais WM, Kleinhofs A (1988) Metabolic activation of the mutagen azide in biological systems. Mutat Res 197:313–323. doi:10.1016/0027-5107(88)90101-7

    Article  PubMed  CAS  Google Scholar 

  • Palotta MA, Warner P, Fox RL, Kuchel H, Jefferies SP, Langridge P (2003) Marker assisted wheat breeding in the southern region of Australia. In: Proceedings of the tenth international wheat genetics symposium, Paestum, 1–6 September 2003, pp 789–791

    Google Scholar 

  • Parry MAJ, Madgwick PJ, Bayon C, Tearall K, Hernandez-Lopez A, Baudo M, Rakszegi M, Hamada W, Al-Yassin A, Ouabbou H, Labhilili M, Phillips AL (2009) Mutation discovery for crop improvement. J Exp Bot 60:2817–2825. doi:10.1093/jxb/erp189

    Article  PubMed  CAS  Google Scholar 

  • Perry JA, Wang TL, Welham TJ, Gardner S, Pike JM, Yoshida S, Parniske M (2003) A TILLING reverse genetics tool and a web-accessible collection of mutants of the legume Lotus japonicus. Plant Physiol 131:866–871. doi:10.1104/pp. 102.017384

    Article  PubMed  CAS  Google Scholar 

  • Raghavan C, Naredo MEB, Wang H, Atienza G, Liu B, Qiu F, McNally KL, Leung H (2006) Rapid method for detecting SNPs on agarose gels and its application in candidate gene mapping. Mol Breed 19:87–101. doi:10.1007/s11032-006-9046-x

    Article  Google Scholar 

  • Rogers C, Wen JQ, Chen RJ, Oldroyd G (2009) Deletion-based reverse genetics in Medicago truncatula. Plant Physiol 151:1077–1086. doi:10.1104/pp. 109.142919

    Article  PubMed  CAS  Google Scholar 

  • Sestili F, Botticella E, Bedo Z, Phillips A (2010) Production of novel allelic variation for genes involved in starch biosynthesis through mutagenesis. Mol Breed 25:145–154. doi:10.1007/s11032-009-9314-7

    Article  CAS  Google Scholar 

  • Shendure J, Ji HL (2008) Next-generation DNA sequencing. Nat Biotechnol 26:1135–1145. doi:10.1038/nbt1486

    Article  PubMed  CAS  Google Scholar 

  • Slade AJ, Knauf VC (2005) TILLING moves beyond functional genomics into crop improvement. Transgenic Res 14:109–115. doi:10.1007/s11248-005-2770-x

    Article  PubMed  CAS  Google Scholar 

  • Slade AJ, Fuerstenberg S, Loeffler D, Steine MN, Facciotti D (2005) A reverse genetic, nontransgenic approach to wheat crop improvement by TILLING. Nat Biotechnol 23:75–81. doi:10.1038/nbt1043

    Article  PubMed  CAS  Google Scholar 

  • Sreelakshmi Y, Gupta S, Bodanapu R, Chauhan VS, Hanjabam M, Thomas S, Mohan V, Sharma S, Srinivasan R, Sharma R (2010) NEATTILL: a simplified procedure for nucleic acid extraction from arrayed tissue for TILLING and other high-throughput reverse genetic applications. Plant Methods 6:3. doi:10.1186/1746-4811-6-3

    Article  PubMed  Google Scholar 

  • Stanssens P, Zabeau M, Meersseman G, Remes G, Gansemans Y, Storm N, Hartmer R, Honisch C, Rodi CP, Böcker S, van den Boom D (2004) High-throughput MALDI-TOF discovery of genomic sequence polymorphisms. Genome Res 14:126–133. doi:10.1101/gr.1692304

    Article  PubMed  CAS  Google Scholar 

  • Stemple DL (2004) TILLING – a high-throughput harvest for functional genomics. Nat Rev Genet 5:145–150. doi:10.1038/nrg1273

    Article  PubMed  CAS  Google Scholar 

  • Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039. doi:10.1073/pnas.76.3.1035

    Article  PubMed  CAS  Google Scholar 

  • Suzuki T, Eiguchi M, Kumamaru T, Satoh H, Matsusaka H, Moriguchi K, Nagato Y, Kurata N (2008) MNU-induced mutant pools and high performance TILLING enable finding of any gene mutation in rice. Mol Genet Genomics 279:213–223. doi:10.1007/s00438-007-0293-2

    Article  PubMed  CAS  Google Scholar 

  • Talamè V, Bovina R, Sanguineti MC, Tuberosa R, Lundqvist U, Salvi S (2008) TILLMore, a resource for the discovery of chemically induced mutants in barley. Plant Biotechnol J 6:477–485. doi:10.1111/j.1467-7652.2008.00341.x

    Article  PubMed  Google Scholar 

  • Till BJ, Reynolds SH, Greene EA, Codomo CA, Enns LC, Johnson JE, Burtner C, Odden AR, Young K, Taylor NE, Henikoff JG, Comai L, Henikoff S (2003) Large-scale discovery of induced point mutations with high-throughput TILLING. Genome Res 13:524–530. doi:10.1101/gr.977903

    Article  PubMed  CAS  Google Scholar 

  • Till BJ, Burtner C, Comai L, Henikoff JG (2004a) Mismatch cleavage by single-strand specific nucleases. Nucleic Acids Res 32:2632–2641. doi:10.1093/nar/gkh599

    Article  PubMed  CAS  Google Scholar 

  • Till BJ, Reynolds SH, Weil C, Springer N, Burtner C, Young K, Bowers E, Codomo CA, Enns LC, Odden AR, Greene EA, Comai L, Henikoff S (2004b) Discovery of induced point mutations in maize genes by TILLING. BMC Plant Biol 4:12. doi:10.1186/1471-2229-4-12

    Article  PubMed  Google Scholar 

  • Till BJ, Zerr T, Comai L, Henikoff S (2006) A protocol for TILLING and Ecotilling in plants and animals. Nat Protoc 1:2465–2477. doi:10.1038/nprot.2006.329

    Article  PubMed  CAS  Google Scholar 

  • Till B, Cooper J, Tai T, Colowit P, Greene E, Henikoff S, Comai L (2007) Discovery of chemically induced mutations in rice by TILLING. BMC Plant Biol 7:19. doi:10.1186/1471-2229-7-19

    Article  PubMed  Google Scholar 

  • Triques K, Sturbois B, Gallais S, Dalmais M, Chauvin S, Clepet C, Aubourg S, Rameau C, Caboche M, Bendahmane A (2007) Characterization of Arabidopsis thaliana mismatch specific endonucleases: application to mutation discovery by TILLING in pea. Plant J 51:1116–1125. doi:10.1111/j.1365-313X.2007.03201.x

    Article  PubMed  CAS  Google Scholar 

  • Triques K, Piednoir E, Dalmais M, Schmidt J, Le Signor C, Sharkey M, Caboche M, Sturbois B, Bendahmane A (2008) Mutation detection using ENDO1: application to disease diagnostics in humans and TILLING and Eco-TILLING in plants. BMC Mol Biol 9:42. doi:10.1186/1471-2199-9-42

    Article  PubMed  Google Scholar 

  • Uauy C, Paraiso F, Colasuonno P, Tran R, Tsai H, Berardi S, Comai L, Dubcovsky J (2009) A modified TILLING approach to detect induced mutations in tetraploid and hexaploid wheat. BMC Plant Biol 9:115. doi:10.1186/1471-2229-9-115

    Article  PubMed  Google Scholar 

  • Wang G-X, Tan M-K, Rakshit S, Saitoh H, Terauchi R, Imaizumi T, Ohsako T, Tominaga T (2007) Discovery of single-nucleotide mutations in acetolactate synthase genes by Ecotilling. Pestic Biochem Physiol 88:143–148. doi:16/j.pestbp.2006.10.006

    Article  CAS  Google Scholar 

  • Wang G-X, Imaizumi T, Li W, Saitoh H, Terauchi R, Ohsako T, Tominaga T (2008a) Self-EcoTILLING to identify single-nucleotide mutations in multigene family. Pestic Biochem Physiol 92:24–29. doi:10.1016/j.pestbp. 2008.05.00

    Article  CAS  Google Scholar 

  • Wang J, Sun JZ, Liu DC, Yang WL, Wang DW, Tong YP, Zhang AM (2008b) Analysis of Pina and Pinb alleles in the micro-core collections of Chinese wheat germplasm by Ecotilling and identification of a novel Pinb allele. J Cereal Sci 48:836–842. doi:10.1016/j.jcs.2008.06.005

    Article  CAS  Google Scholar 

  • Waterhouse PM, Graham MW, Wang M-B (1998) Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA 95:13959–13964. doi:10.1073/pnas.95.23.13959

    Article  PubMed  CAS  Google Scholar 

  • Weil CF (2009) TILLING in grass species. Plant Physiol 149:158–164. doi:10.1104/pp. 108.128785

    Article  PubMed  CAS  Google Scholar 

  • Wienholds E, van Eeden F, Kosters M, Mudde J, Plasterk RHA, Cuppen E (2003) Efficient target-selected mutagenesis in zebrafish. Genome Res 13:2700–2707. doi:10.1101/gr.1725103

    Article  PubMed  CAS  Google Scholar 

  • Winkler S, Schwabedissen A, Backasch D, Bökel C, Seidel C, Bönisch S, Fürthauer M, Kuhrs A, Cobreros L, Brand M, González-Gaitán M (2005) Target-selected mutant screen by TILLING in Drosophila. Genome Res 15:718–723. doi:10.1101/gr.3721805

    Article  PubMed  CAS  Google Scholar 

  • Xin Z, Li Wang M, Barkley N, Burow G, Franks C, Pederson G, Burke J (2008) Applying genotyping (TILLING) and phenotyping analyses to elucidate gene function in a chemically induced sorghum mutant population. BMC Plant Biol 8:103. doi:10.1186/1471-2229-8-103

    Article  PubMed  Google Scholar 

  • Yang B, Wen X, Kodali NS, Oleykowski CA, Miller CG, Kulinski J, Besack D, Yeung JA, Kowalski D, Yeung AT (2000) Purification, cloning, and characterization of the CEL I nuclease. Biochemistry 39:3533–3541. doi:10.1021/bi992376z

    Article  PubMed  CAS  Google Scholar 

  • Yeung AT, Hattangadi D, Blakesly L, Nicolas E (2005) Enzymatic mutation detection technologies. Biotechniques 38:749–758. doi:10.2144/05385RV01

    Article  PubMed  CAS  Google Scholar 

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  1. Faculty of Organic Agricultural Sciences, Department of Organic Breeding and Agrobiodiversity, University of Kassel, Nordbahnhofstr. 1a, 37213, Witzenhausen, Germany

    Gunter Backes

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  1. Dept. Agronomy, Iowa State University, Agronomy Hall 1204, Ames, 50011, Iowa, USA

    Thomas Lübberstedt

  2. ICRISAT, Centre of Excellence in Genomics (CEG), Patancheru, 502 324, Andra Pradesh, India

    Rajeev K. Varshney

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© 2013 Springer Science+Business Media Dordrecht

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Backes, G. (2013). TILLING and EcoTILLING. In: Lübberstedt, T., Varshney, R. (eds) Diagnostics in Plant Breeding. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-5687-8_7

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