Similar articles

Abstract: targeting Induced Local Lesions IN Genomes (TILLING) is a reverse genetics strategy for the high-throughput screening of induced mutations. γ; radiation, which often induces both insertion/deletion (indel) and point mutations, has been widely used in mutation induction and crop breeding. The present study aimed to develop a simple, high-throughput TILLING system for screening γ; ray-induced mutations using high-resolution melting (HRM) analysis. Pooled rice (Oryza sativa) samples mixed at a 1:7 ratio of indel mutant to wild-type DNA could be distinguished from the wild-type controls by HRM analysis. Thus, an HRM-TILLING system that analyzes pooled samples of four M₂ plants is recommended for screening γ; ray-induced mutants in rice. For demonstration, a γ; ray-mutagenized M₂ rice population (n=4560) was screened for mutations in two genes, OsLCT1 and SPDT, using this HRM-TILLING system. Mutations including one single nucleotide substitution (G→A) and one single nucleotide insertion (A) were identified in OsLCT1, and one trinucleotide (TTC) deletion was identified in SPDT. These mutants can be used in rice breeding and genetic studies, and the findings are of importance for the application of γ; ray mutagenesis to the breeding of rice and other seed crops.

基于高分辨率熔解曲线技术的水稻伽玛射线诱发突变的TILLIN体系

[1]Acanda Y, Martínez Ó, Prado MJ, et al., 2014. EMS mutagenesis and qPCR-HRM prescreening for point mutations in an embryogenic cell suspension of grapevine. Plant Cell Rep, 33(3):471-481.

[2]Ahloowalia BS, Maluszynski M, Nichterlein K, 2004. Global impact of mutation-derived varieties. Euphytica, 135(2):187-204.

[3]Botticella E, Sestili F, Hernandez-Lopez A, et al., 2011. High resolution melting analysis for the detection of EMS induced mutations in wheat Sbella genes. BMC Plant Biol, 11:156.

[4]Bovina R, Brunazzi A, Gasparini G, et al., 2014. Development of a TILLING resource in durum wheat for reverse- and forward-genetic analyses. Crop Pasture Sci, 65(1):112-124.

[5]Bush SM, Krysan PJ, 2010. ITILLING: a personalized approach to the identification of induced mutations in Arabidopsis. Plant Physiol, 154(1):25-35.

[6]Colasuonno P, Incerti O, Lozito ML, et al., 2016. DHPLC technology for high-throughput detection of mutations in a durum wheat TILLING population. BMC Genet, 17:43.

[7]Colbert T, Till BJ, Tompa R, et al., 2001. High-throughput screening for induced point mutations. Plant Physiol, 126(2):480-484.

[8]Cousins MM, Donnell D, Eshleman SH, 2013. Impact of mutation type and amplicon characteristics on genetic diversity measures generated using a high-resolution melting diversity assay. J Mol Diagn, 15(1):130-137.

[9]Dong CM, Vincent K, Sharp P, 2009. Simultaneous mutation detection of three homoeologous genes in wheat by high resolution melting analysis and mutation surveyor^®. BMC Plant Biol, 9:143.

[10]Fu HW, Li YF, Shu QY, 2008. A revisit of mutation induction by gamma rays in rice (Oryza sativa L.):implications of microsatellite markers for quality control. Mol Breed, 22(2):281-288.

[11]Gady ALF, Herman FWK, van de Wal MHBJ, et al., 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.

[12]Hofinger BJ, Jing HC, Hammond-Kosack KE, et al., 2009. High-resolution melting analysis of cDNA-derived PCR amplicons for rapid and cost-effective identification of novel alleles in barley. Theor Appl Genet, 119(5):851-865.

[13]Hwang JE, Jang DS, Lee KJ, et al., 2017. Identification of gamma ray irradiation-induced mutations in membrane transport genes in a rice population by TILLING. Genes Genet Syst, 91(5):245-256.

[14]Kumar APK, McKeown PC, Boualem A, et al., 2017. TILLING by sequencing (TbyS) for targeted genome mutagenesis in crops. Mol Breed, 37(2):14.

[15]Li S, Zheng YC, Cui HR, et al., 2016. Frequency and type of inheritable mutations induced by γ rays in rice as revealed by whole genome sequencing. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 17(12):905-915.

[16]Lochlainn SÓ, Amoah S, Graham NS, et al., 2011. High resolution melt (HRM) analysis is an efficient tool to genotype EMS mutants in complex crop genomes. Plant Methods, 7:43.

[17]Lu HP, Zhang HL, Fu HW, et al., 2016. Identification and characterization of a novel lesion mimic mutant in rice. J Nucl Agric Sci, 30(6):1037-1044 (in Chinese).

[18]https://doi.org/10.11869/j.issn.100-8551.2016.06.1037

[19]Mader E, Lukas B, Novak J, 2008. A strategy to setup codominant microsatellite analysis for high-resolution-melting-curve-analysis (HRM). BMC Genet, 9:69.

[20]Matoulkova E, Michalova E, Vojtesek B, et al., 2012. The role of the 3' untranslated region in post-transcriptional regulation of protein expression in mammalian cells. RNA Biol, 9(5):563-576.

[21]McCallum CM, Comai L, Greene EA, et al., 2000. Targeting induced local lesions in genomes (TILLING) for plant functional genomics. Plant Physiol, 123(2):439-442.

[22]Nawaz Z, Shu QY, 2014. Molecular nature of chemically and physically induced mutants in plants: a review. Plant Genet Res, 12(S1):S74-S78.

[23]Nida H, Blum S, Zielinski D, et al., 2016. Highly efficient de novo mutant identification in a Sorghum bicolor TILLING population using the ComSeq approach. Plant J, 86(4):349-359.

[24]Reed GH, Kent JO, Wittwer CT, 2007. High-resolution DNA melting analysis for simple and efficient molecular diagnostics. Pharmacogenomics, 8(6):597-608.

[25]Ririe KM, Rasmussen RP, Wittwer CT, 1997. Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal Biochem, 245(2):154-160.

[26]Rogers C, Wen JQ, Chen RJ, et al., 2009. Deletion-based reverse genetics in Medicago truncatula. Plant Physiol, 151(3):1077-1086.

[27]Sato Y, Shirasawa K, Takahashi Y, et al., 2006. Mutant selection from progeny of gamma-ray-irradiated rice by DNA heteroduplex cleavage using brassica petiole extract. Breed Sci, 56(2):179-183.

[28]Shu QY, Forster BP, Nakagawa H, 2012. Plant Mutation Breeding and Biotechnology. CABI Publishing, Wallingford, UK, p.123-134.

[29]Simko I, 2016. High-resolution DNA melting analysis in plant research. Trends Plant Sci, 21(6):528-537.

[30]Taheri S, Lee Abdullah T, Jain SM, et al., 2017. TILLING, high-resolution melting (HRM), and next-generation sequencing (NGS) techniques in plant mutation breeding. Mol Breed, 37:40.

[31]Tan YY, Yu XM, Shu QY, et al., 2016. Development of an HRM-based, safe and high-throughput genotyping system for two low phytic acid mutations in soybean. Mol Breed, 36:101.

[32]Till BJ, Reynolds SH, Greene EA, et al., 2003. Large-scale discovery of induced point mutations with high-throughput TILLING. Genome Res, 13(3):524-530.

[33]Till BJ, Cooper J, Tai TH, et al., 2007. Discovery of chemically induced mutations in rice by TILLING. BMC Plant Biol, 7:19.

[34]Tsai H, Howell T, Nitcher R, et al., 2011. Discovery of rare mutations in populations: TILLING by sequencing. Plant Physiol, 156(3):1257-1268.

[35]Uraguchi S, Kamiya T, Sakamoto T, et al., 2011. Low-affinity cation transporter (OsLCT1) regulates cadmium transport into rice grains. Proc Natl Acad Sci USA, 108(52):20959-20964.

[36]Wang QZ, Fu HW, Huang JZ, et al., 2012. Generation and characterization of bentazon susceptible mutants of commercial male sterile lines and evaluation of their utility in hybrid rice production. Field Crop Res, 137: 12-18.

[37]Wittwer CT, Reed GH, Gundry CN, et al., 2003. High-resolution genotyping by amplicon melting analysis using LCGreen. Clin Chem, 49(6):853-860.

[38]Yamaji N, Takemoto Y, Miyaji T, et al., 2017. Reducing phosphorus accumulation in rice grains with an impaired transporter in the node. Nature, 541(7635):92-95.

[39]Yoshida S, Forno DA, Cock J, et al., 1976. Laboratory Manual for Physiological Studies of Rice. The International Rice Research Institute, Los Banos, Manila, Philippines.

[40]Zhang HL, Huang JZ, Chen XY, et al., 2014. Competitive amplification of differentially melting amplicons facilitates efficient genotyping of photoperiod-and temperature-sensitive genic male sterility in rice. Mol Breed, 34(4):1765-1776.

[41]Zhao HJ, Liu QL, Ren XL, et al., 2008. Gene identification and allele-specific marker development for two allelic low phytic acid mutations in rice (Oryza sativa L.). Mol Breed, 22(4):603-612.