ArticlePlant methods2011
High Resolution Melt (HRM) analysis is an efficient tool to genotype EMS mutants in complex crop genomes.
Article in Plant methods, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 96 citations in OpenAlex.
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- OsSNDP4, a Sec14-nodulin Domain Protein, is Required for Pollen Development in Rice.Rice (New York, N.Y.) · 2024Article
- OsABA3 is Crucial for Plant Survival and Resistance to Multiple Stresses in Rice.Rice (New York, N.Y.) · 2024Article
- Construction of a Female Sterility Maintaining System Based on a Novel Mutation of the MEL2 Gene.Rice (New York, N.Y.) · 2024Article
- The ribosomal protein P0A is required for embryo development in rice.BMC plant biology · 2023Article
- Is it the end of TILLING era in plant science?Frontiers in plant science · 2023Review
- Construction and analysis of a Noccaea caerulescens TILLING population.BMC plant biology · 2022Article
- Integration of advanced technologies for plant variety and cultivar identification.Journal of biosciences · 2021Review
- Next Generation Sequencing Based Forward Genetic Approaches for Identification and Mapping of Causal Mutations in Crop Plants: A Comprehensive Review.Plants (Basel, Switzerland) · 2020Review
- OsMYB80 Regulates Anther Development and Pollen Fertility by Targeting Multiple Biological Pathways.Plant & cell physiology · 2020Article
- Multiplex exome sequencing reveals genome-wide frequency and distribution of mutations in the 'Micro-Tom' Targeting Induced Local Lesions in Genomes (TILLING) mutant library.Plant biotechnology (Tokyo, Japan) · 2019Article
- The ATP-binding cassette (ABC) transporter OsABCG3 is essential for pollen development in rice.Rice (New York, N.Y.) · 2018Article
- Construction of a male sterility system for hybrid rice breeding and seed production using a nuclear male sterility gene.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Mutagenesis and TILLING to Dissect Gene Function in Plants.Current genomics · 2016Article
- Microspore Induced Doubled Haploids Production from Ethyl Methanesulfonate (EMS) Soaked Flower Buds Is an Efficient Strategy for Mutagenesis in Chinese Cabbage.Frontiers in plant science · 2016Article
- Simultaneous Identification of Multiple Causal Mutations in Rice.Frontiers in plant science · 2016Article
- Molecular Identification of Broomrape Species from a Single Seed by High Resolution Melting Analysis.Frontiers in plant science · 2016Article
- Rapid identification and recovery of ENU-induced mutations with next-generation sequencing and Paired-End Low-Error analysis.BMC genomics · 2015Article
- Article
- Genetical and comparative genomics of Brassica under altered Ca supply identifies Arabidopsis Ca-transporter orthologs.The Plant cell · 2014Article
Corrections and comments
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Authors and funding
12 authors at 5 institutions in 2 countries.
Funding
Abstract
backgroundTargeted Induced Loci Lesions IN Genomes (TILLING) is increasingly being used to generate and identify mutations in target genes of crop genomes. TILLING populations of several thousand lines have been generated in a number of crop species including Brassica rapa. Genetic analysis of mutants identified by TILLING requires an efficient, high-throughput and cost effective genotyping method to track the mutations through numerous generations. High resolution melt (HRM) analysis has been used in a number of systems to identify single nucleotide polymorphisms (SNPs) and insertion/deletions (IN/DELs) enabling the genotyping of different types of samples. HRM is ideally suited to high-throughput genotyping of multiple TILLING mutants in complex crop genomes. To date it has been used to identify mutants and genotype single mutations. The aim of this study was to determine if HRM can facilitate downstream analysis of multiple mutant lines identified by TILLING in order to characterise allelic series of EMS induced mutations in target genes across a number of generations in complex crop genomes.
resultsWe demonstrate that HRM can be used to genotype allelic series of mutations in two genes, BraA.CAX1a and BraA.MET1.a in Brassica rapa. We analysed 12 mutations in BraA.CAX1.a and five in BraA.MET1.a over two generations including a back-cross to the wild-type. Using a commercially available HRM kit and the Lightscanner™ system we were able to detect mutations in heterozygous and homozygous states for both genes.
conclusionsUsing HRM genotyping on TILLING derived mutants, it is possible to generate an allelic series of mutations within multiple target genes rapidly. Lines suitable for phenotypic analysis can be isolated approximately 8-9 months (3 generations) from receiving M3 seed of Brassica rapa from the RevGenUK TILLING service.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.