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High-resolution melting-based TILLING of γ ray-induced mutations in rice

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... Full description

Journal Title: Journal of Zhejiang University-SCIENCE B 2018, Vol.19(8), pp.620-629
Main Author: Li, Shan
Other Authors: Liu, Song-mei , Fu, Hao-wei , Huang, Jian-zhong , Shu, Qing-yao
Format: Electronic Article Electronic Article
Language: eng ; chi
Subjects:
ID: ISSN: 1673-1581 ; E-ISSN: 1862-1783 ; DOI: 10.1631/jzus.B1700414
Link: http://dx.doi.org/10.1631/jzus.B1700414
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recordid: springer_jour10.1631/jzus.B1700414
title: High-resolution melting-based TILLING of γ ray-induced mutations in rice
format: Article
creator:
  • Li, Shan
  • Liu, Song-mei
  • Fu, Hao-wei
  • Huang, Jian-zhong
  • Shu, Qing-yao
subjects:
  • Mutation screening
  • High-resolution melting (HRM) analysis
  • Targeting Induced Local Lesions IN Genomes (TILLING)
  • Mutant
  • Indel
  • γ ray
  • Rice
ispartof: Journal of Zhejiang University-SCIENCE B, 2018, Vol.19(8), pp.620-629
description: 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 M 2 plants is recommended for screening γ ray-induced mutants in rice. For demonstration, a γ ray-mutagenized M 2 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.
language: eng ; chi
source:
identifier: ISSN: 1673-1581 ; E-ISSN: 1862-1783 ; DOI: 10.1631/jzus.B1700414
fulltext: fulltext
issn:
  • 1862-1783
  • 18621783
  • 1673-1581
  • 16731581
url: Link


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titleHigh-resolution melting-based TILLING of γ ray-induced mutations in rice
creatorLi, Shan ; Liu, Song-mei ; Fu, Hao-wei ; Huang, Jian-zhong ; Shu, Qing-yao
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subjectMutation screening ; High-resolution melting (HRM) analysis ; Targeting Induced Local Lesions IN Genomes (TILLING) ; Mutant ; Indel ; γ ray ; Rice
descriptionTargeting 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 M 2 plants is recommended for screening γ ray-induced mutants in rice. For demonstration, a γ ray-mutagenized M 2 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.
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vertitle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
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titleHigh-resolution melting-based TILLING of γ ray-induced mutations in rice, 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
descriptionTargeting 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 M 2 plants is recommended for screening γ ray-induced mutants in rice. For demonstration, a γ ray-mutagenized M 2 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.
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abstractTargeting 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 M 2 plants is recommended for screening γ ray-induced mutants in rice. For demonstration, a γ ray-mutagenized M 2 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.
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doi10.1631/jzus.B1700414
orcidid0000-0002-6454-4731
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date2018-08