CLC number: Q319
On-line Access: 2018-08-02
Received: 2017-08-19
Revision Accepted: 2017-10-22
Crosschecked: 2018-07-10
Cited: 0
Clicked: 5422
Shan Li, Song-Mei Liu, Hao-Wei Fu, Jian-Zhong Huang, Qing-Yao Shu. High-resolution melting-based TILLING of γ ray-induced mutations in rice[J]. Journal of Zhejiang University Science B, 2018, 19(8): 620-629.
@article{title="High-resolution melting-based TILLING of γ ray-induced mutations in rice",
author="Shan Li, Song-Mei Liu, Hao-Wei Fu, Jian-Zhong Huang, Qing-Yao Shu",
journal="Journal of Zhejiang University Science B",
volume="19",
number="8",
pages="620-629",
year="2018",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1700414"
}
%0 Journal Article
%T High-resolution melting-based TILLING of γ ray-induced mutations in rice
%A Shan Li
%A Song-Mei Liu
%A Hao-Wei Fu
%A Jian-Zhong Huang
%A Qing-Yao Shu
%J Journal of Zhejiang University SCIENCE B
%V 19
%N 8
%P 620-629
%@ 1673-1581
%D 2018
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1700414
TY - JOUR
T1 - High-resolution melting-based TILLING of γ ray-induced mutations in rice
A1 - Shan Li
A1 - Song-Mei Liu
A1 - Hao-Wei Fu
A1 - Jian-Zhong Huang
A1 - Qing-Yao Shu
J0 - Journal of Zhejiang University Science B
VL - 19
IS - 8
SP - 620
EP - 629
%@ 1673-1581
Y1 - 2018
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1700414
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.
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