Full Text:   <880>

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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: 2892

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Qing-Yao Shu

https://orcid.org/0000-0002-9201-0593

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Journal of Zhejiang University SCIENCE B 2018 Vol.19 No.8 P.620-629

http://doi.org/10.1631/jzus.B1700414


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


Author(s):  Shan Li, Song-Mei Liu, Hao-Wei Fu, Jian-Zhong Huang, Qing-Yao Shu

Affiliation(s):  National Key Laboratory of Rice Biology, Institute of Crop Sciences, Zhejiang University, Hangzhou 310058, China; more

Corresponding email(s):   qyshu@zju.edu.cn

Key Words:  Mutation screening, High-resolution melting (HRM) analysis, Targeting Induced Local Lesions IN Genomes (TILLING), Mutant, Indel, γ, ray, Rice


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"
}

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%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
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%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
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SP - 620
EP - 629
%@ 1673-1581
Y1 - 2018
PB - Zhejiang University Press & Springer
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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.

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

目的:建立适用于筛选伽马射线诱发突变的、基于高分辨率熔解曲线(high-resolution melting,HRM)技术的高通量定向诱导基因组局部突变技术(Targeting Induced Local Lesions IN Genomes,TILLING)体系.
创新点:建立起了基于HRM技术、适用于伽玛射线诱发的小片段插入/缺失突变的高通量TILLING体系(HRM-TILLING).
方法:通过不同野生型/突变型比例混池DNA的HRM分析,确定HRM检测不同类型插入/缺失突变的能力,确定M2植株突变检测的适宜混池比例,并用一个伽玛诱变M2群体(n=4560)筛选OsLCT1SPDT两个基因的突变体,确定实际效果.
结论:以4株M2植株混样,采用HRM可以有效检出突变.建立的基于HRM的TILLING体系适用于伽玛射线诱发突变的高通量筛选.

关键词:突变筛选;高分辨率熔解曲线(HRM);定向诱导基因组局部突变技术(TILLING);突变体;插入缺失;伽玛射线;水稻

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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