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Research paper | Open Access

Fast mapping of a chlorophyll b synthesis-deficiency gene in barley (Hordeum vulgare L.) via bulked-segregant analysis with reduced-representation sequencing

Dongdong Xua, Dan Sunb, Yanling Diaob, Minxuan Liua, Jia Gaoa, Bin Wua, Xingmiao Yuana, Ping Lua, Zongwen Zhanga, Jing Zhanga, Ganggang Guoa( )
Key Laboratory of Crop Germplasm Resources and Utilization, Ministry of Agriculture, National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Food Crops Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150049, China

Peer review under responsibility of Crop Science Society of China and Institute of Crop Science, CAAS.

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Abstract

Bulked-segregant analysis coupled with next-generation sequencing (BSA-seq) has emerged as an efficient tool for genetic mapping of single genes or major quantitative trait loci controlling (agronomic) traits of interest. However, such a mapping-by-sequencing approach usually relies on deep sequencing and advanced statistical methods. Application of BSA-Seq based on construction of reduced-representation libraries and allele frequency analysis permitted anchoring the barley pale-green (pg) gene on chromosome 3HL. With further marker-assisted validation, pg was mapped to a 3.9 Mb physical-map interval. In the pg mutant a complete deletion of chlorophyllide a oxygenase (HvCAO) gene was identified. Because the product of this gene converts Chl a to Chl b, the pg mutant is deficient in Chl b. An independent Chl b-less mutant line M4437_2 carried a nonsynonymous substitution (F263L) in the C domain of HvCAO. The study demonstrates an optimized pooling strategy for fast mapping of agronomically important genes using a segregating population.

Keywords

BSA-seqBarleyReduced representative sequencingPale-greenChlorophyllide a oxygenase

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The Crop Journal
Volume 7 Issue 1,
February 2019
Pages 58-64
DOI: 10.1016/j.cj.2018.07.002
Cite this article:
Xu D, Sun D, Diao Y, et al. Fast mapping of a chlorophyll b synthesis-deficiency gene in barley (Hordeum vulgare L.) via bulked-segregant analysis with reduced-representation sequencing. The Crop Journal, 2019, 7(1): 58-64. https://doi.org/10.1016/j.cj.2018.07.002

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Received: 07 May 2018
Revised: 04 July 2018
Accepted: 20 July 2018
Published: 04 August 2018
© 2018 Crop Science Society of China and Institute of Crop Science, CAAS.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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