Research Paper

Identification of an EMS-induced causal mutation in a gene required for boron-mediated root development by low-coverage genome re-sequencing in Arabidopsis

Volume 8, Issue 1   January 2013
eLocation ID: e22534
http://dx.doi.org/10.4161/psb.22534
Keywords: Arabidopsis thaliana, boron, low- to medium-coverage sequencing, mutant mapping, next-generation sequencing
Authors: Ryo Tabata, Takehiro Kamiya, Shuji Shigenobu, Katsushi Yamaguchi, Masashi Yamada, Mitsuyasu Hasebe, Toru Fujiwara and Shinichiro Sawa

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Abstract:
Next-generation sequencing (NGS) technologies enable the rapid production of an enormous quantity of sequence data. These powerful new technologies allow the identification of mutations by whole-genome sequencing. However, most reported NGS-based mapping methods, which are based on bulked segregant analysis, are costly and laborious. To address these limitations, we designed a versatile NGS-based mapping method that consists of a combination of low- to medium-coverage multiplex SOLiD (Sequencing by Oligonucleotide Ligation and Detection) and classical genetic rough mapping. Using only low to medium coverage reduces the SOLiD sequencing costs and, since just 10 to 20 mutant F2 plants are required for rough mapping, the operation is simple enough to handle in a laboratory with limited space and funding. As a proof of principle, we successfully applied this method to identify the CTR1, which is involved in boron-mediated root development, from among a population of high boron requiring Arabidopsis thaliana mutants. Our work demonstrates that this NGS-based mapping method is a moderately priced and versatile method that can readily be applied to other model organisms.

Received: September 1, 2012; Accepted: October 12, 2012; Published Online: October 26, 2012

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