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Development of intron-flanking EST markers for the Lolium/Festuca complex using rice genomic information

Tamura, Ken-ichi; Yonemaru, Jun-ichi; Hisano, Hiroshi; Kanamori, Hiroyuki; King, Julie; King, Ian P.; Tase, Kazuhiro; Sanada, Yasuharu; Komatsu, Toshinori; Yamada, Toshihiko


Ken-ichi Tamura

Jun-ichi Yonemaru

Hiroshi Hisano

Hiroyuki Kanamori

Kazuhiro Tase

Yasuharu Sanada

Toshinori Komatsu

Toshihiko Yamada


DNA markers able to distinguish species or genera with high specificity are valuable in the identification of introgressed regions in interspecific or intergeneric hybrids. Intergeneric hybridization between the genera of Lolium and Festuca, leading to the reciprocal introgression of chromosomal segments, can produce novel forage grasses with unique combinations of characteristics. To characterize Lolium/Festuca introgressions, novel PCR-based expression sequence tag (EST) markers were developed. These markers were designed around intronic regions which show higher polymorphism than exonic regions. Intronic regions of the grass genes were predicted from the sequenced rice genome. Two hundred and nine primer sets were designed from Lolium/Festuca ESTs that showed high similarity to unique rice genes dispersed uniformly throughout the rice genome. We selected 61 of these primer sets as insertion-deletion (indel)-type markers and 82 primer sets as cleaved amplified polymorphic sequence (CAPS) markers to distinguish between Lolium perenne and Festuca pratensis. Specificity of these markers to each species was evaluated by the genotyping of four cultivars and accessions (32 individuals) of L. perenne and F. pratensis, respectively. Evaluation using specificity indices proposed in this study suggested that many indel-type markers had high species specificity to L. perenne and F. pratensis, including 15 markers completely specific to both species. Forty-nine of the CAPS markers completely distinguish between the two species at bulk level. Chromosome mapping of these markers using a Lolium/Festuca substitution line revealed syntenic relationships between Lolium/Festuca and rice largely consistent with previous reports. This intron-based marker system that shows a high level of polymorphisms between species in combination with high species specificity will consequently be a valuable tool in Festulolium breeding.


Tamura, K., Yonemaru, J., Hisano, H., Kanamori, H., King, J., King, I. P., …Yamada, T. (2009). Development of intron-flanking EST markers for the Lolium/Festuca complex using rice genomic information. TAG Theoretical and Applied Genetics, 118(8), 1549-1560.

Journal Article Type Article
Acceptance Date Mar 8, 2009
Online Publication Date Mar 27, 2009
Publication Date 2009-05
Deposit Date Dec 6, 2018
Journal Theoretical and Applied Genetics
Print ISSN 0040-5752
Electronic ISSN 1432-2242
Publisher BMC
Peer Reviewed Peer Reviewed
Volume 118
Issue 8
Pages 1549-1560
Public URL
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