Yonemaru Jun-Ichi, Choi Sun Hee, Sakai Hiroaki, Ando Tsuyu, Shomura Ayahiko, Yano Masahiro, Wu Jianzhong, Fukuoka Shuichi
Agrogenomics Research Center, National Institute of Agrobiological Sciences , 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602 , Japan.
Agrogenomics Research Center, National Institute of Agrobiological Sciences , 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602 , Japan ; NARO Institute of Crop Science , 2-1-18 Kannondai, Tsukuba, Ibaraki 305-8518 , Japan.
Breed Sci. 2015 Jun;65(3):249-56. doi: 10.1270/jsbbs.65.249. Epub 2015 Jun 1.
Insertion-deletion (indel) polymorphisms, such as simple sequence repeats, have been widely used as DNA markers to identify QTLs and genes and to facilitate rice breeding. Recently, next-generation sequencing has produced deep sequences that allow genome-wide detection of indels. These polymorphisms can potentially be used to develop high-accuracy polymerase chain reaction (PCR)-based markers. Here, re-sequencing of 5 indica, 2 aus, and 3 tropical japonica cultivars and Japanese elite cultivar 'Koshihikari' was performed to extract regions containing large indels (10-51 bp) shared by diverse cultivars. To design indel markers for the discrimination of genomic regions between 'Koshihikari' and other diverse cultivars, we subtracted the indel regions detected in 'Koshihikari' from those shared in other cultivars. Two sets of indel markers, KNJ8-indel (shared in eight or more cultivars, including 'Khao Nam Jen' as a representative tropical japonica cultivar) and C5-indel (shared in five to eight cultivars), were established, with 915 and 9,899 indel regions, respectively. Validation of the two marker sets by using 23 diverse cultivars showed a high PCR success rate (≥95%) for 83.3% of the KNJ8-indel markers and 73.9% of the C5-indel markers. The marker sets will therefore be useful for the effective breeding of Japanese rice cultivars.
插入缺失(Indel)多态性,如简单序列重复,已被广泛用作DNA标记来鉴定数量性状基因座(QTL)和基因,并促进水稻育种。最近,新一代测序产生了深度序列,使得能够在全基因组范围内检测Indel。这些多态性有可能用于开发基于高精度聚合酶链反应(PCR)的标记。在此,对5个籼稻品种、2个奥氏稻品种、3个热带粳稻品种以及日本优良品种“越光”进行了重测序,以提取不同品种共有的包含大插入缺失(10 - 51 bp)的区域。为了设计用于区分“越光”和其他不同品种基因组区域的Indel标记,我们从其他品种共有的Indel区域中减去了在“越光”中检测到的Indel区域。建立了两组Indel标记,KNJ8 - Indel(在八个或更多品种中共有,包括作为代表性热带粳稻品种的“考南珍”)和C5 - Indel(在五到八个品种中共有),分别有915个和9899个Indel区域。使用23个不同品种对这两组标记进行验证,结果显示83.3%的KNJ8 - Indel标记和73.9%的C5 - Indel标记具有较高的PCR成功率(≥95%)。因此,这些标记组将有助于日本水稻品种的有效育种。