SCAR Markers for Practical Application of Marker-assisted Selection in Persimmon (Diospyros kaki Thunb.) Breeding
SCAR Markers for Practical Application of Marker-assisted Selection in Persimmon (Diospyros kaki Thunb.) Breeding
复制标题
DOI：
10.2503/jjshs1.79.150
复制
发表时间：
2010-04
影响因子：
--
通讯作者：
S. Kanzaki;T. Akagi;T. Masuko;M. Kimura;M. Yamada;A. Sato;N. Mitani;Naoki Ustunomiya;K. Yonemori-K.-Y
中科院分区：
文献类型：
--
作者：
S. Kanzaki;T. Akagi;T. Masuko;M. Kimura;M. Yamada;A. Sato;N. Mitani;Naoki Ustunomiya;K. Yonemori-K.-Y
在以前的研究中，我们已经开发了分子标记连锁的AST基因座，控制果实涩味类型的柿子，然而，这些标记是不可行的实际应用育种计划，因为它们不是完全有效的区分授粉常数和非涩味（PCNA）基因型从非PCNA基因型的后代来自'黑熊'。在这里，我们开发了新的序列特征扩增区（SCAR）标记，使容易和可靠的选择PCNA基因型的育种群体。从“西村wase”、“Jiro”和“Kurokuma”衍生后代的基因组文库中分离出在限制性片段长度多态性（RFLP）分析中显示PCNA和非PCNA基因型之间的多态性片段的探针5 R附近的基因组区域。分离的基因组区域进行了表征，并观察到3个插入/缺失突变之间的ast和AST连接的区域。在Indel-3的侧翼区设计了几个引物，在多重PCR中，显示使用2个正向引物AST-F和PCNA-F以及反向引物5 R3 R是最有用和可靠的引物组。AST连锁的220-bp片段被证明是'Kurokuma'-、'Nisimura-wase'-和'Aizumishirazu'-衍生后代的共同标记。这种多重PCR被认为是最实用的工具，标记辅助选择（MAS），可以提高和加速柿子育种的进展。
In previous studies, we have developed molecular markers linked to the AST locus that controls fruit astringency type in persimmon; however, these markers are not feasible for practical application to breeding programs since they are not fully effective for discriminating the pollination constant and non-astringent (PCNA) genotype from the non-PCNA genotype in a progeny derived from ‘Kurokuma’. Here we developed new sequence characterized amplified region (SCAR) markers that enable easy and reliable selection of the PCNA genotype from breeding populations. Genomic regions adjacent to probe 5R, which showed polymorphic fragments between PCNA and non-PCNA genotypes in restriction fragment length polymorphism (RFLP) analysis, were isolated from the genomic libraries of ‘Nishimura-wase’, ‘Jiro’, and ‘Kurokuma’-derived offspring. The isolated genomic regions were characterized and 3 insertion/deletion mutations were observed between ast- and AST-linked regions. Several primers were designed in the flanking region of Indel-3 and, in multiplex PCR, it was shown that using 2 forward primers, AST-F and PCNA-F and a reverse primer, 5R3R, is the most useful and reliable primer set. The AST-linked 220-bp fragment proved to be a common marker of ‘Kurokuma’-, ‘Nisimura-wase’- and ‘Aizumishirazu’-derived progenies. This multiplex PCR is considered the most practical tool for marker-assisted selection (MAS) and can enhance and accelerate progress in persimmon breeding.