Ueno Saneyoshi, Uchiyama Kentaro, Moriguchi Yoshinari, Ujino-Ihara Tokuko, Matsumoto Asako, Wei Fu-Jin, Saito Maki, Higuchi Yumi, Futamura Norihiro, Kanamori Hiroyuki, Katayose Yuichi, Tsumura Yoshihiko
Department of Forest Molecular Genetics and Biotechnology, Forestry and Forest Products Research Institute, Forest Research and Management Organization, 1 Matsunosato, Tsukuba, Ibaraki 305-8687, Japan.
Graduate School of Science and Technology, Niigata University, 8050, Ikarashi 2-no-cho, Nishi-ku, Niigata 950-2181, Japan.
Breed Sci. 2019 Mar;69(1):19-29. doi: 10.1270/jsbbs.17149. Epub 2019 Feb 15.
is a major forestry tree species in Japan. Male sterility of the species is caused by a recessive gene, which shows dysfunction of pollen development and results in no dispersed pollen. Because the pollen of induces pollinosis, breeding of pollen-free is desired. In this study, single nucleotide polymorphism (SNP) markers located at 1.78 and 0.58 cM to a male sterility locus () were identified from an analysis of RNA-Seq and RAD-Seq, respectively. SNPs closely linked to were first scanned by a method similar to MutMap, where a type of index was calculated to measure the strength of the linkage between a marker sequence and . Linkage analysis of selected SNP markers confirmed a higher efficiency of the current method to construct a partial map around . Allele-specific PCR primer pair for the most closely linked SNP with was developed as a codominant marker, and visualization of the PCR products on an agarose gel enabled rapid screening of male sterile . The allele-specific primers developed in this study would be useful for establishing the selection of male sterile .
是日本的主要造林树种。该物种的雄性不育由隐性基因引起，表现为花粉发育功能障碍，导致花粉不散出。由于该物种的花粉会引发花粉症，因此需要培育无花粉的该物种。在本研究中，分别通过RNA-Seq和RAD-Seq分析，鉴定出位于距雄性不育位点（）1.78和0.58 cM处的单核苷酸多态性（SNP）标记。首先通过类似于MutMap的方法扫描与紧密连锁的SNP，其中计算一种指数来衡量标记序列与之间连锁的强度。对选定SNP标记的连锁分析证实了当前方法在构建围绕的部分图谱方面具有更高的效率。针对与最紧密连锁的SNP开发了等位基因特异性PCR引物对作为共显性标记，并且在琼脂糖凝胶上对PCR产物进行可视化能够快速筛选雄性不育。本研究中开发的等位基因特异性引物将有助于建立雄性不育的选择方法。
