Department of Forest Molecular Genetics and Biotechnology, Forestry and Forest Products Research Institute, Forest Research and Management Organization, Tsukuba, Ibaraki, Japan.
Forestry and Forest Products Research Center, Shizuoka Prefectural Research Institute of Agriculture and Forestry, Hamamatsu, Shizuoka, Japan.
Sci Rep. 2021 Jan 15;11(1):1496. doi: 10.1038/s41598-020-80688-1.
Identifying causative genes for a target trait in conifer reproduction is challenging for species lacking whole-genome sequences. In this study, we searched for the male-sterility gene (MS1) in Cryptomeria japonica, aiming to promote marker-assisted selection (MAS) of male-sterile C. japonica to reduce the pollinosis caused by pollen dispersal from artificial C. japonica forests in Japan. We searched for mRNA sequences expressed in male strobili and found the gene CJt020762, coding for a lipid transfer protein containing a 4-bp deletion specific to male-sterile individuals. We also found a 30-bp deletion by sequencing the entire gene of another individual with the ms1. All nine breeding materials with the allele ms1 had either a 4-bp or 30-bp deletion in gene CJt020762, both of which are expected to result in faulty gene transcription and function. Furthermore, the 30-bp deletion was detected from three of five individuals in the Ishinomaki natural forest. From our findings, CJt020762 was considered to be the causative gene of MS1. Thus, by performing MAS using two deletion mutations as a DNA marker, it will be possible to find novel breeding materials of C. japonica with the allele ms1 adapted to the unique environment of each region of the Japanese archipelago.
在缺乏全基因组序列的情况下，鉴定针叶树繁殖目标性状的致病基因具有挑战性。在这项研究中，我们在日本柳杉中搜索雄性不育基因（MS1），旨在促进雄性不育日本柳杉的标记辅助选择（MAS），以减少日本人工日本柳杉林花粉传播引起的花粉过敏。我们搜索了在雄性球果中表达的 mRNA 序列，发现了编码脂转移蛋白的基因 CJt020762，该蛋白含有雄性不育个体特有的 4-bp 缺失。我们还通过对另一个具有 ms1 的个体的整个基因进行测序发现了一个 30-bp 的缺失。具有等位基因 ms1 的所有 9 个繁殖材料在基因 CJt020762 中都有 4-bp 或 30-bp 的缺失，这两种缺失都可能导致基因转录和功能失常。此外，在石卷自然林的五个人中有三个个体检测到了 30-bp 的缺失。根据我们的发现，CJt020762 被认为是 MS1 的致病基因。因此，通过使用两个缺失突变作为 DNA 标记进行 MAS，就有可能找到具有等位基因 ms1 的日本柳杉新型繁殖材料，这些材料适应日本列岛每个地区的独特环境。
