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Development of diagnostic PCR and LAMP markers for MALE STERILITY 1 (MS1) in Cryptomeria japonica D. Don
bioRxiv - Genetics Pub Date : 2020-05-20 , DOI: 10.1101/2020.05.19.090092 Yoichi Hasegawa , Saneyoshi Ueno , Fu-Jin Wei , Asako Matsumoto , Tokuko Ujino-Ihara , Kentaro Uchiyama , Yoshinari Moriguchi , Masahiro Kasahara , Takeshi Fujino , Shuji Shigenobu , Katsushi Yamaguchi , Takahiro Bino , Tetsuji Hakamata
bioRxiv - Genetics Pub Date : 2020-05-20 , DOI: 10.1101/2020.05.19.090092 Yoichi Hasegawa , Saneyoshi Ueno , Fu-Jin Wei , Asako Matsumoto , Tokuko Ujino-Ihara , Kentaro Uchiyama , Yoshinari Moriguchi , Masahiro Kasahara , Takeshi Fujino , Shuji Shigenobu , Katsushi Yamaguchi , Takahiro Bino , Tetsuji Hakamata
Objective: Due to the allergic nature of the pollen of Cryptomeria japonica, the most important Japanese forestry conifer, a pollen-free cultivar is preferred. Mutant trees detected in nature have been used for the production of a pollen-free cultivar. In order to reduce the time and cost needed for the production and breeding, we aimed to develop simple diagnostic molecular markers for mutant alleles of the causative gene MALE STERILITY 1 (MS1) in C. japonica. The expected function of this gene, its two dysfunctional mutations, and genetic diversity were described recently in a related study. Results: We have developed PCR and LAMP markers to detect mutant alleles and to present experimental options depending on available laboratory equipment. At field stations, where PCR machines are not available, LAMP markers were developed. LAMP only needs heat-blocks or a water bath to perform the isothermal amplification and assay results can be easily seen by eye. Because the causative mutations were deletions, two kinds of PCR markers, amplified length polymorphism (ALP) and allele specific PCR (ASP) markers, were developed. These assays can be carried out by capillary or agarose gel electrophoresis.
中文翻译:
日本柳杉男性不育1(MS1)诊断PCR和LAMP标记的开发
目的:由于日本最重要的林业针叶树日本柳杉的花粉具有过敏性,因此首选无花粉品种。在自然界中检测到的突变树已用于生产无花粉的品种。为了减少生产和育种所需的时间和成本,我们旨在开发简单的诊断分子标记物,以用于日本粳稻致病基因MALE STERILITY 1(MS1)的突变等位基因。最近在相关研究中描述了该基因的预期功能,它的两个功能异常突变和遗传多样性。结果:我们已经开发了PCR和LAMP标记,可以检测突变等位基因并根据可用的实验室设备提供实验选择。在没有PCR仪的现场站,开发了LAMP标记。LAMP仅需加热块或水浴即可进行等温扩增,而且肉眼可轻松看到测定结果。由于致病突变是缺失,因此开发了两种PCR标记,即扩增长度多态性(ALP)和等位基因特异性PCR(ASP)标记。这些测定可以通过毛细管或琼脂糖凝胶电泳进行。
更新日期:2020-05-20
中文翻译:
日本柳杉男性不育1(MS1)诊断PCR和LAMP标记的开发
目的:由于日本最重要的林业针叶树日本柳杉的花粉具有过敏性,因此首选无花粉品种。在自然界中检测到的突变树已用于生产无花粉的品种。为了减少生产和育种所需的时间和成本,我们旨在开发简单的诊断分子标记物,以用于日本粳稻致病基因MALE STERILITY 1(MS1)的突变等位基因。最近在相关研究中描述了该基因的预期功能,它的两个功能异常突变和遗传多样性。结果:我们已经开发了PCR和LAMP标记,可以检测突变等位基因并根据可用的实验室设备提供实验选择。在没有PCR仪的现场站,开发了LAMP标记。LAMP仅需加热块或水浴即可进行等温扩增,而且肉眼可轻松看到测定结果。由于致病突变是缺失,因此开发了两种PCR标记,即扩增长度多态性(ALP)和等位基因特异性PCR(ASP)标记。这些测定可以通过毛细管或琼脂糖凝胶电泳进行。