Development of diagnostic PCR and LAMP markers for MALE STERILITY 1 (MS1) in Cryptomeria japonica D. Don

BMC Research Notes

Asako Matsumoto¹  Fu-Jin Wei¹  Kentaro Uchiyama¹  Saneyoshi Ueno¹  Yoichi Hasegawa¹  Tokuko Ujino-Ihara¹  Takeshi Fujino²  Masahiro Kasahara²  Yoshinari Moriguchi³  Katsushi Yamaguchi⁴  Shuji Shigenobu⁴  Takahiro Bino⁴  Tetsuji Hakamata⁵ 

[1]Forestry and Forest Products Research Institute, Forest Research and Management Organization

[2]Graduate School of Frontier Sciences, The University of Tokyo

[3]Graduate School of Science and Technology, Niigata University

[4]National Institute for Basic Biology

[5]Shizuoka Prefectural Research Institute of Agriculture and Forestry

关键词: ALP;    ASP;    LAMP;    Molecular markers;    Variant detection;    Cryptomeria japonica;   

DOI  :  10.1186/s13104-020-05296-8

来源: DOAJ

【 摘 要 】

Abstract 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 to produce a pollen-free cultivar. In order to reduce the time and cost needed for 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 to rapidly identify pollen-free mutants. Results We developed PCR and LAMP markers to detect mutant alleles and to present experimental options depending on available laboratory equipment. LAMP markers were developed for field stations, where PCR machines are unavailable. The LAMP method only needs heat-blocks or a water bath to perform the isothermal amplification and assay results can be read by the naked eye. Because the causative mutations were deletions, we developed two kinds of PCR markers, amplified length polymorphism (ALP) and allele specific PCR (ASP) markers. These assays can be visualized using capillary or agarose gel electrophoresis.

【 授权许可】

Unknown