CRUNC: a cryopreservation method for unencapsulated gemmae of Marchantia polymorpha

PeerJ

article

Hitomi Takahashi¹  Yutaka Kodama¹ 

[1] Center for Bioscience Research and Education, Utsunomiya University

关键词: Bryophyte;    Cryoconservation;    Liverworts;    Unencapsulation;    Gemma;   

DOI  :  10.7717/peerj.10174

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Genetic modifications such as mutation and transformation are powerful tools to study the function of genes and proteins in the model liverwort Marchantia polymorpha, but maintaining the resulting germplasm requires a practical, reliable method. Cryopreservation methods allow researchers to maintain mutant and transgenic lines of M. polymorpha. To date, two methods have been developed for cryopreservation of M. polymorpha gemmae: in the first method, unencapsulated gemmae are stored in liquid nitrogen at −­196 °C, and in the second method, encapsulated gemmae are stored in liquid nitrogen at −­196 °C or a deep freezer at −80 °C. In the present study, we developed a simple method named CRUNC (cr yopreservation of un en c apsulated gemmae), which can be used to store unencapsulated, dried gemmae of wild-type and transgenic M. polymorpha lines in liquid nitrogen and in freezers at −80 °C and −20 °C. Using the CRUNC method, we observed a high recovery rate (as high as 100%) and successful long-term (5 months) storage of the gemmae. Therefore, the CRUNC method is practical for maintaining valuable M. polymorpha germplasm.

【 授权许可】

CC BY