CRUNC: a cryopreservation method for unencapsulated gemmae of Marchantia polymorpha.

ABSTRACT: 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.

SUBMITTER: Takahashi H 

PROVIDER: S-EPMC7587056 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature