CRUNC: a cryopreservation method for unencapsulated gemmae of Marchantia polymorpha

View article
Loading...
Plant Biology

Main article text

 

Introduction

Materials and Methods

Plant materials and culture condition

Sterilization of silica-gel beads

Calculation of recovery rate and dehydration rate

Results and discussion

Basic CRUNC procedure

A higher dehydration rate gives a higher efficiency of recovery

Gemmae from younger and older gemma cups are not suitable for the CRUNC method

Dehydrated, unencapsulated gemmae can be cryopreserved in liquid nitrogen and frozen at −80 °C and −20 °C

Gemmae cryopreserved by the CRUNC method can tolerate a freeze-thaw-freeze treatment

The recovery rate depends on the transgenic line

Conclusions

Supplemental Information

The recovery rates of gemmae in the CRUNC method with dehydration for 24 h.

A scatter plot to compare between the dehydration rate (%D) and recovery rate (%R).

DOI: 10.7717/peerj.10174/supp-2

Removing a silica-gel bead from the 1.5-mL tube containing gemmae before measurement of the weight of the gemmae.

DOI: 10.7717/peerj.10174/supp-3

Thawing frozen gemmae using sterile ultra-pure water on agar medium for recovery.

DOI: 10.7717/peerj.10174/supp-4

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Hitomi Takahashi performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Yutaka Kodama conceived and designed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Data Availability

The following information was supplied regarding data availability:

The raw measurements are available in the Supplemental File.

Funding

This work was supported by the Japan Society for the Promotion of Science KAKENHI (No. 18H02455). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

4 Citations 1,755 Views 451 Downloads