Cryopreservation of Marchantia polymorpha spermatozoa

Area of Science:

• * Plant reproductive biology
• * Cryobiology
• * Molecular genetics

Background:

• * Marchia polymorpha is a key model organism due to its genetic simplicity and reproductive versatility.
• * Cryopreservation of spermatozoa offers an efficient method for conserving plant genetic resources and reducing maintenance efforts.
• * Preserving fertile spermatozoa eliminates the need for continuous cultivation of male plants, saving time and space.

Purpose of the Study:

• * To establish a reliable protocol for cryopreserving Marchia polymorpha spermatozoa in liquid nitrogen.
• * To assess the viability and motility of cryopreserved spermatozoa.
• * To confirm the functional capacity of cryopreserved spermatozoa in fertilization.

Main Methods:

• * Development of a cryoprotective solution including sucrose, glycerol, and egg yolk.
• * Implementation of controlled cooling and warming procedures for cryopreservation.
• * Assessment of spermatozoal survival rates and motility post-cryopreservation.
• * Fertilization assays using cryopreserved spermatozoa to produce spores.

Main Results:

• * Successful cryopreservation of Marchia polymorpha spermatozoa was achieved using the developed protocol.
• * Post-cryopreservation, spermatozoal survival and motility were maintained at 71% and 54% of pre-cryopreservation levels, respectively.
• * Cryopreserved spermatozoa demonstrated fertilization capability, resulting in the formation of normal spores.

Conclusions:

• * The cryopreservation protocol enables effective long-term storage of Marchia polymorpha spermatozoa.
• * This technique significantly enhances the experimental versatility of Marchia polymorpha as a model organism.
• * The protocol holds potential for broader application in the cryopreservation of plant spermatozoa.