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Zygotic and nucellar embryo survival following dehydration and cryopreservation of citrus intact seeds.

M Lambardi¹, A De Carlo, S Biricolti

¹IIstituto per la Valorizzazione del Legno e delle Specie Arboree, CNR Consiglio Nazionale delle Ricerche, Polo Scientifico, via Madonna del Piano, 50019 Sesto Fiorentino (Firenze), Italy. lambardi@ivalsa.cnr.it

|June 25, 2004

Summary

Cryopreservation of citrus seeds using dehydration and liquid nitrogen improved survival rates for multiple species. This method also favored the germination of zygotic embryos over apomictic ones, enhancing genetic diversity preservation.

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Area of Science:

Plant Science
Cryobiology
Horticulture

Background:

Polyembryonic citrus seeds pose challenges for cryopreservation due to their sensitivity to dehydration and freezing.
Developing effective cryopreservation techniques is crucial for conserving genetic resources of valuable citrus species.

Purpose of the Study:

To develop and evaluate a cryopreservation protocol for polyembryonic citrus seeds involving dehydration and liquid nitrogen immersion.
To investigate the effect of this cryopreservation method on the germination and origin (sexual vs. nucellar) of recovered citrus seedlings.

Main Methods:

Seeds of four polyembryonic Citrus species (C. sinensis, C. aurantium, C. limon, C. deliciosa) were dehydrated to specific moisture contents using sterile airflow.
Dehydrated seeds were directly immersed in liquid nitrogen for cryopreservation.
Germination rates and seedling origin (zygotic vs. apomictic) were assessed, with the latter determined using Inter-Simple Sequence Repeat (ISSR) analysis.

Main Results:

Three citrus species (C. sinensis, C. aurantium, C. limon) tolerated dehydration to 10-16% moisture content with minimal germination loss.
C. deliciosa showed reduced germinability (50%) after drying to 15% moisture.
Cryopreservation significantly improved survival for C. sinensis and C. aurantium (93% germination), with dehydration protecting seeds during liquid nitrogen immersion.
ISSR analysis revealed that the procedure promoted zygotic embryo germination and reduced apomictic seedling production.

Conclusions:

Dehydration followed by liquid nitrogen immersion is an effective cryopreservation strategy for polyembryonic citrus seeds, particularly for C. sinensis and C. aurantium.
This cryopreservation method enhances the recovery of sexually derived (zygotic) seedlings, contributing to the preservation of genetic diversity in citrus.
The protocol offers a promising approach for the long-term conservation of valuable citrus germplasm.