Abstract
Date palm (Phoenix dactylifera L.) is a staple crop across arid and semi-arid regions around the world. Both genebanks and date palm breeders benefit by having clearly defined methods for the long-term preservation of pollen. Methods were developed to cryopreserve pollen from date palm trees in the USDA-ARS National Plant Germplasm System. First, in vitro pollen germination conditions were optimized using five cultivars. Then, date palm pollen equilibrated over saturated salt solutions of Ca(NO3)2 or MgCl2 at 23 °C (46% and 33% RH, respectively) or in a room set at 23% RH and 5 °C. Pollen was kept at these conditions for one to 56 days and then placed into liquid nitrogen vapor (LNV). Viability was assessed for fresh, moisture-equilibrated (non-LNV and LNV-exposed), and pollen stored in LNV for 9 months. The highest level of in vitro pollen germination was observed for date palm cultivar Thoory BC3 (60 ± 2%) when it was rehydrated for 2 h and then incubated at 23 °C on Marquard medium with 15% sucrose. Viability remained stable for pollen that was stored in LNV for 9 months. In addition, pollen also remained viable if it was cooled to and warmed from LNV for up to 60 cycles. These data suggest that date palm pollen can be successfully placed in genebank cryostorage.
Key message
Using optimized in vitro germination conditions, date palm pollen can survive at least 9 months in LNV and can be cooled to and warmed from LNV at least 60 times.
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Abbreviations
- ARS:
-
Agricultural Research Service
- Ca(NO3)2 :
-
Calcium nitrate
- Ca(NO3)2·2H2O:
-
Calcium nitrate dihydrate
- CuSO4·5H2O:
-
Copper sulfate pentahydrate
- DW:
-
Dry weight
- FW:
-
Fresh weight
- H3BO3 :
-
Boric acid
- KNO3 :
-
Potassium nitrate
- LNV:
-
Liquid nitrogen vapor
- MC:
-
Moisture content
- MgCl2 :
-
Magnesium chloride
- MgSO4·7H2O:
-
Magnesium sulfate heptahydrate
- NCGRCD:
-
National clonal germplasm repository for citrus and dates
- NPGS:
-
National Plant Germplasm System
- RH:
-
Relative humidity
- TW:
-
Tare weight
- UC-VARS:
-
University of California Coachella Valley agricultural research station
- USD:
-
United States dollars
- USDA:
-
United States Department of Agriculture
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Acknowledgements
We thank Vince Samons for assistance in date palm spathe collection and orchard maintenance. We acknowledge the Coordination for the Improvement of Higher Education Personnel (CAPES) for granting the doctoral scholarship to A.C.A de Oliveira. USDA is an equal opportunity employer. Any mention of trade names of commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.
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Araújo de Oliveira, A.C., da Silva Lédo, A., Polek, M. et al. Optimization of in vitro germination and cryopreservation conditions for preserving date palm pollen in the USDA National Plant Germplasm System. Plant Cell Tiss Organ Cult 144, 223–232 (2021). https://doi.org/10.1007/s11240-020-01907-1
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DOI: https://doi.org/10.1007/s11240-020-01907-1