Viability Assessment ofGenipa americanaL. (Rubiaceae) Embryonic Axes after Cryopreservation UsingIn VitroCulture

Abstract

Embryonic axes excised from seeds ofGenipa americanaL. desiccated to different water contents were successfully cryopreserved by rapidly plunging seed samples directly into liquid nitrogen. Control and cryopreserved embryonic axes were excised and grown in WPM culture medium for viability assessment. All control embryonic axes (−LN2) excised from fully hydrated seeds (43.89% moisture content) germinated after 21 days of culturein vitro. These high germination percentages persisted even after the water content of the seeds was as low as 6.79%. After freezing in liquid nitrogen high germination percentages, 93%, 96%, and 93%, were observed for embryonic axes excised from seeds dehydrated to 13.26%, 9.57%, and 6.79 moisture content, respectively. The cryopreservation technique described here is recommended for long term conservation ofG. americanagermplasm.

In vitro germination of zygotic embryos excised from cryopreserved endocarps of queen palm (Syagrus romanzoffiana (Cham.) Glassman)

“…The present study reports on a simple cryopreservation technique for S. romanzoffiana, involving partial desiccation of endocarps followed by direct freezing, slow thawing, and aseptic excision and culture in vitro of ZE. Excision and in vitro culture of embryos after cryopreservation is a simple technique that has been applied to many other plant species including C. nucifera (Assy-Bah and Engelmann 1992), V. merrilli and H. fosteriana (Chin et al 1988), and Genipa americana (Santos and Salomão 2016), and it has been helpful in the assessment of viability and the regeneration of plantlets from cryopreserved seed samples (Pritchard et al 2000).…”

“…Embryo culture in vitro has been used in seed conservation as an alternative, rapid method of assessing seed quality or to recover growth of difficult-to-store seeds, and generally, there is a good correlation between embryo growth in vitro and germination percentages (Pritchard et al 2000). Cryopreservation of seeds and zygotic embryos was previously used as an alternative strategy for long-term conservation of many plant species (Pritchard et al 2000;González-Benito et al 2009;Santos and Salomão 2016). In the case of palms, cryopreservation protocols have been developed for seeds and embryos of different species (Grout et al 1983;Chin et al 1988;Assy-Bah and Engelmann 1992;Engelmann et al 1995;Raja et al 2003;Steinmacher et al 2007;N'Nan et al 2012).…”

Long term conservation of embryonic axes of genipap accessions

“…These techniques have been successfully used for different explants in Citrus sp. [13]; Coffea arabica L. [9], Hancornia speciosa Gomes [20,25,26], Genipa americana L. [23,24], Pistacia vera L. [1] and Ananas comosus L.) Merr. [31].…”

“…Many damages can be caused to frozen samples with inadequate water content such as: mechanical disorganization of plant structures, mechanical damage resulting from cell volume reduction and cell membrane injury [24]. Tolerance to dehydration varies from species to species and among tissues of the same plant, and appropriate humidity must be reached to ensure adequate survival of the material [22].…”

Effect of Desiccation Time on Seed Moisture and Regeneration of Mangaba (Hancornia speciosa) Embryos

“…There are no studies in the literature on the cryopreservation of seeds and structures such as zygotic embryos and cotyledonary axes. However, some studies with intermediate species such as jenipapo (Genipa americana L.) [14], cotton (Gossypium hirsutum L.) [9] and coffee trees (Coffea arabica L.) [15] obtained regeneration using embryonic axes.…”