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Abstract Detail

Conservation Biology

Hansel, Nicole [1], Pence, Valerie [2], Tepe, Eric [3].

Comparison of post cryopreservation survival of hyperhydric and more normal phenotypes of Cycladenia humilis var. jonesii, and Crotalaria avonensis.

The physiological disorder in in vitro plants known as hyperhydricity is characterized by an excess of water in the plant tissues. It is thought that this condition is suboptimal for cryopreservation, because the water could crystallize during exposure to liquid nitrogen (LN) and kill the tissues. In this study, two species, Cycladenia humilis var. jonesii (threatened) and Crotalaria avonensis (endangered), were investigated for post-cryopreservation survival of both a more normal (MN) and a hyperhydric (HH) phenotype. Cycladenia humilis var. jonesii was grown on Driver and Kuniyuki Walnut (DKW) medium with 0.5 mg/L of the cytokinin, benzylaminopurine (BAP). The HH phenotype was maintained in tubes with closed caps and the MN phenotype was maintained in tubes with vented caps. The Cr. avonensis HH phenotype was grown on a Murashige and Skoog (MS) medium with 1 mg/L BAP in tubes with closed caps, while the MN phenotype was grown on an MS medium with lower nitrogen (3 mM NO3, 1.5 mM NH4) and 0.1 mg/L BAP and vented caps. Shoot tips were isolated and, after 1-2 days on pre-culture medium with 10 uM of the stress hormone abscisic acid (ABA) and 0.3 M mannitol, shoot tips were exposed to LN using droplet vitrification. Data on survival were taken at two and four weeks measuring both callus and shoot formation. There was no significant difference in post LN exposure survival between the two phenotypes for either species (Tukey-Kramer ANOVA, p >0.05, for all at both 2 and 4 weeks), but there were developmental differences. In both species, the HH form had significantly more callus at 2 weeks than the MN (p<0.05), but shoots did not develop until 4 weeks. In contrast, MN shoot tips for Cy. humilis var. jonesii began developing at 2 weeks, but many were dead by 4 weeks. Thus, while there was no significant difference in survival through cryopreservation between these two phenotypes, there were differences in the development of these two forms during recovery growth.

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Related Links:
CREW Plant Research at the Cincinnati Zoo & Botanical Garden
BryoBioBank at the Cincinnati Zoo & Botanical Garden

1 - University of Cincinnati, Department of Biological Sciences, 614 Rieveschl Hall, Cincinnati, Ohio, 45221, USA
2 - Cincinnati Zoo and Botanical Garden, CREW, 3400 Vine St, Cincinnati, OH, 45220, USA
3 - University Of Cincinnati, Department Of Biological Sciences, 614 Rieveschl Hall, Cincinnati, OH, 45221, United States

Tissue Culture

Presentation Type: Poster
Session: P, Conservation Biology
Location: Grand Ballroom - Exhibit Hall/Mayo Civic Center
Date: Monday, July 23rd, 2018
Time: 5:30 PM This poster will be presented at 5:30 pm. The Poster Session runs from 5:30 pm to 7:00 pm. Posters with odd poster numbers are presented at 5:30 pm, and posters with even poster numbers are presented at 6:15 pm.
Number: PCB013
Abstract ID:835
Candidate for Awards:None

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