PJB-2019-268
CRYOPRESERVATION OF IN VITRO GROWN SHOOT TIPS OF EMMENOPTERYS HENRYI BY DROPLET VITRIFICATION
NIU YANL
Abstract
Emmenopterys henryi. is becoming extremely rare in its natural habitat as a result of some factors for example overexploitation, and a low rate of nature regeneration (due to overexploitation, and a low rate of nature regeneration). In this study, we established a simple and reliable method for cryopreservation of in vitro shoot tips of E. henryi. Shoot tips, about 2 mm in length and with 2-3 leaf primordia, were excised from plantlets cultivated after 3 weeks acclimation. Then the shoot tips were precultured in hormone free Murashige and Skoog (MS) medium containing 0.5 M sucrose for 3 days and incubated in PVS2 for 1 h at 0℃, followed by a direct immersion in liquid nitrogen for 1h. After rapid thawing in a warm water bath at 40℃and rinsed five to six times with sterile-distilled water, the shoot tips were plated on the regeneration medium supplemented with 1.0mg/L TDZ, 1.0mg/L ZT, 0.6g/L PVP, 30g/L sucrose and 6.8g/L agar for one week in dark prior to exposure to normal conditions. The highest survival and regrowth percentage were 88.9% and 79.2%, respectively.
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