Cryopreservation of Zygotic Embryos of Podophyllum Hexandrum Royle, An Endangered Medicinal Plant, by Vitrification and V Cryo-Plate Techniques

Abstract

Background: Podophyllum hexandrum is a highly endangered valuable medicinal plant of the belonging to family Berberidaceae. This plant needs conservation efforts due to the over- and unscrupulous harvesting from the wild because of its ever-increasing demand. To establish a long-term cryopreservation method for Podophyllum hexandrum using techniques: Vitrification and V Cryo-plate. MATERIALS AND METHODS: Zygotic embryos cryopreserved using vitrification and V cryo-plate by optimization of parameters including time, loading time and PVS2 dehydration time. Recovery of zygotic embryos was on different regrowth media for plantlet formation. RESULTS: With V cryo-plate, 90% was obtained as compared to 73.3% with vitrification. V Cryo-plate conditions were pre- of zygotic embryos in 0.3 M sucrose for 4 days, treatment in loading solution with 0.8 M for 20 min, dehydration in PVS2 for 50 min, LN exposure, unloading in 1.2 M sucrose for 20 and transfer of zygotic embryos to regrowth medium for recovery. During recovery, the maximum of shoots (4.2) and highest shoot length (5.1 cm) were observed on regrowth medium with 1.5 BAP and 0.1 mg/l IAA (R 7). CONCLUSION: Zygotic embryos of Podophyllum hexandrum with 90% regrowth using a V cryoplate technique and plantlets were produced cryopreservation.