Investigation on the Protocol for Cryopreservation by Vitrification of Shoot Tips of Taiwan Native Medicinal Herb Limnophila aromaticoides Yang & Yen

• Main Authors: Shang-Chieh Su, 蘇上傑
• Other Authors: 廖松淵
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/42487545262321339083

碩士 === 國立中興大學 === 生命科學系所 === 101 === Limnophila aromaticoides Yang & Yen, a Taiwan native medicinal herb that in vitro-grown was used as experimental material in this study. Excised shoot tips were cryopreserved by vitrification, and investigated for the effect of pretreatment of different precultures with sucrose medium and cryoprotectant duration on survival rate. And then investigate the effect of different precultures on the physiology of bud by physiological analysis. The goal of this study is to find out the most optimal protocols for cryopreservation by vitrification of shoot tips of Limnophila aromaticoides Yang & Yen. 1-2 mm shoot tips excised from 2-3 weeks old plants were precultured, and then treated with different duration of PVS2, LS solution and precultures. The results demonstrated that the shoot tips precultured on 0.3 M sucrose medium for 7 days, treated with LS solution for 90 minutes at 25℃ and dehydrated with PVS2 for 120 minutes at 0℃ under shade for long-term storage. Buds rewarmed rapidly and plated on petri dishes containing 1/2MS medium, supplemented with BA 0.1mg L-1 and NAA 0.01 mg L-1. The survival ratio in this protocol could achieve 86.7%. Physiology analysis revealed that the preculture treatment of Limnophila aromaticoides Yang & Yen decreased relatively water content, water potential and osmotic potential, and increased the content of soluble sugar and soluble protein of shoot tips. The result indicated that the plantlets of Limnophila aromaticoides Yang & Yen regulated the physiology state of buds with osmotic adjustment during the preculture, which increased the ability tolerate the dehydration and freezing, thus enhanced the survival rate.