甘藷莖頂組織超低溫冷凍保存之研究

Study on Cryopreservation for Shoot Tip of Sweet Potato

游佩蓁(Pei-Chen Yu) ； 洪進雄(Chin-Hsiung Hung) ； 羅淑芳(Shu-Fang Lo)

《作物、環境與生物資訊》 9卷4期 (2012/12) Pp. 209-219

https://doi.org/10.30061/CEB.201212.0001

摘要

本文為國內首次以藻膠包埋玻璃質化法或冷凍鋁片法進行超低溫冷凍保存（cryopreservation），使甘藷有再生植株形成的研究報告。試驗以甘藷[ Ipomoea batatas（L.）Lam.]臺農57號、66號、71號及73號莖頂組織為材料，以藻膠包埋玻璃質化法（encapsulation-vitrification）或冷凍鋁片法（aluminum cryo-plate）進行超低溫冷凍保存，結果顯示臺農57號、66號、71號及73號等4個品種，以MS基本鹽類培養基添加0.3 M蔗糖及0.9% Difco agar進行預培養4天，可有效提升莖頂組織經loading solution （LS）溶液及plant vitrification solution 2 （PVS2） 溶液處理後之組織存活率達60-100%。以藻膠包埋玻璃質化法配合LS溶液300 min 及PVS2溶液60 min處理，經液態氮保存後，結果4個品種的植株再生率皆為3.3%，若以plant vitrification solution 3 （PVS3）替代PVS2 溶液配合LS 溶液處理，僅2個品種（即臺農66號及71號）有3.3%的植株再生率。另外，冷凍鋁片技術以LS溶液及PVS2溶液皆處理30 min，經超低溫冷凍處理後結果臺農66號的植株再生率達3.3%，而LS溶液處理30 min及PVS2溶液處理15 min，使臺農71號植株再生率亦為3.3%。冷凍鋁片技術較藻膠包埋玻璃質化法可節省45%的處理時間及減少70%藥品消耗量，此法為頗具潛力的超低溫冷凍保存方法。

關鍵字

甘藷 ； 超低溫冷凍保存 ； 藻膠包埋玻璃質化法 ； 冷凍鋁片法

並列摘要

This is the first report describing the use of encapsulation-vitrification or aluminum cryo-plate technology for cryopreservation of sweet potato and then to regenerate new plants in Taiwan. The shoot tips of sweet potato [Ipomoea batatas (L) Lam.] cultivars Tainung Nos. 57, 66, 71 and 73 were adopted for the experiment. The results showed the survival rate of the shoot tips could increase to 60-100% by pre-culture with MS (Murashige and Skoog) basic salted medium with 0.3 M sucrose and 0.9% Difco agar for 4 d after treatment of loading solution (LS) and plant vitrification solution 2 (PVS2). In the method of encapsulation-vitrification, the shoot tips of four sweet potato cultivars were osmoprotected with LS for 300 min and then dehydrated with PVS2 for 60 min before immersing into liquid nitrogen. The regeneration rate reached 3.3% for all these four cultivars. By replacing PVS2 solution with plant vitrification solution 3 (PVS3) for dehydration and osmoprotection before immersion into liquid nitrogen, both Tainung No. 66 and No. 71 reached 3.3% regeneration rate. With aluminium cryo-plate technology for cryopreservation, the regeneration rate of Tainung No. 66 reached 3.3% when treated with both LS and PVS2 solutions for 30 min each. Tainung No. 71 had the same rate when treated with 30 min LS and 15 min PVS2. By comparing the results obtained from these two technologies, the cryopreservation using aluminum cryo-plate reduced 45% of processing time and 70% of chemical used. Accordingly, aluminum cryo-plate technology seems a better method for cryopreservation of shoot tips of sweet potato.

並列關鍵字

Sweet potato ； Cryopreservation ； Encapsulation-vitrification ； Aluminum cryo-plate