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۱Cryopreservation of Two Egyptian Grape (Vitis vinifera) Cultivars Using Two Steps Vitrification Protocol
نویسنده(ها): ،
اطلاعات انتشار: World Applied Sciences Journal، بيست و هشتم،شماره۲، ۲۰۱۳، سال
تعداد صفحات: ۵
Cryopreservation of tissue–cultured plant, cells, meristems and organ is an important method for longterm storage of plant genetic resources using a minimum of space and maintenance. In this work, shoot tips from two Egyptian cultivars (Black Matrouh and Bez El–Anza) obtained from greenhouse were successfully cryopreserved by two step vitrification protocol. Shoot tips were excised cultured on 1\2 MS +0.16 mM glutathione +0.14 mM ascorbic acid in addition to different concentration 0.1M, 0.2M and 0.3M of sucrose and then treated with a mixture of glycerol and sucrose for 20 min at 25°C. They were then dehydrated with mixture of 1\2X PVS2 and PVS2 for 150–20 min. then plunged into LN for 1 hr. Samples were then warmed rapidly in water at 40°C. Precultured shoot tips with high sucrose concentrations to form Osmo protected shoot tips were showed using 0.3M sucrose that gave the highest recovery before plunging into LN. After pluning into liquid nitrogen the recovery percentage were ranged 12–10 for Black Matrouh and Bez El–Anza, respectively. The recovery of the shoot tips amounted to approximately 40–46% after dehydration with PVS2 and plunging in liquid nitrogen. This method incorporates a simple and reliable approach for providing high levels of growth recovery. It also appears promising for the cryopreservation of Vitis germplasm.
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