Keywords

Soybeans, Chickpeas, Protoplasts, Isolation Systems, Subcellular Localization

Abstract

Using the young leaves of Xiangdou No. 3 soybean and Kabuli type chickpea as materials, the effects of enzymatic hydrolysis types and ratios, mannitol concentration in the enzymatic hydrolysis solution, pH of the enzymatic hydrolysis solution and enzymatic hydrolysis time on the young leaves of the two leguminous plants were studied. Effects on protoplast yield and survival rate of young leaves. The results show that Xiangdou No. 3 soybean leaves contain 0.5% cellulase Onozuka R-10, 0.8% hemicellulase, 0.8% segregase Macerozyme R-10, 0.4% pectolyase Y-23, 0.1% In the enzymatic hydrolysis solution of 2-(N-morpholine)ethanesulfonic acid (MES) and 10% mannitol (pH 6.0), the enzyme was hydrolyzed for 6 hours in a constant temperature water bath (45 r/min) under dark conditions at 27 ¡æ, and then isolated. The protoplast yield and survival rate are the highest; Kabuli type chickpea leaves contain 0.5% cellulase Onozuka R-10, 0.8% hemicellulase Hemicellulase, 0.8% hemicellulase Macerozyme R-10, 0.1% MES and 10% In mannitol enzymatic hydrolyzate (pH 4.8), the protoplasts obtained were isolated with the highest yield and survival rate in a constant temperature shaking water bath (45 r/min) at 27¡ãC in the dark for 7¨C8 h. The protoplasts from the young leaves of Xiangdou No. 3 soybean and Kabuli type chickpea isolated through the above-mentioned methods have the best effect for subcellular localization.

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Original research done by Wang Zhankui, Shu Yingjie, Chen Ming, Tao Yuan, Ma Hao, Huang Liyan

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