The Optimization of Isolation Method for Mesophyll Protoplast from Common Wheat（Triticum aestivum）Seedlings

doi:10.13560/j.cnki.biotech.bull.1985.2016.04.008

Abstract

Abstract: By orthogonal test，we analyzed the effects of concentrations of cellulose，macerozyme，and mannitiol as well as the time of enzyme digestion on the productivity and vitality during the isolation of protoplast from 7 days of wheat seedling mesophyll. Further，the transient expression technology was employed to analyze the effects of PEG concentration on the transformation efficiency by PEG-Ca2+-mediated transformation of mesophyll. The results showed that the optimal condition for isolating the mesophyll from wheat leaves was：1.5% of cellulase，0.75% of macerozyme，0.4 mol/L mannitol，and 3 h of enzyme digestion；the protoplast yield was 7.2×106 cells/g·FW，the vitality was more than 95%. The results from the transient expression technology revealed that transformation efficiency was the highest under 25% PEG.

Key words: common wheat, protoplast, isolation, transient expression

XI Hai-xiu, AI Ke-jun, TONG Shao-ming. The Optimization of Isolation Method for Mesophyll Protoplast from Common Wheat（Triticum aestivum）Seedlings[J]. Biotechnology Bulletin, 2016, 32(4): 68-73.

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