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| Isolation and Culture of Protoplasts in Eelgrass Zostera marina |
| CUI Cui-ju,LIU Yan-ling,WANG Na,ZHANG Li-nan,TIAN Ping-ping,LI Xiao-jie,LUO Shi-ju,LI Yan |
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Abstract The influences of hydrolase combination ,solution pH ,and enzymolysis time on isolation and preparation of protoplast in eelgrass Zostera marina were studied ,and the conditions for separation of pro-toplasts were optimized from the eelgrass aseptic seedlings .It was found that high protoplast at a rate of 3 .90 × 106 individuals/g eelgrass was obtained in 1 .0 g aseptic seedling leaves which were digested in 5ml enzyme solution containing 2% cellulase ,0 .5% pectinase and 0 .5mol/L sorbitol under dark conditions of pH 4 .0 for 16 hours at 17 ℃ .Neutral red staining revealed that these protoplasts had viability of 85 .0% , and the conditions of the protoplast culture were also discussed in the eelgrass .
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Received: 25 August 2014
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