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Isolation and Purification of Protoplasts in Roots and Leaves of Soybean Seedlings(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2009年04期
Page:
697-701
Research Field:
Publishing date:

Info

Title:
Isolation and Purification of Protoplasts in Roots and Leaves of Soybean Seedlings
Author(s):
ZHANG Xiao-ke1; YU Bing-jun1; 2
1.College of Life Sciences，Nanjing Agricultural University，Nanjing 210095；
2.Soybean Research Institute，Nanjing Agricultural University；National Center for Soybean Improvement，Ministry of Agriculture，Nanjing 210095，Jiangsu，China
Keywords:
Soybean; Root; Leaf; Protoplast; Isolation
PACS:
S565.1
DOI:
10.11861/j.issn.1000-9841.2009.04.0697
Abstract:
The methods for isolation and purification of protoplast in roots and leaves of soybean seedling were investigated together in this study.The results showed that，the suitable enzyme species and concentrations for isolation of root protoplasts were CPW-13M+13%(W/V) mannotol+3% cellulose R-10+1.1% macerozyme R-10+1.0% hemicellulase，pH5.8；those for leaf were 0.15% CaCl22H2O+9% mannitol+1% cellulase R-10+0.20% pectolaseY-23，pH 5.8，and the temperature was 28℃.The reasonable enzymatic times for isolation of root and leaf protoplasts were 16 and 4 h，respectively；and the higher protoplast yield and viability were accordingly 1.46×105protoplastsg-1?FW and 57.8%，1.74×106protoplastsg-1FW and 70.3%.The better purification for root protoplasts was the sinking method of 23% sucrose mixed with CPW-18M，that for leaf was the rising method of 25% sucrose.

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Last Update: 2014-09-22