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On-line Access: 2024-08-27
Received: 2023-10-17
Revision Accepted: 2024-05-08
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Xiao-yu Gao, Xing-cheng Liao, Ruo-lai Wu, Ting Liu, Hai-xing Wang, Ling-li Lu. Purified isolation of vacuoles from Sedum alfredii leaf-derived protoplasts[J]. Journal of Zhejiang University Science B, 2017, 18(1): 85-88.
@article{title="Purified isolation of vacuoles from Sedum alfredii leaf-derived protoplasts",
author="Xiao-yu Gao, Xing-cheng Liao, Ruo-lai Wu, Ting Liu, Hai-xing Wang, Ling-li Lu",
journal="Journal of Zhejiang University Science B",
volume="18",
number="1",
pages="85-88",
year="2017",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1600138"
}
%0 Journal Article
%T Purified isolation of vacuoles from Sedum alfredii leaf-derived protoplasts
%A Xiao-yu Gao
%A Xing-cheng Liao
%A Ruo-lai Wu
%A Ting Liu
%A Hai-xing Wang
%A Ling-li Lu
%J Journal of Zhejiang University SCIENCE B
%V 18
%N 1
%P 85-88
%@ 1673-1581
%D 2017
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1600138
TY - JOUR
T1 - Purified isolation of vacuoles from Sedum alfredii leaf-derived protoplasts
A1 - Xiao-yu Gao
A1 - Xing-cheng Liao
A1 - Ruo-lai Wu
A1 - Ting Liu
A1 - Hai-xing Wang
A1 - Ling-li Lu
J0 - Journal of Zhejiang University Science B
VL - 18
IS - 1
SP - 85
EP - 88
%@ 1673-1581
Y1 - 2017
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1600138
Abstract: This study aims to develop a method for isolating and purifying protoplasts/vacuoles from fresh leaves of the Cd hyperaccumulator plant species, Sedum alfredii. The results revealed that preheating cellulase and macerozyme at 50 °C for 5 min significantly accelerated the cell wall degradation. For the most optimal conditions for mesophyll protoplast isolation, the mixture of fresh leaves and cell lysates was followed by a 2-h–long vibration. The protoplast lysate for vacuole isolation was diluted, and 0.675 mmol/L was identified as the most appropriate 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonic acid (CHAPS) level, in which S. alfredii large vacuoles are characterized by a high metal and malic acid content. For the best vacuole purification results, we established that 0.8 mol/L was the most optimal mannitol level in the vacuole buffer in terms of vacuole protection during centrifugation, whereas a Ficoll concentration of 0.10 g/ml was adopted in the density-gradient centrifugation.
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