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Isolation and regeneration of protoplasts from Penicillium digitatum

Published online by Cambridge University Press:  12 February 2007

Song Ai-Huan ,
Li Hong-Ye  and
Liu Xiao-Hong
Song Ai-Huan
Affiliation:
Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou,Zhejiang 310029, China Shandong Marine Culture Institute, Qingdao, Shandong 266002, China
Li Hong-Ye*
Affiliation:
Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou,Zhejiang 310029, China
Liu Xiao-Hong
Affiliation:
Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou,Zhejiang 310029, China
*
*Corresponding author: E-mail: hyli@zju.edu.cn
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Abstract

The effects of some factors on the isolation of protoplasts from Penicillium digitatum were studied, including the appropriate material (young mycelia and generating spores), the concentrations of enzyme and osmotic pressure stabilizers, reaction time and reaction temperature. Results demonstrated that germinating spores were an ideal material resource for the isolation of P. digitatum protoplasts. Highest yield and quality of P. digitatum protoplasts were obtained by shaking germinating spores suspended in a solution of 10 mg/ml Lywallzyme™ dissolved in 0.7 M NaCl as osmotic pressure stabilizer at 80 rev/min and 30°C for 3.0–3.5 h. The regeneration rate of the isolated protoplasts was as high as 24.9% on double-layer Czapek medium containing 0.7 M NaCl. Additionally, observation of the protoplast release pattern showed that the protoplasts of P. digitatum were released primarily from the hyphal apex and occasionally from the subapical or original sites of germinating tubes. The protoplasts of P. digitatum were regenerated in a direct manner of either yeast-like cell development or mycelium formation.

Keywords

Penicillium digitatumprotoplast isolationprotoplast regeneration
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 1 , Issue 3 , December 2004 , pp. 197 - 202
Copyright
Copyright © China Agricultural University and Cambridge University Press 2004

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