Help! making protoplasts

Issei Kobayashi issei at BIO.MIE-U.AC.JP
Fri Apr 14 02:12:58 EST 1995


Dear All,

We are tring to make protoplasts from Rhizoctonia solani AG2-2. We are using
standard procedure to make protoplast from other AGs (AG 1,4 and 5).
But, the efficiency is very low, so we could get very small no of parotoplasts
in this moment. We are using Novozyme234 as wall degrading enzyme and NaCl or
mannitol for stabilizer. We actually got huge numbers of protoplasts from 
AG4 with this method, but not from AG2-2. What is difference in cell wall 
between AG2-2 and Ag4?
If someone has experience to make protoplasts from AG 2-2 or some information
about differences in cell wall in different AGs, please let me know.

Thanks in advance.



          Issei Kobayashi
          Lab of Plant Pathology, Fac of Bioresources,
          Mie University, Tsu city, Mie, 514, Japan
          TEL: 0592-32-1211(X3559)
          [International: +81-592-32-1211(X3559)]
          FAX: 0592-31-9634
          [International: +81-592-31-9634]
          Email: issei at bio.mie-u.ac.jp



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