I am concerned about the effect that freezing v. refrigeration will have on soil samples that i will be analyzing for EMF % colonization using a gridline intersect method. my university lab has limited space to store 40+ samples in the refrigerator. these samples will be tested for pH as well but only after thawing. If anyone has thoughts regarding how freezing might affect the fine roots and hartig nets of EMF roots, i would appreciate some insight.
Why should you freeze the soil sample for the study of root colonization? If you are interested in spore density studies, better you keep the soil air dried and preserve. There is no need of keeping soil in a freezer for the study of EMF (better use the term AMF).
Please follow standard procedures for root colonization study.
For the study of root colonization, it is always better to use the fresh root samples of specific plants collected from the field itself. Preservation of roots, if is to be done, it is usually done in 10% KOH. But soft roots will be liquefied if kept in KOH for a long time.
Maceration of the fresh or preserved roots are done by boiling (two minutes) in 10% KOH and afterwards in 1% HCl (1 minute); then stain in Trypan Blue stain prepared in Lactophenol. After proper staining, the root colonization can be easily observed under a microscope.
Thank you for the advice. I was able to secure space in the refrigerator. The procedure for EMF (Ectomycorrhizae) does not require staining as it is a visual inspection using gridline intersect methods adapted from AMF studies (Newman, 1966; Giovanetti & Mosse,1980). There was a concern with desiccation of the hartig net and fine root tips without refrigeration. Again, this is a study of Ectomycorrhizae not Arbuscular mycorrhizae.
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