2011 Inoculation of WBP Seeds with Spores of Native Ectomycorrhizal Fungi

Project: Inoculation of whitebark pine seeds with spores of native ectomycorrhizal fungi

AttachmentsCripps_2011_Lonergan paper        Cripps- 2011WBPprogressRPT final

Agency/Forest or Park/District: Montana State University

Project coordinator: Dr. Cathy L. Cripps

Contact: Dr. Cathy L. Cripps, Plant Sciences & Plant Pathology Dept., Montana

State University, Bozeman, MT 59717, 406-994-5226, ccripps@montana.edu


John Schwandt and Holly Kearns, Kent Eggleston, USDA FS. We are also doing applied research with Parks Canada and Glacier National Park which may benefit from this proposed project.

Source of funding /amount

FHP: $12,600

Supplemental funding: $3,500 in kind one month Cripps salary; $700 Greenhouse rental and supplies donated by MSU and Parks Canada via another project focusing on limber pine.

Dates of restoration efforts

January-August 2012 with an official extension until May 2013.


The primary objective of the current project is to examine and develop various methods for the application of ectomycorrhizal basidiospores (spore powder, spray slurry, chips) to whitebark pine seeds that are to be directly planted in the fieldl [not nursery inoculation of seedlings]. The goal is enhanced germination, enhanced survival and/or enhanced/timely mycorrhizal colonization of germinants inoculated with native ectomycorrhizal fungi specific to 5-needle pines. A second objective is to continue to develop various types of inoculum from native ectomycorrhizal fungi to be available for use in the various whitebark pine projects of other researchers, land managers and greenhouse personnel.

Acres/ha treated  N/A


The individual objectives of this project are to: 1) Evaluate effectiveness of various methods for encapsulation of whitebark pine seeds with spores of native ectomycorrhizal fungi specific for 5 needle pines, and 2) evaluate effectiveness of various chip formulations (spores of native ectomycorrhizal fungi and hydromulch) added to whitebark pine seeds at planting. Trials will take place in the MSU Plant Growth center under typical greenhouse conditions; prime considerations are: soil type, soil pH, spore concentration levels, adhesion and release of spores, and maintaining minimal depth of spore coating.

General Methods:

1. Whitebark pine seeds will be warm stratified (Schwandt et al. 2007; Burr et al. 2001).

2. Seeds will be encapsulated with basidiospores using: dry powder, spray slurry dried on, or other formulations (Marx et al. 1984, Theodorou & Bowen 1973, 1983). Concentration of spore levels to be tested will be around 1 million spores/seed.

3. Chips containing various concentrations of spores (around 1 M/ml) will be added to uninoculated seeds at planting. Various formulations will be tested (Beckjord et al. 1984, 1986, Marx et al. 1979).

4. Un-inoculated seeds will be used as controls and planted without addition of spores.

5. Seeds will be germinated under greenhouse conditions in a previously selected media type (Cripps & Grimme 2011).

6. Assessment will be for % germination, % mycorrhizal colonization and % survival at time intervals. Graphing or statistical analysis will be used as applicable (Cripps & Grimme 2011).

7. Effective methods will be selected for further examination and use with whitebark pine seeds to be out-planted in the field (as in Schwandt et al. 2007).

8. Continue to develop various types of inoculum to be available for whitebark pine projects.

Planting? If so, source of seedlings? Resistance?

No, but the source of our seedlings for greenhouse experiences are putatively rust resistant seedlings from the USDA FS Coeur D’Alene Nursery, Idaho.

Outcome Summary

We have several ongoing experiments for which we are waiting for final results and two completed experiments:

1. Development of Basidiospore chips: the goal of this research is to develop a ‘chip’ type of inoculum that can be added to whitebark pine seedlings or seeds at the time of out-planting.

Results showed a very low colonization and unacceptable mycorrhizal colonization rate using the basidiospore chips. This could be a result of the hydrocolloid component which might have desiccated or attached to spores. Alternatively, the dried inoculum might have been ineffective before being used in the basidiospore chips. This second hypothesis is currently being tested by using the spores without the other ‘chip’ components. If spores are otherwise viable, then the “basidiospore chip” line of research will be terminated.

2. Inoculation (using liquid slurry) of directly planted seeds after warm stratification and prior to vernalization. This experiment was intended to mimic the direct out-planting of warm stratified seeds in Fall. [This experiment also tests viability of spores used in basidiospore chip experiment] Results for this experiment are pending.

3. Large experiment to determine if seedlings already colonized with mycorrhizal fungi can then be subjected to a final fertilization treatment (before out-planting). The goal is to determine if the mycorrhizal condition is maintained after the fertilization treatment. This would allow for both mycorrhizal inoculation and fertilization before out-planting. Results for this experiment showed no effects for slurry type or inoculation method so data was pooled for the graph shown below to highlight the effects of fertilization and container length on mycorrhizal colonization of seedlings.

Monitoring since completion of the project

Monitoring does not apply to our greenhouse experiments however this research has led to other projects funded by other sources. Our inoculated seedlings were used in a large project in the International Peace Park, and inoculated seedlings were planted on a variety of site conditions in Glacier and Waterton National Parks. These seedlings were monitored in 2010 and 2012, and results from this project are in an upcoming paper to be published in Forest Science.

Will outcome meet goals? The outcomes do meet our overall goal of researching the potential of inoculation with native ectomycorrhizal fungi to benefit the survival of out-planted whitebark pine seedlings.  We have gained information on how to promote ectomycorrhizal colonization in the greenhouse with particular fertilizer regimes. We also used inoculated seedlings for the project at the Yellowstone Club.

Future actions/follow up?  We continue to research how native fungi can benefit nursery grown whitebark pine seedlings with greenhouse and field studies.

Miscellaneous comments: Results from our field project (not funded by WPRP) have shown the potential for inoculation of whitebark pine seedlings with native ectomycorrhizal fungi to enhance seedling survival on some types of sites (site dependent). We are awaiting analysis of further results from the out-plantings.