Location: Crop Germplasm Research
Project Number: 6202-21000-032-06
Start Date: Jul 01, 2009
End Date: Jun 30, 2012
One–year-old woody seedlings (1-0 nursery stock or its equivalent) of J. californica, J. nigra, J. regia, J. ailantifolia, J. cinerea, J. major, J. hindsii, J. microcarpa, Paradox rootstocks, C. illinoinensis, C. aquatic, C. ovata, and C. texana approximately 0.5-1 cm diameter at ground level will be used for pathogenicity tests. J. major is included in the study as a control, since it is a host for P. juglandis but is not usually killed by Geosmithia. Dormant trees will be planted in 3.8-liter pots in a commercial nursery mix and placed in a greenhouse at Colorado State University. Inoculations will be made after the trees resume growth and leaves have fully emerged. Two isolates each of Geosmithia will be grown for 10 days on 1/2 strength PDA. Inoculations will be made by slicing down through the bark with a sterile scalpel at three sites on each stem. Resulting wounds are approximately 0.5-1.0 cm wide and 1 cm long with the flap of bark still attached to the stem at the base of the wound. A plug of sterile 1/2 strength PDA approximately 0.5 cm2 will be inserted under the bark flap and against the wood on the middle wound on each tree. An agar plug of similar size but colonized by one of the fungal isolates will then be inserted under the bark flap on the top and bottom wound. Four to 18 trees of each taxa will be inoculated with each isolate. All wounds will be sealed with Parafilm(R) and the trees will be randomly placed on a greenhouse bench. The Parafilm(R) will be removed after 3 wk. After 8 wk, all trees will be harvested and the outer bark shaved from the wounds with a sterile scalpel to expose the extent of bark colonization. The length and area of discolored tissue will be recorded and compared to cankers on the susceptible J. nigra and the control species J. major. If there are space, labor, or time constraints in the completion of the research, the cooperator will consult with the chair of the Juglans CGC and the other PIs to determine how many trees of each species to inoculate and evaluate. The goal will be to obtain clean, replicated data from as many taxa as possible, with priority given to those genotypes currently in NCGR repositories.