Brief Summary of Minutes of Annual Meeting

The business meeting was conducted by Chair Sandra Anagnostakis.

1. John Anderson, NE-140 Administrative Advisor was present and reminded the group that the current project terminates September 30, 2003.
   
2. A committee was formed to begin rewriting the new proposal that will be due for review October, 2002, with a letter of intent given to John Anderson by February 2002. Anderson reminded committee members that the renewal re-write should include these major objectives:
   
   
   
   1. The need for research as indicated by stakeholders.
      
   2. The need to include cutting edge research.
      
   3. Importance of the work, and what the consequences will be if the work is not done.
      
   4. The technical feasibility of the work.
      
   5. Advantages of doing the work as a multi-state effort.
      
   6. What the likely impacts will be from successfully completing the work.
      
   7. How cooperation between participating groups enhances the overall research.
   
   The project should not exceed 15 pages of text, excluding bibliography. Committee members include Michael Gold, Bradley Hillman, Bill MacDonald, Fred Hebard, and Don Nuss. The first task for the committee will be to write a request to write a proposal. This request is not to exceed 4,000 characters. It should be done by February and sent to John Anderson.
   
3. NE-140 now has a web page completed by Ruby Meiz. The URL is: www.agnr.umd.edu/userforms/nera/projects Ruby has set the home page up to receive all project submissions, reviews etc.
   
4. Bill MacDonald reported on the deaths of two former NE-140 members, Dale F. Hindal from West Virginia University and John E. Elliston (Connecticut Agricultural Experiment Station).
   

1. To better understand the interactions and ecology of the host/pathogen/parasite system at the molecular, organismal and environmental levels in order to develop effective biological controls for chestnut blight.
   
   
   
   1. McKeen reported that hypovirulence in Ontario is not prolonging the life of chestnut trees due to the use of a hypovirulence type that is not aggressive, and timing of its application.
      
   2. Fulbright has continued work on characterizing the nature of the hypovirulence caused by plasmid DNA in the mitochondria of C. parasitica Some mutants exhibit reduced or abnormal growth and reduced production of conidia. Others enhance senescence and reduce sporulation or attenuate virulence. Pathogenicity tests are underway to explore the potential of 2 specific mutants, a mitochondrial insertion, InC9, and a plasmid residing in the mitochondria, pCRY1.
      
   3. Median-Mora sampled cankers on American chestnuts in Hamden, CT, Rockford, MI, and West Salem, WI and tested C. parasitica isolates for the mt insertion InC9 and the mt plasmid, pCRY1. She reported a high incidence InC9 and pCRY1 in the CT isolates. The plasmid pCRY1 has not been found at either Rockford or West Salem, but the prevalence of InC9 at Rockford, MI leads her to believe it may have originated there.
      
   4. Tattar has treated killing chestnut blight cankers with slurry mixtures containing spores of Bacillus megaterium and soil. Many trees are still surviving, but the success rate is not 100%.
      
   5. Hillman has reported that the virus SR2 may be the most common Cryphonectria hypovirus species in North America. It is unique, but is most closely related to the GH2 type dsRNA, CHV3. It is present in a much lower titer than other hypoviruses, and has little effect on pathogen virulence. He also reported on Crypt2, a class III transposon found in all C. parasitica isolates examined so far. He has also found that C. nitschkei occupies the same habitat as C. parasitica and contains the same genetic elements.
      
   6. Powells group has developed American chestnut stem-expression sequence tags. Fifty-five have been sequenced and sent to GeneBank. They have developed a tissue culture system for American chestnut as well as transformation protocols, and field testing of non-transgenic somatic embryo derived plantletts has begun.
      
   7. Hebard reported that the AFLP mapping has been completed. Working with support from a TACF grant, Steve Rogstad at the University of Cincinnati has developed a new method for extraction of DNA from chestnut and is working with mini satellite DNA. Based on Hebards continuing observation of naturally occurring hypovirulence, he believes that where disease remission is occurring in the U.S., it may ultimately reach European levels.
      
   8. Anagnostakis reported on a CT chestnut plot treated between 1983-1986 with Ep747, a CHV1 containing C. parasitica strain. Many of the larger chestnuts in the plot have died as a result of a drought in 1999-2000 but many remaining trees are infected with blight strains similar to the original Ep747.
      
   9. Nuss reported that the CHV1-Ep713 transgenic strains used in the Meshomasic State Forest study are ecologically unfit. The cDNA derived hypovirus used at that site can be transmitted through ascospores, but has not spread outside to the control plots. A new field trial is underway in West Virginia that will test more ecologically fit transgenic C. parasitica strains.
      
   10. Jarosz reported on the evaluation of C. parasitica and the effect of hypovirus introduced in plots in MI in 1992. The trees have been GPS mapped, but the fungus is so prevalent that it is impossible to sample every canker. New plots were established in 2001 to better assess the spread of the fungus and the hypovirus.
       
   11. MacDonald reminded the group that there will be a symposium at the West Salem, WI chestnut site during the APS meeting in July of 2002. TACF is reimbursing the owners of the land for preserving this site, where CHV1 continues to be the most common hypovirus, but CHV3 still persists. Hypovirus spreads most easily from infected cankers to uninfected on the same tree. New vegetative compatibility types have also been discovered.
       
   12. Double reported on the long-term dissemination studies in cleared and cut-over areas. Conclusions: hypovirus recovery was best from trees in non-cleared plots that received hypovirus inoculum. CHV3 reduced canker growth more than CHV1. Hypovirus was most easily recovered from the outer growth ring of cankers in April.
       
   13. Kazmierczak reported that deletion of the mf1-1 pheromone of C. parasitica results in male sterility. Expression studies show that the pheromone it is not expressed in conidia, but is enhanced under conditions of starvation. She also reported evidence that symptom expression of viral infection of C. parasitica is caused by perturbation of protein secretion.
       
   
   
2. To improve chestnut trees for timber and nut production, and determine the cultural requirements of chestnut seedlings in nursery and natural settings.
   
   1. Gold has determined that Willamette?and Qing?are good cultivars for commercial chestnut production, producing good size nuts with an early ripening date. A new orchard was established in MO in October 2001 to examine pruning treatments, irrigation and fertilizer regimes, and tolerance to blight in 3 cultivars: ‘Peach,‘ ‘Qing,‘ and ‘Willamette.‘
      
   2. Maddox reported that tillage had no significant effect on American chestnut tree survival, tree height, or root length after four years. Using cover crops, survival and growth of trees was better with crimson clover than alfalfa. Yellow sweet clover grown as a cover crop reduced survival and growth. Nursery source did not generally effect survival after 3 years.
      
   3. Rhoades examined establishment of chestnut seedlings in clear-cuts, shelterwood harvested areas, and midstory sites with both mesic and xeric conditions. He found that insect herbivory was heaviest in the midstory. He also found that conditions for seedlings were best on high, light xeric sights and that ectomycorrhizae had little effect except on poor sandy soils.
      
   4. Carlson is working on American chestnut silviculture in conjunction with Tim Phelps and Kim Steiner. Their objective is to restore blight resistant American chestnuts to forest settings. He has developed an in situ hybridization protocol for selection against Chinese chestnut genetic backgrounds in the ACF backcross program.
      
   5. Dale reported that Canada is starting a chestnut restoration project that will assess the current situation in the Province of Ontario. They have begun a blight resistance breeding program, hoping to breed a blight resistant chestnut suitable for Canadas conditions.
      
   6. Gordon presented his thesis that the current research will do little to restore the American chestnut, as it will probably recover on its own. In order to hasten this process, he suggests that we institute widespread planting of American chestnut in riparian zones.
      
   7. Anagnostakis reported that BC2 and BC3 hybrids, American, and Japanese chestnuts were dug up, their heights and roots were measured, and the hybrids were planted in 2000 in two forest clear-cuts in central CT, and in a nursery. Two additional plots were established in 2001, in sites with a mature canopy. She reported good growth, even at a site where the soil pH is 3.6. A 6th plot will be established in 2002 in a partially harvested forest area. She has found no correlation between soil and bark manganese levels and blight resistance.
      

Impacts:
The group determined that it has accomplished one of its proposed milestones for the year 2001, namely new forests plantings in Connecticut, Tennessee, North Carolina, Pennsylvania, Illinois and New York. Underway, but not yet completed, is the characterization of putative genetically transformed American chestnut embryogenic cultures and the field evaluation of confirmed transgenic chestnut trees containing single-transgene constructs.

1. Anagnostakis, S.L. 2001. American chestnut sprout survival with biological control of the chestnut-blight fungus population. Forest Ecology and Management 152:225-233.
   
2. Connors, B.J., Maynard, C.A., and Powell, W.A. 2001. Expression sequence tags from stem tissue of American chestnut. Biotechnology Letters 23:1407-1411.
   
3. Dawe, A.L. and Nuss, D.L. 2001. Hypoviruses and chestnut blight: exploiting viruses to understand and modulate fungal pathogenesis. Annu. Rev. Genet. 35:1-29.
   
4. Groome, P.C., Tattar, T.A., and Mount, M.S. 2001. Bacillus megaterium: a potential biocontrol agent against chestnut blight. J. American Chestnut Foundation 15:45-49.
   
5. Linder-Basso, D., Foglia, R., Zhu, P., and Hillman, B.I. 2001. Crypt1, an active Ac-like transposon from the chestnut blight fungus, Cryphonectria parasitica. Molecular Genetics and Genomics. 265:730-738.
   
6. Rhoades, C.C. 2001. Pre-blight abundance of American chestnut in Kentucky. J. of the American Chestnut Foundation 15:36-44.
   
7. Van Heerden, S.W., Geletka, L.M., Preisig, O. Nuss, D.L., Wingfield, B.D., and Wingfield, M.J. 2001. Characterization of South African Cryphonectria cubensis isolates infected with a C. parasitica hypovirus. Phytopathology 91:628-632/.
   
8. Yuan, W. and Hillman, B.I., 2001. In vitro translational analysis of genomic, defective, and satellite RNAs of Cryphonectria hypovirus 3-GH2. Virology 28:117-123.