Brief Summary of Minutes of Annual Meeting

The meeting was called to order by Chairman Fulbright at 9:00 am on October 17, 2008 at Chimney Corners Resort in Frankfort, MI. Fulbright acknowledged the fact that the NE-1015 technical committee was honored with the 2008 Award for Excellence in Multistate Research, given by the Northeastern Regional Association of State Agricultural Experiment Station Directors. Acknowledgement was given to Michael Vayda, the projects administrative advisor, for submitting the project for the award. OBJECTIVE 1. To improve chestnut trees for reestablishment in forest ecosystems, and chestnut cultivars for nut production by selection, breeding, and marketing, and determine the cultural criteria of all chestnuts for successful production in nurseries, orchards, and/or natural settings. Dennis Fulbright, Michigan State University; Microbial decay of fresh chestnuts and its management in Michigan. Michigan has found chestnuts that have a long shelf life when compared to others. Swollen/superficial cankers. Fulbright showed chestnut stems with swollen/superficial cankers and he asked the group, what causes swellings such as these? Many in the group responded, hypovirus. The stems he showed were part of the irradiation study in Maryland planted in Michigan. There is still the disease triangle, the host, fungus and environment. The stems that he showed will die because they do not have sufficient resistance to keep them alive. The County Line site was sufficient to keep the trees alive. If we think of hypovirulence as just the reduction of aggressiveness, then we are not thinking of biocontrol. If we think about swollen cankers, we arent thinking about biocontrol. It is everything together that gives us biocontrol. Carmen Medina-Ferrer, Understanding pollination in Michigan orchards. A problem in Michigan chestnut orchards is low yields. Maximum yield potential will only be realized when all nuts in a bur are pollinated. Medinas goal is to improve the understanding of chestnut reproductive phenology, pollination, and fruit-set using genetic analysis of parents and progeny. DNA was collected from the 50 nuts from each cross; for the DNA fingerprinting, she used a universal primer (GTG)5 and the SSRs CsCAT1; CsCAT 16; EMCs15; EMCs3; and, ssrQrZAG96. Sandra Anagnostakis, Connecticut Agricultural Experiment Station. Back-crossed trees. Four families of back-crossed trees with resistance from Chinese chestnut (BC2s) and from Japanese chestnut (BC3s) were planted in three forest clearcuts and an old tobacco field. The four families of trees all had larger dbh in the field plantings than in the forest, but survival was better for all families in the forest sites. Nutrients in Chestnuts. Nuts from 10 crosses were sent to a commercial laboratory for nutrient analyses (one analysis was $100). Four pollen parents were crossed with Colossal in Michigan. Fatty acid contents in the nuts from the CT hybrid and from Colossal were different depending on which pollen parent was used. Michele R. Warmund, University of Missouri-ColumbiaFlowering Sequence. Primary burs (1o) are formed, in Missouri, by 30 June and secondary (2o) burs are formed by late July or early August. Primary burs are generally responsible for the crop of nuts. The disadvantages of 2o burs are: limb breakage; and, adverse effect on subsequent 1o bur production. Warmund commented that there are many chestnut growers in the Midwest, and a late spring frost may be common; thus, Midwestern chestnut growers may want to consider cultivars that have abundant 2o burs as they may be able to harvest a marketable crop. Increasing Chinese Chestnut Primary Nut Weight and Bur Production by Hand Removal of Secondary Burs. Removal of 2o burs enhanced 1o nut weight per shoot at harvest and the number of shoots bearing 1o burs in the subsequent growing season on Orrin trees. Characterization of Secondary Flowering and Fruiting of Willamette Chinese Chestnut Trees. In 2007 and 2008, the number of 2o burs produced and retained on Willamette trees were recorded at weekly intervals from 1 Aug. to 1 Nov. In 2007, as many as 30, 2o burs per shoot were produced from 26 July to 16 Sept. The greatest number were recorded on 16 Sept. Chip Budding of AU-Super Chinese Chestnut Scions on AU-Cropper and Qing Seedling Rootstocks. A study was conducted to ascertain the anatomical structure of grafted and non-grafted Qing and AU-Super Chinese chestnut stems, as well as the optimal time of chip budding AU-Super on Qing and AU-Cropper seedling rootstocks. Budding success increased from 15% on 21 July to 65% and 75% for Qing and AU-Cropper rootstocks, respectively, on 19 Sept. Whip and tongue grafts can give 95% success rate for grafts done in the spring. Pam Kazmierczak, UC-DavisChestnut inventory. She listed the grafted cultivars at the Foundation Plant Services/UC-Davis. These trees will be outplanted in 2009. These are original imports to California from 2004. Fred Hebard, The American Chestnut Foundation, MeadowviewChina trip. Preliminary conclusions from the trip were: Cryphonectria was confirmed by isolation from C. mollissima and C. henryi, and cankers or lesions that appeared to be blight were common. However, the trees were thriving despite the presence of the disease. C. mollissima in native forest habitats was somewhat more tree-like than regarded in the United States. Castanea henryi is the largest and most forest-tree-like of the three native chestnuts, but all became timber trees. C. seguinii was larger than is reported in the literature. Backcrossing. The backcross technique was first begun with American chestnut in 1989. Hebard commented that resistance is not holding up as well as he thought. Environmental factors can have a large effect on tree condition. In Chinese chestnut, the phelloderm becomes lignified. This facilitates the sloughing of cankers. Lignification of wound periderm phelloderm occurs in Chinese but not American chestnut. It could be an important component of blight resistance, reducing penetration of wound periderm and facilitating sloughing of cankers. We may not be selecting for it with our current screen for blight resistance. Paul Sisco, Southern Pests and Pathogens: Breeding for Resistance to Phytophthora cinnamomi. Joe James of the Carolinas Chapter and Steve Jeffers, Clemson, continue their experiment to determine which backcross families have resistance to P. cinnamomi, Conclusions from are as follows: The resistance from Chinese chestnut is controlled by more than one gene; The resistance genes are not completely dominant. F1s have high survival but their roots have lesions; It may be difficult to get American-looking hybrids that have resistance to both P. cinnamomi and C. parasitica. Southern Pests and Pathogens: Phosphites to protect seedlings against P. cinnamomi. Steve Barilovits of the Carolinas Chapter conducted a backyard experiment this summer to test the efficacy of various phosphite products to protect first-year chestnut seedlings against P. cinnamomi. Steve experimented with Aliette WDG (aluminum phosphite), Agri-Fos (potassium phosphite), and Prudent-44 (urea phosphite). His first-year results were that both Agri-Fos and Prudent-44 provided a good level of protection, but that Aliette, was relatively ineffective. The Asian Ambrosia beetle, Xylosandrus crassiusculus, has a wide host range and causes damage by introducing pathogenic fungi into the interior of the stem. Bob Paris, Establishment and growth of chestnut on reclaimed mine lands. The TACF and its cooperators have been planting American chestnut and backcross chestnut on reclained mined lands in four states (KY, IN, PA, and OH), since 2002. After two seasons of growth, chlorosis was evident on most seedlings. With the exception of nitrogen-fixing species, chestnut has the ability to grow as well as, or better than, other tree species over a four state area spanning the ARRI region. Leila Pinchot, University of Tennessee, KnoxvilleContainerized chestnut shadehouse study. The objective of this study was to study the effects of light on early growth and development on American chestnut seedlings germinated in 10-gallon spin out coated pots. Data on this experiment is forthcoming. Chestnut morphology study. The objective of this study is to compare the morphological traits of the ACCFs American, TACFs American, Chinese, F1, BC3F3, BC2F3 and BC1F1 and CAES Japanese, BC1F1 and BC2F1 chestnut seedlings in order to determine how closely later generations of backcross chestnuts resemble American chestnuts. Daniel Boone National Forest Silvicultural Study. The objective of this study was to test the effects of three silvicultural treatments on growth, survival and competitive ability of planted BC2F3 seedlings. The long-term goal is to provide forest managers with appropriate silvicultural prescriptions for American chestnut reintroduction once blight-resistant seedlings become available. Field testing of blight resistant B3F3 chestnuts. The objective of this study is to evaluate TACF BC3F3 chestnut material in the field including growth performance, survival, morphological characteristics and blight resistance in order to define critical silvicultural parameters for successful restoration. John Carlson, Pennsylvania State UniversityPA-TACF activities. The PA Chapter is continuing work on many already established initiatives. Investigations into the use of cytoplasmic male sterility (CMS), various silvicultural methods, biomass and coppicing plantings, and plantings of various generations of breeding material continued in full force.. One of the most exciting developments in Pennsylvania was the harvesting of the first B3F3 generation seed to be produced in the state. These seed were collected from Meadowview, TACF's main breeding farm in southwestern VA. The seed were distributed to two locations for testing purposes. One exciting find by a Chapter member in northwest Pennsylvania was an orchard that was originally established by Mr. Haun in the early 1980s. In 2007, a student from Grove City College took several initial measurements on the planting. Of an original planting of 500 trees, 250 still stand. The clumps appear to be doing quite well. These trees offer a chance to investigate the life of an American chestnut orchard faced with incredible amounts of blight infection. These trees also demonstrate the value of planting trees with little to no blight-resistance. Besides conserving local genetic reserves, these trees offer great reservoirs of seed production, are an attraction to wildlife, and offer easily accessible breeding material. Mike Marshall, Shippensburg UniversityInoculations of C. parasitica on various fruits and vegetables. Previous researchers have employed apples of several varieties to assess virulence of C. parasitica and C. cubensis isolates. Curiosity led to the inoculation of other fruits and vegetables. C. parasitica was able to grow on most of the inoculated materials, the potato tubers being a notable exception. The best growth and sporulation was seen on strawberries, pear, pepino melon, pomegranate, and avocado, in that order. OBJECTIVE 2. To evaluate and integrate multiple approaches for the biological control of the chestnut blight fungus and other pathogens and pests that threaten chestnut, by investigating host/pathogen/parasite relationships from the molecular to the ecological level. Sandra Anagnostakis, Connecticut Agricultural Experiment StationTransgenic study. An experiment with Don Nuss and co-workers to introduce genetically engineered hypovirulent strains of C. parasitica into a natural population of the fungus was started in 1997 in a clearcut in the Meshomasic State Forest. The dbh of all sprouts >2.5 cm was measured in the winter of 2007. The sprouts in treated clumps are surviving longer than sprouts in the control group. Pam Kazmierczak, UC-DavisCHV1 disrupts secretion of cryparin in C. parasitica. Cryparin is a cell surface class II hydophobin. In liquid shake culture, she has found cryparin to comprise 30% of the mRNA and up to 1% of the protein. Cryparin was chosen for study because it is very abundant. When cryparin protein was deleted, it was found to be essential for eruption of the fruiting bodies of C. parasitica through the outer bark layer of the chestnut tree. Because of cryparins abundance, both as mRNA and protein, Kazmierczak chose to follow it as a marker during secretion in the presence of CHV1. Cryparin is rapidly secreted into the culture fluid where it is bound to the cell wallthe virus-infected fungus is greatly reduced in cryparin protein accumulation. Don Nuss, University of Maryland Biotechnology InstituteCommunity Sequencing Program Update. The project title, Genome sequencing of the chestnut blight fungus C. parasitica was submitted in March, 2006 and approved June, 2006. Genomic DNA was prepared from C. parasitica strain, EP155. Expectations from JGI include the generation of approximately 400Mb of raw sequencing reads required for 8  10 X coverage of the estimated 40-Mb C. parasitica genome, sequence assembly and initial automated annotations. An 8.54x assembly of the C. parasitica v1.0 assembly was released in 2008 for initial annotation efforts. RNA silencing as an antiviral defense mechanism in fungi. Nuss posed the question: Does C. parasitica have antiviral defense mechanisms to counter hypovirus modulation of fungal-plant interaction? There are two mechanisms: (1) vegetative incompatibility on a population level is one method that prevents viral spread; and, (2) RNA silencing another method. In C. parasitica are two dicers that are very similar to the dicers in Neurospora, Dicer-1 and Dicer-2. They knocked them both out and there is no phenotype that they can identify in the absence of virus infection. There are five virus families that have been characterized that that replicate in Cryphonectria. They have very different replication mechanisms so they will be useful in looking at a large number of basic questions on how these viruses interact in the RNA silencing pathway. They found the first evidence for a role of RNA silencing in viral RNA recombination. Hypovirus papain-like protease p48 is required for initiation but not for maintenance of virus RNA propagation in the chestnut blight fungus C. parasitica. p48 is required for initiation of CHV1-EP713 RNA infection in C. parasitica. The ?p48 mutant virus can be rescued when p48 is provided in trans. Demonstrated by transfection, transformation and anastomosis; p48 is not required for hypovirulence, but does contribute to viral RNA accumulation levels; surprisingly, the ?p48 mutant virus retains replication competence in the apparent absence of p48 following transmission to wild-type C. parasitica and successive sub-culturing; Novel hit and run mechanism may involve p48-mediated conformation change, priming, or recruitment of other factors.Transcription factors. It is remarkable that two of three transcription factor genes that were identified as being hypovirus-responsive turned out to be required for three major biological processes that are altered as a result of hypovirus infection. Many transcription factors have been mapped and identified. Michael Milgroom, Cornell UniversityMilgroom limited his discussion to two topics: (1) Worldwide population genetic structure (with Cyril Dutech and Cécile Robin, INRA, Bordeaux, France; and, (2) biological control with hypovirulence in Macedonia (Kiril Sotirovski, University Ss. Kiril I Metodij). Alice Churchill, Characterization of a secondary metabolite pathway in C. parasitica. C. parasitica produces a family of orange and yellow aromatic polyketide pigments (anthraquinones) that color the mycelium and spores of the fungus when grown both in vitro and in planta. Production of this family of pigments is suppressed in strains of C. parasitica that contain European hypovirus. They cloned, sequenced, and disrupted a C. parasitica polyketide synthase gene PKS1 by Agrobacterium tumefaciens-mediated transformation (ATMT) and began to characterize several targeted gene knockout (KO) strains that resulted from these experiments. Analyses of genome sequencing data confirmed that PKS1 is a single copy gene. In combination with our gene disruption data and gene expression analyses, the results to date suggest that PKS1 does not encode an anthraquinone synthase. Another notable observation was that multiple strains of Agrobacterium tumefaciens appear to have a nonspecific effect on virulence of C. parasitica transformants in chestnut stems and reduce aerial mycelium production in vitro in transformants created by ATMT. Bradley Hillman, Rutgers Universityreported the complete characterizations of two C. parasitica viruses and their genome sequences: RCI (Michigan); and, JS13 (Japan); Genomic and proteomic analysis of interactions between Lysobacter enzymogenes and C. parasitica and between two reoviruses and C. parasitica; Examination of interactions between C. parasitica hypoviruses and reoviruses. Jo Anne Crouch, DNA transposons in the C. parasitica EP155 genome. Crouch hypothesizes that there are four species of DNA transposons in C. parasitica strain EP155, Crypt1, Crpyt2, Crpyt3 and Crpyt4. Crypt1 is part of the haT super family of transposons, while the other three species are from the Tc1/mariner (aka fot1-like) family DNA transposons. Crypt1 was the first element identified in C. parasitica a few years ago. It is a Class II DNA transposon of the hAT family. Crypt1 was identified as moderately repetitive probe by Milgroom in the early 1990s. It has 8-20 copies in most C. parasitica isolates and it is a 3.6 kb element. Crypt2 was the second element identified. It is also a Class II DNA transposon but of the Tc1/mariner family. With regard to the populations of Crypt2, Crypt3 and Crypt4 in the EP155 genome, Crypt4 is likely an extinct species. John Carlson, Pennsylvania State UniversityGenomic Tool Development for the Fagaceae. The goal is to identify the genes in the Chinese chestnut genome for blight resistance. The bulk of sequencing was conducted on Chinese and American chestnut cDNA libraries. In comparing the transcriptomes of Chinese chestnut and American chestnut cankers versus healthy stem tissues the following was found: The transcriptomes of American and Chinese chestnut are highly similar in gene function and diversity; The 454 cDNA sequences from chestnut cover all biological and molecular functions.; Genes involved in response to biotic and abiotic stress/stimuli are perhaps more highly expressed in Chinese chestnut than in American chestnut. Linkage mapping studies. Deitrick conducted a small linkage mapping study with 17 RAPD markers that were previously identified by bulk segregant analysis (BSA) as differentiating pooled DNA from blight-resistant versus blight-sensitive trees. Eight of the markers were associated with blight sensitivity and 9 with resistance DNA pools. Deitrick was able to map six markers using segregation analyses of in the F2 family in which the genetic linkage map and QTL were mapped previously. None of the new loci appeared to be closely linked to the QTL loci on the LG A and LG F linkage groups. However, the marker orders differed from the original F2 map. This may have been caused by use of a smaller data set, and a different mapping program. Tom Kubisiak, USDA-Forest Service Southern Institute of Forest Genetics, Gulfport, MSDNA marker (SSRs) development for various members of the Fagaceae. Kubisiak summarized the anticipated goals and presented the why behind using genetic marker mapping in Chinese chestnut. He also presented some of the work being done to help finish the C. parasitica genome. Genetic Mapping of Chinese Chestnut. A total of 16 SSR primer pairs developed from project 454-based sequence data were chosen for parentage analysis. Based on this data only one Vanuxem x Nanking progeny is definitively a pollen contaminant and only 5 of the Mahogany x Nanking are possibly pollen contaminants. Only five of the Vanuxem x Nanking progeny are dead and only four of the Mahogany x Nanking progeny are dead. Genome Sequence-Based SSRs for C. parasitica. The goals of the SSR marker work were to: identify additional SSR markers in C. parasitica using the full genome sequence; design primers and identify polymorphic SSR markers near the ends of physical scaffolds; use recombinational data to suggest genetic linkage between/among markers on different physical scaffolds to infer scaffolds that are likely to reside on the same chromosome; identify additional SSR markers in C. parasitica using the full genome sequence; design primers and identify polymorphic SSR markers near the ends of physical scaffolds; use recombinational data to suggest genetic linkage between/among markers on different physical scaffolds to infer scaffolds that are likely to reside on the same chromosome. A total of 141 primer pairs were designed from SSRs as close to the ends of scaffolds as possible. Where previously mapped markers were found to be more terminal then no primer pairs were developed. All 141 primer pairs were screened on the parents (JA17 and X17.8) of the mapping cross to identify segregating markers. Paul Sisco, The American Chestnut Foundation, Southern Appalachian Regional Science CoordinatorCytological comparison of the chromosome structure of C. dentata and C. mollissima. There might be a reciprocal translocation because there are 11 linkage groups. Sisco showed slides that indicated reciprocal translocation between American and Chinese chestnut. Laura Georgi has developed BACs from most of the linkage groups, and she has a good DNA marker for one of the linkage groups that may be involved in translocation based on their mapping. Faridi is attempting to align the physical map with the genetic map. Loci on both maps are in the same order but they are not the same distance. Evidence for greater diversity of American chestnut in the South. It is clear from their studies that chestnut and chinkapin have interbred in the past. Mark Double, West Virginia UniversityIntroduction of hypoviruses at West Salem, WI. Hypovirus introduction was conducted from 1992-1997. Beginning in 2003, hypoviruses were reintroduced. In 2001, twelve permanent plots were established in three regions of the stand (Disease Center; Front and Beyond the Front). Hypoviruses are recovered most readily from treated cankers followed by non-treated cankers on treated trees. Hypoviruses have spread less effectively to non-treated trees. In June 2008, a study examined the spread of hypoviruses to non-inoculated American chestnut trees outside the permanent plots. Hypovirulent isolates were recovered from 82% of the trees on the perimeter of the Disease Center plots (an area with the longest treatment history), 20% from of the trees outside the Front plots; and 11% from perimeter trees in Beyond the Front plots (the most recently infected portion of the stand). The biological control potential of C. parasitica strains containing an infectious cDNA copy of the hypovirus CHV1-Euro7 .This study was designed to evaluate whether transgenic C. parasitica strains containing a cDNA transgene encoding the viral genome of CHV1-Euro7 show greater potential to biologically control chestnut blight than their cytoplasmically-infected counterparts. Tree mortality, as of July 2008, was greatest in the virulent plots (62%), followed by CH plots (40%) and TG plots (22%). William MacDonald, Infectivity of virulent and transgenic hypovirulent inocula of C. parasitica, and the influence of inoculation methodology on canker development. The objectives of this study were to: (1) compare the infectivity of TG and V ascospores, conidia, and a mixture of mycelia and conidia; and, (2) examine the effect(s) of strain, inoculum type, wound size, and the delivery medium on the development of artificially initiated cankers. Strain, inoculum, wound size were the most significant factors contributing to infection. For all inoculation methods, those performed with V inocula resulted in the greatest percent infection (55.6%) and were significantly greater than inoculations utilizing TG inocula (24.3%). Linear growth of cankers that resulted from infection was most significantly influenced by strain, delivery medium, and wound size. Strain, delivery medium, and wound size were the most important effects between or among their associated interactions. Saprophytic activity of C. parasitica on American Chestnut Trees. This was a study to determine the potential of hypovirulent strains of C. parasitica to grow saprophytically on chestnut stems and produce hypovirulent inoculum. There was no significant difference for growth of cankers when initiated on any of the four chestnut substrate treatments when inoculations were made in June, due to the high amount of variability. Susan Howard, Michigan State UniversityWest Salem. For hypovirus to work in a real biological setting, it must be able to move within the tree and within the forest. What is happening outside the twelve permanent plots that were established in the West Salem chestnut stand in 2001? Her findings: If a trunk is treated, sprouts are not necessarily protected. Euro-7 hypovirus has spread beyond expanded plots at West Salem. Swelling cankers don't necessarily have detectible hypovirus. Josh Springer, Matrix demography. Matrix demography suggests that 1-10 cm trees will respond best to hypovirulence. To test this prediction, he will introduce a local hypovirus, Grand Haven 2 (GH2) into three populations in Michigan. Trends in tree growth: a 12 year retrospective. Jarosz has conducted a census of six MI populations since 1996. Conclusions were: growth (and survivorship) of recovering populations is similar to or better than initially healthy or diseased populations; and successful vertical and horizontal transmission of hypovirulence is key to ecological recovery. Angus Dawe, New Mexico State UniversityIdentification of proteins altered in accumulation by hypovirus infection using 2D electrophoresis. Changes in protein production of hypovirus infected strains has been explored by separating total proteins on 17 cm isoelectric focusing strips (pH range 3-10) and then running these on large formal SDS-PAGE gels. In this manner, they identified important enzymes in glycolysis as being influenced by hypovirus infection. Thus, glycolysis is affected by hypovirus infection. A potential controlled expression system for C. parasitica. They would like to have a method to efficiently control ectopic gene expression. In order to develop a new tool for gene analysis in C. parasitica, they cloned a region upstream of a putative copper transporter. Based on observations in the fungus Cryptococcus neoformans, this sequence controls transcription of the transporter gene in the presence (off) or absence (on) of copper ions. Using the C. parasitica genome information, they created a reporter construct using GFP and assayed for the ability to control GFP production. Using these conditions, the production of GFP was reduced to near-zero when copper was in excess, but was highly expressed when copper was not available. Rachel Acuña. BDM-1 Phosphorylation and consequences of Casein Kinase 2 (CK2) inhibition. BDM-1 is a phosphoprotein that is required for G-protein signaling in C. parasitica and is functionally associated with the G² subunit CPGB-1. In 2008, 24 site-directed mutants affecting the putative targets for phosphorylation by casein kinase 2 (CK2) were made. Findings suggest that in fungi, phosducin-like proteins may be regulated differently than in mammalian systems in which CK2-mediation phosphorylation plays an important role. Hilary Boyer, Cis-regulator regions controlling expression in bdm-1 and cpgb-1. A reporter system based on GFP was designed to examine genomic regions required for the control of expression of these two genes. They now have six reporter constructs and validated expression controlled by regions of 1726, 1243 and 818 bp upstream of bdm-1. Boyer has generated five constructs to similarly analyze the putative promoter region of cpgb-1. She has identified one possible enhance region between 555 and 800 bp upstream of the start codon. Gloricelys Rivera, Identifying Circadian-Clock Genes in C. parasitica. She has successfully identified putative orthologues of light response genes in C. parasitica showing an average identity of 59% indicating a conservation between species. Her results suggested a response to light stimulus during growth of C. parasitica on solid medium. Via RT-PCR, she was able to demonstrate transcription of the genes wc-1, wc-2, frq, ccg-6, vvd, pkc and nop-1 in C. parasitica. Business MeetingSisco was elected Chair-elect. Hillman will chair the 2009 meeting, and agreed to host the meeting at the New Jersey shore in late September/early October. As of October 1, 2008, the project officially became NE-1033. Fulbright thanked MacDonald and Double for their work on the NE-1033 extension (2009-2013). At the IV International Chestnut Conference held in China in September 2008, the consensus of the conference attendees was that the V International Meeting, to be held in 2012, should be held in the U.S. When provided with options for a meeting site in the U.S . most of the attendees chose the Washington, DC metro area. Discussion centered on the National Conservation Training Center located in Shepherdstown, WV as a possible site. MacDonald and Nuss agreed to work on the meeting arrangements. Following the meeting, Fulbright and Jarosz provided a tour of two chestnut sites.

2004 Milestones Accomplished: " Nutritional analyses of nuts was conducted from orchard selections and cultivars of chestnut.(Anagnostakis) " Hypovirulent strains of C. parasitica were developed and deployed for blight control on native chestnut trees at each of three clear-cut forest areas and one nursery are planted with hybrid chestnut trees (Anagnostakis) " Site was selected for release of Euro 7 transgenic strains in West Virginia (MacDonald and Nuss) " Breeding program begun for selecting improved protein content, peelability, and blight resistance in high yielding orchard-based chestnut cultivars (FulbrightEndowed chestnut experiment station, Rogers Reserve Farm, MI). " Nuts collected for nutritional analyses and size comparison from orchard selections and cultivars of chestnut (Anagnostakis). " Hypovirulent strains of C. parasitica developed and deployed for blight control on native chestnut trees at each of three clear-cut forest areas and one nursery area planted with hybrid chestnut trees (Anagnostakis). " Scion wood collected from WI American chestnut trees surviving well with hypovirulence in the C. parasitica population, grafted onto root stocks in MI (Fulbright and Jarosz). " Site selected for release of new transgenic-hypovirulent strains of C. parasitica in WV (MacDonald and Nuss). 2005 Milestones Accomplished: " Market research analyses were completed and the findings reported (MO, MI). New selections of experimental lines from MO, CT, TN and MI are being established. (Gold) " Orchard of advanced backcross chestnut for assessment of host resistance with hypovirulence was established in WV, albeit the planting failed due to raccoon predation of nuts (MacDonald and Hebard) " Characterization of the role of hypovirus p29 in virus RNA accumulation in C. parasitica and virus transmission through conidia of the fungus (Nuss) " Generation of polyclonal antibodies against 5 overlapping regions of hypovirus ORF B and construction of a C. parasitica database (Nuss) " Evaluation and comparison of improved methods of hypovirus introduction into cankers under field conditions. Sequencing and genetic structure analyses of a gene cluster for anthraquinone pigment biosynthesis in C. parasitica completed, and targeted gene knockout strains constructed for the PKS gene and other genes to determine gene function, and studies conducted of pigment gene knockout mutants for possible phenotypic changes (Churchill). " More American chestnut scions collected in WI (as in 2004) and grafted on trees in MI (Fulbright and Jarosz). " Orchard established in WV with advanced, back-cross chestnut trees from VA for assessment of host resistance with hypovirulence in the C. parasitica population (MacDonald and Hebard). " Cryparin, the type II hydrophobin of Cryphonectria parasitica were shown to be essential for stromal pustule eruption through the bark of the host, Castanea dentate (VanAlfen). 2006 Milestones Accomplished: " Publication of a C. parasitica EST database containing approximately 2500 ESTs (Nuss) " Demonstration that hypovirus p29 suppresses RNA silencing in C. parasitica and in heterologous plant system, the first report of a mycovirus-encoded suppressor of RNA silencing (Nussnot proposed) " A proposal to sequence the C. parasitica genome was approved by the Department of Enegy Community Sequencing Program. A draft of the C. parasitica genome sequence should be available to the research community in 2007 (Nuss, Churchill, Milgroom). " A proposal to develop more genetic markers and a better genetic map for the Fagaceae was approved by the National Science Foundation's Plant Genome Research Program. The results of the four-year project will be posted at (Carlson, Sisco, Hebard, Anagnostakis) " Chestnut market analyses completed and findings reported (Gold) " New processed chestnut products introduced (Fulbright, Gold) " New chestnut cultivars established in several cooperating locations (Fulbright, Gold) " First transgenic American chestnut was outplanted (Powell). " Role of canker age and vegetative compatibility on the perpetuation of hypoviruses determined, following their introduction into forest chestnut trees (MacDonald). " Cloned Cpkk1 and Cpkk2, mitogen activated protein kinase kinases from C. parasitica. Raised polyclonal antibodies to Cpkk1 and Cpkk2. Cpkk1 found to be expressed under conditions of active mycelial growth. Cpkk1 was more abundant in the CHV1 infected strains, and persisted longer in the growth cycle of the fungus than it did in unifected strains (VanAlfen). " CHV1 protease p29 found to associate with vesicle membranes of C. parastica and behaves as an integral membrane protein of the vesicular fraction derived from the fungal trans-Golgi network (TGN). The viral dsRNA, helicase, polymerase and p29 copurify with the TGN, suggesting that the virus utilizes the fungal TGN for replication. Fractionation analysis of p29 deletion variants showed that sequences in the C terminal of the protein mediate membrane association. The C-terminal portion of the protein (Met-135-Gly-248) is sufficient for membrane association and is enough to direct p29 to the TGN vesicles in the absence of other viral elements (VanAlfen). 2007 Milestones Accomplished: " Demonstrated that RNA silencing serves as an antiviral defense mechanism in C. parasitica (first example for any fungus) against hypoviruses and mycoreoviruses (Nuss). " Orchard established in WV with advanced, back-cross chestnut trees from VA for assessment of host resistance with hypovirulence in the Cryphonectria parasitica populationreplanted in 2007 (MacDonald and Hebard). " Study to assess the saprophytic activity of Cryphonectria parasitica was initiated (MacDonald). " Study using two chinquapin species to look at resistance to chestnut blight and gall wasp resistance was initiated (Anagnostakis and Schlarbaum). " Treatments to produce high quality somatic embryos of American chestnut were determined (Merkle). " Study to estimate the number of American chestnuts in Ontario Canada was completed (Boland, Dale). " A breeding program, with the goal of having American chestnuts that are 75% Canadian origin in 20 years, was established (Dale). " A new chestnut cultivar, Labor Day, was developed for the cold midwest climate (Fulbright). " The father tree program was instituted utilizing pollen from chestnut in southern states (Craddock, Hebard and Paris). " Corn, tobacco and grass plots were established to assess the success of American chestnut when outplanted in areas other than forests (Paris). " Open pollinated progeny tests were established to reveal requirement necessary for reestablishment of American chestnut (Schlarbaum). " A tool for the rapid assessment of foliar nitrogen and for the diagnosis of nitrogen deficiency of Qing Chinese chestnut trees was identified (Gold, Warmund and Hunt). " DNA microarray analysis of hypovirus-mediated alterations of fungal gene expression initiated (Nuss). " Pigment knockout mutants of C. parasitica used to assess role(s) of pigments in fitness and competitive abilities in chestnut, and gene knockout strains for other PKS genes cloned (Churchill). " Hypoviruses found in native C. parasitica strains analyzed for evidence for spontaneous infection or long-distance transmission based on molecular sequence. " MI grafted (5-year-old) American chestnut selections from WI inoculated with C. parasitica to determine reaction to various strains (Fulbright and Jarosz). " Experiments initiated to assess roles of other C. parasitica PKS genes in virulence, fungal development, and chemical defense (VanAlfen). " Naturally occurring chestnut bark microorganisms analyzed for their antagonism toward C. parasitica. " Silencing of Kex2 found to significantly diminish the virulence of C. parasitica (VanAlfen). " A dwarf chestnut tree with unknown germplasm (Asian) was identified and grafted. It is male sterile, produces dwarf offspring but at an unknown ratio to normal offspring (Fulbright). " Hypovirulent strains were isolated from non-lethal cankers found at the Crystal Lake (Benzie County) site that treated with the GH2 hypovirulent strain in the 1980's (Fulbright). 2008 Milestones Accomplished: " The optimum time (19 Sept.) for budding Chinese chestnut was elucidated. Unpublished studies indicate that low tissue moisture content is related to successful graft union formation. Propagation of Chinese chestnut trees by grafting (whip and tongue method) in the spring resulted was successful (90% formed unions) (Warmund, Gold and Hunt). " Results from experiments demonstrated that removal of secondary burs enhanced 1o nut weight per shoot at harvest and the number of shoots bearing 1o burs in the subsequent growing season on Orrin trees. A marketable crop from 2o burs can be produced on Willamette trees after 1o flowers were eliminated by a spring freeze. Secondary flowering was evaluated on Chinese chestnut trees in the repository at the Horticulture and Agroforestry Research Center (HARC), New Franklin, MO ) (Warmund, Gold and Hunt). " Trees were propagated for a ten-year trial to evaluate three sources of dwarfing rootstocks for chestnut at HARC. " Chestnut production of 65 cultivars planted at HARC was evaluated ) (Warmund, Gold and Hunt). " In mid-Missouri there are two distinct adult Curculio sayi emergence periods, one occurring in the spring and the other in the fall. The latter period had never been reported before) (Warmund, Gold and Hunt). " Orchard established in WV with advanced, back-cross chestnut trees from VA for assessment of host resistance with hypovirulence in the Cryphonectria parasitica populationreplanted in 2008 (MacDonald, Double, Hebard and Paris). " Study to assess the saprophytic activity of Cryphonectria parasitica was completed (MacDonald and Double).

NE-1015 Publication List 2004-2008 Alexander, M.T., L.W. Worthen and J.H. Craddock. 2005. Conservation of Castanea dentata germplasm of the southeastern United States. III International Congress, Claves, Portugal, Oct. 20-23, 2004, Acta Hort (ISHS) 693:485-490. . Alexander, S., M. Alexander and J.H. Craddock. 2006. Host pathogen interactions in a segregating population of BC2F2 hybrid Castanea dentata after exposure to hypovirus-containing and hypovirus-free strains of Cryphonectria parasitica. In: Restoration of American Chestnut to Forest Lands: Proc. Of a Conference and Workshop. May 4-6, 2004. The North Carolina Arboretum., . Allen, T.D. and D.L. Nuss. 2004. Linkage between mitochondrial hypovirulence and viral hypovirulence in the chestnut blight fungus revealed by cDNA microarray analysis. Eukaryotic Cell 3:1227-1232. Allen, T.D. and D.L. Nuss. 2004. Specific and common alterations in host gene transcript accumulation following infection of the chestnut blight fungus by mild and severe hypovirus. J. Virol. 78:4145-4155. Allen, T.D., A.L. Dawe and D.L. Nuss. 2003. Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonectria parasitica to infection by virulence-attenuating hypovirus. Eurkaryotic Cell 2:1253-1265. Anagnostakis, S. L. 2007. Effect of shade on growth of seedling American chestnut trees. Northern Journal of Applied Forestry 24:317-318. Anagnostakis, S., J. Carlson, F.V.Hebard, T.L. Kubisiak, D. Nielsen, W. Powell, R. Sederof, P. Sisco,, C. Smith, J. Tomkins, and N. Wheeler. 2007. Genomic tool development for the Fagaceae. Abstract P505. Plant and Animal Genome XV, San Diego, CA, Jan. 13  17. Anagnostakis, S.L. 2005. Chestnuts in the United States for food and timber. Acta Hort. (ISHS) 693:41-46. . Anagnostakis, S.L. 2007. Effect of male parent on nut weight in chestnut. Norther Nut Growers Association. Nov. 7, 2007. . Anagnostakis, S.L. and P. Devin. 2004. Nutrients in chestnut. Journal of The American Chestnut Foundation 18(1):38-41. Andrade, G.M. and S.A. Merkle. 2005. Enhancement of American chestnut somatic seedling production. Plant Cell Reports 24:326-334. Andrade, G.M., C. J. Nairn, H.T. Le and S.A. Merkle. 2006. Transgenic American chestnut trees: a novel approach for the restoration of the species. In Plant Biology 2006 Program and Abstracts, August 5-9, 2006, Boston, MA. Abstract No. P46017. Biella S, M.L. Smith, J.R. Aist, P. Cortesi and M.G. Milgroom. 2002. Programmed cell death correlates with virus transmission in a filamentous fungus. Proc. Royal Soc. London, Series B-Biological Sciences 269 (1506): 2269-2276. (not included in previous report). Buck, K.W., R. Eestban and B.I. Hillman. 2005. Family Narnaviridae. In Virus Taxonomy: Eighth Report of the International Committee for the Taxonomy of Viruses. C.M. Fauquet et al., etds. Elsevier Academic Press, San Diego, CA. Carbone, I., Y.-C. Liu, B.I. Hillman and M.G. Milgroom. 2004. Recombination and migration of Cryphonectria hypovirus 1 as inferred from gene genealogies and the coalescent . Genetics 166:1611-1629. Carlson, J.C. 2006. Biological confinement of genetically engineered organisms. Pages 151-158 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Cernusca, M.M., M.A. Gold and L.D. Godsey. 2008. Influencing consumer awareness through the Missouri chestnut roast. Missouri Journal of Extension (in press). Churchill, A.C.L., Milgroom, M.G., and D.L. Nuss. 2007. Genomic sequencing of the chestnut blight fungus Cryphonectria parasitica. Joint Genome Institute User's Group Meeting, Walnut Creek, CA, March 28-30, 2007 (abstract). Cooper, W.R. and L.K. Rieske. 2007. Community associates of an exotic gallmaker, Dryocosmus kuriphilus, (Hymenoptera: Cynipidae) in eastern North America. Annals of the Entomological Society of America 100. 236-244. Cooper, W.R. and L.K. Rieske. 2007. Community associates of an exotic gallmaker, Dryocosmus kuriphilus, in eastern North America. Annals of the Entomol Soc America 100, 236-244. Cooper, W.R. and L.K. Rieske. 2008. Differential responses in American and Chinese chestnut to foliar application of jasmonic acid. Chemoecology 18, 121-127. Cooper, W.R., and L.K. Rieske. Jasmonic acid treatment on chestnut benefits a gall forming wasp, Dryocosmus kuriphilus. Entomological Society of America National Meeting, San Diego, CA. December 2007. Craddock, J.H. 2006. Chestnut breeding in the United States. Pages 109-128 in: Tree Breeding Researchers: for the next half century. Proc. International Symposium for the 50th Anniversary of KFGR and the 20th Anniversary of the late Dr. S.K. Hyun. June 15-16, 2006, Forest Seed Research Center, Suanbo, Korea. KFRI. Craddock, J.T., M.T. Alexander, S.H. Alexander, J.L. Bramblett and L.M. Worthen. 2005. Chestnut cultivar evaluations in Tennessee: orchard establishment and early germplasm characterization, III International Congress, Claves, Portugal, Oct. 20-23, 2004, Acta Hort (ISHS) 693:465-470, . Davelos, A.L. and A.M. Jarosz. 2004. Demography of American chestnut populations: efforts of pathogen and a hyperparasite. J. Ecology 92:675-685. Davis, D. 2006. Historical significance of American chestnut to Appalachian culture and ecology. Pages 53-60 in: Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Davis, J.E., T.L. Kubisiak and M.G. Milgroom. 2005. Polymorphic sequence-characterized codominant loci in the chestnut blight fungus, Cryphonectria parasitica. Mol. Ecol. Notes 5 (2):195-197. Dawe, A.L., G.C. Segers, T.D. Allen, V. C. McMains and D.L. Nuss. 2004. Microarray analysis of Cryphonectria parasitica G±- and G²³-signaling pathways reveals extensive modulation by hypovirus infection. Microbiology 150:4033-4043. Dawe, A.L., V.C. McMains, M. Panglao, S. Kasahara, B. Chen and D.L. Nuss. 2003. An ordered collection of expressed sequences from Cryphonectria parasitica and evidence of genomic microsynteny with Neurospora crassa and Magnaporthe grisea. Microbiology 149:2373-2384. Deng, Fuyou and D.L. Nuss. 2008. Hypovirus papain-like protease p48 is required for initiation but not for maintenance of virus RNA propagation in the chestnut blight fungus Cryphonectria parasitica. J. Virology 82:6369-6378. Deng, Fuyou, T.D. Allen and D.L. Nuss. 2007. Ste12 transcription factor homologue CpST12 is down-regulated by hypovirus infection and required for virulence and female fertility of the chestnut blight fungus, Cryphonectria parasitica. Eukaryotic Cell 6:235-244. Deng, Fuyou, T.D. Allen, B.I. Hillman and D.L. Nuss. 2007. Comparative analysis of alterations in host phenotype and transcript accumulation following hypovirus and mycoreovirus infections of the chestnut blight fungus, Cryphonectria parasitica. Eukaryotic Cell 6: 1286-1298. Diskin, M. and K.C. Steiner. 2006. Rate of recovery of the American chestnut phenotype through backcross breeding of hybrid trees. Pages 129-131 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Enderton, D.J. 2007. Increasing Chinese primary nut weight and bur production by hand removal of secondary burs. M.S. Thesis, University of Missouri-Columbia. 30 pp. Finley, J. and K.C. Steiner. 2006. Summary of facilitated workshop of restoration of American chestnut to National Park system lands. Pages 227-230 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Fulbright, D. W. 2007. Hypovirulence of chestnut blight. In: Mighty Giants An American Chestnut Anthology, C. Bolgiano and G. Novak, editors. Pages 189  200. American Chestnut Foundation, Bennington Vermont. Fulbright, D. W., J.Jacobs, S. Stadt, M. Catal, A.M. Vettraino, and A. Vannini. 2006 . Phytophthora root rot of Abies and Pinus in Michigan. In Progress in Research on Phytophthora Diseases of Forest Trees (Brasier C., Jung T. and Osswald W., eds) pp. 104-108. Forest Research, Farnham, UK. Gold, M.A. and M. Hall (eds). 2007. The Chestnut Grower. Newsletter of the Chestnut Growers of America. Vol. 8, No. 3-4, Summer, Fall 2007. Gold, M.A. and M. Hall (eds). 2008. The Chestnut Grower. Newsletter of the Chestnut Growers of America. Vol. 10, No. 1-4, Winter, Spring,Summer, Fall 2008. Gold, M.A. and R. McCoy (eds). 2004. The Western Chestnut. Newsletter of the Chestnut Growers of America. Vol. 6 No. 4, Fall 2004. Gold, M.A. and R. McCoy (eds). 2005. The Chestnut Grower. Newsletter of the Chestnut Growers of America. Vol. 7 No. 1-4, Winter, Spring, Spring, Summer, Fall 2005. Gold, M.A. and R. McCoy (eds). 2006. The Chestnut Grower. Newsletter of the Chestnut Growers of America. Vol. 8, No. 1-4, Winter, Spring, Spring, Summer, Fall 2006. Gold, M.A. and R. McCoy (eds). 2007. The Chestnut Grower. Newsletter of the Chestnut Growers of America. Vol. 9, No. 1-2, Winter, Spring 2007. Gold, M.A., M.M. Cernusca and L.D. Godsey. 2004. Comparing consumer preferences for chestnuts with Eastern black walnuts and pecans. HortTechnology 14:583-589. Gold, M.A., M.M. Cernusca and L.D. Godsey. 2005. Chestnut market analysis producers perspective. UMCA Publication. Gold, M.A., M.M. Cernusca and L.D. Godsey. 2005. Competitive market analysis chestnut producers. In: Brooks, K.N. and P.F. Ffolliott (eds.) Moving agroforestry into the mainstream. The 9th North American Agroforestry Conference Proceedings, June 12-15, 2005, St. Paul, MN Department of Forest Resources, University of Minnesota, St. Paul, MN 16 p. Gold. M.A., M.M. Cernusca and L.D. Godsey. 2005. Update on consumers preferences for chestnuts. HortTechnology 15: 904-906. Gold. M.A., M.M. Cernusca and L.D. Godsey. 2006. Competitive market analysis: chestnut producers. HortTechnology 16:360-369. Golinski, M.R., W. J. Boecklen and A.L. Dawe. 2008. Two-dimentional fractal growth properties of the filamentous fungus Cryphonectria parasitica: the effects of hypovirus infection. J. Basic Microbiol. 48:426-429. Griffin, G. J. Elkins, D. McCurdy and L. Griffin. 2006. Integrated use of resistance, hypovirulence and forest management to control blight on American chestnut. Pages 97-107 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Haiyan Lin, Xiuwan Lan, Hong Liao, T.B. Parsley, D.L.Nuss, and Baoshan Chen. 2007. Genome Sequence, Full-Length Infectious cDNA Clone, and Mapping of Viral Double-Stranded RNA Accumulation Determinant of Hypovirus CHV1-EP721. Journal of Virology 81:1813-1820. Hebard, F.V. 2004. Notes from Meadowview research farms 2003-2004. Journal of The American Chestnut Foundation 18(2):38-45. Hebard, F.V. 2004. Research objectives of the American Chestnut Foundation, 2004-2014-Summary of TACFs 10-year plan. Journal of The American Chestnut Foundation 18 (2):13-19. Hebard, F.V. 2005. The backcross breeding program of the American Chestnut Foundation. J. Am. Chest. Found. 19:55-78. Hebard, F.V. 2006. The backcross breeding program of The American Chestnut Foundation. Pages 61-77 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Hebard, Frederick V., Mark L. Double and William L. MacDonald. 2007 A Pathogen without Rival. In: Mighty Giants: An American Chestnut Anthology, Pages 171-177. C. Bolgiano and G. Novak, eds. American Chestnut Foundation, Bennington, VT. Hebard. F.V. 2006. Notes from Meadowview 2005-2006. J. Am. Chestnut Found. 10:18-25. Hillman, B. I. 2004. Plant dsRNA Virus Diseases. in: Encyclopedia of Plant and Crop Science. R. M. Goodman, Ed., Marcell Dekker, Inc., New York. Hillman, B. I. and N. Suzuki. 2004. Viruses of Cryphonectria parasitica. Advances in Virus Research 63:423-472. Hillman, B.I. 2008. Mycoreoviruses in Encyclopedia of Virology. 3rd Edition, Elsevier Press, NY. Hillman, B.I., S. Supyani,, H. Kondo and N. Suzuki. 2004. A reovirus of the fungus Cryphonectria parasitica that is infectious as particles and related to the Coltivirus genus of animal pathogens. J. Virology 78:892-898. Hunt, K. L., M. A. Gold, and M. R. Warmund. 2008. Chinese chestnut cultivar performance in Missouri. Annual. Rpt. Northern Nut rowers Assn. 95: (in press). Hunt, K. M.A. Gold, W. Reid and M. Warmund. 2005. Growing Chinese chestnuts in Missouri. Agroforestry in action. Univ. of Missouri Center for Agroforestry, 23 Jan. 2006. . Hunt, K.L., M.A. Gold and M.R. Warmund. 2005. Chinese chestnut cultivar performance in Missouri. III International Congress, Claves, Portugal, Oct. 20-23, 2004, Acta Hort. (ISHS) 693:145-148, . Islam-Faridi, N., C.D. Nelson, B. Harshawardhan, M. Abdul Majid, T.L. Kubisiak, F.V. Hebard, P.H. Sisco, R.L. Paris, and R. L. Phillips. 2008. Cytogenetic analysis of a reciprocal translocation in F1 hybrid between American and Chinese chestnuts. Abstract W341. Plant and Animal Genome XVI, San Diego, CA, Jan. 12  16. Islam-Faridi, N., D. Nelson, P. Sisco, T. Kubisiak, F. Hebard, R. Paris, and R. Phillips. 2008. Chestnut cytogenetics: physical location of ribosomal DNA and telomere repeat sites, and reciprocal translocation in chestnut. Abstract S3-04 and talk at the Fourth International Chestnut Symposium, Beijing, China, Sept. 23  25. Jacob-Wilk, D., M. Turina, P. Kazmierczak and N.K. Van Alfen. 2007. Silencing of Kex2 significantly diminishes the virulence of Crypyonectria parasitica. Phytopathology 97:S50. Jacob-Wilk, D.M. Turina and N.K. Van Alfen. 2006. Mycovirus Crphonectria hypovirus 1 elements cofractionate with trans-golgi network membranes of the fungal host Cryphonectria parasitica. J. Virol. 80:6588-6596. Jakobi, Steven R. 2005. The Effects of hypovirulent Cryphonectria parasitica inoculum, developmental stages of cankers, and time of year of wounding on the survival of the American chestnut, Dissertation, West Virginia University, Available: . Jeffers, S.N., J.B. James and P.H. Sisco. 2007. Screening for resistance to Phytophthora cinnamomi in hybrid seedlings of American chestnut. 4th IUFRO Phytophthoras in Forest and Natural Ecosystems, Asilomar, CA. . Jiménez-Gasco, M. M., M.G.Milgroom and R.M. Jiménez-Díaz. 2004. Stepwise evolution of races in a plant pathogenic fungus inferred from fingerprinting with repetitive DNA sequences. Phythopathology 94:228-235. Kazmierczak, P., D.H. Kim, M. Turina and N. K. Van Alfen. 2005. A hydrophobin of the chestnut blight fungus, Cryphonectria parasitica, is required for stromal pustule eruption. Eukaryotic Cell. 4: 931-936. Kazmierczak, Pam, P. McCabe, M. Turina, D. Jacob-Wilk, and N.K. Van Alfen. 2008. The mycovirus CHV1 disrupts secretion of developmentally regulated proteins in Cryphonectria parasitica. MMPI (in press). Keesey I.W. and Bruce A. Barrett. 2006. Preliminary data on the seasonal occurrence and oviposition preferences of the lesser chestnut weevil, Curculio sayi, in mid-Missouri. Entomological Society of America, Annual Meeting; Indianapolis, IN; 11 December 2006. Oral presentation. Keesey, I.W and B. Barrett. 2008. Seasonal occurrence and soil distribution of the lesser chestnut weevil, Curculio sayi (Coleoptera: Curculionidae) in mid-Missouri. Journal of the Kansas Entomological Society (in press). Keesey, I.W. 2007. Seasonal occurrence, soil distribution and flight characteristics of Curculio sayi in Mid-Missouri. M.S. Thesis, University of Missouri-Columbia. 74 pp. Kellogg, S.K. and L.K. Rieske. 2005. Resistance to a fungal pathogen influences resistance to generalist herbivores. Entomologia experimentalis et applicata 117: 209-219. Kim, M.-J., B.-R. Kwon, S.-M. Park, H.-J Chung, M.-S. Yang, A.C.L. Churchill, N.K. VanAlfen and D.-H. Kim. 2008. Promoter analysis of the cell surface-abundant and hypoviral-regulated cryparin gene from Cryphonectria parasitica. Mol. Cells (in press). Kobayashi, D. Y. and B.I. Hillman. 2005. Fungi, bacteria, and viruses as pathogens of the fungal community. In: The Fungal Community, 3rd Edition. J. Dighton, P. V. Oudemans, and J. F. White, eds. CRC Press, Boca Raton, FL, 960 pp. Kubisiak, T.L. and J. Roberds. 2006. Genetic structure of American chestnut populations based on neutral DNA markers. Pages 1009-122 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Kubisiak, T.L. and M.G. Milgroom. 2006. Markers lined to vegetative incompatibility (vic) loci and a region of reduced recombination near the mating type locus (MAT) in Cryphonectria parasitica. Fungal. Genet. Biol. 43:453-463. Kubisiak, T.L. C. Dutech and M.G. Milgroom. 2007. Fifty-three polymorphic microsatellite loci in the chestnut blight fungus, Cryphonectria parasitica. Mol. Ecol. Notes 7 (3):428-432. Lan, X., Z. Yao, Y. Zhou, J. Shang, H. Lin, D.L. Nuss and B. Chen. 2008. Deletion of the cpku80 gene in the chestnut blight fungus, Cryphonectria parasitica, enhances gene disruption efficiency. Curr. Genet. 53:59-66. Lang, P. F. Dane and T.L. Kubisiak. 2006. Phylogeny of Castanea (Fagaceae) based on chloroplast trnT-L-F sequence data. Tree Genetics and Genomes 2:132-139. Lang, P., F. Dane, T.L. Kubisiak, and H.W. Huang. 2007. Molecular evidence for an Asian origin and a unique westward migration of species in the genus Castanea via Europe to North America. Molecular Phylogenetics and Evolution 43: 49-59. Liang, H, H. Gao, C.A. Maynard and W.A. Powell. 2005. Expression of a self-processing putative pathogen resistance-enhancing gene construct in Arabidopsis. Biotech. L. 27:435-442. Linder-Basso, D., J.N. Dynek and B.I. Hillman. 2005. Genome analysis of Cryphonectria hypovirus 4, the most common hypovirus species in North America. Virology 337:192-303. Liu, S. and J.C. Carlson. 2006. Selection for Chinese vs American genetic material in blight-resistant backcross progeny using genomic DNA. Pages 133-150 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Liu, Y.-C. and M.G. Milgroom. 2007. Diversity of viruses in Cryphonectria parasitica and C. nitschkei in Japan and China, and partial characterization of a new chrysovirus species. Mycological Research 111:433-442. Liu, Y.-C. and M.G. Milgroom. 2007. High diversity of vegetative compatibility types in Cryphonectria parasitica in Japan and China. Mycologia. 99:279-284. Liu, Y.-C., D. Linder-Basso, B.I. Hillman, S. Kaneko and M.G. Milgroom. 2003. Evidence for interspecies transmission of viruses in natural populations of filamentous fungi in the genus Cryphonectria. Mol. Ecol. 12:1619-1628. Liu, Y.-C., J.N. Dynek, B.I. Hillman and M.G. Milgroom. 2007. Diversity of Cryphonectria parasitica and C. nitschkei in Japan and China, and partial characterization of a new chrysovirus species. Mycol. Res. 111:433:442. MacDonald, W.L. and M.L. Double. 2006. Hypovirulence: use and limitations as a chestnut blight biological control. Pages 87-95 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Marra, R. E., P. Cortesi, M. Bissegger and M.G. Milgroom. 2004. Mixed mating in natural populations of the chestnut blight fungus, Cryphonectria parasitica. Heredity 86:134-143. Maynard, C.A., L.D. Polin, S. LaPierre, R.E. Rothrock and W.A. Powell. 2006. American chestnut (Castanea dentata (Marsh.) Borkh.). Pages 239-0251. In K. Wang, Ed. Agrobacterium Protocols: (2nd Edition) Methods in Molecular Biology Book Series #344, Humana Press, Inc. Totowa, NJ. Maynard, C.A., W.A. Powell, L.D. Polin-McGuigan, A.M. Viéitez, A. Ballester, E. Corredoira, S.A. Merkle and G.M. Andrade (2008) Chestnut. In: Kole, C., Hall, T.C. (eds.) A Compendium of Transgenic Crop Plants: Forest Tree Species, Blackwell Publishing, Oxford, UK, 2008. McGuire, I. C., R.E. Marra and M.G. Milgroom. 2004. Mating-type heterokaryosis and selfing in Cryphonectria parasitica. Fung. Genet. Biol. 41:521-533. McGuire, I.C., J.E. Davis, M.L. Double, W.L. MacDonald, J.T. Rauscher, S. McCawley and M.G. Milgroom. 2005. Heterokaryon formation and parasexual recombination between vegetatively incompatible lineages in a population of the chestnut blight fungus, Cryphonectria parasitica. Mol. Ecol. 14: 3657-3669. Merkle, S.A., G.M. Andrade, C.J. Nairn, W.A. Powell and C.A. Maynard, C.A. 2007. Restoration of threatened species: A noble cause for transgenic trees. Tree Genetics and Genomes 3: 111-118. Mertens, P., C. Wie and B.I. Hillman. 2005. Genus Mycoreovirus. In: Virus Taxonomy: Eighth Report of the International Committee for the Taxonomy of Viruses. C. M. Fauquet et al., eds. Academic Press, NY. Milgroom. M.G., K. Sotirovski, D. Spica, J.E. Davis, M.T. Brewer, M. Milev and P. Cortesi. 2008. Clonal population structure of the chestnut blight fungus in expanding ranges in southeastern Europe. Mol. Ecology 17:4446-4458. Newhouse, Andrew E., Nicholas S. Kaczmar, Charles A. Maynard. American Elm, in Kole, C. and Hall, T. C. (eds.), Compendium of Transgenic Crop Plants: Transgenic Forest Tree Species, Blackwell Publishing, Oxford, UK, 2008, pp 241-262. Nuss, D. L., B.I. Hillman, D. Rigling and N. Suzuki. 2005. Family Hypoviridae. In: Virus Taxonomy: Eighth Report of the International Committee for the Taxonomy of Viruses. C. M. Fauquet et al., eds. Academic Press, NY. Nuss, D.L. 2005. Hypovirulence: Mycoviruses and the fungal-plant interface. Nature Reviews Microbiology 3:632-642. Papazova-Anakieva, I., K. Sotirovski, P. Cortesi and M.G. Milgroom. 2007. Horizontal transmission of hypoviruses between vegetative compatibility types of Cryphonectria parasitica in Macedonia. Eur. J. Plant Pathology 120:35-42. Phelps, T. K.C. Steiner, C.C. Chen and J. Zacek. 2006. Planting trials of American chestnut in central Appalachian forests. Pages 161-166 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Polin, L.D., H. Liang, R.E. Rothrock, M. Nishii, D.L. Diehl, A.E. Newhouse, C.J. Nairn, W.A. Powell and C.A. Maynard. 2006. Agrobacterium-Mediated Transformation of American Chestnut (Castanea dentata (Marsh.) Borkh.) Somatic Embryos. Plant Cell Tissue and Organ Culture 84:69-79 Powell, W.A., C.A. Maynard, B. Boyle and A. Seguin. 2006. Fungal and bacterial resistance in transgenic trees. Pages 235-252. In: M. Fladung and D. Ewald, Eds., Tree Transgenics, Recent Developments. Springer, Berlin Heidelberg, Germany. 357 p. Powell, W.A., S.A. Merkle, H. Liang, and C.A. Maynard. 2005. Blight Resistance Technologies: Transgenic Approaches. In, proc. of conf. on restoration of American chestnut to forest lands, Steiner, K.C. and J.E. Carlson (eds.). Powell. W.A., P. Morley, M. King and C.A. Maynard. Small stem chestnut blight resistance assay. J. Am. Chest. Found. 21:36-40. Rhoades, C.C., D.L. Loftis and J. Lewis. 2005. Silvicultural treatment and topographic controls on American chestnut (Castanea dentata) seedling establishment and initial growth in eastern Kentucky, USA. Project Final Report. Agreement SRS 02-CA-11220134-194, USDS Forest Service, Southern Research Station. Rieske, L.K. 2007. Success of an exotic gallmaker, Dryocosmus kuriphilus, on chestnut in the USA: a historical account. EPPO Bulletin 37, 172-174. Rieske, L.K. 2007. Success of an exotic gallmaker, Dryocosmus kuriphilus, on chestnut in the USA: a historical account. European & Mediterranean Plant Protection Org. Bulletin 37, 172-174. Rieske, L.K. Woody plant gall source-sink relations: Reducing source strength influences gallmaker success. Entomological Society of America National Meeting. Reno. Nov. 2008 (pending). Rieske, L.K., C.C. Rhoades and S.P. Miller. 2003. Foliar chemistry and gypsy moth herbivory on pure American chestnut, Castanea dentata (Fam: Fagaceae) and a disease-resistant hybrid. Environmental Entomology 32:359-365. Rittenour, W.R. 2005. The biological control potential of Cryphonectria parasitica strains containing an infectious cDNA copy of the hypovirus CHV1-Euro7. Masters Thesis, West Virginia University, Available: . Robichaud, R.L., V.C. Lessard and S.A. Merkle. 2004. Treatments affecting maturation and germination of American chestnut embyos. J. Plant Physiol. 161:957-969. Root, C., C.J. Balbalian, R. Bierman, L.M. Geletka, S.L. Anagnostakis, W.L. MacDonald, M.L. Double and D.L. Nuss. 2005. Multiseasonal field release and spermatization trials of transgenic hypovirulent strains of Cryphonectria parasitica containing cDNA copies of hypovirus CHV1-EP713. For. Path. 35:277-297. Rothrock, R.E., L.D. Polin-Mcguigan, A.E. Newhouse, W.A. Powell and C.A. Maynard. 2007. Plate flooding as an alternative Agrobacterium-mediated transformation method for American chestnut somatic embryos. Plant Cell, Tissue and Organ Culture 88: 93-99. Schlarbaum, S., S. Brosi, and S. Anagnostakis. 2006. Feasibility of large-scale reintroduction of chestnut to National Park Service lands. Pages 195-202 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Sederoff, R., D. Nielsen, C. Smith, J. Tomkins, M. Atkins, B. Blackmon, M. Staton, S. Ficklin, F. Hebard, P. Sisco, J. Carlson, S. DiLoreto, A. Barakat, W. Powell, K. Baier, S. Anagnostakis, T. Kubisiak and N. Wheeler. 2008. Genomic tool development for the Fagaceae. Abstract P494. Plant and Animal Genome XVI, San Diego, CA, Jan. 12  16. Segers, G.C., J.C. Regier and D.L. Nuss. 2004. Evidence for a role of the regulator of G-protein signaling protein CPRGS-1 in G± subunit CPG-1-mediated regulation of fungal virulence, conidiation and hydrophobin synthesis in the chestnut blight fungus Cryphonectria parasitica. Eukaryotic Cell 3:1454-1463. Segers, G.C., R. van Wezel, X. Zhang, Y. Hong and D.L. Nuss. 2006. Hypovirus papain-like protease p29 suppresses RNA silencing in the natural fungal host and in a heterologous plant system. Eukaryotic Cell 5: 896-904. Segers, G.C., X. Zhang, F. Deng, Q. Sun and D.L. Nuss. 2007. Evidence that RNA silencing functions as an antiviral defense mechanism in fungi. PNAS 104: 12902-12906. Shang, J., X. Wu, X. Lan, Y. Fan. H. Dong, Y. Deng, D.L. Nuss and B. Chen. 2008. Large-scale expressed sequence tag analysis for the chestnut blight fungus Cryphonectria parasitica. Fungal Genetics and Biology 45:319-327. Sisco, P.H. 2004. Breeding blight-resistant American chestnut trees. Journal of The American Chestnut Foundation 18(1):12-16. Sisco, P.H. 2006. An update on chestnut DNA projects: Part I. The genes for blight resistance from Chinese chestnut. J. Am. Chestnut Found. 20(1):21-29. Sisco, P.H. 2007. Breeding blight-resistant American chestnut trees. Abstract W149 and Invited Talk. Plant and Animal Genome XV, San Diego, CA, Jan 13  17. Sisco, P.H. 2008. Chestnut cytogenetics: Faridi, Burnham, and McClintock. Compass, Issue 11, p. 13 Sisco, P.H. 2008. The United States National Science Foundation Project on developing genetic tools for the study of the Fagaceae: Castanea, Quercus, and Fagus. Abstract 3-14 for a plenary talk at the Fourth International Chestnut Symposium, Beijing, China, Sept. 23  25. Sisco, P.H. 2004. Designing chestnut orchards to control pollen without hand-pollinationthe use of self-incompatibility and cytoplasmic male sterility. Journal of The American Chestnut Foundation 18(2):46-53. Sisco, P.H. 2006. An update on chestnut DNA projects: Part II. Other uses of molecular markers in the TACF breeding program. J. Am. Chestnut Found. 20(2):35-43. Sisco, P.H. and M. Jordan. 2008. American chestnut, a premium hardwood. Hardwood Matters 77:18-19. Sisco, P.H., T.L. Kubisiak, M. Casasoli, T. Barreneche, A. Kremer, R.R. Sederoff, F.V. Hebard and F. Villani. 2005. An improved genetic map for Castanea mollissima/Castanea dentata and its relationship to the genetic map of Castanea sativa. pp. 491-495. In C.G. Abreu, E. Rosa, and A.A. Monteiro (eds). Proc. 3rd Intl. Chestnut Congress, Chaves, Portugal. Acta Hort. 693. Smith, M.L. C.C. Gibbs and M.G. Milgroom. 2006. Heterokaryon incompatibility function of barrage-associated vegetative incompatibility genes (vic) in Cryphonectria parasitica. Mycologia 98:43-50. Smith, M.L., M.G. Milgroom, D. Rigling and U. Heineger. 2006. Occurrence of Cryphonectria hypovirus 1 in the chestnut blight fungus in Macedonia. Forest Pathology 36:136-143. Sotirovski, K., I. Papazova-Anakieva, N.J. Grünwald, N. J. and M.G. Milgroom. 2004. Low diversity of vegetative compatibility types and mating type in Cryphonectria parasitica in the southern Balkans. Plant Pathol 53:325-333. Sotirovski, K., M.G. Milgroom, D. Rigling and U. Heineger. 2006. Occurrence of Cryphonectria hypovirus 1 in the chestnut blight fungus in Macedonia. Forest Pathology 36:136-143. Steiner, K.C. 2006. Regional adaptation in American chestnut. Pages 123-128 in: Steiner K.C. and J.E. Carlson, eds. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Steiner, K.C. and J.E. Carlson, eds. 2006. Restoration of American Chestnut To Forest Lands-Proceedings of a Conference and Workshop. May 4-6, 2004, The North Carolina Arboretum, Natural Resources Report NPS/NCR/CUE/NRR-2006/001, National Park Service, Washington, DC. Strasser, Robert. 2006. Of place and time. J. Am. Chestnut Found. 10:12-15. Sudo, K., McLane, H.L., Krasnoff, S.B., Anagnostakis, S.L., Gibson, D.M., and A.C.L. Churchill. 2007. Characterization of a polyketidesynthase gene of the chestnut blight fungus by Agrobacterium tumefaciens-mediated targeted gene disruption. American Phytopathological Society Annual Meeting, San Diego, CA, July 28-August 1, 2007 (abstract) Sudo, K., McLane, H.L., Krasnoff, S.B., Anagnostakis, S.L., Gibson, D.M., and A.C.L. Churchill. 2007. Agrobacterium tumefaciens-mediated disruption of a polyketide synthase gene in Cryphonectria parasitica. Fung. Genet. Newsl. 54, #260 (abstract). http://www.fgsc.net/asil2007/xxivFGCposterAbs.htm], XXIV Fungal Genetics Conference, Pacific Grove, CA, March 20-25, 2007. Sun, L., D.L. Nuss and N. Suzuki. 2006. Synergism between a mycovirus and a hypovirus mediated by the papain-like protease p29 of the prototypic hypovirus CHV1-EP713. J. General Virology 87:3703-3714. Supyani, S., B.I. Hillman and N. Suzuki. 2007. Baculovirus expression of the 11 Mycoreovius-1 genome segments and identification of the guanylyltransferase-encoding segment. J. Gen. Virol. 88:342-350. Suzuki, N., K. Maruyama, M. Moriyama and D.L. Nuss. 2003 Hypovirus paparin-like protease p29 functions in trans to enhance viral double-stranded RNA accumulation and vertical transmission. J. Virol. 77:11697-11707. Suzuki, N., S. Supyani, K. Maruyama, and B.I. Hillman. 2004. Complete genome sequence of Mycoreovirus 1/Cp9B21, a member of a new genus in the family Reoviridae isolated from the chestnut blight fungus, Cryphonectria parasitica. Journal of General Virology 85:3447-3448. Turina, M, P. Kazmierczak and N.K. Van Alfen. 2003. Intracellular processing and secretion of the fungal hydrophobin cryparin. Phytopathology 93:S85. Turina, M., A. Prodi and N. K. Van Alfen. 2003. Role of Mf1-1 pheromone precursor gene of the filamentous ascomycete Cryphonectria parasitica. Fungal Genetics and Biology 40: 242-251. Turina, M., L. Zhang and Neal K. Van Alfen. 2006. Effect of Cryphonectria hypovirus 1 (CHV1) infection on Cpkk1, a mitogen-activated protein kinase kinase of the filamentous fungus Cryphonectria parasitica. Fungal Genetics and Biology 43:764-774. Turina, M., P.M. McCabe, A. Prodi and N.K. Van Alfen. 2001. Characterization of vesicle subsets in CHV1 infected Cryphonectria parasitica. Phytopathology 91:S90. University of Missouri Center for Agroforestry. 2004. Why chestnuts: Nutrition and your health. Univ. of Missouri Center for Agroforestry, 23 Jan. 2006. Vettraino, A.M., Vannini, A., Flamini, L., Lagnese, R., Pizzichini, L., Talevi, S. and Fulbright, D.W. 2005. A new transmissible syptomology on sweet chestnut in Italy. Acta Hort. (ISHS) 693:547-550, . Warmund, M.R. 2008. Fertilizer applications after foliar nitrogen Chinese chestnut growth and yield. Northern Nut Growers 98th Ann. Rept. (In press). Warmund, M.R. 2008. Primary and secondary fruiting of Chinese chestnut trees. Northern Nut Growers 99th Ann. Rept. (submitted). Warmund, M.R., K.L. Hunt, and M.A. Gold. 2005. Removal of secondary burs increases average nut weight from primary burs of 'Armstrong', 'Orrin' and 'Willamette' Chinese chestnuts. III International Congress on Chestnut, Claves, Portugal, Oct. 20-23, 2004, Acta Hort. (ISHS) 693:149-152, Welch, A.J., C.A. Maynard, A.J. Stipanovic, and W.A. Powell. 2007. The effects of oxalic acid on transgenic Castanea dentata callus tissue expressing oxalate oxidase. Plant Science 172:488-496 Worthen, L.M. 2005. Expression of hypovirulence in a segregating population of BC2F2 Castanea dentata hybrids. Departmental Honors Thesis, University of Tennessee at Chattanooga. Zhang, X. and D.L. Nuss. 2008. A host dicer is required for defective viral RNA production and recombinant virus vector RNA instability for a positive sense RNA virus. PNAS 105: 1674916754. Zhang, Xuemin, G.C. Segers, Q. Sun, F. Deng and D.L. Nuss. 2008. Characterization of hypovirus-derived small RNAs generated in the chestnut blight fungus by an inducible DCL-2-dependent pathway. J. Virology 82:2613-2619.