MOLECULAR AND MORPHOLOGICAL SYSTEMATICS OF PLANT PATHOGENIC FUNGI

• Sponsoring Institution: Agricultural Research Service/USDA
• Project Status: COMPLETE
• Funding Source: USDA INHOUSE
• Reporting Frequency: Annual
• Accession No.: 0406816
• Project Start Date: Jan 12, 2003
• Project End Date: Mar 8, 2007
• Program Code: [(N/A)]- (N/A)

Recipient Organization
AGRICULTURAL RESEARCH SERVICE
RM 331, BLDG 003, BARC-W
BELTSVILLE,MD 20705-2351

Performing Department
(N/A)

Non Technical Summary
(N/A)

Animal Health Component

25%

Research Effort Categories

Basic

75%

Applied

25%

Developmental

0%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
212	4020	1060	100%

Knowledge Area
212 - Pathogens and Nematodes Affecting Plants;

Subject Of Investigation
4020 - Fungi;

Field Of Science
1060 - Biology (whole systems);

Keywords

molecular biology

plant morphology

systematics

fungi

diaporthe

phomopsis

tilletia

ascomycetes

pathogen identification

fungus diseases (plants)

phylogeny

taxonomy

introns

fungus genetics

elongation

calmodulin

blight (castanea)

canker

smut

actin

anthracnose

teliospores

plant pathogens

grain

wild plants

grasses

gene analysis

dna sequences

Goals / Objectives
1. Use molecular and morphological characteristics to develop a phylogeny of bunt fungi and related groups on cultivated grains and wild grasses. 2. Analyze relationships among species of Diaporthe and Phomopsis and the sexual and asexual states of related canker-causing fungi in the chestnut blight order and determine species and species-groups based on morphological and molecular characteristics.

Project Methods
Species of Tilletia will be collected from around the world. Each specimen will be characterized morphologically. Molecular data will be obtained for each isolate. The LSU nrDNA, the EF1-alpha, actin, and RPB1 and RPB2 genes will be sequenced and analyzed. Species concepts will be defined based on both morphological and molecular data. Species will be described and illustrated in monographic publications and keys prepared for the on-line identification of species of Tilletia and related genera. Strains of Phomopsis-Diaporthe will be obtained through isolation from recently collected material. Specimens and isolates will be examined to determine morphological characteristics of microscopic and colony features. For determining species relationships within Phomopsis-Diaporthe and relationships to other genera in the Diaporthales, the conserved regions of the LSU nrDNA DNA, EF1-alpha, and RPB1 and RPB2 genes will be sequenced. Morphological and molecular data will be analyzed to determine species and genera concepts. Taxonomic monographs will be published and made available as on-line resources.

Progress 01/12/03 to 03/08/07

Outputs
Progress Report Objectives (from AD-416) 1. Use molecular and morphological characteristics to develop a phylogeny of bunt fungi and related groups on cultivated grains and wild grasses. 2. Analyze relationships among species of Diaporthe and Phomopsis and the sexual and asexual states of related canker-causing fungi in the chestnut blight order and determine species and species-groups based on morphological and molecular characteristics. Approach (from AD-416) Species of Tilletia will be collected from around the world. Each specimen will be characterized morphologically. Molecular data will be obtained for each isolate. The LSU nrDNA, the EF1-alpha, actin, and RPB1 and RPB2 genes will be sequenced and analyzed. Species concepts will be defined based on both morphological and molecular data. Species will be described and illustrated in monographic publications and keys prepared for the on-line identification of species of Tilletia and related genera. Strains of Phomopsis-Diaporthe will be obtained through isolation from recently collected material. Specimens and isolates will be examined to determine morphological characteristics of microscopic and colony features. For determining species relationships within Phomopsis- Diaporthe and relationships to other genera in the Diaporthales, the conserved regions of the LSU nrDNA DNA, EF1-alpha, and RPB1 and RPB2 genes will be sequenced. Morphological and molecular data will be analyzed to determine species and genera concepts. Taxonomic monographs will be published and made available as on-line resources. Accomplishments Three distinct fungi cause destructive bark canker of beech Beech bark canker has been known in North America since that late 1800�s but recently this disease has spread westward as far as Michigan and southward to Tennessee. For about thirty years it has been known that two different species of fungi are involved in beech bark canker but the distinction between is not clear. In this research the fungi that cause beech bark canker are differentiated based on their microscopic characteristics as well as a comparison of their genes. It was determined that one fungal species on beech causes cankers only in North America and infects only beech trees. A second fungal species attacks a wide range of trees including beech, birch, apple, maple, and tulip poplar in both North America and Europe. A third species is known only on beech in Europe. This research will be used by forest pathologists who are breeding for beech trees that are resistant to beech bark canker and other diseases of hardwood trees. In addition it will be used by plant quarantine officials to prevent the introduction of these disease- causing fungi from Europe to North America. This research is related to NP 303 Component 1. New species of anthracnose fungi on specialty crops Fungi cause serious diseases of crop plants and trees. Many of these fungi do not occur in the United States and have potential as invasive species. In order to prevent the entry of invasive fungi, it is critical to know which species exist both inside and outside the United States. Fungi in a group that causes anthracnose leaf diseases were examined using characters visible with the microscope as well as genetic characters. Anthracnose fungi on the plant family that includes American aloe, New Zealand flax, and lucky bamboo were characterized, described and illustrated. A total of five fungal species were found on these plants. Of these, two species are present in the United States while three species do not occur in the U.S. Thus, it is important to be able to tell the difference between these closely related fungi. One of the three species not found in the U.S., one is new to science and was named, described and illustrated. A key to all species of anthracnose fungi on this plant family was presented. This research will be used by plant quarantine officials and plant pathologists to identify these fungi and to prevent entry of these potentially invasive species into the United States. This research is related to NP 303 Component 1. Life cycle of fungus causing disease of birch tree Fungi cause serious diseases of hardwood trees throughout the world. An asexual or clonal fungus causes a disease of birch trees in the summer and fall while the sexual state of a fungus occurs on birch leaves in the spring. In this study, it was discovered that these two seemingly different fungi actually represent two states in the life cycle of the same fungus. This paper also shows that that the spores from either state can cause disease. Using DNA sequence data the fungus on birch trees in Europe was determined to be related to a similar fungus causing a disease on butternut trees in North America. This information will be used by forest pathologists and horticulturalists to identify fungi causing diseases of birch trees. This research is related to NP 303 Component 1. Rust fungi evolved from insect and dead plant fungi Fungi cause billions of dollars damage to agricultural crops each year. In addition, they can greatly hinder the export of agriculturally commodities from the U.S. to other counties. In order to control diseases caused by fungi both in the U.S. and abroad it is necessary to identify and characterize fungal species that cause diseases of crops. In this paper a number of rust fungi are characterized and compared in order to determine how closely related they are to each other. These comparisons are made using portions of the genome that are sequenced. It was determined that most of the rust fungi are relatively closely related to each other especially those that grow on living plants. Within the rust fungi several small groups are related to the rusts that grow on living plants but occur on insects and dead plant material. Knowledge of the characteristics and relationships of rust fungi to each other is important and will be used by plant pathologists who breed for resistance to the diseases caused by rust fungi. This research is related to NP 303 Component 1. Soybean rust in Mexico confirms model pathway Soybean rust causes a serious disease of soybean that has been considered a threat to the United States for at least two decades. Once this disease moved from Africa to South America, scientists predicted that soybean rust could be blown northward into the U.S. by hurricane winds. This fungus was discovered in a soybean field in Louisiana in November, 2004 following Hurricane Ivan. Since then, scientists have searched for soybean rust in Central America to determine how widespread is this pathogen. Exploration for rust fungi in Mexico revealed the existence of soybean rust in the northeastern state of San Lu�s de Potos�. This is along the path predicted for spread of soybean rust and confirms the model pathway. This research will be used to determine the spread of invasive rust fungi and to predict their possible future path. This research is related to NP 303 Component 1. Technology Transfer Number of New CRADAS and MTAS: 1 Number of Active CRADAS and MTAS: 1 Number of Non-Peer Reviewed Presentations and Proceedings: 5 Number of Newspaper Articles,Presentations for NonScience Audiences: 3

Impacts
(N/A)

Publications

• Castlebury, L.A., Rossman, A.Y., Sheer Hyten, A.M. 2006. Phylogenetic Relationships of Neonectria/Cylindrocarpon on Fagus in North America. Canadian Journal of Botany. 84:1417-1433.
• Farr, D.F., Aime, M.C., Rossman, A.Y., Palm, M.E. 2006. The Species of Colletotrichum on Agavaceae. Mycological Research. 110:1395-1408.
• Green, S., Castlebury, L.A. 2006. Connection of Gnomonia Intermedia to Discula Betulina and its relationship to other taxa in the Gnomoniaceae. Mycological Research. 111:62-69.
• Aime, M.C., Hibbett, D., Matheny, B.P., Henk, D.A., Frieders, E.M., Szabo, L.J. 2007. An overview of the higher-level classification of urediniomycotina based on combined analyses of nuclear large and small subunit rDNA sequences. Mycologia. 98:896-905.
• Rodriguez, A.C., Rios, J.A., Hernandez, J. 2006. First report of Asian Soybean Rust caused by Phakopsora Pachyrhizi, from Mexico. Plant Disease. 90:1260.
• Chlebicki, A., Aime, M.C. 2006. New distribution and host records for Puccinia Atrofusca and other notable Puccinia species (Uredinales) from Central Asia. Polish Botanical Journal. 51:87-91.
• Deadman, M.L., Al Sa'Di, A., Al Maqbali, Y., Klod, A., Aime, M.C. 2007. First report of Puccinia Triticina on Wheat in Oman. Plant Disease. 91:113.
• Deadman, M.L., Al Sadi, A.M., Al Maqbali, Y.M., Al Subhi, A., Al Yahya, R., Aime, M.C. 2007. First Report of Rust Caused by Tranzschelia Discolor on Peach in Oman. Plant Disease. 91:638.
• Rossman, A.Y., Farr, D.F., Castlebury, L.A. 2007. A Review of the Phylogeny and Biology of the Diaporthales. Mycoscience. 48:135-144.
• Zhang, N., Sung, G., Castlebury, L.A., Seifert, K.A., Rossman, A.Y., Rogers, J.D.,Miller, N.,Huhndorf, S.M., Schoch, C.L., Kohlmeyer, J., Volkmann-Kohlmeyer, B., 2007. An overview of Molecular Phylogeny of the Sordariomycetes. Mycologia. 98:1076-1087.
• James, T.Y., Kauff, F., Schoch, C., Matheny, B., Hofstetter, V., Cox, C.J., Celio, G., Guiedan, C., Fraker, E., Miadlikowska, J., Lumbsh, T., Rauhut, A., Reeb, V., Arnold, A., Amtoft, A., Stajich, J.E., Hosaka, K., Sung, G., Johnson, D., O'Rourke, B., Crockett, M., Binder, M., Curtis, J.M., Slot, J. C., Wang, Z., Wilson, A.W., Schueller, A., Longcore, J.E., O Donnell, K., Mozley-Standridge, S., Porter, D., Letcher, P.M., Powell, M.J., Taylor, J. W., White, M.M., Griffith, G.W., Davies, D.R., Humber, R.A., Morton, J.B., Sugiyama, J., Rossman, A.Y., Rogers, J.D., Pfister, D.H., Hewitt, D., Hansen, K., Hambleton, S., Shoemaker, R.A., Kohlmeyer, J., Volkmann- Kohlmeyer, B., Spotts, R.A., Serdani, M., Crous, P.W., Hughes, K.W., Matsuura, K., Langer, E., Langer, G., Untereiner, W.A., Lucking, R., Budel, B., Geiser, D.M., Aptroot, D.M., Diederich, P., Schmitt, I., Schultz, M., Yahr, R., Hibbett, D.S., Lutzoni, F., Mclaughlin, D.J., Spatafora, J.W., Vilgalys, R. 2006. Reconstructing the early evolution of Fungi using a six- gene phylogeny. Nature. 443:818-822.
• Hibbett, S.A., Binder, M., Bischoff, J. F., Blackwell, M., Cannon, P. F., Eriksson, O. E., Huhndorf, S., James, T., Kirk, P. M., L�cking, R., H. Lumbsch, T., Lutzoni , F., Matheny, P. B., Mclaughlin, D. J., Powell, M. J. , Redhead , S., Schoch, C. L., Spatafora, J. W., Stalpers, J. A., Vilgalys, R., Aime, M. C., Aptroot, A., Bauer, R., Begerow, D., Benny, G. L., Castlebury, L. A., Crous, P. W., Dai, Y.-C., Gams, W., Geiser, D. M., Griffith, G. W., Gueidan, C., Hawksworth, D. L., Hestmark, G., Hosaka, K., Humber, R. A. , Hyde, K. D., Ironside, J. E., KL�ljalg, U., Kurtzman, C. P. , Larsson, K.-H., Lichtwardt, R., Longcore, J., Mi�dlikowska, J., Miller, A. Moncalvo, J.-M., Mozley-Standridge, S., Oberwinkler, F., Parmasto, E., Reeb, V., Rogers, J. D., Roux, C., Ryvarden, L., Sampaio, J. P., Sch��ler, A., Sugiyama, J., Thorn, R. G., Tibell, L., Untereiner, W. A., Walker, C., Wang, Z., Weir, A., Weiss, M., White, M. M., Winka, K., Yao, Y.-J., and Zhang, N. 2007. A higher-level phylogenetic classification of the Fungi. Mycological Research. 111:509-547.
• Matheny, B.P., Curtis, J.M., Hofstetter, V., Aime, M.C., Moncalva, J. M., and others. 2007. Major Clades of Agaricales: A Multilocus Phylogenetic Overview. Mycologia. 98:982-995.
• Crozier, J., Thomas, S.E., Aime, M.C., Evans, H.C., Holmes, K.A. 2006. Molecular Characterisation of Fungal Endophytic Morphospecies Isolated from Stems and Pods of Theobroma Cacao. Plant Pathology. 55:783-791.
• Rossman, A.Y., Palm, M.E. 2006. Why aren�t Phytophthora and other Oomycota true Fungi? Outlooks in Pest Management. 17:217-219.

Progress 10/01/05 to 09/30/06

Outputs
Progress Report 1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? Agricultural and forest plants are threatened by fungal pathogens native to the United States and invasive species from other countries. In addition, some disease-causing fungi have potential for use as biocontrol agents for noxious and invasive weeds. This research addresses the classification and characterization, or systematics, of difficult groups of disease-causing fungi specifically bunt fungi of cereals and canker- causing fungi of fruit crops. Accurate knowledge about their classification provides the means for their identification. This knowledge is also essential for making sound plant quarantine decisions to prevent the introduction of new pathogen species and for developing effective disease and weed management strategies. Results of this research include keys for identification and characteristics of fungi that cause bunt and canker diseases as well as invasive and potential biocontrol fungi. The project has two specific objectives: 1) to use molecular and morphological characteristics to develop a phylogeny of bunt fungi and related groups on cultivated grains and wild grasses; 2) to analyze relationships among species of Diaporthe and Phomopsis and the sexual and asexual states of related canker-causing fungi in the chestnut blight order and determine species and species-groups based on morphological and molecular characteristics. The research undertaken falls under National Program 303 Plant Diseases and addresses Action Plan Component I Identification and Classification of Pathogens (70%) and National Program 304 Crop Protection and Quarantine and addresses Action Plan Component IX Biological Control of Weeds (30%). Economic loss to agricultural and horticultural crops due to disease- causing fungi is estimated at $20 billion per year in the United States. In addition, fungi often limit U.S. agricultural exports, a significant factor in the U.S. balance of trade. Karnal bunt and similar fungi threatened the U.S. wheat export market. This problem was solved using systematic knowledge about Karnal bunt and its close relatives. Inadequate knowledge of the classification and characterization of Phomopsis limits the management of the diseases on fruit and woody crops and causes export-import problems for these commodities. Results of this research are essential for plant pathologists, extension agents, and policy makers in managing diseases, maintaining the crop export market, and preventing the introduction of harmful fungi into the United States. In addition to these specific objectives, an additional objective is to respond to problems arising due to invasive fungi. 2. List by year the currently approved milestones (indicators of research progress) Year 1 (FY 2003) Development and/or screening of gene primers for T. indica and T. walkeri. Molecular and morphological characterization of Phomopsis strains from Vitis in the U.S. and determine if more than one species is involved in Phomopsis canker of grapes. Development and/or screening of PCR primers for multiple genes for species-level grouping of Diaporthe-Phomopsis isolates. Update online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera. Year 2 (FY 2004) Sequencing of multiple gene regions for T. indica and T. walkeri cultures to determine utility of various genes and regions for identification and phylogenetic analysis of these closely related species of bunt fungi. Expand online resources for Tilletia species in North America. Sequencing of multiple gene for Diaporthe-Phomopsis strains from soybean and cucurbits to determine species-level groupings and relationships of pathogens and symptomless isolates from these and unrelated hosts. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales. Year 3 (FY 2005) Phylogenetic analysis of multi-gene sequence data for T. indica and T. walkeri isolates to determine relationships of isolates from diverse geographic areas. Development of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation is present in available isolates. Development and/or screening of PCR primers for multiple protein-coding genes for Tilletia species phylogeny to determine if morphological characters and hosts are indicative of phylogenetic relationships. Continued expansion of online resources for Tilletia to all of North America. Collection and DNA sequencing for strains of Diaporthe-Phomopsis from juniper and sunflower. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales. Year 4 (FY 2006) Screening and analysis of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation exists with these loci. DNA sequence and morphological characterization of existing cultures for Tilletia species phylogeny. Expansion of online resources for Tilletia to worldwide. Collection and DNA sequencing of Diaporthe-Phomopsis strains from Ericaceae to determine identity of non-Phomopsis vaccinii isolates and their relationships to P. vaccinii. Molecular and morphological characterization of new cultures of Diaporthe-Phomopsis species. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales. Year 5 (FY 2007) Phylogenetic analysis of multi-gene sequence data for species of Tilletia and related genera in the Tilletiales. Expand online resources for Tilletia to worldwide. Characterization of strains from Rosaceae. Phylogenetic analysis of multi-gene sequence data for all available species in the genera Diaporthe and Phomopsis to determine connections and relationships among sexual and asexual species. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales. 4a List the single most significant research accomplishment during FY 2006. Fungal pathogens causing cacao diseases determined to be close relatives Cocoa is a tropical tree crop that forms the basis for the multibillion dollar chocolate industry in the United States and elsewhere. Two devastating fungal diseases have caused the recent plummet of cocoa production in tropical America. These diseases are known as witches broom and frosty pod rot. Despite their importance, relatively little known about either of the fungi that cause these diseases. One fungus produces a mushroom fruiting body while the other develops only a thin coating of spores on the cocoa fruit. By analyzing and comparing genomic data, the relatedness of these two fungal pathogens to each other and their placement in the fungal family tree was determined. This research proves that, despite their very different outward appearances, these two pathogens of cocoa are closely related to each other and are placed in the same genus. This research has important implications for cocoa breeding programs and for efforts to fight the diseases. Because these two disease-causing fungi are closely related, it will be possible to breed for resistance to both diseases. Importantly, disease control measures that are currently working against witches broom may also be effective against frosty pod rot. Plant breeders and plant pathologists will use this research to develop effective control measures for these serious diseases of cocoa that threaten the U.S. chocolate industry. This research is related to NP 303 Component 1. 4b List other significant research accomplishment(s), if any. Bunt fungi determined to be related based on cereal hosts Bunt fungi cause serious diseases of cereal crops in the United States some of dire quarantine importance. Knowledge of the relationships among bunt fungi is essential for their accurate identification and controlling the diseases they cause. At present, disagreement exists about how to define species of bunt fungi and how they are related to each other. In this paper molecular sequence data are used to determine which morphological characteristics can be used to accurately identify these fungi and how closely related they are to each other. Results indicated that species recognized in different genera were actually all members of the genus Tilletia. Species infecting grass hosts in the same subfamily as wheat, including Karnal bunt, dwarf bunt and common bunt, were found to be more closely related to one another than to species on hosts in other subfamilies such as rice and switchgrass. This information will be used by wheat and other cereal grain pathologists and breeders to more effectively manage bunt diseases, as well as by quarantine and regulatory officials to accurately identify bunt spores in import or export shipments. This research is related to NP 303 Component 1. 5. Describe the major accomplishments to date and their predicted or actual impact. Canker disease of grape in U.S. widespread A canker disease of grape is devastating this crop throughout the eastern and midwester United States but the identity of the causal organism was not well known. Diseased grape vines from Ohio, New York and Michigan were studied and the causal agent was identified as Phomopsis viticola using DNA sequence data. Specialization within host tissue, with some isolates causing more disease in fruits and some causing more disease in leaves, was noted for a number of isolates. Two additional species of Phomopsis were isolated and characterized but caused little or no disease. This information will be used to identify the disease-causing Phomopsis viticola and distinguish it from nondisease- causing species to insure that proper control measures are used. This research is related to NP 303 Component 1. Cause of new disease on Pacific madrone identified Pacific madrone, a broadleaf evergreen tree native to western North America, has been suffering a decline for the past thirty years. A fungus has been isolated and verified as the cause of cankers on dying madrone trees. The fungus was determined to be a previously unknown species belonging to a group that commonly causes canker diseases of trees. The fungus causing the canker disease of madrone was characterized, described, illustrated, and named as a new species. Evidence suggests that this fungus has been present in western North America since 1968. Prior to this research, plant pathologists had misidentified the causal fungus. This research allows forest pathologists to accurately determine the cause of this disease in order to develop control measures. This research is related to NP 303 Component 1. Distinction defined between Karnal bunt on wheat and lookalike bunt on ryegrass Karnal bunt is a quarantine significant disease of wheat that has not been clearly defined nor has it been distinguished from closely related fungi. DNA sequence data from intron regions from protein-coding genes provided additional molecular support for the distinction of Tilletia indica and Tilletia walkeri as separate species. Relationships of a number of wild-grass infecting Tilletia species will be resolved with reference to the wheat bunts. This information will aid in the correct identification of these important quarantine fungi and their morphologically similar relatives and the development of KB resistant wheat. This research is related to NP 303 Component 1. Rapid identification for bunt fungi on cereal crops in United States Cereal crops such as wheat, rice and oats are a valuable export commodity for the United States. Bunt fungi infecting these cereal crops can hinder their export to other countries. In order to allow rapid identification of bunt fungi, on-line identification system was developed for species that occur in the United States. Based on teliospores the system is easy to use because of the easily determined characteristics and numerous illustrations. This system was used to persuade plant pathologists in Algeria that a shipment of wheat from Minnesota was not infected with Karnal bunt thus saving millions of dollars for farmers. The identification system is used by plant pathologists and plant quarantine officials to determine the accurate species of bunt fungi. This research is related to NP 303 Component 1. Species of canker-causing fungi characterized Canker diseases in the genus Diaporthe-Phomopsis are caused by fungi that are difficult to identify. Some of these fungi especially those on soybean are of plant quarantine significance. Species of canker-causing fungi have been characterized and distinguished using both morphological and molecular characteristics. Fungi on agriculturally important crops such as soybeans, grapes and melons have been studied. This will be extended to junipers, sunflowers, blueberries and its relatives and tree fruits. Based on the accurate characterization of these canker-causing fungi, plant pathologists will be able to rapidly identify these fungi especially using molecular characteristics. This research is related to NP 303 Component 1. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? An interactive key for the identification of teliospores of bunt fungi in the United States is available over the Web at http://nt.ars-grin. gov/TaxaDescriptions/Tilletia/Index2.cfm. This system allows non- specialists such as plant quarantine officials and extension agents to rapidly identify these fungi. A resource page for systematics of the Diaporthales is available at http://nt.ars-grin. gov/taxadescriptions/diaporthales/HomePage/Intro.cfm. This information summarizes current family level relationships of fungi in this order, including Diaporthe-Phomopsis, dogwood anthracnose, and chestnut blight. Additionally DNA sequences used to analyze relationships of or identify plant pathogens have been submitted to GenBank, a publicly available database, for use by other researchers. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Fungal Threats to Cacao Share an Ancient History. Agricultural Research Magazine. October 2005 Vol. 53, No. 10. Article featured research on relationships of cacao pathogens by Dr. M.C. Aime. Also, as news release on October 12, 2005. The MatrixPlant and Fungal. Agricultural Research Magazine. June 2006 - Vol. 54, No. 6. Article featured coffee and chocolate endophyte work by Dr. M.C. Aime. ARS Experts as Firefighters. Agricultural Research Magazine. May 2006 - Vol. 54, No. 5. Article featured work by Dr. L.A. Castlebury concerning a shipment of wheat to Algeria. Dr. A. Y. Rossman was interviewed on National Public Radio about mold in New Orleans following Hurricane Katrina. Dr. A. Y. Rossman was interviewed on National Public Radio about a fungus implicated in eye infection in contact lens wearers. Invited presentation by Dr. A.Y. Rossman at Harvard University.

Impacts
(N/A)

Publications

• Aime, M.C. 2005. Book review of: smut fungi (ustilaginomycetes p.p. and microbotryales, basidiomycota). Inoculum. 56:13.
• Castlebury, L.A., Carris, K. Vanky, 2005. Phylogenetic analysis of Tilletia and allied genera in order Tilletiales (Ustilaginomycetes; Exobasidiomycetidae) based on large submit nuclear RDNA sequences. Mycologia 97(4):888-900.
• Sogonov, M.V., Castlebury, L.A., Farr, D.F. Rossman, A.Y., White, J.F. 2005. The type species of the genus Gnomonia, G. gnomon, and the closely related G. setacea. Sydowia 57(1):102-120.
• Carris, L.M., L.A. Castlebury, and B.J. Goates. 2006. Nonsystemic Bunt Fungi Tilletia indica and T. horrida: A Review of History, Systematics, and Biology. Annual Review of Phytopathology 44:5.1-5.21. (doi:10. 1146/annurev.phyto.44.070505.143402)
• Aime, M.C. 2006. Toward resolving family-level relationships in rust fungi (Uredinales). Mycoscience 47(3):112-122.
• Deadman, M.L., Al Sadi, A.M., Al Maqbali, Y.M., Livingston, S., Aime, M.C. 2006. First report of puccinia sorghi on maize in oman.. Plant Disease. 90:826.
• Henkel, T.W., James, T.Y., Miller, S.L., Aime, M.C., Miller, Jr., O.K. 2006. The mycorrhizal status of Pseudotulostoma volvata (Elaphomycetaceae, Eurotiales, Ascomycota). Mycorrhiza 16(3):241-244.
• Phillips, W., Coutino, A., Ortiz, C.F., Lopez, A.P., Hernandez, J., Aime, M.C. 2006. First report of Moniliophthora roreri causing frosty pod rot (moniliasis disease) of cocoa in Mexico. Plant Pathology 55:584. (Doi: 10. 1111/j.1365-3059.2006.01418.x)
• Scholler, M., Aime, M.C. 2006. On some rust fungi (Uredinales) collected in an Acacia koa-Metrosideros polymorpha woodland, Mauna Loa Road, Big Island, Hawaii. Mycoscience 47:159-165.
• Aime, M.C. 2006. Systematics and molecular variability of bean rusts [abstract]. Annual Report of the Bean Improvement Cooperative 49:45-46.
• Aime, M.C., Phillips-Mora, W. 2006. The causal agents of witches broom and frosty pod rot of cocoa (chocolate, Theobroma cacao) form a new lineage of Marasmiaceae. Mycologia. 97(5):1012-1022.
• Van Rensburg, J.C.J., Lamprecht, S.C., Groenewald, J.Z., Castlebury, L.A., Crous, P.W. 2006. Characterisation of Phomopsis spp. associated with die- back of rooibos (Aspalathus linearis) in South Africa. Studies in Mycology 55:65-74.
• Osterbauer, N., Trippe, A., French, K., Butler, T., Aime, M.C., Bruckart, W., Peerbolt, T., Kaufman, D. 2005. First report of Phragmidium violaceum infecting Himalaya and Evergreen Blackberries in North America. Plant Health Progress doi:10.1094/PHP-2005-0923-01-BR.
• Hernandez, J., Eboh, D.O., Rossman, A.Y. 2005. New Reports of Rust Fungi (Uredinales) from Nigeria. Caldasia. 27(2):213-221.
• Henkel, T.W., Meszaros, R., Aime, M.C., Kennedy, A. 2005. New clavulina species from the pakaraima mountains of guyana. Mycological Progress. 4(4) :343-350.
• Hernandez, J., Aime, M.C., Henkel, T.W. 2005. The rust fungi (uredinales) of guyana. Sydowia. 57(2):189-222.
• Tunali, B., Yildirim, A., Aime, M.C., Hernandez, J., Berner, D. 2005. First report of the rust fungus coleosporium tussilaginis on tussilago farfara in turkey. Plant Disease. 89:1131
• Aime, M.C., Miller, O.K. 2006. Proposal to conserve the name Chroogomphus against Brauniellula (Gomphidiaceae, Agaricales, Basidiomycota). Taxon. 55(1):227-228.
• Bruckart, W.L., Eskandari, F., Littlefield, J., Becktell, M., Aime, M.C., Bean, D., Sands, D., Pilgeram, A. 2006. Puccinia acroptili on russian knapweed in colorado, montana and wyoming. Plant Disease. 90:971.
• Deadman, M.L., Al Sadi, A.M., Al Maqbali, Y.M., Al Jahdhami, S., Patzelt, A., Aime, M.C. 2006. First report of the rust Melampsora euphorbiae on Euphorbia heterophylla in Oman. Journal of Plant Pathology. 88(2):121.
• Tunali, B., Yildirim, A., Aime, M.C., Hernandez, J. 2006. First report of rust disease caused by Uromyces galegae on Galega officinalis in Turkey. Plant Disease. 90:525.
• Hernandez, J.R. 2006. First Report of Puccinia puta on Ipomoea carnea subsp. fistulosa from Puerto Rico. Plant Disease 90(2):245.
• Avanzato, M.V., Borges da Silveira, R.M., Hernandez, J.R. 2006. First report of Ravenelia spegazziniana on Acacia farnesiana from Brazil. Plant Disease 90(8):1109.
• Crozier, J., Thomas, S.E., Aime, M.C., Evans, H.C., Holmes, K.A. 2006. Molecular characterization of endophytic morpho-species isolated from cocoa (Theobroma cacao): a potential source of biocontrol agents? [abstract]. Proceedings of the 1st International Conference on Plant- Microbe Interactions: Endophytes and biocontrol agents, Saariselka, Lapland, Finland, April 18-22.
• Sogonov, M.V., Castlebury, L.A., Mejia, L.C., Rossman, A.Y., White, J. F. 2006. Generic trends in the Gnomoniaceae. [Abstract]. 8th International Mycological Congress, Cairns, Australia, Abstracts Book 1, p. 20.
• Mejia, L. C., Castlebury, L.A., Rossman, A.Y., White, Jr., J.F. 2006. Clarification of the nomenclature and relationships of the genera Cryptosporella, Ophiovalsa and Winterella (Gnomoniaceae) [abstract]. Inoculum 57(4):29.
• Castlebury, L.A., Mengistu, A. 2006. Phylogenetic distinction of Diaporthe/Phomopsis isolates from soybeans[abstract]. Inoculum 57(4)13.
• Hughes, K.W., Mata, J.L., Cifuentes, J., Aime, M.C., Henkel, T., Kovalenko, A., Psurtseva, N., Petersen, R. 2006. The hunt for Megacollybia: One name fits all? [abstract]. Inoculum 57(4):22.
• Vega, F.E., Posada, F., Aime, M.C., Peterson, S.W., Rehner, S.A. 2006. Coffee endophytes [abstract]. Inoculum 57(4):38.
• Henk, D.A., Pastor-Corrales, M., Aime, M.C. 2006. Selection acting on infection specific genes in the common bean rust, Uromyces appendiculatus [abstract]. Inoculum 57(4):20.
• Aime, M. C. 2006. New phylogenies: Revelations on cacao pathogens and the diseases they cause [abstract]. Phytopathology 96:S138.
• Phillips, W., Cawich, J., Garnett, W., Aime, M.C. 2006. First report of frosty pod rot (= moniliasis disease) caused by Moniliophthora roreri on cacao in Belize. Plant Pathology 55:584. (Doi:10.1111/j.1365-3059.2006. 01378.x)

Progress 10/01/04 to 09/30/05

Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Agricultural and forest plants are threatened by fungal pathogens native to the United States and invasive species from other countries. In addition, some disease-causing fungi have potential for use as biocontrol agents for noxious and invasive weeds. This research addresses the classification and characterization, or systematics, of difficult groups of disease-causing fungi specifically bunt fungi of cereals and canker- causing fungi of fruit crops. Accurate knowledge about their classification provides the means for their identification. This knowledge is also essential for making sound plant quarantine decisions to prevent the introduction of new pathogen species and for developing effective disease and weed management strategies. Results of this research include keys for identification and characteristics of fungi that cause bunt and canker diseases as well as invasive and potential biocontrol fungi. The project has two specific objectives: 1) to use molecular and morphological characteristics to develop a phylogeny of bunt fungi and related groups on cultivated grains and wild grasses; 2) to analyze relationships among species of Diaporthe and Phomopsis and the sexual and asexual states of related canker-causing fungi in the chestnut blight order and determine species and species-groups based on morphological and molecular characteristics. The research undertaken falls under National Program 303 - Plant Diseases and addresses Action Plan Component I - Identification and Classification of Pathogens (70%) and National Program 304 - Crop Protection and Quarantine and addresses Action Plan Component IX - Biological Control of Weeds (30%). Economic loss to agricultural and horticultural crops due to disease- causing fungi is estimated at $20 billion per year in the United States. In addition, fungi often limit U.S. agricultural exports, a significant factor in the U.S. balance of trade. Karnal bunt and similar fungi threatened the U.S. wheat export market. This problem was solved using systematic knowledge about Karnal bunt and its close relatives. Inadequate knowledge of the classification and characterization of Phomopsis limits the management of the diseases on fruit and woody crops and causes export-import problems for these commodities. Results of this research are essential for plant pathologists, extension agents, and policy makers in managing diseases, maintaining the crop export market, and preventing the introduction of harmful fungi into the United States. In addition to these specific objectives, an additional objective is to respond to problems arising due to invasive fungi. 2. List the milestones (indicators of progress) from your Project Plan. Year 1 (FY 2003) Development and/or screening of gene primers for T. indica and T. walkeri Molecular and morphological characterization of Phomopsis strains from Vitis in the U.S. and determine if more than one species is involved in Phomopsis canker of grapes Development and/or screening of PCR primers for multiple genes for species-level grouping of Diaporthe-Phomopsis isolates Update online resources with existing information for agriculturally important Diaporthe-Phomopsis spp. and related genera Year 2 (FY 2004) Sequencing of multiple gene regions for T. indica and T. walkeri cultures to determine utility of various genes and regions for identification and phylogenetic analysis of these closely related species of bunt fungi Expand online resources for Tilletia species in North America Sequencing of multiple gene for Diaporthe-Phomopsis strains from soybean and cucurbits to determine species-level groupings and relationships of pathogens and symptomless isolates from these and unrelated hosts Continued updating of online resources with existing information for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 3 (FY 2005) Phylogenetic analysis of multi-gene sequence data for T. indica and T. walkeri isolates to determine relationships of isolates from diverse geographic areas Development of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation is present in available isolates Development and/or screening of PCR primers for multiple protein-coding genes for Tilletia species phylogeny to determine if morphological characters and hosts are indicative of phylogenetic relationships Continued expansion of online resources for Tilletia to all of North America Collection and DNA sequencing for strains of Diaporthe-Phomopsis from juniper and sunflower Continued updating of online resources with existing information for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 4 (FY 2006) Screening and analysis of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation exists with these loci. DNA sequence and morphological characterization of existing cultures for Tilletia species phylogeny Collection and DNA sequencing of Diaporthe-Phomopsis strains from Ericaceae to determine identity of non-Phomopsis vaccinii isolates and their relationships to P. vaccinii Molecular and morphological characterization of new cultures of Diaporthe-Phomopsis species Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 5 (FY 2007) Phylogenetic analysis of multi-gene sequence data for species of Tilletia and related genera in the Tilletiales. Expand online resources for Tilletia to worldwide Characterization of strains of Phomopsis from Rosaceae Phylogenetic analysis of multi-gene sequence data for all available species in the genera Diaporthe and Phomopsis to determine connections and relationships among sexual and asexual species. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales 3a List the milestones that were scheduled to be addressed in FY 2005. For each milestone, indicate the status: fully met, substantially met, or not met. If not met, why. 1. Phylogenetic analysis of multi-gene sequence data for T. indica and T. walkeri isolates to determine relationships of isolates from diverse geographic areas and if significant genetic variation exists in one or both species. Milestone Substantially Met 2. Development of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation is present in available isolates. The focus of this research has switched to distinguishing species of Tilletia currently being found in grass seed and wheat exports that are being confused with dwarf bunt. Milestone Not Met Redirection of Research focus due to change in priorities 3. Development and/or screening of PCR primers for multiple protein-coding genes for Tilletia species phylogeny to determine if morphological characters and hosts are indicative of phylogenetic relationships. Milestone Fully Met 4. Continued expansion of online resources for Tilletia to all of North America. Milestone Fully Met 5. Collection and DNA sequencing for strains of Diaporthe-Phomopsis from juniper and sunflower. Isolates from soybeans continue to be sequenced as unanticipated diversity has been encountered. Milestone Not Met Redirection of Research focus due to change in priorities 6. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales. Milestone Fully Met 3b List the milestones that you expect to address over the next 3 years (FY 2006, 2007, and 2008). What do you expect to accomplish, year by year, over the next 3 years under each milestone? Year 4 (FY 2006) Screening and analysis of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation exists with these loci. DNA sequence and morphological characterization of existing cultures for Tilletia species phylogeny Collection and DNA sequencing of Diaporthe-Phomopsis strains from Ericaceae to determine identity of non-Phomopsis vaccinii isolates and their relationships to P. vaccinii Molecular and morphological characterization of new cultures of Diaporthe-Phomopsis species Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 5 (FY 2007) Phylogenetic analysis of multi-gene sequence data for species of Tilletia and related genera in the Tilletiales. Expand online resources for Tilletia to worldwide Collection and characterization of Diaporthe-Phomopsis strains from fruit crops in the plant family Rosaceae. Phylogenetic analysis of multi-gene sequence data for all available species in the genera Diaporthe and Phomopsis to determine connections and relationships among sexual and asexual species. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales FY 2008 Project expires in 2007. Anticipated milestones for the next project plan include the following: Molecular characterization of rust fungi on rosaceous fruit crops. Molecular characterization of newly encountered disease-causing species of Diaporthe-Phomopsis. Characterization of smut fungi, including Tilletia, Sporisorium and Urocystis on grasses. 4a What was the single most significant accomplishment this past year? A study of diseased grape vines from Ohio, New York and Michigan identified the causal agent as Phomopsis viticola using DNA sequence data. Specialization within host tissue, with some isolates causing more disease in fruits and some causing more disease in leaves, was noted for a number of isolates. Two additional species of Phomopsis were isolated and characterized but caused little or no disease. This information will be used to identify the disease-causing Phomopsis viticola and distinguish it from nondisease-causing species to insure that proper control measures are used. 4b List other significant accomplishments, if any. Pacific madrone, a broadleaf evergreen tree native to western North America, has been suffering a decline for the past thirty years. A fungus has been isolated and verified as the cause of cankers on dying madrone trees. The fungus was determined to be a previously unknown species belonging to a group that commonly causes canker diseases of trees. The fungus causing the canker disease of madrone was characterized, described, illustrated, and named as a new species. Evidence suggests that this fungus has been present in western North America since 1968. Prior to this research, plant pathologists had misidentified the causal fungus. This research allows forest pathologists to accurately determine the cause of this disease in order to develop control measures. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. A study of diseased grape vines from Ohio, New York and Michigan identified the causal agent as Phomopsis viticola using DNA sequence data. Specialization within host tissue, with some isolates causing more disease in fruits and some causing more disease in leaves, was noted for a number of isolates. Two additional species of Phomopsis were isolated and characterized but caused little or no disease. This information will be used to identify the disease-causing Phomopsis viticola and distinguish it from nondisease-causing species to insure that proper control measures are used. Pacific madrone, a broadleaf evergreen tree native to western North America, has been suffering a decline for the past thirty years. A fungus has been isolated and verified as the cause of cankers on dying madrone trees. The fungus was determined to be a previously unknown species belonging to a group that commonly causes canker diseases of trees. The fungus causing the canker disease of madrone was characterized, described, illustrated, and named as a new species. Evidence suggests that this fungus has been present in western North America since 1968. Prior to this research, plant pathologists had misidentified the causal fungus. This research allows forest pathologists to accurately determine the cause of this disease in order to develop control measures. A rust pathogen was discovered on safflower (Carthamus tinctorius), an important crop in Oman. Morphological and DNA analyses identified the causal agent as Puccinia carthami, one of a complex of rust species infecting members of the Cardueae, which also includes plants such as artichokes and thistles. Additionally, DNA sequence data has identified several host-specific rust species in this group. This information will be used by plant pathologists to identify disease-causing Puccinia species on beneficial Cardueae and to identify potential biocontrol strains for invasive species of thistles and related plants. Confusion about relationships of bunt fungi on cereal crops has been cleared up. A bunt fungus called Neovossia iowensis was thought to be a close relative of the Karnal bunt and rice kernel smut fungi. Research showed that the Neovossia bunt fungus is more closely related to wild- grass infecting Tilletia species than to Karnal bunt or rice kernel smut. This research also confirms that Karnal bunt and rice kernel smut are true bunt fungi in the genus Tilletia. This information will be used by plant pathologists to correctly identify these disease-causing fungi. In addition, this research will ensure that plant quarantine policy is effective at preventing the entry of fungi that most directly threaten U. S. agriculture. Karnal bunt is a quarantine significant disease of wheat that has not been clearly defined nor has it been distinguished from closely related fungi. DNA sequence data from intron regions from protein-coding genes provided additional molecular support for the distinction of Tilletia indica and Tilletia walkeri as separate species. Relationships of a number of wild-grass infecting Tilletia species will be resolved with reference to the wheat bunts. This information will aid in the correct identification of these important quarantine fungi and their morphologically similar relatives and the development of KB resistant wheat. Bunt fungi cause diseases of cereal crops. Being able to recognize and distinguish the various bunt fungi is essential for effective plant quarantine as well as breeding resistance to these fungi. Molecular techniques such as sequencing multiple genes will be used to characterize the bunt fungi worldwide. This research will be used by plant quarantine officials to determine which cereal crops can be safely allowed entry into the U.S. and to prevent the entry of invasive fungi. A study of canker-causing fungi using DNA sequence data revealed that a species on celery-relatives, Diaporthopsis angelicae, was a species of Diaporthe. It was determined that this species is closely related to a serious pathogen of sunflowers, Diaporthe helianthi, and a seedborne soybean pathogen, Phomopsis longicolla. This information will be useful for distinguishing disease-causing from harmless species of Diaporthe and its asexual state Phomopsis as well as for developing breeding strategies for disease resistance to the soybean pathogen. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? A shipment of wheat from the United States (Minnesota and North Dakota) valued at $4.4 million was being held at the port of entry in Algeria because of the alleged presence of Karnal bunt (Tilletia indica). Dr. Lisa Castlebury, an expert systematist in bunt fungi, traveled to Algeria to work with the Algerian plant pathologists. Dr. Castlebury reviewed slides prepared by the lab and concluded that Karnal bunt was not present. She explained to Algerian lab technicians and administrators why the specimens were not Karnal bunt. As a result of her visit, this shipment was released and the export of wheat from the U.S. to Algeria valued at $80 million remains open. An interactive key for the identification of teliospores of bunt fungi in the United States is available over the Web at http://nt.ars-grin. gov/TaxaDescriptions/Tilletia/Index2.cfm. This system allows non- specialists such as plant quarantine officials and extension agents to rapidly identify these fungi. A resource page for systematics of the Diaporthales is available at http://nt.ars-grin. gov/taxadescriptions/diaporthales/HomePage/Intro.cfm. This information summarizes current family level relationships of fungi in this order, including Diaporthe-Phomopsis, dogwood anthracnose, and chestnut blight. Additionally DNA sequences used to analyze relationships of or identify plant pathogens have been submitted to GenBank, a publicly available database, for use by other researchers. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Fungal Threats to Cocoa Share an Ancient History, Agriculture Research Magazine, Feb. 2005. Article about research by Dr. C. Aime. Correct Identification of Fungus Saves Wheat Export Market. Article about research of Dr. L. Castlebury published in: Davies, H., N. King and R. Smith (eds). 2004. Taxonomy: targeting invasives. BioNet International. Also, http://www.bionet-intl.org/case_studies/case8.htm Systematics of Invasive Fungi and the U.S. National Fungus Collections (BPI). Invited presentation by Dr. A. Rossman at Harvard University, November 5, 2004. How to identify mushrooms 101. Presentation by Dr. C. Aime to Mycological Association of Washington, Bethesda, MD on June 7, 2005.

Impacts
(N/A)

Publications

• Hernandez, J.R., Aime, M.C., Newbry B. 2004. Aecidium kalanchoe, a new species of rust on Kalanchoe blossfeldiana (Crassulaceae). Mycological Research 108(7):846-848.
• Schneider, W., Hollier, C.A., Whitam, H.K., Palm, M.E., Mckemy, J.M., Hernandez, J., Levy, L., Devries-Paterson, R. 2005. First report of soybean rust caused by Phakopsora pachyrhizi in the continental United States. Plant Disease. 89:774.
• Kapooria, R.G. Aime, M.C. 2005. First report of Olivea scitula on Vitex doniana in Zambia. Plant Disease 89:431.
• Deadman, M.L., Al Saadi, A.M., Al Johdami, S., Aime, M.C. 2004. First report of rust on safflower in Oman. Plant Disease 89:208.
• Gams, W., Rossman, A.Y. 2005. What is Oospora oryzetorum Sacc.? Mycotaxon 92:339-340.
• Berner, D.K., Eskandari, F., Rossman, A.Y., Aime, M.C., Kashefi, J. 2004. First report of anthracnose of crupina vulgaris caused by colletotrichum cf. gloeosporioides. Plant Disease. 88:1161.
• Aime, M.C., Vilgalys, R. and Miller, O.K. 2005. The Crepidotaceae (Basidiomycota, Agaricales): phylogeny and taxonomy of the genera and revision of the family based on molecular evidence. American Journal of Botany 92(1):74-82.
• Erincik, O., Castlebury, L.A., Schilder, A., Rossman, A.Y., Ellis, M.A. 2005. Characterization of Phomopsis spp. infecting grapesvines in the Great Lakes region of North America. Plant Disease 89:755-762.
• Farr, D.F., Elliott, M., Rossman, A.Y., Edmonds, R. 2005. Fusicoccum arbuti sp. nov. causing cankers on Pacific madrone in western North America with notes on Fusicoccum dimidiatum, the correct name for Scytalidium dimidiatun and Nattrassia mangiferae. Mycologia. 97(3):730-741.
• Rossman, A.Y., Aime, M.C., Farr, D.F., Castlebury, L.A., Peterson, K., Leahy, R. 2004. The coelomycetous genera Chaetomella and Pilidium, a newly discovered lineage of inoperculate discomycetes. Mycological Progress. 3(4) :275-290.
• Castlebury, L.A., Rossman, A.Y., Sheer Hyten, A.M., Sung, G., Spatafora, J. W. 2004. Multigene phylogeny reveals new lineage for Stachybotrys chartarum, the indoor air fungus. Mycol. Res. 108: 864-872.
• Palm, M.E., Rossman, A.Y. Invasion pathways of terrestrial plant- inhabiting fungi. 2004. Invasive Plants Global Issues Local Challenges. p. 31-43
• Ripkova, S., Aime, M.C., Lizon, P. 2005. Crepidotus crocophyllus includes C. nephrodes. Mycotaxon. 91:397-403.
• Henkel, T.W., Aime, M.C., Chin, M., Andrew, C. 2004. Edible mushrooms from Guyana's Pakaraima Mountains. The Mycologist 18(3):104-110.
• Sun, H.J., Depriest, P.T., Gargas, A., Rossman, A.Y., Friedman, I. 2003. Pestalotiopsis maculans is a parasymbiontic fungus in North American lichens. Symbiosis 33(2):215-226.
• Lutzoni, F., Kauff, F., Cox, C.J., Rossman, A.Y. 2004. Assembling the fungal tree of life: progress, classification, and evolution of subcellular traits? American Journal of Botany 91(10):1446-1480.
• Aime, M.C., Henkel, T.W. 2004. Ectomycorrhizal and saprotrophic fungal diversity in western guyana [Abstract]. Proceedings of the Association for Tropical Biology and Conservation, July 2004, Miami, FL, pp 38-39.
• Van Niekerk, J., Groenwald, J., Farr, D.F., Fourie, P., Halleen, F., Crous, P. 2005. Reassessment of Phomopsis species on grapevines. Australasian Plant Pathlogy Society. 34(1):27-39
• Krupinsky, J.M., Berdahl, J.D., Schoch, C.L., Rossman, A.Y. 2004. A new leaf spot disease on switchgrass (panicum virgatum)caused by bipolaris oryzae. Canadian Journal of Plant Pathology 26:371-378.
• Kennedy, A., Aime, M.C. 2005. Cultural studies of Moniliopthora roreri, an important cacao pathogen. Inoculum. 56(4):30-31.
• Bennett, C., Newcombe, G., Aime, M.C. 2005. Regional studies of Melampsora on Salix in the Pacific Northwest. Inoculum. 56(4):9.
• Aime, M.C., Henkel, T.W. 2005. Strategies for bio-inventory: Lessons from Guyana. Inoculum. 56(4):6.
• Aime, M.C., Chin, M., Andrew, C. 2005. Mycophagy of the Patamona Amerindians of Western Guyana. Inoculum. 56(4):24-25.
• Henkel, T.W., James, T.Y., Miller, S.L., Aime, M.C., Miller, Jr., O.K. 2005. The mycorrhizal status of Pseudotulostoma volvata (Elaphomycetaceae, Eurotiales, Ascomycota). Inoculum. 56(4):25.
• Carris, L.M., Castlebury, L.A. 2005. Identification of species of Tilletia in U.S. wheat and grass seed exports. Inoculum. 56:12.
• Sogonov, M., Castlebury, L.A., Rossman, A.Y., White, Jr., J.F. 2005. Towards monophyletic genera in the Gnomoniaceae. Inoculum. 56:54.
• Aime, M.C. 2005. Molecular systematics of Uredinales. Inoculum. 56(4):6.
• Castlebury, L.A. 2005. The Diaporthe vaccinii complex of fruit pathogens. Inoculum. 56:12.
• Mayor, J.R., Henkel, T.W., Aime, M.C. 2005. Ectomycorrhizae accelerate calcium acquisition from leaf litter in a monodominant Dicymbe corymbosa (Caesalpiniaceae) forest in Guyana. Inoculum. 56(4):40.

Progress 10/01/03 to 09/30/04

Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Agricultural and forest plants are threatened by fungal pathogens native to the United States and invasive species from other countries. In addition, some disease-causing fungi have potential for use as biocontrol agents for noxious and invasive weeds. This research addresses the classification and characterization, or systematics, of difficult groups of disease-causing fungi specifically bunt fungi of cereals and canker- causing fungi of fruit crops. Accurate knowledge about their classification provides the means for their identification. This knowledge is also essential for making sound plant quarantine decisions to prevent the introduction of new pathogen species and for developing effective disease and weed management strategies. Results of this research include keys for identification and characteristics of fungi that cause bunt and canker diseases as well as invasive and potential biocontrol fungi. The project has two specific objectives: 1) to use molecular and morphological characteristics to develop a phylogeny of bunt fungi and related groups on cultivated grains and wild grasses; 2) to analyze relationships among species of Diaporthe and Phomopsis and the sexual and asexual states of related canker-causing fungi in the chestnut blight order and determine species and species-groups based on morphological and molecular characteristics. The research undertaken falls under National Program 303 - Plant Diseases and addresses Action Plan Component I - Identification and Classification of Pathogens (70%) and National Program 304 - Crop Protection and Quarantine and addresses Action Plan Component IX - Biological Control of Weeds (30%). Economic loss to agricultural and horticultural crops due to disease- causing fungi is estimated at $20 billion per year in the United States. In addition, fungi often limit U.S. agricultural exports, a significant factor in the U.S. balance of trade. Karnal bunt and similar fungi threatened the U.S. wheat export market. This problem was solved using systematic knowledge about Karnal bunt and its close relatives. Inadequate knowledge of the classification and characterization of Phomopsis limits the management of the diseases on fruit and woody crops and causes export-import problems for these commodities. Results of this research are essential for plant pathologists, extension agents, and policy makers in managing diseases, maintaining the crop export market, and preventing the introduction of harmful fungi into the United States. In addition to these specific objectives, an additional objective is to respond to problems arising due to invasive fungi. 2. List the milestones (indicators of progress) from your Project Plan. Year 1 (FY 2003) Development and/or screening of gene primers for T. indica and T. walkeri Molecular and morphological characterization of Phomopsis strains from Vitis in the U.S. and determine if more than one species is involved in Phomopsis canker of grapes Development and/or screening of PCR primers for multiple genes for species-level grouping of Diaporthe-Phomopsis isolates Update online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera Year 2 (FY 2004) Sequencing of multiple gene regions for T. indica and T. walkeri cultures to determine utility of various genes and regions for identification and phylogenetic analysis of these closely related species of bunt fungi Expand online resources for Tilletia species in North America Sequencing of multiple gene for Diaporthe-Phomopsis strains from soybean and cucurbits to determine species-level groupings and relationships of pathogens and symptomless isolates from these and unrelated hosts Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 3 (FY 2005) Phylogenetic analysis of multi-gene sequence data for T. indica and T. walkeri isolates to determine relationships of isolates from diverse geographic areas Development of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation is present in available isolates Development and/or screening of PCR primers for multiple protein-coding genes for Tilletia species phylogeny to determine if morphological characters and hosts are indicative of phylogenetic relationships Continued expansion of online resources for Tilletia to all of North America Collection and DNA sequencing for strains of Diaporthe-Phomopsis from juniper and sunflower Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 4 (FY 2006) Screening and analysis of microsatellite primers and loci or other fragment-based molecular data for T. indica and T. walkeri to determine if geographic variation exists with these loci. DNA sequence and morphological characterization of existing cultures for Tilletia species phylogeny Expansion of online resources for Tilletia to worldwide Collection and DNA sequencing of Diaporthe-Phomopsis strains from Ericaceae to determine identity of non-Phomopsis vaccinii isolates and their relationships to P. vaccinii Molecular and morphological characterization of new cultures of Diaporthe-Phomopsis species Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales Year 5 (FY 2007) Phylogenetic analysis of multi-gene sequence data for species of Tilletia and related genera in the Tilletiales. Expand online resources for Tilletia to worldwide Characterization of strains from Rosaceae Phylogenetic analysis of multi-gene sequence data for all available species in the genera Diaporthe and Phomopsis to determine connections and relationships among sexual and asexual species. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales 3. Milestones: The milestones listed below were scheduled to be completed. All milestones were completed or substantially met. Sequencing of multiple gene regions for T. indica and T. walkeri cultures to determine utility of various genes and regions for identification and phylogenetic analysis of these closely related species of bunt fungi. Expand online resources for Tilletia species in North America. Sequencing of multiple gene for Diaporthe-Phomopsis strains from soybean and cucurbits to determine species-level groupings and relationships of pathogens and symptomless isolates from these and unrelated hosts. Continued updating of online resources with existing info for agriculturally important Diaporthe-Phomopsis spp. and related genera in the Diaporthales 4. What were the most significant accomplishments this past year? Confusion about relationships of bunt fungi on cereal crops has been cleared up. A bunt fungus called Neovossia iowensis was thought to be a close relative of the Karnal bunt and rice kernel smut fungi. Research showed that the Neovossia bunt fungus is more closely related to wild- grass infecting Tilletia species than to Karnal bunt or rice kernel smut. This research also confirms that Karnal bunt and rice kernel smut are true bunt fungi in the genus Tilletia. This information will be used by plant pathologists to correctly identify these disease-causing fungi. In addition, this research will ensure that plant quarantine policy is effective at preventing the entry of fungi that most directly threaten U. S. agriculture. B. Other Significant Accomplishments: A study of canker-causing fungi using DNA sequence data revealed that a species on celery-relatives, Diaporthopsis angelicae, was a species of Diaporthe. It was determined that this species is closely related to a serious pathogen of sunflowers, Diaporthe helianthi, and a seedborne soybean pathogen, Phomopsis longicolla. This information will be useful for distinguishing disease-causing from harmless species of Diaporthe and its asexual state Phomopsis as well as for developing breeding strategies for disease resistance to the soybean pathogen. C. Significant Accomplishments/Activities that Support Special Target Populations: None. D. Progress report opportunity to submit additional programmatic information to your Area Office and NPS: None. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Confusion about relationships of bunt fungi on cereal crops has been cleared up. A bunt fungus called Neovossia iowensis was thought to be a close relative of the Karnal bunt and rice kernel smut fungi. Research showed that the Neovossia bunt fungus is more closely related to wild- grass infecting Tilletia species than to Karnal bunt or rice kernel smut. This research also confirms that Karnal bunt and rice kernel smut are true bunt fungi in the genus Tilletia. This information will be used by plant pathologists to correctly identify these disease-causing fungi. In addition, this research will ensure that plant quarantine policy is effective at preventing the entry of fungi that most directly threaten U. S. agriculture. Karnal bunt is a quarantine significant disease of wheat that has not been clearly defined nor has it been distinguished from closely related fungi. DNA sequence data from intron regions from protein-coding genes provided additional molecular support for the distinction of Tilletia indica and Tilletia walkeri as separate species. Relationships of a number of wild-grass infecting Tilletia species will be resolved with reference to the wheat bunts. This information will aid in the correct identification of these important quarantine fungi and their morphologically similar relatives and the development of KB resistant wheat. Bunt fungi cause diseases of cereal crops. Being able to recognize and distinguish the various bunt fungi is essential for effective plant quarantine as well as breeding resistance to these fungi. Molecular techniques such as sequencing multiple genes will be used to characterize the bunt fungi worldwide. This research will be used by plant quarantine officials to determine which cereal crops can be safely allowed entry into the U.S. and to prevent the entry of invasive fungi. An on-line identification system for bunt fungi is needed so that plant pathologists throughout the world can readily identify these fungi. At present an identification system exists for bunt fungi in the U.S. based on teliospores. This system will be expanded to include additional characters including sequence data. Canker diseases in the genus Diaporthe-Phomopsis are caused by fungi that are difficult to identify. Some of these fungi especially those on soybean are of plant quarantine significance. Species of canker-causing fungi will be characterized and distinguished using both morphological and molecular characteristics. Fungi on agriculturally important crops such as soybeans, grapes and melons will be studied initially. This will be extended to junipers, sunflowers, blueberries and its relatives and tree fruits. Based on the accurate characterization of these canker- causing fungi, plant pathologists will be able to rapidly identify these fungi especially using molecular characteristics. A study of canker-causing fungi using DNA sequence data revealed that a species on celery-relatives, Diaporthopsis angelicae, was a species of Diaporthe. It was determined that this species is closely related to a serious pathogen of sunflowers, Diaporthe helianthi, and a seedborne soybean pathogen, Phomopsis longicolla. This information will be useful for distinguishing disease-causing from harmless species of Diaporthe and its asexual state Phomopsis as well as for developing breeding strategies for disease resistance to the soybean pathogen. An on-line identification resource will be developed to allow users to rapidly identify canker-causing and related fungi. This resource will include descriptions, illustrations, accurate scientific names, host and geographic ranges as well as signature molecular sequence data. This system will allow plant pathologists and plant quarantine officials to determine which fungi are serious pathogens of crops in the U.S. in order to prevent their entry. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? An interactive key for the identification of teliospores of bunt fungi in the United States is available over the Web at http://nt.ars-grin. gov/TaxaDescriptions/Tilletia/Index2.cfm. This system allows non- specialists such as plant quarantine officials and extension agents to rapidly identify these fungi. A resource page for systematics of the Diaporthales is available at http://nt.ars-grin. gov/taxadescriptions/diaporthales/HomePage/Intro.cfm. This information summarizes current family level relationships of fungi in this order, including Diaporthe-Phomopsis, dogwood anthracnose, and chestnut blight. Additionally DNA sequences used to analyze relationships of or identify plant pathogens have been submitted to GenBank, a publicly available database, for use by other researchers. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. Peabody, E. March 2004. Mycologists Help Guard U.S. Borders. Agricultural Research Magazine. Peabody, E. March 10, 2004. ARS Scientists Help Experts Recognize Foreign Fungi. ARS News Service. Peabody, E. June 18, 2004. Indoor "Black Mold" Fungus Has Unexpected Relatives. ARS News Service. Ellison, Robert. 2004. Interviewed and filmed for AgDay Television Program. USDA Office of Communications.

Impacts
(N/A)

Publications

• Elliott, M., Rossman, A.Y., Farr, D.F., Edmonds, R.L. A new species of fusicoccum causing a canker disease of pacific madrone. Phytopathology.Vol. 94(6)P.527,2004.
• Rossman, A.Y., Aime, M.C., Castlebury, L.A., Farr, D.F. Revealing undiscovered lineages in the ascomycetes. Inoculum. Vol.55 p.33,2004.
• Rossman, A.Y. Chester ray benjamin, 1923-2002. Mycologia. Vol.95(3)pp.557- 558,2003.
• Gregory, N.F., Mulrooney, R.P., Rossman, A.Y., Castlebury, L.A. First report of discula fraxinea on chinese fringe tree in the united states and on white ash in delaware. Plant Disease.Vol. 88(4),P.427,2004.
• Rossman, A.Y., Castlebury, L.A., Putnam, M. Ash anthracnose caused by discula fraxinea reported from oregon. Plant Disease. Vol. 88(2)p.222,2004.
• Rossman, A.Y., Farr, D.F. Dwiroopa, a coelomycetous genus with two species. Mycoscience. Vol.44(6)pp.443-446,2003. Vol.44(6)pp.443-446,2003.
• Rossman, A.Y., Schauff, M.E. 2004. Systematics and biological pest control. Encyclopedia of Pest Management.
• Sogonov, M.V., Castlebury, L.A., Farr, D.F., White, J.F. 2004. The taxonomy of the genus gnomonia: distinction between g. gnomon and g. setacea. Inoculum. v. 55 (4) p. 35.
• Pastor-Corrales, M.A., Aime, M.C. 2004. Differential cultivars and molecular markers reveal two distinct groups of isolates of uromyces appendiculatus, that correspond to the andean and middle american gene pools of their host.. Phytopathology. v. 95 p. S82.
• Pastor-Corrales, M.A., Steadman, J.R., Aime, M.C. 2004. Host gene-pool specialization and species concepts in uromyces appendiculatus and other pathogens of common bean. Phytopathology. v. 95 p. S124.
• Aime, M. C., Pastor-Corrales, M.A. 2004. Host gene specialization in Uromyces appendiculatus [abstract]. Inoculum. 55 (4) p.6
• Aime, M.C. 2004. Intercompatibility tests and phylogenetic analysis in the crepidotus sphaerula group complex: concordance between icgs and nuclear rdna sequences highlight phenotypic plasticity within appalachian species. Fungi in Forest Ecosystems: Diversity, Systematics, and Ecology. pp. 71-80.
• Henkle, T.W., Roberts, P., Aime, M.C. 2004. Sebacinoid species from the pakaraima mountains of guyana. Mycotaxon. v.89(2) pp.433-439.2004

Progress 10/01/02 to 09/30/03

Outputs
1. What major problem or issue is being resolved and how are you resolving it? Agricultural and forest plants are threatened by fungal pathogens native to the United States as well as invasive fungi from other countries. In addition, the export of agricultural crops from the United States to other countries may be influenced by the presence/absence of fungi of plant quarantine importance. This research uses molecular and morphological approaches to classify and characterize difficult groups of disease-causing fungi. The fungi to be studied are the bunt fungi of cereals and canker fungi that attack a wide range of crop plants. Knowledge about their characterization and classification is essential for accurate identification and disease control. The objectives of this project are to develop a phylogeny of bunt fungi and related groups on cultivated grains and wild grasses. Species of Diaporthe and Phomopsis, a group of canker-causing fungi, will be defined based on morphological and molecular characteristics. The results of this research will be to define and characterize these fungi in order to accurately identify disease and quarantine pests and to control the diseases they cause. The on-line, interactive identification system for Tilletia will be expanded and one will be developed for Phomopsis-Diaporthe and related canker-causing fungi. This will allow rapid identifications based on unusual morphological or cultural characteristics as well as DNA sequences. This knowledge is critical for making sound plant quarantine decisions to prevent the introduction of new plant pathogens and for developing effective disease management strategies. 2. How serious is the problem? Why does it matter? Economic loss to agricultural and horticultural crops due to disease- causing fungi is estimated at $20 billion per year in the United States. In addition, fungi often limit U.S. agricultural exports, a significant factor in the U.S. balance of trade. The wheat export market was threatened by Karnal bunt and similar fungi. This problem was solved using systematic knowledge about Karnal bunt and its close relatives. Inadequate knowledge of the classification and characterization of Phomopsis limits the management of the diseases these fungi cause on fruit and woody crops and causes export-import problems for these commodities. Results of this research are directly relevant to plant pathologists, extension agents, and policy makers, helping them to better manage diseases, maintain or expand the crop export market, and prevent the introduction of harmful fungi into the United States. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? This research contributes to control of plant diseases as part of National Program 303, Plant Diseases (100%) in the program component on Identification and Classification of Pathogens. This research allows scientists to accurately identify plant pathogenic fungi and develop effective disease management strategies. The crops most directly affected by this research are wheat and other grain crops, soybeans, tree fruits such as peaches, pears and citrus, and small fruits including cranberries, blueberries, and melons. 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2003 year: A new, unknown invasive fungus was threatening Christmas trees grown in the midwestern United States. Using molecular and morphological approaches scientists at the Systematic Botany and Mycology Laboratory, Beltsville, Maryland, identified this fungus as a species identified known only from France. They provided a modern description, illustrations, accurate host and geographic range, and determined the close relatives of the new pathogen. This research allows plant pathologists and tree breeders to manage this disease and breed for resistance to provide healthy Christmas trees at a reasonable cost. B. Other Significant Accomplishments: Phylogenetic analysis revealed that Diapothopsis angelicae, a weed- inhabiting fungus, is closely related to Diaporthe helianthi, a serious pathogen of sunflowers, and Phomopsis longicolla, a seedborne soybean pathogen. This research will be useful for identification of both pathogenic and non-pathogenic species of Diaporthe and its asexual state Phomopsis. It will also be used to determine strategies for breeding disease resistance and making accurate quarantine decisions that facilitate commodity exchange while safeguarding U.S. agriculture. C. Significant Accomplishments/Activities that Support Special Target Populations: None. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Species relationships were determined among a subgroup of bunt fungi (Tilletia spp.) infecting grasses in the subfamily Pooideae. DNA sequence data from two protein coding genes provided additional molecular support for the distinction of Tilletia indica and Tilletia walkeri as separate species. The relationship of wild grass-infecting Tilletia species to wheat bunts was resolved. This research will aid in the correct identification of these important quarantine fungi and their morphologically similar relatives. It will also be used to determine strategies for breeding disease resistance and making accurate quarantine decisions that facilitate commodity exchange while safeguarding U.S. agriculture. 6. What do you expect to accomplish, year by year, over the next 3 years? FY 2004: Development of PCR primers and DNA sequencing of isolates of Tilletia indica (Karnal bunt) and related bunt species will be completed following the development and application of gene specific primers. Phomopsis on soybeans and melons will be accurately defined using both morphological and molecular characteristics based on multiple genes. Expansion of existing on-line resources for both Tilletia and Phomopsis known from North America will continue. FY 2005: Molecular characteristics will be used to define species of Tilletia including kernel smut of rice and related grasses. Isolates of Phomopsis on Juniperus and sunflower will be related to reference names in the sexual state genus Diaporthe. Expansion of existing on-line resources for both Tilletia and Phomopsis will continue. FY 2006: The genetic variability of the Karnal bunt fungus will be determined using microsatellite markers. Phomopsis on grapes and other fruit crops will be accurately defined using both morphological and molecular characteristics based on multiple genes. Existing on-line resources for Tilletia and Phomopsis will be expanded for world-wide species. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Talks were presented to daylily growers groups about daylily rust, its introduction, management and control in Pennsylvania and Maryland. 8. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: This does not replace your peer-reviewed publications listed below). Castlebury, L., Rossman, A. Molecular markers for plant pathogenic and biocontrol fungi. 2003. April 15. Poster for BARC Technology Showcase.

Impacts
(N/A)

Publications

• Rossman, A.Y., Farr, D.F., Castlebury, L.A., Shoemaker, R., Mengistu, A. Setomelanomma holmii (Pleosporales, Phaeosphaeriaceae) on living spruce twigs in Europe and North America. Canadian Journal of Botany. 2002. v. 80. p.1209-1215.
• Camara, M.P.S., Ramaley, A.W., Castlebury, L.A.,Palm, M.E. Neosphaeria and Phaeosphaeriopsis, segregates of Paraphaeosphaeria. Mycological Research. 2003. v. 107. p. 516-522.
• Castlebury, L.A., Rossman, A.Y., Farr, D.F. Phylogenetic placement of the anamorph genus Sirococcus. Inoculum. 2003. v. 54. Abstract p. 15.
• Carpenter-Boggs, L., Carris, L.M., Castlebury, L.A. Multi-locus phylogenetic analysis of Tilletia species infecting hosts in grass subfamily Pooideae. Inoculum. 2003. v. 54. Abstract p. 15.
• Castlebury, L.A., Farr, D.F., Rossman, A.Y., Jaklitsch, W. Diaporthe angelicae, comb. nov., a modern description and placement of Diaporthopsis in Diaporthe. Mycoscience. 2003. v. 44. p. 203-208.