349 CONTENTS Abstract …………………………………………………………..…………….………… 353 1. Introduction ……………………………………………………….……….…….. 354 2. History of the Cryphonectriaceae and its members ……………………............... 356 2.1 Early taxonomic history of Cryphonectria and Endothia ……….............. 356 2.2 Typification of Cryphonectria …………………………………………… 357 2.3 Newly described species and genera …………………………………….. 358 2.4 Description of a new family ………………………………………............ 359 2.5 Anamorph states …………………………………………………………. 360 3. Diseases and ecology ……………………..………………………………............. 361 3.1 Chestnut blight ……………………………………………………............ 362 3.1.1 The disease ………………………………………………………. 362 3.1.2 Etiology ……….…………………………………………………. 366 3.1.3 Control ……………………………………………………........... 366 3.1.4 Population genetics ……………………………………………… 374 3.2 Chrysoporthe canker of Eucalyptus ………………………………............ 376 3.2.1 The disease ………………………………………………………. 376 3.2.2 Control ……………………………………………………........... 378 3.2.3 Hosts other than Eucalyptus …………………………………….. 380 3.2.4 Population genetics ……………………………………………… 383 3.2.5 Other species in Chrysoporthe ……………………………........... 386 3.3 Canker of Eucalyptus caused by Holocryphia eucalypti …………............ 387 3.4 Canker caused by Microthia havanensis …………………………............ 388 3.5 Die-back caused by Aurapex penicillata …………………………............ 389 3.6 Cryphonectria gyrosa stem canker ………..………………………............ 390 3.7 Cankers on Eucalyptus caused by Cryphonectria spp. …………………... 391 3.8 Celoporthe canker ………………………………………………………... 391 3.9 Canker of Terminalia caused by Rostraureum tropicale …………............ 392 350 3.10 Root cankers on Elaeocarpus ……………………………………………. 393 3.11 Endothia canker or pin oak blight ………………………………………... 393 4. Pathogen identification …………………………………………………………… 394 5. Working with the Cryphonectriaceae …………………………………………….. 408 5.1 Isolates and specimens …………………..…….......................................... 408 5.1.1 Observations in the field ...………………………………………. 408 5.1.2 Isolations from bark ………………………………………........... 409 5.1.3 Growth in culture …………………………………………........... 411 5.1.4 Morphological studies …………………………………………... 413 5.1.5 Microscopic examination of herbarium specimens ……………... 413 5.1.6 Colony growth …………………………………………………... 415 5.2 Phylogenetic circumscription through DNA sequence comparisons ……. 417 5.2.1 DNA sequence comparisons …………………………………….. 417 5.2.2 Restriction Length Polymorphisms (RFLPs) ……………………. 419 5.2.3 Methodology …………………………………………………….. 420 6. Identification of genera and species ………………………………………............. 424 6.1 Morphological characteristics and terminoloy…………………………… 424 6.1.1 Stromata …………………………………………………………. 424 ` 6.1.2 Ascostromata ……………………………………………………. 425 6.1.3 Conidiomata ………………………………………………........... 425 6.1.4 Occurrence of the teleomorph and anamorph ...…………………. 427 6.1.5 Spore morphology ………………………………………………. 427 6.1.6 Conidiophores and paraphyses ………………………………….. 428 6.1.7 Morphological characteristics used for family, generic and species delimitation ………………………………………………………………. 428 6.2 Identification using phylogenetic analyses.………..................................... 438 6.2.1 Materials and methods ……………………………………........... 438 6.2.2 Results of the LSU sequence analysis …………………………... 440 351 6.2.3 Results of the ITS/β-tubulin sequence analysis ……………......... 441 6.3 Dichotomous key to genera .……....…………………..……..…………... 449 6.4 Synoptic key to genera ………………………..……………….................. 450 6.5 Descriptions ……………………………………………..……………….. 453 Cryphonectriaceae Gryzenh. & M.J. Wingf. ...…………………………................ 453 Cryphonectria (Sacc.) Sacc. .………………………………..…………................. 454 Endothiella Sacc. gen. anam., emend. Gryzenh. & M.J. Wingf. ............................ 455 1. Cryphonectria parasitica (Murrill) M.E. Barr .....…......………................. 458 2. Cryphonectria radicalis (Schwein.: Fr.) Fr. ……..……………................. 462 3. Cryphonectria japonica (Tak. Kobay. & Kaz. Itô) Gryzenh. & M.J. Wingf., comb. nov. ……………………………….....……………......................... 469 4. Cryphonectria macrospora (Tak. Kobay. & Kaz. Itô) M.E. Barr ….......... 475 Microthia Gryzenh. & M.J. Wingf. ...…………………………..…..…………….. 479 5. Microthia havanensis (Bruner) Gryzenh. & M.J. Wingf. ........................... 481 6. Microthia coccolobae (Vizioli) Gryzenh. & M.J. Wingf. ..…….………... 486 Endothia Fr. ...…………………………………………………………………….. 490 7. Endothia gyrosa (Schwein.: Fr.) Fr. ..……..………………….………….. 491 8. Endothia singularis (Syd. & P. Syd.) Shear & N.E. Stevens .…………… 497 Holocryphia Gryzenh. & M.J. Wingf. ...………………...………..…..…………... 501 9. Holocryphia eucalypti (M. Venter & M.J. Wingf.) Gryzenh. & M.J. Wingf. ………………………………………………………...………………….. 502 Amphilogia Gryzenh., Glen & M.J. Wingf. ...…………………………...………... 506 10. Amphilogia gyrosa (Berk. & Broome) Gryzenh., Glen & M.J. Wingf. …………………………………………………………..………............... 507 11. Amphilogia major Gryzenh., Glen & M.J. Wingf. ..…………................... 512 Rostraureum Gryzenh. & M.J. Wingf. .....………………………………............... 515 12. Rostraureum tropicale Gryzenh. & M.J. Wingf. ...……............................. 516 13. Rostraureum longirostre (Earle) Gryzenh. & M.J. Wingf. ........................ 520 352 Ursicollum Gryzenh. & M.J. Wingf. …………...……...……………..................... 523 14. U. fallax Gryzenh. & M.J. Wingf. ...….……………………....………….. 524 Aurapex Gryzenh. & M.J. Wingf. ...………………………………….....………… 527 15. Aurapex penicillata Gryzenh. & M.J. Wingf. ...……………….…………. 528 Chrysoporthe Gryzenh. & M.J. Wingf. ...……………............................................ 532 Chrysoporthella Gryzenh. & M.J. Wingf. anam. gen. ……………………………. 534 16. Chrysoporthe cubensis (Bruner) Gryzenh. & M.J. Wingf. ………............. 539 17. Chrysoporthe austroafricana Gryzenh. & M.J. Wingf. …......................... 546 18. Chrysoporthe doradensis Gryzenh. & M.J. Wingf. ……….……............... 551 19. Chrysoporthe inopina Gryzenh. & M.J. Wingf. ……………..................... 555 20. Chrysoporthella hodgesiana Gryzenh. & M.J. Wingf. ……....................... 559 Celoporthe Nakab., Gryzenh., Jol. Roux & M.J. Wingf. ……………..................... 562 21. Celoporthe dispersa Nakab., Gryzenh., Jol. Roux & M.J. Wingf. ............. 564 7. Species excluded or of questionable validity ………………………........………... 568 7.1 Excluded species …………………………………………………………. 568 7.2 Species of questionable validity …………………………………………. 569 Acknowledgements .…………………………………………….……………..................... 570 References ………………………………………...…………………………...................... 571 List of abbreviations ………………………………………..……………………................ 598 353 Taxonomy, phylogeny and ecology of Cryphonectria species and other members of the Cryphonectriaceae Marieka Gryzenhout1, Brenda D. Wingfield2 & Michael J. Wingfield1 1Department of Microbiology and Plant Pathology, 2Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, 0002, South Africa. Abstract: The Cryphonectriaceae includes some of the most important pathogens of trees and shrubs. Well-known examples are the chestnut blight pathogen, Cryphonectria parasitica, and the Eucalyptus canker pathogen, Chrysoporthe cubensis. The family includes all genera in the Diaporthales with orange stromatic tissue that turns purple in KOH and yellow in lactic acid. These include Cryphonectria, Endothia, Chrysoporthe, Amphilogia, Rostraureum, Microthia, Holocryphia and Celoporthe. The mitotic genera Aurapex and Ursicollum are also included as they have been shown to reside in the Cryphonectriaceae based on DNA sequence data. This monograph presents details of the pathology, ecology and morphological characteristics defining these genera, the majority of which have been described relatively recently. Comprehensive phylogenetic analyses are presented that include all taxa in the Cryphonectriaceae for which cultures are available. Endothiella, the anamorph genus currently recognized for both Cryphonectria and Endothia, is specifically assigned to Cryphonectria. A new combination, Cryphonectria japonica, is also suggested for Cryphonectria nitschkei. 354 Taxonomic novelties: Cryphonectria japonica (Tak. Kobay. & Kaz. Itô) Gryzenh. & M.J. Wingf. comb. nov., Endothiella Sacc. emend. Gryzenh. & M.J. Wingf. Key words: canker, Cryphonectriaceae, Diaporthales, forestry, phylogeny 1. INTRODUCTION The Cryphonectriaceae Gryzenh. & M.J. Wingf. nom. prov. represents a group of fungi that include some of the world’s most important pathogens of trees. The best known of these pathogens is Cryphonectria parasitica (Murrill) M.E. Barr, the causal agent of chestnut blight that has all but destroyed the American chestnut (Castanea dentata) in its North American natural habitat. Other important examples of species that cause serious canker diseases on Eucalyptus spp., particularly those grown in plantations, are Chrysoporthe cubensis (Bruner) Gryzenh. & M.J. Wingf. and Chrysoporthe austroafricana Gryzenh. & M.J. Wingf., previously known as Cryphonectria cubensis (Bruner) Hodges. The Cryphonectriaceae has recently been erected to contain Cryphonectria (Sacc.) Sacc., Endothia Fr. and various genera that have been relatively recently described. Many of these new genera include newly discovered taxa or existing species of Cryphonectria that have been shown as distinct based on DNA sequence comparisons for a number of variable regions of the genome. These phylogenetic groupings are supported by morphological characteristics not previously recognized to represent generic differences.